Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170

Contents lists available at ScienceDirect

Environmental Nanotechnology, Monitoring

& Management
journal homepage: www.elsevier.com/locate/enmm

Analysis of multiclass antibiotic residues in urban wastewater in Tunisia T

a b a b,⁎
Malek Harrabi , Saulo Varela Della Giustina , Fatma Aloulou , Sara Rodriguez-Mozaz ,
Damià Barcelób,c, Boubaker Elleucha
a
National School of Engineering Sfax, GEET, Tunisia
b
Catalan Institute for Water Research (ICRA), H2O Building, Scientific and Technological Park of the University of Girona, Emili Grahit 101, 17003 Girona, Spain
c
Water and Soil Quality Research Group, Department of Environmental Chemistry, IDAEA-CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain

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

Keywords: The present work describes the occurrence of antibiotic residues in urban wastewaters discharged into southern
Antibiotics Sfax wastewater treatment plant (WWTP). Thirteen out of the fifty-six antimicrobial compounds analyzed were
Environment pollution detected in the wastewater samples with concentration ranging from 23.30 (for cefalexin) to 690.50 ng/L (for
WWTP spiramycin) in WWTP influent and from 7.50 (for cefalexin) to 370.04 ng/L (for spiramycin) in WWTP effluent.
Wastewater
The highest removal efficiency was observed for trimethoprim (88%), while the lowest one was observed for
Water reuse
sulfapyridine (33%).The potential impact of the regenerated waters in the environment and pubic health is also
discussed.

1. Introduction
Antibiotics are a group of pharmaceuticals extensively used in
human medicine (Schwartz et al., 2003). A global increase in antibiotic
consumption from current 63.2 to 105.3 thousand tons by 2030 is
foreseen by World Health Organization (INC, 2016). In Tunisia, the
consumption of antibiotics increased by 38% between 2005 and 2013,
and further rise is foreseen in the next years (The MSI Data Base
"SIAMED") (INC, 2016). This increase of antibiotic consumption has
been reported to be alligned with an increment in their irrational use,
from 28 to 65% in the last few years (Erbay et al., 2005; Tünger et al.,
2000). In this regard, a study recently performed in Tunisia showed that
61% of consumers were obtaining antibiotics directly from the phar-
macist, without a medical prescription (INC, 2016). This uncontrolled
use of antibiotics can have a direct effect on the health of consumers
because human body becomes resistant to antibiotics (Carlet et al.,
2012) and indirect problems also on the environment. The later is re-
lated to the considerable quantities of antibiotics that enter into the
ecosystem through effluent from urban wastewater treatment plants.
Several studies have shown that wastewater from municipal conven-
tional WWTPs could be a significant source of contamination of aquatic
environment by antibiotics from human consumption since WWTPs are
not designed to remove them (Adams et al., 2002; Gobel et al., 2007;
Rodriguez-Mozaz et al., 2015; Vieno et al., 2007; Watkinson et al.,
2009). Therefore, antibiotics have been detected in the natural en-
vironment, namely ground water (Batt and Aga, 2005; Negreanu et al.,
2012), surface water (Proia et al., 2016; Schwartz et al., 2003), soil and
sediment samples (Gobel et al., 2007; Kim and Carlson, 2007) as well as
in aquatic organisms (Álvarez-Muñoz et al., 2015; Homem and Santos,
2011; Serra-Compte et al., 2017). Furthermore, applying treated was-
tewater to agricultural fields for irrigation may contaminate agri-
cultural soils (Negreanu et al., 2012) and lead to uptake of antibiotics
by plants (Wu et al., 2015). In summary, antibiotics show pseudo-
persistent behavior in the environment (Hernando et al., 2006;
Richardson et al., 2005) and their accumulation in different environ-
mental compartments can threaten aquatic and terrestrial ecosystems
and may have an impact on human and animal health (Homem and
Santos, 2011). In fact, antibiotic residues present in the ecosystem
provide an ideal setting for the acquisition and spread of antibiotic
resistance genes, resulting in serious environmental problems (Pruneau
et al., 2011; Rodriguez-Mozaz et al., 2015).
The objective of this study was to monitor the presence of 56 an-
tibiotics in wastewater effluents in a WWTP located in a highly in-
dustralized area of southern Sfax, Tunisia (Fig. 1). So far there is only a
study in Tunisia that evaluated the presence of 14 aminoglycosides and
phenicol antibiotics in municipal wastewater, sea water and in a
pharmaceutical company effluents (Tahrani et al., 2016). Removal ef-
ficiencies of antibiotics in Sfax WWTP were evaluated and discussed. To
the authors’ knowledge, our work is the first study on the presence and
elimination of such broad set of antibiotics in Tunisia.

⁎
Corresponding author.
E-mail address: srodriguez@icra.cat (S. Rodriguez-Mozaz).

https://doi.org/10.1016/j.enmm.2018.05.006
Received 27 November 2017; Received in revised form 30 March 2018; Accepted 10 May 2018
2215-1532/ © 2018 Elsevier B.V. All rights reserved.
M. Harrabi et al. Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170

Fig. 1. Southern SFAX WWTP location.

2. Materials and methods
2.1. Sampling site and sample collection
Southern Sfax WWTP is located in a industrialized area (Fig. 1) and
receives both domestic and treated industrial wastewater. Domestic
source accounts for approximately 75% of influent while industrial
sources account for 25%. The WWTP serves a population of 526.800
inhabitants and is designed to purify urban wastewater with a daily
average flow rate of 49.500 m3 d−1, with an hydraulic retention time
(HRT) of 48 h and a sludge retention time (SRT) of 22 days (Belhaj
et al., 2015). The treatment process of the plant are outlined in Fig. 2.
Raw wastewater passes first through the mechanical screen and the
aerated grit chamber and is further evacuated into the primary sedi-
mentation tanks. Secondary treatment consists of three alternative
anoxic/aerobic bioreactors aimed at the elimination of nitrogen and
phosphorus. After settling in the secondary sedimentation tanks final
effluent is either discharged into the sea or used to irrigate plants and
trees. Decanted sludge is recirculated to the bioreactors and the re-
maining portion is pumped into the sludge storage tank.
Three different sampling campaigns were executed during March
2016 in dry weather conditions, (average temperature was 28 °C).
Although the collection of 24 h composite samples is recommended in
order to take representative samples, due to instrumental limitations
(no autosampler available), composite samples were manually collected
during a 6 hour period at the WWTP: one sample every hour, from 7 h
to 13 h, which is the daytime when the WWTP the receives the highest
load of wastewater at an average flow rate of 50 L/s (personal com-
munication of the staff at Sfax WWTP). The influent sample was col-
lected before the primary treatment (influent) and the effluent sample

Fig. 2. Wastewater treatment steps in southern SFAX WWTP.

164
M. Harrabi et al.
was collected at the outlet of the secondary treatment after 48 h (which
is the hydraulic retention time (HRT) of the WWTP, in order to calcu-
late the corresponding removal efficiencies. Information about sam-
pling dates is provided in Table S1. Amber glass bottles rinsed with
ultra-pure water were used for sample collection. Wastewater was fil-
tered through 1 μm glass fibre filters followed by 0.45 μm glass fibre
filters (Whatman, U.K.) and kept in the dark at −20 °C in stable con-
ditions until the anlysis within the 1st week after the sampling, as it is
recommended in a previous study (Llorca et al., 2014).
2.2. Reagents and chemicals
All antibiotic standards were of high purity grade (> 90%) and
purchased from Sigma–Aldrich: tetracycline, chlortetracyline, oxyte-
tracycline and lincomycin were purchased as hydrochloride salts, ox-
acillin, cefazolin, cefotaxime and cefapirin were acquired as sodium
salts, penicillin V and penicillin G as potassium salt, amoxicillin and
ampicillin were purchased as trihydrate salts in contrast, tylosin and
doxycycline were acquired as tartrate and hyclate salts, respectively.
Isotopically labeled compounds, used as internal standards, were,
ofloxacin-d3, ciprofloxacin-d8 (as hydrochloride hydrate salt), eri-
thromycin-N, N-dimethyl13C, ampicillin-15N and ronidazole-d3, pur-
chased from Sigma–Aldrich, and azithromycin-d3, sulfamethoxazole-d4
and lincomycin-d3, which were purchased from Toronto Research
Chemicals (Ontario, Canada). On the other hand, sulfadimethoxine-d6
and sulfadoxine-d3, which were used as surrogate standards, were
purchased from Sigma–Aldrich.
Individual stock standards solutions as well as isotopically labeled
internal standard and surrogate standard solutions were prepared at a
concentration of 1000 mg/L. After preparation, standards were stored
at −20 °C (Gros et al., 2013). Standard solutions containing the anti-
biotics mixtures were prepared in methanol/water (50:50, v/v) just
before the analysis; they were preparedby mixing appropriate amounts
of the intermediate standard solutions. Separate mixtures of isotopically
labeled internal standards were prepared in methanol, except ampi-
cillin-15N, which was diluted in acetonitrile/water solution (50:50, v/
v). Further diluted solutions were prepared in a methanol/HPLC water
mixture (50:50, v/v).
Glass fiber filters (1 μm) and glass fiber filters (0.45 μm) from
Whatman (UK) were used for filtration of the samples. OASIS
Hydrophilic-Lipophilic-Balanced (HLB) cartridges (60 mg, 3 mL) from
Waters Corporation (Milford, MA, U.S.A.) were used for solid phase
extraction.
HPLC grade methanol, acetonitrile, formic acid 98% and water
(Lichrosolv) were purchased by Merck (Darmstadt, Germany).
Ammonium hydroxide, hydrochloric acid 37% and the nitrogen for
drying was purchased by Abelló Linde S.A. (Spain).
2.3. Analytical method for antibiotic determination
The analytical method developed by Gros was used to quantify the
selected antibiotics in the wastewater samples (Gros et al., 2013).
Briefly, an appropriate volume of ethylenediaminetetraacetic acid dis-
odium salt dehydrate (Na2EDTA) solution having a concentration of
0.1 M was added to the various types of filtered water to get a final
concentration equal to 0.1% (g solute/g solution). Water samples were
further adjusted to pH 2.5 with hydrochloric acid. The cartridges were
conditioned with 6 mL methanol followed by 6 mL water (HPLC grade).
25 mL of influent and 50 mL of filtered effluent were loaded onto the
SPE cartridge. After preconcentration of the sample, the cartridges were
rinsed with 6 mL HPLC grade water and further air dried for about
5 min to remove excess water. The analytes were eluted from the car-
tridge with 6 mL of methanol. The extracts were then evaporated to
dryness underneath a flow of nitrogen and reconstituted with 1 mL of
mixture of methanol and water (50:50, v/v). Finally, 10 μL of a stan-
dard 1 ng/mL mixture containing all isotopically labelled standards was
165
Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170
added to the extract as internal standards. The volume of sample in-
jected each time was 5 μL.
Chromatographic separation was carried out with an ultra-perfor-
mance liquid chromatography system (Waters Corp. Mildford, MA,
USA), using an Acquity HSS T3 column (50 mm × 2.1 mm i.d., 1.7 μm
particle size) for the compounds analyzed under positive electrospray
ionization (PI) and an Acquity BEH C18 column (50 mm × 2.1 mm i.d.,
1.7 μm particle size) for the ones analyzed under negative electrospray
ionization (NI), both from Waters Corporation. The UPLC instrument
was coupled to 5500 QqLit, triple quadrupole–linear ion trap mass
spectrometer (5500 QTRAP, Applied Biosystems, Foster City, CA, USA)
with a Turbo V ion spray source. Two MRM transitions per compound
were recorded by using the Scheduled MRM™ algorithm. All data were
acquired and processed using Analyst 1.6.3 software.
2.4. Environmental risk assessment (ERA)
In order to assess the environmental risk of WWTP efflluents, the
hazard quotient (HQ)were calculated according to the European
Community guidelines (European C, 2003):
HQ = (MEC)/(PNEC)
where MEC is the “measured concentration” for each antibiotic in the
WWTP effluent (Table S2) and PNEC is its “Predicted No Effect
Concentration” calculated following the European Committee method
(European C, 2003), which establishdes that PNECs is 1000 times lower
than the toxicity concentration value found for the most sensitive
species assayed:
PNEC = (EC50 or LC50)/1000
where LC50 (half maximal lethal concentration) or EC50 (half maximal
effective concentration) values from each antibiotic were obtained from
international databases and from the literature (Table S3) or using
ECOSAR software (Meylan and Howard, 1998; Rincon et al., 2014;
Sanderson, 2003).
3. Results and discussion
3.1. Occurrence of antibiotics in raw wastewater
Thirteen antimicrobial compounds, out of fifty-six analyzed were
detected in the raw wastewater samples collected over the three sam-
pling campaigns (Table S2 and Fig. 3a). Fuoroquinolones, a group of
antibiotics used to treat a high number of human infections, was the
most abundant in WWTP influent, achieving the highest concentration
for enrofloxacin (400.20 ng/L), followed by ciprofloxacin (330.33 ng/L)
and ofloxacin (175.01 ng/L). Fluoroquinolones have also been found at
very high concentrations ina previous study covering the same target
antibiotics in raw domestic wastewater in Spain: higher levels were
found in Spain of ciprofloxacin (639 ng/L) and ofloxacin (529 ng/L)
whereas enrofloxacin was not even detected (Rodriguez-Mozaz et al.,
2015). The high levels encountered in Sfax WWTP of enrofloxacin (a
veterinary antibiotic) might be attributed to the presence of certain
veterinary practices and a pharmaceutical industry discharging in the
urban WWTP. Similar concentration was detected for enrofloxacin in
U.S.A WWTPs (270 ng/L) (Karthikeyan and Meyer, 2006). Up to
100 μg/L for ciprofloxacin and 10 μg/L for ofloxacin have even been
detected wordwide (Verlicchi et al., 2010).
Despite the high consumption of penicilin antibiotics (INC, 2016),
neither penicillin G and V nor amoxicillin were detected in raw was-
tewater, probably because of their chemical instability; they are easily
transformed through hydrolysis (Knapp et al., 2011). In contrast, am-
picillin was found at values up to 75.40 ng/L, although much lower
than those found by other authors, for example Papageorgiou et al.
detected ampicillin up to 1805 ng/L in influent wastewater of central
M. Harrabi et al.
Fig. 3. Antibiotics concentration (ng/L) in WWTP influent (a) and effluent (b)
during the three sampling campaigns (S.C I, S.C II and S.C III).
Greece WWTP (Papageorgiou et al., 2016).
The levels of sulfapyridine and sulfamethoxazole were also high in
all sampling campaigns up to 365.50 and 126.70 ng/L, respectively but
lower than levels found in other countries such as Korea and Czech
where sulfapyridine was found in WWTP influent as high as 921 ng/L
and 660 ng/L repectively (Kim et al., 2012) (Golovko et al., 2014).
Sulfametoxazole was found at high levels in several WWTPs in China,
up to 959.13 ng/L (Wu et al., 2016) and in Spain, up to 417.4 ng/L
(Rodriguez-Mozaz et al., 2015). In contrast, trimethoprim, commonly
used combined with sulfametoxazole (Batt and Aga, 2005; Gros et al.,
2013), was found at lower concentration ranging between 56.60 and
86.67 ng/L.
Tetracycline and metronidazole were detected in some occasions
during sampling campaign, but always below limit of quantification
(MQL), probably due to its low consumption in Tunisia (INC, 2016).
Incontrast, higher concentration of antibiotics belonging to the group of
macrolides, such as spiramycin and azithromycin,were detected all
sampling days, as high as 690.50 ng/L and 135.45 ng/L, respectivey.
In regard to lincosamides group, lincomycin antibiotic was detected
in all influent samples ranging between 100.33 and 150.08 ng/L, higher
values than those found in Greece, as high as 66.7 ng/L (Papageorgiou
et al., 2016), but much lower than those reported in Australia, up to
500 ng/L) (Watkinson et al., 2009).
Finally, cefalexin was the only cephalosporines detected ranging
from 23.30 ng/L to 33.33 ng/L. These concentrations were higher than
those found in Taiwan (up to 4.36 ng/L) (Lin et al., 2009). However,
values of cefalexin reported in five WWTPs in Australia were much
166
Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170
Fig. 4. Average removal efficiencies of antibiotics by primary and secondary
treatmentsin the WWTP.
higher (2800–64000 ng/L) (Watkinson et al., 2009).
3.2. Antibiotic removal during conventional wastewater treatment
The removal efficiencies of the antibiotics detected in southern Sfax
WWTP during the sampling campaign are summarized in Fig. 4. Max-
imum removal was achieved for trimethroprim (88% average), similar
to those observed in a WWTP in New Mexico, USA (50–100%) (Brown
et al., 2006), Spain (55–110%) (Collado et al., 2014; Rodriguez-Mozaz
et al., 2015) and in Greece (up to 90%) (Papageorgiou et al., 2016).
This high removal may be due to its tendency to adsorb in sewage
sludge (Huang, 2001). Ampicillin removal rangied between 60 and 69%
during the three samlping days, a bit lower than the removal rate re-
ported in Greece (78–90%) (Papageorgiou et al., 2016). Similar re-
moval rate was detected for cefalexin (63% average), lower than that
reported in Taiwan (up to 99.8%) (Lin et al., 2009). Removal efficiences
between 41 and 59% were obtained for enroflaxacin, ciprofloxacin,
ofloxacin, lincomycin and sulfamethoxazole, lower than those reported
in a Japan WWTP: 74%, 83% and 57% for enrofloxacin, ciprofloxacin
and lincomycin, respectively (Ghosh et al., 2009). In addition, 100%
removal rate was reported for lincomycin in Greece (Papageorgiou
et al., 2016). For sulfamethoxazole similar removal values were ob-
tained (up to 60%) in Czech (Golovko et al., 2014) and in Spain
(40–45%). In fact, a high removal variability was observed for sulfa-
methoxazole between the 3 different campaings (ranging from 23% in
the first sampling day to 61% in the third day) compared to other an-
tibiotics removal efficiencies, much more homogeneous in the 3 sam-
pling campaigns. As regard ofloxacin, similar removal values (40–66%)
were reported by Gros (Gros et al., 2010) though lower than those re-
ported by Rodriguez-Mozaz et al. (79–89%) (Rodriguez-Mozaz et al.,
2015), both in Spain. The other sulfonamide antibiotic detected in
WWTP, sulfapyridine, showed one of the lowest removal efficiency
(33%), in agreement with those removal rates observed in Tudela
WWTP in Spain (2008) (29%) (García-Galán et al., 2011).
Concerning macrolide antibiotics, which have been reported to be
more persistent than other antibiotics (Huang, 2001), azythromycin
and spiramycin were both detected in raw and treated water. Spir-
amycin showed a medium removal rate (38% average), a bit lower than
that found in Italy (52%) (Petrovic and Verlicchi, 2014). Finally, the
level of azithromycin increased after wastewater treatment in the 3
campaigns. Similar behavior was occasionally observed in other
WWTPs in Spain (Rodriguez-Mozaz et al., 2015). A plausible explana-
tion for this observation, is that conjugated metabolites of azy-
thromycin in raw wastewater could be de-conjugated during the
treatment process back to the parent compound (Miao et al., 2002). In
addition, desorption from particulate matter can lead to the increase of
the pollutant concentration during the treatment and thus negative
removal was calculated (Miao et al., 2002; Zhang et al., 2015).
Nevertheless, low removal of azithromycin has been observed in
Switzerland WWTP (10–33%) (Gobel et al., 2007) and in Japan
M. Harrabi et al.
Table 1
Hazard quotient (HQ) values of antibiotics detected in WWTP effluent.
Chemical group Compounds Campaign 1
Fluoroquinolones Ofloxacin 6,25
Ciprofloxacin 36,00
Enrofloxacin 0,03
Penicillins Ampicillin 0,00
Cephalosporins Cefalexin 0,00
Macrolides Azithromycin 7,89
Spiramycin 0,07
Lincosamides Lincomycin 0,00
Sulfonamides Sulfamethoxazole 3,33
Sulfapyridine 0,00
Dihydrofolate reductase inhibitors Trimethoprim 1,44
(6–12%) (Ghosh et al., 2009).
3.3. Environmental risk assessment (ERA)
Hazard quotients (HQ) values were calculated for each antibiotic
present in WWTP effluent (Table 1) based on their abundance (Fig. 3b
and Table S2) and PNEC values (Table S3).
According to the results (Table 1), it can be asserted that the most
hazardous antibiotics in the WWTP effluent are the fluoroquinolones:
ciprofloxacin and ofloxacin and the macrolide azithromycin. These re-
sults can be attributed to the very low PNEC values of these antibiotics:
ofloxacin (0.016 μg/L), ciprofloxacin (0.005 μg/L) and azithromycin
(0.019 μg/L). Similar HQ for ofloxacin (6.18) and azithromycin (7.90)
have been reported in other study in Spain (7.38 and 7.53 respectively
(Lucas et al., 2016)), whereas HQ value for ciprofloxacin (33.56) was
higher than those found in another WWTP Spain (Gros et al., 2010). In
the same WWTP in Spain, HQ values for sulfamethoxazole (22.96) was
higher than the one calculated in the present work (up to 3.33).
3.4. Potential risks of water reuse in Sfax area
Although 25% of southern Sfax WWTP treated effluent is used to
irrigate crops, c.a. 75% flows into a channel where, after being mixed
with industrial effluents, it is discharged into the Mediterranean Sea,
thus contributing to coastline pollution. In fact, Houda et al. reported
that Sfax coast is characterized by high values of mineral and organic
loads, exceeding more than 10 and 37 folds the maximum permitted
concentrations in sea water of DCO and DBO5 respectively, established
by the Tunisian authorities (NT 106.002 1989) (Houda et al., 2011). In
addition, several studies detected a high level of heavy metals (Zn, Cu,
Cr, Cd, Pb Fe) in the surface marine sediments, higher than American
marine sediment quality standards (SQS) and the Chinese Marine Se-
diment Quality Standards (GB 18668-2002) (Gargouri et al., 2011;
Houda et al., 2011; Mkawar et al., 2007). Furthermore, non-aromatic
hydrocarbons were detected at high concentrations compared to other
Mediterranean sites, in the range of 310–1406 ng/g dry weight (Louati
et al., 2001; Zaghden et al., 2005).This contamination can definitely
threaten the biodiversity of coastal fauna and flora living in this area as
some studies have described, such as malformation in bivalves
(Maatoug et al., 2009), high proliferation of some toxic microalgae,
reduction of the diversity indexes of phytoplankton, ciliate and copepod
communities (Salem et al., 2015) as well ashigh levels of pathogenic
bacteria like Aeromonas (Maalej et al., 2003). There are no studies re-
porting about the levels of antibiotics in Sfax marine environment, but
it is expected that their presence due to the loads of effluents such as the
one from Sfax WWTP could also influence the organisms living in this
coastal area. Several studies have reported the presence of pharma-
ceuticals, including antibiotics in water, sediments and biota from the
167
Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170
Campaign 2 Campaign 3 Average
6,88 5,42 6,18
32,00 32,67 33,56
0,02 0,03 0,03
0,00 0,00 0,00
0,00 0,00 0,00
8,25 7,54 7,90
0,07 0,06 0,07
0,00 0,00 0,00
2,35 1,85 2,51
0,00 0,00 0,00
1,72 1,29 1,49
coastal areas worldwide (Arpin-Pont et al., 2016; Rodriguez-Mozaz
et al., 2017). Antibiotics have been measured in marine organisms in
France, as high as 1 and 1.5 mg/L for oxytetracycline and oxolinic acid,
respectively in blue mussel viscera (Le Bris and Pouliquen, 2004), China
(up to 9.45 ng/g, dry weight in Mollusks of oxytetracycline (Na et al.,
2013) or Belgium (up to 16.69 ng/g, sulfamerazine and 65 ng/g oflox-
acin) (Wille et al., 2011). This is particularly worrisome since the ac-
cumulation of antibiotics can impact not only marine organisms
themselves but also human health (as a results of consumption of sea-
food).
About 25% of southern Sfax WWTP treated effluents are used to
irrigate an area of 536 halocated in El Hajeb Town, planted with olive
trees (80%), cereals, vegetable crops and fodder. Wastewater reuse in
agriculture is widespread throughout the world, especially in arid and
semi-arid areas in order to fight water scarcity (Angelakis and Durham,
2008; Travis et al., 2010). Several studies have shown that the perfor-
mance of Sfax WWTP is not fully satisfactory in terms of water reuse;
Chemical Oxygen Demand (COD), Biological Oxygen Demand (BOD5)
and Nitrogen (N) values in the treated water exceeded the maximum
values recommended by the Tunisian authorities (NT 106.03 1989)
concerning wastewater reuse in crop irrigations in several studies
(Belaid et al., 2012; Belhaj et al., 2014; Ouali et al., 2009). In addition,
although there is no regulation about micropollutants, as is the case for
other general parameters, their presence in wastewater effluents can
also impact the receiving soils and groundwater reservoirs and even-
tually the plants and crops growing in this area. Among unregulated
micropollutants, antibiotics have been reported in many occasions in
natural environment. Kinney et al. detected erythromycin in waste-
water irrigated soils at concentrations between 0.02–15 μg/kg (Kinney
et al., 2006). Moreover, Chen detected three antibiotics (oxytetracy-
cline, tetracycline and trimethoprim) in the soils of Hebei, China which
has been used for wastewater irrigation for about 50 years (Chen et al.,
2011). Other antibiotics residues found in crops irrigated with waste-
water effluents are norfloxacin (4.6–23.6 μg/kg), chloramphenicol
(2.6–22.4 μg/kg) and tetracycline (4.0–10.1 μg/kg) (Pan et al., 2014).
Similar results were found by Hu in 2010 (Hu et al., 2010).
Not only antibiotic residues but antibiotic-resistant bacteria and
their genes can be transported by wastewater and deposit in soil as it
has been reported in some recent study (Becerra-Castro et al., 2015).
Even though the values of antibiotic concentrations are not extremely
high, antibiotic-resistant bacteria and/or their corresponding resistance
genes have been shown to contaminate humans via the food chain
(Boehme et al., 2004). To his respect, many studies have proved that
the combination of low levels of antibiotics and heavy metals present in
polluted environment could be an important factor in the selection of
multidrug resistance plasmids (Cabral et al., 2016; Gullberg et al.,
2014; Teixeira et al., 2016). Concentration of heavy metal in Sfax
WWTP effluents exceeded the limits set by the Tunisian Authorities (NT
M. Harrabi et al.
106.03 1989) as reported in previous studies (Belhaj et al., 2016; Ben
Fredj et al., 2014). Thus irrigation of crops by these WWTP effluents
means an exposure pathway to food consumers, posing a risk for human
health.
Furthermore, this treated water is characterized by high salinity
(Belaid et al., 2012; Belhaj et al., 2016). The electrical conductivity (a
salinity value) sometimes exceeds 5000 μS cm−1 (Belaid et al., 2012),
well above the limits established by Food and Agriculture Organization
of the United Nations (FAO) for the reuse of wastewater in agriculture
(Ayers and Westcot, 1985). In fact, prolonged irrigation with these
waters will lead to an accumulation of salts and their concentration in
the root zone of the crops, which could cause secondary salinization of
the soil in this area and in the long term, could lead to structural de-
gradation.
In summary, irrigation with Sfax treated wastewaters can impact
environment and human health and therefore, some farmers mix the
effluent with freshwater before irrigating their fields. It has been re-
commended to reduce salinity problems by using a 50% dilution of
effluent with fresh water (Belhaj et al., 2016). Other approaches to
improve the quality of treated effluent have been suggested like the
application of tertiary treatments, in this sense oxidation using hy-
drogen peroxide was tested in Sfax WWTP and proved to reduce COD
(Chemical Oxygen Demand) including refractory organic substances,
the unpleasant smell, the foam, sulfides as well as to achieve high
bacterial inactivation (Ksibi, 2006). Another study in the same WWTP
investigated the combination of electro-Fenton system (anaerobic di-
gestion) and ultrafiltration (UF) treatment and removals up to 52.6% of
total COD, 83.8% of TSS (Total Suspended Solids), 78% of polyphenols
and more than 93% of lipids were reported (Khoufi et al., 2009).
4. Conclusion
The presence of 56 antibiotic residues and their removal in the
WWTP of southerm Sfax was assessed in this study. Thirteen antibiotics
were detected with concentrations ranging between 23 to 690 ng/L in
the influent and from 7.5 to 370 ng/L in the effluent. The compound
detected in treated wastewater posing the highest risk to the environ-
ment was ciprofloxacin followed by azythromycin, ofloxacin and sul-
famethoxazol. Removal efficiencies presented variation between the
different therapeutic groups with the highest removal observed for
trimethoprim (88%) and the lowest for sulfapyridine (35%). These re-
sults urge us to seek a solution to improve WWTP treatment through the
development and application of appropriate tertiary treatments in order
to improve the quality of the treated water and reduce antibiotic pol-
lution. Such approaches would permit a safer use of regenerated waters
for irrigation of neighboring agricultural land.
Acknowledgments
This study has been supported by the Mınıstry of Higher Educatıon
and Scientific Research-Tunisia, by the Spanish Ministry of Economy,
Industry and Competitiveness (through project StARE, JPIW2013-089-
C02-02), and by the Generalitat de Catalunya (Consolidated Research
Groups: 2017 SGR 1404 and ICRA-ENV 2017 SGR). Dr. SRM ac-
knowledges her Ramón y Cajal fellowship (RyC-2014-16707) from the
Spanish Ministry of Economy, Industry and Competitiveness. Best ac-
knowledgments for the operators and staff at the southern Sfax WWTP
(Tunisia) for their assistance. We wish to thank Mr. Jamil JAOUA,
Founder and Head of the English Teaching Unit at the Sfax Faculty of
Science, for revising the paper.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in the
online version, at https://doi.org/10.1016/j.enmm.2018.05.006.
168
Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170
References
Adams, C., Wang, Y., Loftin, K., Meyer, M., 2002. Removal of antibiotics from surface and
distilled water in conventional water treatment processes. J. Environ. Eng. 128,
253–260. http://dx.doi.org/10.1061/(ASCE)0733-9372(2002)128:3(253).
Álvarez-Muñoz, D., Rodríguez-Mozaz, S., Maulvault, A.L., Tediosi, A., Fernández-Tejedor,
M., Van den Heuvel, F., Kotterman, M., Marques, A., Barceló, D., 2015. Occurrence of
pharmaceuticals and endocrine disrupting compounds in macroalgaes, bivalves, and
fish from coastal areas in Europe. Environ. Res. 143, 56–64. http://dx.doi.org/10.
1016/j.envres.2015.09.018.
Angelakis, A.N., Durham, B., 2008. Water recycling and reuse in EUREAU countries:
trends and challenges. Desalination 218, 3–12. http://dx.doi.org/10.1016/j.desal.
2006.07.015.
Arpin-Pont, L., Bueno, M.J.M., Gomez, E., Fenet, H., 2016. Occurrence of PPCPs in the
marine environment: a review. Environ. Sci. Pollut. Res. 23, 4978–4991. http://dx.
doi.org/10.1007/s11356-014-3617-x.
Ayers, R.S., Westcot, D.W., 1985. Water Quality for Agriculture, FAO Irrigation and
Drainage Paper. Food and Agriculture Organization of the United Nations, Rome.
Batt, A.L., Aga, D.S., 2005. Simultaneous analysis of multiple classes of antibiotics by ion
trap LC/MS/MS for assessing surface water and groundwater contamination. Anal.
Chem. 77, 2940–2947.
Becerra-Castro, C., Lopes, A.R., Vaz-Moreira, I., Silva, E.F., Manaia, C.M., Nunes, O.C.,
2015. Wastewater reuse in irrigation: A microbiological perspective on implications
in soil fertility and human and environmental health. Environ. Int. 75, 117–135.
http://dx.doi.org/10.1016/j.envint.2014.11.001.
Belaid, N., Neel, C., Lenain, J.F., Buzier, R., Kallel, M., Ayoub, T., Ayadi, A., Baudu, M.,
2012. Assessment of metal accumulation in calcareous soil and forage crops subjected
to long-term irrigation using treated wastewater: case of El Hajeb-Sfax, Tunisia.
Agric. Ecosyst. Environ. 158, 83–93. http://dx.doi.org/10.1016/j.agee.2012.06.002.
Belhaj, D., Baccar, R., Jaabiri, I., Bouzid, J., Kallel, M., Ayadi, H., Zhou, J.L., 2015. Fate of
selected estrogenic hormones in an urban sewage treatment plant in Tunisia (North
Africa). Sci. Total Environ. 505, 154–160. http://dx.doi.org/10.1016/j.scitotenv.
2014.10.018.
Belhaj, D., Jaabiri, I., Ayadi, H., Kallel, M., Zhou, J.L., 2014. Occurrence and removal of
steroidal estrogens in Centre Eastern Tunisia municipal sewage treatment plant.
Desalin. Water Treat. 52, 2330–2339. http://dx.doi.org/10.1080/19443994.2013.
804448.
Belhaj, D., Jerbi, B., Medhioub, M., Zhou, J., Kallel, M., Ayadi, H., 2016. Impact of treated
urban wastewater for reuse in agriculture on crop response and soil ecotoxicity.
Environ. Sci. Pollut. Res. 23, 15877–15887. http://dx.doi.org/10.1007/s11356-015-
5672-3.
Ben Fredj, F., Wali, A., Khadhraoui, M., Han, J., Funamizu, N., Ksibi, M., Isoda, H., 2014.
Risk assessment of heavy metal toxicity of soil irrigated with treated wastewater
using heat shock proteins stress responses: case of El Hajeb, Sfax, Tunisia. Environ.
Sci. Pollut. Res. 21, 4716–4726. http://dx.doi.org/10.1007/s11356-013-2411-5.
Boehme, S., Werner, G., Klare, I., Reissbrodt, R., Witte, W., 2004. Occurrence of anti-
biotic-resistant enterobacteria in agricultural foodstuffs. Mol. Nutr. Food Res. 48,
522–531. http://dx.doi.org/10.1002/mnfr.200400030.
Brown, K.D., Kulis, J., Thomson, B., Chapman, T.H., Mawhinney, D.B., 2006. Occurrence
of antibiotics in hospital, residential, and dairy effluent, municipal wastewater, and
the Rio Grande in New Mexico. Sci. Total Environ. 366, 772–783. http://dx.doi.org/
10.1016/j.scitotenv.2005.10.007.
Cabral, L., Júnior, G.V.L., Pereira de Sousa, S.T., Dias, A.C.F., Lira Cadete, L., Andreote,
F.D., Hess, M., de Oliveira, V.M., 2016. Anthropogenic impact on mangrove sedi-
ments triggers differential responses in the heavy metals and antibiotic resistomes of
microbial communities. Environ. Pollut. 216, 460–469. http://dx.doi.org/10.1016/j.
envpol.2016.05.078.
Carlet, J., Rambaud, C., Pulcini, C., 2012. Alliance contre les bactéries multirésistantes :
sauvons les antibiotiques !. Ann. Fr. Anesth. Réanimation 31, 704–708. http://dx.doi.
org/10.1016/j.annfar.2012.07.003.
Chen, F., Ying, G.-G., Kong, L.-X., Wang, L., Zhao, J.-L., Zhou, L.-J., Zhang, L.-J., 2011.
Distribution and accumulation of endocrine-disrupting chemicals and pharmaceu-
ticals in wastewater irrigated soils in Hebei, China. Environ. Pollut. 159, 1490–1498.
http://dx.doi.org/10.1016/j.envpol.2011.03.016.
Collado, N., Rodriguez-Mozaz, S., Gros, M., Rubirola, A., Barceló, D., Comas, J.,
Rodriguez-Roda, I., Buttiglieri, G., 2014. Pharmaceuticals occurrence in a WWTP
with significant industrial contribution and its input into the river system. Environ.
Pollut. 185, 202–212. http://dx.doi.org/10.1016/j.envpol.2013.10.040.
Erbay, A., Bodur, H., Akıncı, E., Çolpan, A., 2005. Evaluation of antibiotic use in intensive
care units of a tertiary care hospital in Turkey. J. Hosp. Infect. 59, 53–61. http://dx.
doi.org/10.1016/j.jhin.2004.07.026.
European C, 2003. Technical Guidance Document on Risk Assessment in Support of
Commission Directive 93/67/EEC on Risk Assessment for New Notified Substances
and Commission Regulation (EC) 1488/94 on Risk Assessment for Existing
Substances and Directive 98/8/EC of the European Parliament and of the Council
Concerning the Placing of Biocidal Products on the Market. Technical Report EUR
20418 EN/2European Commissions Joint Research Centre, Ispra, Italy.
García-Galán, M.J., Díaz-Cruz, M.S., Barceló, D., 2011. Occurrence of sulfonamide re-
sidues along the Ebro river basin: Removal in wastewater treatment plants and en-
vironmental impact assessment. Environ. Int. 37, 462–473. http://dx.doi.org/10.
1016/j.envint.2010.11.011.
Gargouri, D., Azri, C., Serbaji, M.M., Jedoui, Y., Montacer, M., 2011. Heavy metal con-
centrations in the surface marine sediments of Sfax Coast, Tunisia. Environ. Monit.
Assess. 175, 519–530. http://dx.doi.org/10.1007/s10661-010-1548-7.
Ghosh, G.C., Okuda, T., Yamashita, N., Tanaka, H., 2009. Occurrence and elimination of
M. Harrabi et al.
antibiotics at four sewage treatment plants in Japan and their effects on bacterial
ammonia oxidation. Water Sci. Technol. 59, 779. http://dx.doi.org/10.2166/wst.
2009.067.
Gobel, A., Mcardell, C., Joss, A., Siegrist, H., Giger, W., 2007. Fate of sulfonamides,
macrolides, and trimethoprim in different wastewater treatment technologies. Sci.
Total Environ. 372, 361–371. http://dx.doi.org/10.1016/j.scitotenv.2006.07.039.
Golovko, O., Kumar, V., Fedorova, G., Randak, T., Grabic, R., 2014. Seasonal changes in
antibiotics, antidepressants/psychiatric drugs, antihistamines and lipid regulators in
a wastewater treatment plant. Chemosphere 111, 418–426. http://dx.doi.org/10.
1016/j.chemosphere.2014.03.132.
Gros, M., Petrović, M., Ginebreda, A., Barceló, D., 2010. Removal of pharmaceuticals
during wastewater treatment and environmental risk assessment using hazard in-
dexes. Environ. Int. 36, 15–26. http://dx.doi.org/10.1016/j.envint.2009.09.002.
Gros, M., Rodríguez-Mozaz, S., Barceló, D., 2013. Rapid analysis of multiclass antibiotic
residues and some of their metabolites in hospital, urban wastewater and river water
by ultra-high-performance liquid chromatography coupled to quadrupole-linear ion
trap tandem mass spectrometry. J. Chromatogr. A 1292, 173–188. http://dx.doi.org/
10.1016/j.chroma.2012.12.072.
Gullberg, E., Albrecht, L.M., Karlsson, C., Sandegren, L., Andersson, D.I., 2014. Selection
of a multidrug resistance plasmid by sublethal levels of antibiotics and heavy metals.
mBio 5http://dx.doi.org/10.1128/mBio.01918-14. e01918-14-e01918-14.
Hernando, M., Mezcua, M., Fernandezalba, A., Barcelo, D., 2006. Environmental risk
assessment of pharmaceutical residues in wastewater effluents, surface waters and
sediments. Talanta 69, 334–342. http://dx.doi.org/10.1016/j.talanta.2005.09.037.
Homem, V., Santos, L., 2011. Degradation and removal methods of antibiotics from
aqueous matrices − a review. J. Environ. Manage. 92, 2304–2347. http://dx.doi.
org/10.1016/j.jenvman.2011.05.023.
Houda, B., Dorra, G., Chafai, A., Emna, A., Khaled, M., 2011. Impact of a mixed “in-
dustrial and domestic” wastewater effluent on the southern coastal sediments of Sfax
(Tunisia) in the Mediterranean sea. Int. J. Environ. Res. 5 (3), 691–704. http://dx.
doi.org/10.22059/ijer.2011.376.
Hu, X., Zhou, Q., Luo, Y., 2010. Occurrence and source analysis of typical veterinary
antibiotics in manure, soil, vegetables and groundwater from organic vegetable
bases, Northern China. Environ. Pollut. 158, 2992–2998. http://dx.doi.org/10.1016/
j.envpol.2010.05.023.
Huang, J., 2001. Global analysis of growth phase responsive gene expression and reg-
ulation of antibiotic biosynthetic pathways in Streptomyces coelicolor using DNA
microarrays. Genes Dev. 15, 3183–3192. http://dx.doi.org/10.1101/gad.943401.
INC (Institut National de la Consommation), 2016. Rapport scientifique de la journée
mondiale 2016 des droits des consommateurs « Aliments sans ATB » ou « Du mésusage
des ATB ».
Karthikeyan, K.G., Meyer, M.T., 2006. Occurrence of antibiotics in wastewater treatment
facilities in Wisconsin. USA. Sci. Total Environ. 361, 196–207. http://dx.doi.org/10.
1016/j.scitotenv.2005.06.030.
Khoufi, S., Aloui, F., Sayadi, S., 2009. Pilot scale hybrid process for olive mill wastewater
treatment and reuse. Chem. Eng. Process. Process Intensif. 48, 643–650. http://dx.
doi.org/10.1016/j.cep.2008.07.007.
Kim, J.W., Yoon, S.M., Lee, S.J., Narumiya, M., Nakada, N., Han, I.S., Tanaka, H., 2012.
Occurrence and fate of PPCPs wastewater treatment plants in Korea. In: 2nd
International Conference on Environment and Industrial Innovation. Singapore. pp.
57–61.
Kim, S.-C., Carlson, K., 2007. Temporal and spatial trends in the occurrence of human and
veterinary antibiotics in aqueous and river sediment matrices. Environ. Sci. Technol.
41, 50–57.
Kinney, C.A., Furlong, E.T., Werner, S.L., Cahill, J.D., 2006. Presence and distribution of
wastewater-derived pharmaceuticals in soil irrigated with reclaimed water. Environ.
Toxicol. Chem. 25, 317. http://dx.doi.org/10.1897/05-187R.1.
Knapp, C.W., McCluskey, S.M., Singh, B.K., Campbell, C.D., Hudson, G., Graham, D.W.,
2011. Antibiotic resistance gene abundances correlate with metal and geochemical
conditions in archived Scottish soils. PLoS One 6, e27300. http://dx.doi.org/10.
1371/journal.pone.0027300.
Ksibi, M., 2006. Chemical oxidation with hydrogen peroxide for domestic wastewater
treatment. Chem. Eng. J. 119, 161–165. http://dx.doi.org/10.1016/j.cej.2006.03.
022.
Le Bris, H., Pouliquen, H., 2004. Experimental study on the bioaccumulation of oxyte-
tracycline and oxolinic acid by the blue mussel (Mytilus edulis). An evaluation of its
ability to bio-monitor antibiotics in the marine environment. Mar. Pollut. Bull. 48,
434–440. http://dx.doi.org/10.1016/j.marpolbul.2003.08.018.
Lin, A.Y.-C., Yu, T.-H., Lateef, S.K., 2009. Removal of pharmaceuticals in secondary
wastewater treatment processes in Taiwan. J. Hazard. Mater. 167, 1163–1169.
http://dx.doi.org/10.1016/j.jhazmat.2009.01.108.
Llorca, M., Gros, M., Rodríguez-Mozaz, S., Barceló, D., 2014. Sample preservation for the
analysis of antibiotics in water. J. Chromatogr. A 1369, 43–51. http://dx.doi.org/10.
1016/j.chroma.2014.09.089.
Louati, A., Elleuch, B., Kallel, M., Saliot, A., Dagaut, J., Oudot, J., 2001. Hydrocarbon
contamination of coastal sediments from the Sfax Area (Tunisia), Mediterranean Sea.
Mar. Pollut. Bull. 42, 444–451. http://dx.doi.org/10.1016/S0025-326X(00)00179-X.
Lucas, D., Barceló, D., Rodriguez-Mozaz, S., 2016. Removal of pharmaceuticals from
wastewater by fungal treatment and reduction of hazard quotients. Sci. Total
Environ. 571, 909–915. http://dx.doi.org/10.1016/j.scitotenv.2016.07.074.
Maalej, S., Mahjoubi, A., Elazri, C., Dukan, S., 2003. Simultaneous effects of environ-
mental factors on motile Aeromonas dynamics in an urban effluent and in the natural
seawater. Water Res. 37, 2865–2874. http://dx.doi.org/10.1016/S0043-1354(03)
00117-9.
Maatoug, J., El Ayari, T., Kefi, F.J., Abdallah, L.G.B., El Menif, N.T., 2009. Premiere
description de malformation du pied chez le mollusque bivalve ruditapes decussatus
169
Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170
(linne 1758) preleve sur les cotes tunisiennes.
Meylan, W.M., Howard, P.H., 1998. User’s Guide for the ECOSAR Class Program. MS-
Window Version 099. Syracuse Res. Corp., North Syracuse NY.
Miao, Z., Xu, D., Ouyang, J., Guo, G., Zhao, X., Tang, Y., 2002. Electrochemically induced
sol-gel preparation of single-crystalline TiO2 nanowires. Nano Lett. 2, 717–720.
http://dx.doi.org/10.1021/nl025541w.
Mkawar, S., Azri, C., Kamoun, F., Montacer, M., 2007. Impact sur les biophases marines
des rejets anthropiques, notamment des métaux lourds rejetés sur le littoral nord de la
ville de Sfax (Tunisie). Tech. Sci. Méthodes 71–85. http://dx.doi.org/10.1051/tsm/
200710071.
Na, G., Fang, X., Cai, Y., Ge, L., Zong, H., Yuan, X., Yao, Z., Zhang, Z., 2013. Occurrence,
distribution, and bioaccumulation of antibiotics in coastal environment of Dalian,
China. Mar. Pollut. Bull. 69, 233–237. http://dx.doi.org/10.1016/j.marpolbul.2012.
12.028.
Negreanu, Y., Pasternak, Z., Jurkevitch, E., Cytryn, E., 2012. Impact of treated waste-
water irrigation on antibiotic resistance in agricultural soils. Environ. Sci. Technol.
46, 4800–4808. http://dx.doi.org/10.1021/es204665b.
Ouali, A., Azri, C., Medhioub, K., Ghrabi, A., 2009. Descriptive and multivariable analysis
of the physico-chemical and biological parameters of Sfax wastewater treatment
plant. Desalination 246, 496–505. http://dx.doi.org/10.1016/j.desal.2008.04.058.
Pan, M., Wong, C.K.C., Chu, L.M., 2014. Distribution of antibiotics in wastewater-irri-
gated soils and their accumulation in vegetable crops in the Pearl River Delta,
Southern China. J. Agric. Food Chem. 62, 11062–11069. http://dx.doi.org/10.1021/
jf503850v.
Papageorgiou, M., Kosma, C., Lambropoulou, D., 2016. Seasonal occurrence, removal,
mass loading and environmental risk assessment of 55 pharmaceuticals and personal
care products in a municipal wastewater treatment plant in Central Greece. Sci. Total
Environ. 543, 547–569. http://dx.doi.org/10.1016/j.scitotenv.2015.11.047.
Petrovic, M., Verlicchi, P., 2014. Water treatment plants and pharmaceutical residues in
Catalonia and Italy. Contrib. Sci. 135–150.
Proia, L., von Schiller, D., Sànchez-Melsió, A., Sabater, S., Borrego, C.M., Rodríguez-
Mozaz, S., Balcázar, J.L., 2016. Occurrence and persistence of antibiotic resistance
genes in river biofilms after wastewater inputs in small rivers. Environ. Pollut. 210,
121–128. http://dx.doi.org/10.1016/j.envpol.2015.11.035.
Pruneau, M., Mitchell, G., Moisan, H., Dumont-Blanchette, É., Jacob, C.L., Malouin, F.,
2011. Transcriptional analysis of antibiotic resistance and virulence genes in multi-
resistant hospital-acquired MRSA. FEMS Immunol. Med. Microbiol. 63, 54–64.
http://dx.doi.org/10.1111/j.1574-695X.2011.00830.x.
Richardson, B.J., Lam, P.K.S., Martin, M., 2005. Emerging chemicals of concern:
Pharmaceuticals and personal care products (PPCPs) in Asia, with particular re-
ference to Southern China. Mar. Pollut. Bull. 50, 913–920. http://dx.doi.org/10.
1016/j.marpolbul.2005.06.034.
Rincon, R.F., Fatoyinbo, T., Ranson, K.J., Osmanoglu, B., Sun, G., Deshpande, M., Perrine,
M., Toit, C.D., Bonds, Q., Beck, J., Lu, D., 2014. The ecosystems SAR (EcoSAR) an
airborne P-band polarimetricInSAR for the measurement of vegetation structure,
biomass and permafrost. IEEE 1443–1445. http://dx.doi.org/10.1109/RADAR.2014.
6875827.
Rodriguez-Mozaz, S., Chamorro, S., Marti, E., Huerta, B., Gros, M., Sànchez-Melsió, A.,
Borrego, C.M., Barceló, D., Balcázar, J.L., 2015. Occurrence of antibiotics and anti-
biotic resistance genes in hospital and urban wastewaters and their impact on the
receiving river. Water Res. 69, 234–242. http://dx.doi.org/10.1016/j.watres.2014.
11.021.
Rodriguez-Mozaz, S., Alvarez-Muñoz, D., Barcelo, D., 2017. Pharmaceuticals in Marine
Environment: Analytical Techniques and Applications. In: Barrera, Tamara García,
Gómez Ariza, José Luis (Eds.), Environmental Problems in Marine Biology:
Methodological Aspects and Applications. Taylor & Francis Publisher. CRC Press,
6000 Broken Sound Parkway NW, Suite 300, Boca Raton, FL 33487-2742, pp.
268–316.
Salem, Z.B., Drira, Z., Ayadi, H., 2015. What factors drive the variations of phyto-
plankton, ciliate and mesozooplankton communities in the polluted southern coast of
Sfax, Tunisia? Environ. Sci. Pollut. Res. 22, 11764–11780. http://dx.doi.org/10.
1007/s11356-015-4416-8.
Sanderson, H., 2003. Probabilistic hazard assessment of environmentally occurring
pharmaceuticals toxicity to fish, daphnids and algae by ECOSAR screening. Toxicol.
Lett. 144, 383–395. http://dx.doi.org/10.1016/S0378-4274(03)00257-1.
Schwartz, T., Kohnen, W., Jansen, B., Obst, U., 2003. Detection of antibiotic-resistant
bacteria and their resistance genes in wastewater, surface water, and drinking water
biofilms. FEMS Microbiol. Ecol. 43, 325–335. http://dx.doi.org/10.1111/j.1574-
6941.2003.tb01073.x.
Serra-Compte, A., Álvarez-Muñoz, D., Rodríguez-Mozaz, S., Barceló, D., 2017. Multi-re-
sidue method for the determination of antibiotics and some of their metabolites in
seafood. Food Chem. Toxicol. 104, 3–13. http://dx.doi.org/10.1016/j.fct.2016.11.
031.
Tahrani, L., Van Loco, J., Ben Mansour, H., Reyns, T., 2016. Occurrence of antibiotics in
pharmaceutical industrial wastewater, wastewater treatment plant and sea waters in
Tunisia. J. Water Health 14, 208–213. http://dx.doi.org/10.2166/wh.2015.224.
Teixeira, P., Tacão, M., Alves, A., Henriques, I., 2016. Antibiotic and metal resistance in a
ST395 Pseudomonas aeruginosa environmental isolate: a genomics approach. Mar.
Pollut. Bull. 110, 75–81. http://dx.doi.org/10.1016/j.marpolbul.2016.06.086.
Travis, M.J., Wiel-Shafran, A., Weisbrod, N., Adar, E., Gross, A., 2010. Greywater reuse
for irrigation: effect on soil properties. Sci. Total Environ. 408, 2501–2508. http://dx.
doi.org/10.1016/j.scitotenv.2010.03.005.
Tünger, Ö., Dinç, G., Özbakkaloglu, B., Atman, Ü.C., Algün, Ü., 2000. Evaluation of ra-
tional antibiotic use. Int. J. Antimicrob. Agents 15, 131–135. http://dx.doi.org/10.
1016/S0924-8579(00)00158-8.
Verlicchi, P., Galletti, A., Petrovic, M., Barceló, D., 2010. Hospital effluents as a source of
M. Harrabi et al.
emerging pollutants: an overview of micropollutants and sustainable treatment op-
tions. J. Hydrol. 389, 416–428. http://dx.doi.org/10.1016/j.jhydrol.2010.06.005.
Vieno, N.M., Härkki, H., Tuhkanen, T., Kronberg, L., 2007. Occurrence of pharmaceu-
ticals in river water and their elimination in a pilot-scale drinking water treatment
plant. Environ. Sci. Technol. 41, 5077–5084. http://dx.doi.org/10.1021/es062720x.
Watkinson, A.J., Murby, E.J., Kolpin, D.W., Costanzo, S.D., 2009. The occurrence of an-
tibiotics in an urban watershed: from wastewater to drinking water. Sci. Total
Environ. 407, 2711–2723. http://dx.doi.org/10.1016/j.scitotenv.2008.11.059.
Wille, K., Kiebooms, J.A.L., Claessens, M., Rappé, K., Vanden Bussche, J., Noppe, H., Van
Praet, N., De Wulf, E., Van Caeter, P., Janssen, C.R., De Brabander, H.F., Vanhaecke,
L., 2011. Development of analytical strategies using U-HPLC-MS/MS and LC-ToF-MS
for the quantification of micropollutants in marine organisms. Anal. Bioanal. Chem.
400, 1459–1472. http://dx.doi.org/10.1007/s00216-011-4878-6.
Wu, M., Que, C., Tang, L., Xu, H., Xiang, J., Wang, J., Shi, W., Xu, G., 2016. Distribution,
170
Environmental Nanotechnology, Monitoring & Management 10 (2018) 163–170
fate, and risk assessment of antibiotics in five wastewater treatment plants in
Shanghai, China. Environ. Sci. Pollut. Res. 23, 18055–18063. http://dx.doi.org/10.
1007/s11356-016-6946-0.
Wu, X., Dodgen, L.K., Conkle, J.L., Gan, J., 2015. Plant uptake of pharmaceutical and
personal care products from recycled water and biosolids: a review. Sci. Total
Environ. 536, 655–666. http://dx.doi.org/10.1016/j.scitotenv.2015.07.129.
Zaghden, H., Kallel, M., Louati, A., Elleuch, B., Oudot, J., Saliot, A., 2005. Hydrocarbons
in surface sediments from the Sfax coastal zone, (Tunisia) Mediterranean Sea. Mar.
Pollut. Bull. 50, 1287–1294. http://dx.doi.org/10.1016/j.marpolbul.2005.04.045.
Zhang, Y., Tian, Z., Liu, M., Shi, Z.J., Hale, L., Zhou, J., Yang, M., 2015. High con-
centrations of the antibiotic spiramycin in wastewater lead to high abundance of
ammonia-oxidizing archaea in Nitrifying Populations. Environ. Sci. Technol. 49,
9124–9132. http://dx.doi.org/10.1021/acs.est.5b01293.