Bioresource Technology 245 (2017) 1238–1244

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Review

Anaerobic treatment of pharmaceutical wastewater: A critical review T

⁎
Xueqing Shi, Kwok Yii Leong, How Yong Ng
Centre for Water Research, Department of Civil and Environmental Engineering, National University of Singapore, 1 Engineering Dr. 2, Singapore 117576, Singapore

G RA P H I C A L AB S T R A C T

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

Keywords: Pharmaceutical wastewaters are usually produced by chemical-synthetic process, and thus contain high levels of
Anaerobic technology organic pollutants, biotoxicity and salinity. Anaerobic technology is a viable option for treating pharmaceutical
Antibiotics wastewater owing to its advantages of withstanding high organic-loading, less sludge production and lower
Microorganism operating cost as compared with conventional activated sludge process. In this paper, several types of modern
Pharmaceutical wastewater
anaerobic or hybrid systems were reviewed on their pollutant reduction performance and operating conditions
for treating pharmaceutical wastewater. Meanwhile, the typical predominant microbial populations found in
anaerobic process treating pharmaceutical wastewater were summarized. Moreover, the environmental impact
of antibiotic residues and health risk of spreading of antibiotic resistant genes (ARGs) were also assessed to offer
an in-depth understanding of the growing concern on the discharge of treated pharmaceutical effluent.

1. Introduction
Pharmaceutical manufacturing processes can be divided into five
categories, namely, fermentation, extraction, chemical synthesis, for-
mulation and packaging (Ince et al., 2002). Among them, chemical
synthesis and fermentation processes generate larger amount of was-
tewater which usually contains high levels of spent solvents, re-
calcitrant organics, residue pharmaceuticals as well as salts (Chen et al.,
2008). Table 1 lists the general characterization of pharmaceutical
wastewater that generated from several manufacturing plants
according to the existing literature. It was found that although most of
the wastewaters contain high amounts of COD, the variation between
different manufacturing activities can still be huge, where the raw
wastewater COD ranges between 4410 and 40000 mg/L (Cetecioglu
et al., 2015; Oktem et al., 2006). In addition, high levels of nitrogenous
compounds are commonly found in the pharmaceutical wastewaters,
which could be explained by the frequent use of nitrogen-containing
organics as raw material for the manufacturing process (Shi et al.,
2014). Wastewater-derived dissolved organic nitrogen (DON) is very
complex in nature, with potential toxicity that may inhibit biological

⁎
Corresponding author.
E-mail address: howyongng@nus.edu.sg (H.Y. Ng).

http://dx.doi.org/10.1016/j.biortech.2017.08.150
Received 6 July 2017; Received in revised form 22 August 2017; Accepted 23 August 2017
Available online 30 August 2017
0960-8524/ © 2017 Elsevier Ltd. All rights reserved.
X. Shi et al. Bioresource Technology 245 (2017) 1238–1244

Nomenclature MBR membrane bioreactor

MBBR moving bed biofilm bioreactor
6-APA 6-Aminopenicillanic acid NHAR novel micro-aerobic hydrolysis acidification reactor
ABR anaerobic baffled reactor O3 ozonation
AF anaerobic filter OFX ofloxacin
AFFBR anaerobic fixed film fixed bed reactor OLR organic loading rate (kg COD/m3d)
AMCBR anaerobic multi-chamber bed reactor OTC oxytetracycline
AnBEMR anaerobic bio-entrapped membrane reactor PAD psychrophilic anaerobic digestion
AnMBR anaerobic membrane bioreactor PAH polycyclic aromatic hydrocarbon
ANOVA analysis of variance PW pharmaceutical wastewater
AnSBR anaerobic sequencing membrane bioreactor PWWTP pharmaceutical wastewater treatment plant
AOP advanced oxidation process SBA sequential biocatalysts addition
ARG antibiotic resistance gene SBR sequencing batch reactor
BASR biofilm airlift suspension reactor SMA specific methanogenic activity
BCOT biological contact oxidation tank SMX sulfamethoxazole
CASS cyclic activated sludge system SO24− sulphate
CSTR continuous stirred tank reactor SRT sludge retention time (h or day)
COD chemical oxygen demand (mg/L) TCOD total chemical oxygen demand (mg/L)
DO dissolved oxygen (mg/L) TDS total dissolved solids (mg/L)
DOM dissolved organic matter (mg/L) TKN total kjeldahl nitrogen (mg/L)
DON dissolved organic nitrogen (mg/L) TN total nitrogen (mg/L)
EC electrocoagulation TP total phosphorus (mg/L)
EC50 half maximal effective concentration TPAD two-phase anaerobic digestion
EGSB expended granular sludge blanket TSS total suspended solids (mg/L)
FQ fluoroquinolone UASB upflow anaerobic sludge blanket
H2O2 hydrogen peroxide UASR upflow anaerobic stage reactor
HA hydrolysis/acidification VFA volatile fatty acid
HRT hydraulic retention time (h or day) VSS volatile suspended solids (mg/L)
IC50 half maximal inhibitory concentration WOS waste organic solvent
IWS intertidal wetland sediment WWTP waste water treatment plant

treatment efficiency (Hu et al., 2017). On the other hand, high salinity
level could be another major challenge in treating pharmaceutical
wastewater. The salinity inhibition on microbial activity is attributed to
the unbalanced osmotic potential across cell wall, which in turn cause
water loss and eventually kill the cell (Shi et al., 2015).
Moreover, one growing concern in recent years with pharmaceutical
wastewater treatment and discharge is the health risk of emission of
antibiotic residues into natural environments, since common biological
treatment has limited removal efficiency on antibiotics due to their
antimicrobial activity (Aydin, 2016). Given the high organic-strength
nature, anaerobic technology is a preferred treatment option for phar-
maceutical wastewater considering its advantages such as withstanding

Table 1
Characterization of pharmaceutical wastewaters.

Type of pharmaceutical wastewater COD (mg/L) TN/TKN (mg/L) TP (mg/L) TSS (mg/L) TDS (mg/L) pH References

Antibiotics waste 15365 ± 1214 1422 ± 173 1763 ± 36.6 388 ± 87 22168 ± 3757 7–8 Ng et al. (2014)
Herbal pharmaceutical factory 5000 ± 80000 135–1250 30–120 900–18800 – 4.2–4.5 Nandy and Kaul (2001)
Chemical synthesis-based pharmaceutical 40000 ± 60000 800–900 3–6 0.6–0.7 900–1000 5.5 ± 0.1 Oktem et al. (2006,
factory 2007)
Antibiotic production factory 10000–43000 – – 120–580 – – Deǧirmentaş and Deveci
(2004)
Generated from synthetic pharmaceutical waste 4400 – – 81.2 ± 33.8 6.8–7.2 Cetecioglu et al. (2015)
Chemical synthesis-based pharmaceutical waste 39000 ± 60000 1010–1575 3–6 800–1000 – 7–8 Ince et al. (2002)
Fermentation wastewater 20.140 2570 420 – – 6.42 Chen et al. (2014)
Antibiotic waste 16249 ± 714 1612 ± 353 188 ± 29 99 ± 59 29450 ± 1209 7.02 Ng et al. (2015)
Penicillin G pharmaceutical industry 12500 ± 1070 1250 ± 25 38 ± 1.9 871 ± 87 – 7.5 ± 0.3 Rodríguez-Martinez et al.
(2005)
Brewery waste pharmaceutical industry 7000 ± 800 364 ± 50 – – – 5.2–6.8 Chelliapan et al. (2006,
2011)
Product manufacturing and equipment cleaning 20000 364 – 765 – 7.4 Chen et al. (2011 a)
Antibiotic waste 15476 ± 1614 1472 ± 453 – – 26450 ± 1732 7.02 Ng et al. (2016)
Antibiotics waste from manufacturing and 16547 ± 1827 1568 ± 314 – 285 ± 175 24899 ± 1758 7.26 Shi et al. (2014)
equipment cleaning industry
Bulk drug pharmaceutical industry 13000–15000 120–170 100–120 2800–3000 8500–9000 7.0–7.5 Sreekanth et al. (2009)
Antibiotic pharmaceutical waste 3000 ± 450 98–135 15–20 3400–4100 – 6.99–7.59 Sponza and Demirden
(2010)
Chemical synthesis pharmaceutical wastewater 16000 ± 23000 32–36 0.5–22 – – 3.3–3.7 Kaya et al. (2017)
Bulk drug manufacturing industry 34400 ± 2000 370 ± 50 – 6250 ± 200 15000 7.2 ± 0.3 Deshpande et al. (2010,
2012)
Product manufacturing and equipment cleaning 5000–60000 560–980 51.41–120.4 600–2000 – 4.3–7.2 Chen et al. (2008)
Fermentation-based pharmaceutical wastewater 6800.5 251.8 – 188.3 – 6–7 Xing et al. (2014)

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 X. Shi et al.

Table 2
Summary of anaerobic and hybrid technologies for pharmaceutical wastewater treatment.

Type of pharmaceutical wastewater Technology COD (mg/L) Target pharmaceutical compound OLR (kg COD/ HRT SRT COD removal Target pharmaceutical References
m3d) (%) compound removal (%)

Product manufacturing and equipment UASB 20000 6-APA (192 mg/L); Amoxicillin 2.7–7.2 32–38.5 h – 52.2 6-APA: 26.3; Amoxicillin: 21.6 Chen et al. (2011a)
cleaning (real) (92 mg/L)
Brewery waste pharmaceutical UASR 7000 ± 800 Tylosin (20–200 mg/L); 0.43–3.73 2–4 d – 93 Tylosin: 95 Chelliapan et al.
industry (real) Avilamycin (2006)
(not determined)
Synthetic antibiotic waste (synthetic) ABR 2500 Erythromycin (0.1–2.5 mg/L); 2.5 2.5 d 30 d 68 Erythromycin: 40; Aydin et al. (2014)
Sulfamethaxazole Sulfamethaxazole: 37
(0.5–25 mg/L)
Synthetic antibiotic pharmaceutical ABR+CSTR 3000 Kemicetine (32, 49, 125 mg/L) 3.6 0.83 d 20 d 98 Kemicitine: 100 Sponza and
waste (synthetic) Demirden (2010)
Synthetic pharmaceutical wastewater AMCBR+CSTR 3000 ± 45 Oxytetracycline (< 350 mg/L) 2 × 10−4 1.2–2 d 45–98 d 95 Oxytetracycline: 99 Sponza and Ҫelebi
(synthetic) (2011)
6-APA and amoxicillin waste(real) UASB+NHAR+CASS 63–13093 Amoxicillin (92.2 mg/L) – 1.35–23.5 h – 97 Amoxicillin: 97.2 Chen et al. (2011b)
+BCOT+Clarifier
Antibiotics waste from manufacturing UASB+MBR/SBR 16547 ± 1827 Penicillin (with different salinity 4–11 36–96 h – 94.7 – Shi et al. (2014)
and equipment cleaning industry concentration from 25 g/L)
(real)
Synthetic pharmaceutical wastewater UASB+CSTR 3000 Sulfamerazine (10–90 mg/L) 3.6 0.83 20 d 71–84 97–100 Sponza and

1240
(synthetic) Demirden, (2007)
Ampicillin- and aureomycin-bearing ABR+BASR 1300–14000 Ampicillin(3.2 mg/L); – 1.25–12.5 d – 89.6 Ampicillin: 31.3; Zhou et al. (2006)
pharmaceutical wastewater (real) Aureomycin Aureomycin: < 10
(1.0 mg/L)
Chemical synthesis pharmaceutical Ozonation+AnMBR 16000–23000 Etodolac (50–215 mg/L) – – – 90 Etodolac: 99 Kaya et al. (2017)
wastewater (real)
Synthetic antibiotic pharmaceutical AnSBR – Sulfamethoxazole Erythromycin 2.5 – – – 85–94 Aydin (2016)
waste (synthetic) Tetracycline
Antibiotic waste (real) AnMBR 16249 ± 714 Penicillin 13.0 ± 0.6 30.6 h 100 d 46.1–60.3 – Ng et al. (2015)
Penicillin G pharmaceutical industry UASB 12500 ± 1070 Penicillin G 0.07–2.09 7d – 85–90 – Rodríguez-Martinez
(real) et al. (2005)
Brewery waste pharmaceutical AnSBR 17200 Erythromycin (1–500 mg/L) 0.4–3 6–42 d – 94.7–99.4 – Amin et al. (2006)
industry (synthetic)
Antibiotic synthetic waste (synthetic) AnSBR – Sulfamethaxazole (0.5–40 mg/L); – – – – – Aydin et al.
Erythromycin (2015 c)
(0.1–4 mg/L); Tetracycline
(0.1–4 mg/L)
Synthetic pharmaceutical waste EGSB 5000 Propanol (30–150 mM); 2.5–20 6–48 h – 60–70 – Enright et al.
(synthetic) Methanol (2005)
(30–3750 mM); Acetone
(30–750 mM)
Bioresource Technology 245 (2017) 1238–1244
X. Shi et al.
of high organic-loading, less sludge production and lower operating
cost as compared with conventional activated sludge process (Ji et al.,
2013a, b). In this paper, application potential of various anaerobic and
anaerobic-hybrid technologies is reviewed, and pharmaceutical waste-
water associated with environmental issues were documented to offer
an in-depth understanding of treatment of this specific industrial ef-
fluent.
2. Anaerobic technologies for pharmaceutical wastewater
treatment
Several modern anaerobic technologies have shown their cap-
abilities in efficient treatment of pharmaceutical, including anaerobic
membrane bioreactor (AnMBR), upflow anaerobic sludge blanket
(UASB), anaerobic sequencing batch reactor (AnSBR), moving bed
biofilm reactor (MBBR) and other hybrid technologies. Table 2 shows a
summary of anaerobic and hybrid technologies for pharmaceutical
wastewater treatment.
2.1. Anaerobic membrane bioreactor (AnMBR)
The development of AnMBR provides a promising solution because
AnMBR coupled with the anaerobic process and the membrane filtra-
tion to produce a solid-free effluent by the complete retention of bio-
mass (Ng et al., 2014). AnMBR also contributes an important role in
energy generation due to its ability to produce methane from the uti-
lization of a large fraction of organics in wastewaters (Skouteris et al.,
2012). Furthermore, footprint and space can be reduced while produ-
cing an effluent of highly-improved quality (Ng et al., 2010).
For the treatment of chemical synthetic-based pharmaceutical
wastewater, a comparison was made between AnMBR and anaerobic
bio-entrapped membrane reactor (AnBEMR) at different organic
loading rates (OLRs) and membrane fouling behavior (Ng et al., 2014).
Despite the many advantages offer by the AnMBR, membrane fouling is
still a major concern especially with high biomass concentration in the
AnMBR. The entrapped biomass has shown to offer great advantage
such as being able to reduce membrane fouling due to lower production
of abundant soluble organics and suspended biomass concentration in
the system. Decreasing the hydraulic retention time (HRT) or increasing
the OLR resulted in lower organic removal efficiencies and methane
yields from the AnMBR and AnBEMR. The high salinity of the chemical
synthetic-based pharmaceutical wastewater also attributed to the
overall lower treatment performances for the AnMBR and AnBEMR.
Overall, the results show that the AnBEMR has a better treatment
performance by having 5–10% total chemical oxygen demand (TCOD)
removal and around 15% higher methane yield as compared to that of
the AnMBR.
2.2. Upflow anaerobic sludge blanket (UASB)
Upflow anaerobic sludge blanket (UASB) is by far the most widely
used high-rate anaerobic system for domestic and industrial wastewater
treatment due to its high biomass concentration, microbial diversity,
low cost, flexibility and ability to withstand fluctuation of pH and
temperature (Chen et al., 2014).
For the treatment of diluted pharmaceutical fermentation waste-
water, it was found that increasing the OLR from 2.7 to 7.2 kg COD/
m3/d led to an increase in COD removal from 83% to 91% for a con-
tinuous operation of 140 days (Chen et al., 2014). However, for phar-
maceutical penicillin G producing wastewater, the COD removal was
negatively affected when the OLR was increased from 1.5 to
2.09 kg COD/m3/d. The overall reactor performance was affected as
well. It was discovered that at 2.09 kg COD/m3, accumulation of sul-
fides was observed that hampered the methanogenic activity due to
having pH ≥8, which allows sulfate reducing bacteria to thrive over the
methanogens in the UASB (Rodríguez-Martinez et al., 2005). Similar
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Bioresource Technology 245 (2017) 1238–1244
observation was also reported by Li et al. (2015) in the treatment of
chemical synthesis-based pharmaceutical wastewater. They also high-
lighted the importance of the COD to sulphate SO24− ratio that could
explain the ups and downs of the treatment performance of the UASB.
As the ratio of COD/SO24− decreased from 5 to 1.5, significant reduc-
tions in the removal of COD and methane content production were
observed.
Not only UASB is used for the treatment of pharmaceutical waste-
water operated in lab-scale or bench-scale, but also as pre-treatment of
pharmaceutical wastewater. This pre-treatment step was applied for
pharmaceutical wastewater containing 6-aminopenicillanic acid (6-
APA) and amoxicillin under high OLRs ranging from 12.57 to
21.02 kg COD/m3/d and sharp pH fluctuations between 5.57 and 8.26.
The results show that the UASB was effective in the removal of COD, 6-
APA and amoxicillin, and able to keep pH at a narrow range of
7.18–7.22 (Chen et al., 2011a). Yi et al. (2017) used hydrolysis as their
pretreatment for antibiotic production wastewater before subjecting to
the UASB system. The initial batch test studies showed that the sup-
pression of anaerobic digestion activity by tetracycline could be re-
moved after the enhanced hydrolysis, which was evident for using
hydrolysis as a pretreatment approach for the biological treatment of
antibiotic production wastewater.
Expended Granular Sludge Blanket (EGSB) reactor is a family of the
UASB reactor. The definitive feature of EGSB reactor is the rapid upflow
velocity which enables the reactor to separate the dispersed sludge from
mature granules out of the reactor. Low temperature or psychrophilic
(< 20 °C) anaerobic digestion (PAD) using EGSB were carried out by
treating synthetic pharmaceutical wastewater (propanol, methanol and
acetone) at 15 °C (Enright et al., 2005). They found that the COD re-
moval efficiencies was about 60–70% with OLRs of 5–20 kg COD/m3/d
during the 450-day trial. Apart of that, a full-scale oxytetracycline
production wastewater treatment system in factory in Shijiazhuang
city, Hebei Province, China using EGSB and aerobic reactors in suc-
cession were also operated (Yi et al., 2017).
Meanwhile, the upflow anaerobic stage reactor (UASR) is similar in
design and application to the anaerobic baffled reactor (ABR) which
consists of a number of UASB reactor connected in series. The design of
UASR allows the separation of acidogenesis and methanogenesis, which
has potential benefits for reactor performance. UASR could help reduce
biomass washout (i.e., retaining a high active biomass content), allow
quick recovery from hydraulic and organic shock loads and do not re-
quire special gas or sludge separation equipment. Initially, the feasi-
bility of UASR was carried out to test this system as a pretreatment
option for pharmaceutical wastewater containing tylosin and avila-
mycin antibiotics (Chelliapan et al., 2006). Effects of different OLR on
the performance of UASR system were also tested to comprehend the
toleration of the methanogenic sludge and assessed the stability of the
reactor (Chelliapan et al., 2011). It was found that good COD removal
efficiencies (70–75%) were obtained at low OLRs. However, with in-
creasing OLRs up to 3.73 kg COD/m3/d and reduction of HRT from 4 to
2 days, the COD removal efficiency was found to plummet to 45%.
2.3. Anaerobic sequencing batch reactor (AnSBR)
The anaerobic sequencing batch reactor (AnSBR) is a type of acti-
vated sludge process whereby five stages of process occur in a single
reactor, namely fill, react, settle, decant and idle in respective time ratio
without the presence of oxygen under dark condition. The effect of
different doses of antibiotic sulfamethoxazole (SMX) on the utilization
of volatile fatty acids (VFA) and other parameters were studied using
AnSBR (Cetecioglu et al., 2015). Their findings suggested that anae-
robic treatment was suitable for pharmaceutical wastewater with con-
centration up to 40 mg/L of SMX. Higher SMX levels exerted toxic ef-
fects on the microbial community, causing the inhibition of substrate/
COD utilization and biogas production and leading to failure of reactor
operations. In addition, the effect of different doses of antibiotic SMX
X. Shi et al.
was investigated against the composition of microbial community using
AnSBR (Cetecioglu et al., 2016). It was concluded that the microbial
community maintains the system stability under high antibiotic con-
centration and long-term operation by homoacetogenesis coupled with
hydrogenotrophic methanogenesis.
2.4. Hybrid anaerobic + aerobic systems
Although single anaerobic system can generally achieve efficient
COD reduction of pharmaceutical wastewater, the performance varies
greatly. The difference in the anaerobic treatment performance could
be explained by the different wastewater characteristics, which include
salinity, toxicity and refractory compounds that are hardly biodegrad-
able under anaerobic conditions. Therefore, a combination of anaerobic
and aerobic process is found to be a viable and promising option for
more complete oxidation and stable treatment performance. Basically, a
sequential anaerobic-aerobic process consists of an anaerobic stage
where most of the organic pollutants are removed, and an aerobic stage
as the polishing step to further remove the organic residues in the
anaerobic effluent, as well as achieving ammonia reduction through
nitrification. According to the literature, sequential UASB + aerobic
continuous stirred tank reactor (CSTR) is found as a popular combi-
nation for pharmaceutical wastewater treatment. As reported by
Sponza and Demirden (2007), the treatability of sulfamerazine in a
sequential UASB and CSTR achieved COD removal efficiency of 97%
with an initial sulfamerazine concentration of 90 mg/L. A few years
later, they also found that COD originating from the readily degradable
organics did not limit the anaerobic biodegradation process, while the
CODs originating from the slowly degradable organics and from the
inert microbial products significantly decreased the anaerobic ABR re-
actor performance. The statistical ANOVA test also revealed that there
was a strong linear correlation between acute toxicity, CODs originating
from the slowly degradable organics and inert microbial products,
while weak correlation was found between acute toxicity and CODs
originating from the inert compounds was detected (Sponza and
Demirden, 2010).
A further extension of CSTR combined with UASB-AF, which is
known as the two-phase anaerobic digester, was studied by Demirer
and Chen (2005). The group investigated different growth rates and
optimum pH required for acidogenic and methanogenic microorgan-
isms and different requirements regarding reactor conditions using this
hybrid UASB system. However, to meet the discharge requirements, a
subsequent aerobic process is desired and MBR was found to be efficient
for meeting the discharge limit. Hence, TPAD followed by sequential
MBR were carried out on pharmaceutical wastewater with COD as high
as 58000 mg/L, and as a result, almost all the COD (∼99%) was re-
moved by this hybrid system (Chen et al., 2008). Similar results were
obtained by Shi et al. (2014), whom investigated the treatment of high
saline pharmaceutical wastewater by combining the anaerobic (UASB)
and sequential aerobic treatment (MBR or SBR). COD removal effi-
ciencies for single UASB, UASB+MBR and UASB+SBR were reported
to be 41.3%, 94.7% and 91.8%, respectively.
2.5. Hybrid anaerobic + chemical/physical systems
Apart of hybrid anaerobic systems which employ the biological
processes, other hybrid anaerobic systems would also take into account
of processes other than biological such as chemical and physical pro-
cesses to achieve enhanced treatment performance. For the purpose of
simultaneous removal of multiple pollutants, a novel combined process
comprised of a UASB, a novel micro-aerobic hydrolysis acidification
reactor (NHAR), a cyclic activated sludge system (CASS) and a biolo-
gical contact oxidation tank (BCOT) was investigated to treat both or-
ganics and ammonia in chemical synthesis-based pharmaceutical was-
tewater (Chen et al., 2011b). It was found that the removal of COD,
ammonia and amoxicillin were all > 90%, and the major amoxicillin
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Bioresource Technology 245 (2017) 1238–1244
removal mechanism was proposed as biodegradation, adsorption and
hydrolysis. Meanwhile, the addition of sequential biocatalyst to the
UASB (SBA-UASB) was found to be effective in nitrogen removal as
compared to treatment with a single UASB system. It was found that the
one with biocatalyst addition was able to remove nitrogen of up to
9.4 kg N/m3/d as compared to the one without biocatalyst addition that
was able to remove up to 0.178 kg N/m3/d with initial nitrogen loading
of up to 10.4 kg N/m3/d (Tang et al., 2011). Moreover, electro-
coagulation (EC) was applied as pretreatment in some research studies
to remove suspended solids in pharmaceutical wastewater, and hence
the pretreated wastewater was found to be with higher bio-treatability
in the subsequent anaerobic treatment processes (Deshpande et al.,
2010, 2012).
3. Key microbial populations involved in anaerobic treatment of
pharmaceutical wastewater
Anaerobic biodegradation is known as a fermentation process,
which generally consists of four stages, namely hydrolysis, acidogen-
esis, acetogenesis and methanogenesis (Ji et al., 2013a). This process is
carried out by two main microbial taxa, one is the bacteria that is re-
sponsible of hydrolyzing polymeric organics into monomers and further
break down of these monomers into smaller organic fragments (i.e.,
alcohols and volatile fatty acids); while the other one is the archaea
(usually refer as methanogen) that utilizes the fermentation metabolites
and converts these organics into their end form – methane gas. During
anaerobic degradation, complex interaction network occurs among the
various microbial groups, where any unbalance of microbial activity
could possibly cause accumulation of metabolic intermediates and in
turn, leads to system failure. Therefore, besides proper design of the
system operating parameters, a balanced microbial community struc-
ture is also crucial for anaerobic treatment process.
The prevalence of microbial populations found in pharmaceutical
treatment plants has been widely reported. For bacterial populations,
Bacteroidetes, Firmicutes and Proteobacteria were found to be the pre-
dominant phyla for most of the reported anaerobic systems, where
Clostridia (belongs to phylum Firmicutes) and β-Proteobacteria (belongs
to phylum Proteobacteria) are the dominant classes which are widely
reported in biological processes, and thus are expected to play vital
roles in the fermentation process. On the other hand, class
Methanomicrobia, Thermoplasmata and Methanobacteria were found as
the predominant archaeal groups, members of which are known for
their metabolic versatility in converting anaerobic digestion metabo-
lites into methane gas as well as their relatively high toxicity resistance
(Shi et al., 2015).
4. Acute toxicity evaluation of antibiotics and other organic
pollutants
Pharmaceutical wastewater contains several types of inhibitory or-
ganics including antibiotic residues, spent solvents, dissolved organic
nitrogen (DON), etc., which may pose strong inhibition to microbial
degradation even at low concentrations (Ng et al., 2015). Therefore, it
is crucial to investigate the acute toxicity of these potential toxicants in
both raw pharmaceutical wastewaters as well as treated effluents, in
order to maintain efficient organic removal performance of WWTPs and
evaluate environmental risk regarding treated effluent discharge. Con-
ventionally, specific methanogenic activity (SMA) based toxicity assay
is widely applied in anaerobic process (Amin et al., 2006; Enright et al.,
2005; Oktem et al., 2007) to understand the wastewater inhibition on
methanogenic populations, which as mentioned is a key group of mi-
croorganisms in the anaerobic process. Since 1990s, luminescent bac-
teria based bioluminescence toxicity assay has gained increasing in-
terest due to its high sensitivity and rapid response (Hu et al., 2017;
Sponza and Demirden, 2010). Table 3 summarizes the existing litera-
tures on acute toxicity of pharmaceutical production relevant water
X. Shi et al. Bioresource Technology 245 (2017) 1238–1244

Table 3 carbon as their energy source, and thus is considered as potential

Acute toxicity of pharmaceutical production relevant water samples via bioluminescence seeding source for treating organic pollutants from industrial saline
assay.
effluents. To date, a number of halo-tolerant/halo-philic strains has
Indicator strain Sample category Concentration Toxic unit References been isolated from saline environments and to be further applied in the
with acute wastewater treatment field. Their capability in degradation of various
toxicity recalcitrant organics such as polycyclic aromatic hydrocarbons (PAHs),
phenol and azo dyes, etc., has been widely studied in recent years
Vibrio fischeri Amoxicillin 3.99 g/L 15 min- Ji et al.
Ciprofloxacin 5.63 g/L IC50 (2013 b) (Arulazhagan and Vasudevan, 2011; Xu et al., 2016; Jiang et al., 2015).
Lincomycin 4.32 g/L In addition, Shi et al. (2015) was the first that reported on the lab-scale
Kanamycin 5.11 g/L application of intertidal wetland sediment (IWS) as inoculation source
Vibrio fischeri Aureomycin 12.06 mg/L IC50 Ji et al. for the treatment of real pharmaceutical wastewater using AnSBR sys-
Chloromycetin 429.90 mg/L (2013 a) tems. Their results showed that the IWS reactor achieved 71.4% of COD
Polymyxin 6.24 mg/L reduction, which was more than doubled as compared with conven-
Acetate 20.71 g/L
tional digested sludge reactor of 32.3%. Moreover, Ng et al. (2016)
Butyrate 12.17 g/L
Ethanol 19.40 g/L operated a novel anaerobic bio-entrapped membrane bioreactor (An-
propionate 10.47 g/L BEMR) for pharmaceutical wastewater treatment using marine micro-
Photobacterium Kemicetine 95 mg/L IC50 Sponza and organisms, who also found that the bio-augmented reactor achieved
phos- Demirden significant superior organic removal performance over the conventional
phoreum (2010) seeding biomass.
Photobacterium Raw wastewater 3.46% *
Relative Tang et al. Pharmaceutical (especially antibiotic compounds) residues in the
phos- from colistin and luminosity (2011) raw wastewater as well as treated effluents serve as an environment
phoreum kitasamycin selection factor that aid in the proliferation of ARGs. In recent years,
(T3 production
ARGs have been detected in various environments including soil,
mutation)
groundwater and sediment (Tao et al., 2016), where improper dis-
Photobacterium Dissolved organic 5.3–86 mg/L EC50 Hu et al.
charge of bio-solids from biological WWTPs was found as one of the
phos- matter (DOM) (2017)
phoreum(T3 extracted from
major contributors on the spreading of ARGs (Munir et al., 2011). Tao
mutation) pharmaceutical et al. (2016) investigated the diversity and abundance of ARGs in the
plant sludge collected from both sewage treatment plants (STPs) and
PWWTPs. The results showed that although ARGs were detected in all
* Relative Luminosity (%) = Lsample/Lblank × 100%.
the sludge samples, remarkably higher abundance of ARGs was found in
PWWTPs. Concerns on health risk have been raised with regards to the
samples using bioluminescence toxicity assay. It was found that the fact that ARGs can be rapidly transferred across different species
team of Ji has assessed the acute toxicity over a number of antibiotic (known as horizontal gene transfer) through plasmids, integrons and
compounds as well as anaerobic digestion intermediates, and the results transposons (Aydin et al., 2015a,b), which in turn can result in the
showed that acute toxicity of different compounds varied significantly development of antibiotic resistant pathogen populations in the natural
from only 6.24 mg/L (polymyxin) to 20.71 g/L (acetate) (Ji et al., environments. Therefore, removal of antibiotic residues from pharma-
2013a, b). Moreover, acute toxicity assessments on raw pharmaceutical ceutical wastewater prior to biological treatment or proper handling on
wastewater samples have also been reported in the last decade, where the biosolids generated especially from PWWTPs are of vital im-
in some cases the acute toxicity of real pharmaceutical wastewater portance on the prevention of spreading of ARGs. Advanced oxidation
could be even higher than antibiotics (Tang et al., 2011; Hu et al., processes (AOPs) have been proven to effectively degrade antibiotic
2017). These experimental values are of vital importance in both design compounds (Li et al., 2008). However, the co-existing of bulk pollutants
and maintenance of pharmaceutical wastewater treatment plants in the pharmaceutical wastewater burden its application feasibility due
(PWWTPs), and also provides valuable reference information for to the high cost of the AOP treatment (Yi et al., 2017). On the other
trouble-shooting in case of sudden failures of the biological WWTPs. hand, micro-algae as alternative biological pretreatment has been re-
ported in recent years to effectively degrade antibiotic residues in the
5. Future perspectives pharmaceutical wastewater (Guo and Chen, 2015) with significant
economic advantage, while the robustness of this treatment concept has
As discussed in the previous sections, anaerobic process offers a to be further evaluated with extensive future studies.
viable option for the treatment of high-strength pharmaceutical was-
tewater. However, regarding the increasing pharmaceutical manu- 6. Conclusions
facturing activities and accelerating progress on novel pharmaceutical
products globally, some potential technologies may attract intensive Pharmaceutical wastewaters are feasible to be treated by the
research interests in the near future, in order to meet the ever-tigh- anaerobic process. A number of modern anaerobic systems have been
tening discharge requirements as well as the growing public concern on reported to achieve efficient organic removal for pharmaceutical was-
the environmental impact of micro-pollutants in the treated effluent. tewater. Meanwhile, typical predominant microbial populations found
To address the issue of salinity in pharmaceutical wastewater which in pharmaceutical wastewater treatment systems were summarized in
inhibits the performance of biological treatment, application of mi- this paper. Moreover, with growing concern on the antibiotic residues
croorganisms isolated from saline environments could be a potential and spreading of ARGs in the surrounding environment, advanced
solution. Comparing with common microbial populations found in the treatment technology on destruction of antibiotic active compounds in
conventional biological WWTPs, microorganisms isolated from saline the anaerobically treated effluent is worthy of receiving special atten-
environments such as ocean or salt lakes are able to withstand high tion in future studies.
saline conditions. This special feature is contributed either by having
specific enzyme systems to maintain functioning under high osmotic Appendix A. Supplementary data
pressure, or to actively synthesis organic inside their cell to balance
osmotic potential from the outer environment (Ventosa et al., 1998). Supplementary data associated with this article can be found, in the
Among these unique populations, the heterotrophs utilize organic online version, at http://dx.doi.org/10.1016/j.biortech.2017.08.150.

1243
X. Shi et al.
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