Water: Influence of Pre-Hydrolysis On Sewage Treatment in An Up-Flow Anaerobic Sludge BLANKET (UASB) Reactor: A Review
Water: Influence of Pre-Hydrolysis On Sewage Treatment in An Up-Flow Anaerobic Sludge BLANKET (UASB) Reactor: A Review
Water: Influence of Pre-Hydrolysis On Sewage Treatment in An Up-Flow Anaerobic Sludge BLANKET (UASB) Reactor: A Review
Review
Influence of Pre-Hydrolysis on Sewage Treatment
in an Up-Flow Anaerobic Sludge BLANKET (UASB)
Reactor: A Review
Rajinikanth Rajagopal 1, *, Mahbuboor Rahman Choudhury 2 , Nawrin Anwar 2 ,
Bernard Goyette 1 and Md. Saifur Rahaman 2
1 Sherbrooke Research and Development Center, Agriculture and Agri-Food Canada, 2000 College Street,
Sherbrooke, QC J1M 0C8, Canada; [email protected]
2 Department of Building, Civil, and Environmental Engineering, Concordia University,
1515 Saint-Catherine St W, Montreal, QC H3G 2W1, Canada; [email protected] (M.R.C.);
[email protected] (N.A.); [email protected] (M.S.R.)
* Correspondence: [email protected]; Tel.: +1-819-780-7132
Received: 18 December 2018; Accepted: 15 February 2019; Published: 21 February 2019
Abstract: The up-flow anaerobic sludge blanket (UASB) process has emerged as a promising high-rate
anaerobic digestion technology for the treatment of low- to high-strength soluble and complex
wastewaters. Sewage, a complex wastewater, contains 30–70% particulate chemical oxygen demand
(CODP ). These particulate organics degrade at a slower rate than the soluble organics found in
sewage. Accumulation of non-degraded suspended solids can lead to a reduction of active biomass
in the reactor and hence a deterioration in its performance in terms of acid accumulation and poor
biogas production. Hydrolysis of the CODP in sewage prior to UASB reactor will ensure an increased
organic loading rate and better UASB performance. While single-stage UASB reactors have been
studied extensively, the two-phase full-scale treatment approach (i.e., a hydrolysis unit followed by
an UASB reactor) has still not yet been commercialized worldwide. The concept of treating sewage
containing particulate organics via a two-phase approach involves first hydrolyzing and acidifying
the volatile suspended solids without losing carbon (as methane) in the first reactor and then treating
the soluble sewage in the UASB reactor. This work reviews the available literature to outline critical
findings related to the treatment of sewage with and without hydrolysis before the UASB reactor.
Keywords: anaerobic digestion; hydrolysis; methanogens; sewage; up-flow anaerobic sludge blanket
reactor; volatile fatty acids
1. Introduction
Domestic sewage is considered a complex wastewater, as it contains both particulate and dissolved
organics. About 30–70% of the particulate chemical oxygen demand (CODP ) of domestic sewage is
found in the form of organic polymers such as carbohydrates, lipids, and proteins. These particulate
organics, which have slow degradation kinetics, can diminish the performance of treatment processes.
Treatment of sewage by conventional approaches, including primary sedimentation and secondary
aerobic biological treatment, is very effective. However, this efficiency comes at high capital
and operational costs, as well as advanced technology requirements [1,2]. High-rate anaerobic digestion
biotechnology has proven to be an excellent process and is considered by many authors to be the core
of sustainable waste management techniques [1,3–7].
The up-flow anaerobic sludge blanket (UASB) digester is one such high-rate anaerobic system
that has been extensively studied and adopted all over the world in laboratory, pilot-scale,
and full-scale implementations [8]. Compared to aerobic processes, high-rate anaerobic sewage treatment
processes offer: (a) high removal efficiency in the system, even at high organic loading rates (OLR)
and short hydraulic retention times (HRT); (b) simpler reactor construction and operation; (c) flexibility in
terms of design scale; (d) a net-positive energy producing process through the production of high quality
renewable fuel in the form of biogas; (e) lower sludge production rates with well stabilized sludge
production for final disposal with good dewatering characteristics (due to the slow growth rate of
anaerobic microorganisms); and (f) low nutrient and chemical requirements [3,5,6,9,10]. While these
comparisons show the positive aspects of anaerobic process, it should be kept in mind that comparison
of aerobic and anaerobic processes should be based on the type of wastewater. Anaerobic treatment
processes have been found to be advantageous for very high strength sewage treatment. Despite all these
advantages, there exist some drawbacks to the application of high-rate anaerobic treatment processes.
These limitations include: long solids retention time in the reactors, long start up time requirement,
impure biogas generation possibly leading to bad odors, incomplete or insufficient removal of organic
matter, pathogens and nutrients in the final effluent, and necessity of further post-treatment to meet
discharge or reuse standards [3,5,6,9,10].
Anaerobic digestion of complex wastewater is a multistep process involving microorganisms
and occurring in the absence of oxygen. The interactions between the microbial community takes
place in a series and parallel reactions that degrade complex polymers like carbohydrates, proteins,
nucleic acids, and lipids, into methane (CH4 ) and carbon dioxide (CO2 ) [11]. At a molecular level, these
steps occur in sequence; however, in a reactor they appear to progress simultaneously. Several groups
of microorganisms present in reactors catalyze reactions occurring in the anaerobic digestion process.
These are fermentative bacteria, hydrogen producing acetogenic microorganisms, hydrogen-consuming
acetogenic microorganisms, CO2 reducing methanogens, and aceticlastic methanogens. A schematic of
the processes of anaerobic digestion is presented in Figure 1, which shows the reaction pathways in
anaerobic digestion along with the catalytic microorganisms.
Figure 1. Anaerobic digestion of organic polymeric materials indicating sub-processes and involved
bacterial groups (Figure adapted from Seghezzo, 2004 [11]).
Water 2019, 11, 372 3 of 27
Due to the limitations of anaerobic treatment and UASB technology in the treatment of sewage,
researchers are continuously working to enhance the performance of anaerobic digesters by changing
reactor configurations [1,6,12]; enhancing start-up and granulation processes in the reactors by using
multivalent cations [13–16], natural polymers [17,18], or synthetic polymers [19,20]; and incorporating
complementary post-treatment schemes such as activated sludge [21,22] or sequencing batch
reactors [23,24] to treat the effluent produced by UASB. Various studies have shown that accumulation
of suspended solids in sewage adversely affects the anaerobic digestion process [3,11,25–27].
These solids decrease sludge activity due to adsorption and entrapment, limit substrate transfer,
lead to the formation of a “scum” layer, inhibit granulation, and increase sludge volume, which in turn
requires frequent emptying of the reactor [1]. The relatively slow digestion of organic solids results
in high accumulation in the reactors, especially at lower temperatures (<20 ◦ C, i.e., psychrophilic
range). As a result, the solid retention time is decreased and the performance of UASB reactors is
also reduced [4,27–30]. Both gas production rate and chemical oxygen demand (COD) removal drop
significantly when the temperature drops to the psychrophilic range [31]. Several reactor modification
strategies have been adopted to incorporate pre-hydrolysis of wastewater prior to UASB, which should
increase the digester OLR and improve the performance of UASB. Such reactor modifications involve
applying a two-stage anaerobic process, which results in the entrapment of particulate organic
matter and its partial hydrolysis into soluble compounds in the first stage and these pre-hydrolyzed
organics are then digested in the second stage. The removal efficiency of suspended solids in the first
reactor will be higher than that of organic matter and excess sludge needs to be discharged regularly.
As a result, of that, the sludge age remains relatively low in this reactor, hindering the development of
the slow-growing methanogens and reducing methanogenesis to a minimum. Two examples of such
first-stage reactors include Hydrolysis Up-flow Sludge Blanket (HUSB) reactors and Up-flow Anaerobic
Solids Removal (UASR) reactors. Incorporation of HUSB reactor achieved over 50% hydrolysis of
the removed suspended solids at higher ambient temperatures (exceeding 19 ◦ C) [32]. The HUSB
reactor can be considered as a relatively highly loaded UASB system for the removal and hydrolysis of
suspended COD. The hydraulic retention time in the HUSB reactor is very similar to that applied in
primary sedimentation tanks, but the removal efficiencies of COD, BOD, and suspended solids are
considerably higher [32]. In UASR, only suspended solids removal is obtained, as in normal settling
tanks [33], while in HUSB reactors, hydrolysis also takes place. Therefore, more sludge has to be
discharged from UASR than from HUSB reactors [5].
The objective of this study was to review the effect of pre-hydrolysis on the treatment of complex
sewage in an UASB reactor. The review reports in the performance of UASB reactors (with and without
pre-hydrolysis) from the literature and makes a comparison between the treatment efficiencies of
the anaerobic digestion systems in terms of solids reduction, removal of various COD fractions,
and sludge production. Finally, the review provides some perspectives for future research requirements
regarding complex wastewater treatment in a two-phase hydrolysis-UASB approach.
(b) Fermentation, (c) Acidogenesis, (d) Acetogenesis/Dehydrogenation, and (e) Methanogenesis [37–40].
In an initial exoenzyme-catalyzed reaction, composite biopolymers are hydrolyzed to soluble mono, di,
or oligomers [41]. The complex particulate materials are converted into carbohydrates, proteins,
and lipids, and later these complex organic compounds are transformed into corresponding monomers
(like amino acids, sugars, and long chain fatty acids, etc.). This step, commonly referred to as ‘hydrolysis’,
is the rate-limiting step in the overall anaerobic treatment processes for wastes containing lipids and/or
substantial amount of particulate matter [42,43]. Intracellular enzymes in acidogenic microorganisms
induce fermentation of these soluble fractions to reduced organic compounds like short chain fatty
acids, alcohols, and lactate in a process known as acidogenesis [38]. The hydrolyzed amino acids,
sugars, and long chain fatty acids are taken up by the acidogenic microorganisms and fermented,
resulting in the production of formate, propionate, butyrate, lactate, etc. If fatty acid isomers are
produced, they are mainly derived as a result of hydrolysis of lipids and amino acids, which are
produced as a result of protein hydrolysis. Acetogenic microorganisms further oxidize fatty acids
and the resulting cleavage products (CO2 /H2 formed by hydrogen-forming microorganisms or acetate
formed by acetate-forming microorganisms) can be taken up by methanogens and be converted
to CH4 and CO2 . Lactate is oxidized to pyruvate, which is decarboxylated to yield acetate, CO2 ,
and H2 . If ethanol is present, it is oxidized to acetate and hydrogen, and the hydrogen is used for CO2
reduction [41]. Acetate can also be formed via the CO2 /H2 pathway in a step called homoacetogenesis.
The methanogens are able to directly use substrates like H2 , acetate, formate and methanol to produce
CH4 [38]. To optimize the overall anaerobic digestion process, the rate-limiting hydrolysis process
must be improved. To accommodate the rate-limiting hydrolysis of particulates in complex wastewater,
a longer SRT is required depending on the applied process temperature [34]. The increased SRT
ensures retention of slowly growing organisms even at relatively shorter HRTs, which ensures high
OLR [1]. Different high-rate anaerobic systems have been developed, including the anaerobic filter [44],
the up-flow anaerobic sludge blanket [45], the fluidized and expanded bed reactors [46], the down flow
stationary fixed film reactor [47], and the anaerobic baffled reactors [48].
Several researchers have introduced and investigated optimized versions of the UASB system.
These modified systems include the expanded granular sludge bed (EGSB) reactor [49], the UASB-septic
tank reactor [50], the hydrolysis up-flow sludge bed (HUSB) reactor [32], the thermophilic up-flow
staged sludge bed (USSB) reactor [51], the up-flow anaerobic solids removal (UASR) reactor [33],
the hybrid EGSB-fixed bed reactor [52], the anaerobic bioreactor with a fixed-structure bed
(ABFSB) [53], and the two-stage anaerobic filter/anaerobic hybrid (AF/AH) system [54]. While UASB
and EGSB reactors have been identified as the most effective anaerobic treatment system for low
strength wastewater [5], the modified anaerobic baffled reactor has also shown improved treatment
performance [55].
when seed sludge is not available due to low growth rate of methanogenic microorganisms [61].
Generally, UASBRs are inoculated with a suitable seed source to shorten the startup time [62].
Figure 2. Schematic showing different components and zones of a UASBR system. (Figure reproduced
following van Haandel and Lettinga 1994, [63]).
A dense sludge bed, having high settling properties, develops either with granular or flocculent
features. In the case of sewage, which is a low- to medium-strength wastewater, flocculent sludge
forms in the reactor. Above the dense sludge bed, a sludge blanket zone develops with a diffuse
growth pattern and lower particle settling velocities [1]. Biological reactions take place in the sludge
bed and sludge blanket region. As the wastewater passes through the biomass in the dense sludge
bed and the sludge blanket region, the dissolved substrate is digested and the particulate organic
matter is retained and digested to form biogas [61,64]. The produced biogas (which consists mostly
CH4 and CO2 ) in the digestion section is captured by the phase separator setup so the gas bubbles
cannot interfere with the solid settling. An air/gas pocket is located under the phase separator element
to prevent solids from escaping through the gas outlet. As a result, sludge particles on the phase
separator fall back into the digestion zone, and a large sludge mass is retained in the reactor [56,58].
Finally, an effluent with less suspended solids is discharged from the settling zone.
In comparison with other high-rate anaerobic digesters, UASB processes have the advantage of
not requiring any support media for biomass attached growth or immobilization yet achieve high
removal of COD through utilization of available granular or flocculent sludge [6,65]. The biomass of
good settling properties is not prone to washout from the system under practical reactor conditions.
The natural turbulence caused by influent up-flow and gas production helps to maintain adequate
contact between biomass and wastewater. Like all other modern high-rate reactors, the UASBR is able
to separate SRT/HRT through the use of the sludge blanket that develops as a result of granulation [66].
single-stage UASB process for sewage treatment in different laboratory- and pilot-scale settings.
The pilot-scale UASB plant built in Cali, Columbia in 1982–1983 with support from the Netherlands was
claimed to be the first of its kind in the world [69]. A 64-m3 reactor was operated at an average sewage
temperature of 25 ◦ C. Diluted digested cow manure was used as inoculum, and the plant was fully
operational after 6 months at an HRT of 8 h. COD and BOD removal efficiencies higher than 75% were
observed while SS removal was about 70%. Tables 1 and 2 outline selected key operating parameters
and performance of laboratory-scale (reactor size range 2–35 liters) and pilot-scale (reactor size range
55–2000 liters) UASB systems as studied by different researchers.
viable sludge in the reactor due to reduced SRT, reduction in sludge activity due to accumulation
of non-viable solids, sludge lifting and sludge washout, and required disposal of non-viable sludge
from filter beds. There exist few, if any, differences between flocculent and granular sludge bed
UASB reactors with respect to the applicable OLRs, when a high total suspended solids (TSS) removal
efficiency should be accomplished [58]. Lower OLR is recommended for high TSS removal efficiency,
as it aids stabilization of the accumulated solid substrate ingredients, particularly in low ambient
temperature conditions [58]. Prasanth et al. [79] showed that presence of CODP in synthetic wastewater
minimizes the biodegradability rate constant, substrate biodegradability, anaerobic degradability,
substrate activity, and sludge activity.
Water 2019, 11, 372 8 of 27
Table 1. Selected key operating parameters and performance of laboratory-scale (reactor size range 2–35 L) UASB systems studied by different researchers.
Influent Characteristics OLR (kg HRT Temperature % Removed Methane
Inocula (CH4 ) Collection Remarks References
CODT CODS COD/m3 day) (h) (◦ C) CODT SS (L/g COD Removed)
310 — — 12 15 48 44 —
• Low biogas production could not serve for
Anaerobically digested an effective mixing of sludge. Mechanical stirrer
municipal sludge installed in the reactor for sludge-wastewater contact [82]
310 — — 12 9 37 38 — did not lead to an improvement in removal efficiency.
15 76 ± 10
Flocculent sludge 10 79 ± 4
from municipal anaerobic — 8 83 ± 7 — — • Reactor was inoculated with 13.5 g VSS/L of
digester 6 92 ± 5 flocculent or granular sludge.
50–70% of 4 89 ± 4 • Synthetic domestic wastewater (made from a mixture
700–1000 25 ± 1 of dog food, clay, sucrose, and peptone) was used. [60]
CODT
15 81 ± 11
• The composition of these materials was chosen to
10 84 ± 14
Granular sludge from UASBR maintain the ratio of organic constituents in
— 8 86 ± 8 — —
treating brewery wastewater the sewage (protein 50%, carbohydrates 40%, fat 10%).
6 91 ± 4
4 87 ± 3
Note: (1) “—” = Information not provided. (2) All values are in mg/L unless otherwise mentioned.
Water 2019, 11, 372 9 of 27
Table 2. Selected key operating parameters and performance of laboratory-scale (reactor size range 55–6000 L) UASB systems studied by different researchers.
Influent OLR Methane (CH4 )
Temperature % Removed
Inocula Characteristics (kg-COD/m3 HRT (h) Collection (L/g COD Remarks References
(◦ C)
CODT CODS day) CODT SS Removed)
Granular Sludge 100–900 — — 9–16 10–18 46–60 — — • Reactor volume: 6000 L. [85]
500 3 4
• BOD reductions = 90–92%.
(For 200 day)
Digested sludge from an anaerobic — 4 3 20–35 83–88 — 0.141 • Due to the presence of a high concentration of active
300 [86]
digester treating sewage 2 6 granular sludge in the lower portion of the reactor,
(For 60 day)
an 80% reduction in COD occurred within the bed
1.2 6 level itself.
Note: (1) “—” = Information not provided. (2) All values are in mg/L unless otherwise mentioned.
Water 2019, 11, 372 10 of 27
Table 3. Selected key operating parameters and performance of pilot to full-scale (reactor size range 20–30,000 m3 ) UASB systems studied by different researchers.
Influent Methane (CH4 )
Temperature % Removed
Inocula Characteristics OLR (kg-COD/m3 day) HRT (h) Collection (L/g COD Remarks References
(◦ C)
CODT CODS CODT SS Removed)
• VSS/TSS ratio: 50.
Digested cow manure 267 95 — 6-8 25 75–82 70–80 — • Start-up period: 6 months. [90]
Granular sludge grown on paper • 54 L of seed material was used with standard
mill wastewater from full scale 391 164 — 2–7 ≥13 16–34 — — acetotrophic methanogenic activity 0.17 g [57]
UASB CH4 -COD/g VSS.d at 30◦ C.
16
(Winter • Total BOD removal: 80%.
average) • The plant operated at a low inlet flow, which is only
Digested sludge added 10% (V/V) 436 402 — 7 74 87 — 17% of the design flow. [92]
23
(Summer • No requirement of any excess sludge discharge.
average)
Table 3. Cont.
Influent Methane (CH4 )
Temperature % Removed
Inocula Characteristics OLR (kg-COD/m3 day) HRT (h) Collection (L/g COD Remarks References
(◦ C)
CODT CODS CODT SS Removed)
3.6–5.0 & 2.9–4.6 kg
COD/m3 d for stage I
and stage II,
respectively • Single stage UASBR operated at OLR of 1.5–1.8 kg
8–10 62 60 COD/m3 day (average HRT = 24 h) resulted in
(during 1st year).
(I stage) (summer) (summer) 0.439 87–93% COD removal. [94]
Without inoculum 1419–1650 — Only stage I was 18–25
5–6 51 55 0.249 • CH4 content was 75% and 65% during summer
operated
(II stage) (winter) (winter) and winter, respectively.
as a single-stage
UASBR at half of
the previous loading
rate (during 2nd yr).
375 ± 97.6 246 ± 50 9.9 27 ± 6 ~46.7 ~44.4
403 ± 66.2 179 ± 62 9.6 28 ± 6 ~45.4 ~42.5 • Performance of five full-scale UASB-based STP were
— 390 ± 64 157 ± 54 — 9.8 29 ± 6 ~38.5 ~35.7 — [95]
performed with the reactors size ranging
443 ± 101.9 213 ± 58 10.3 29 ± 6 ~41.3 ~48.4 from 11,200–30,000 m3 .
318 ± 101.9 213 ± 58 9.4 24 ± 6 ~62.3 ~34.0
— — — — — [75]
• Plant capacity 90,000 m3 /day.
549 8 67 70 • reactors each with a volume of 2705 m3 .
• Up-flow velocity 0.44 m/h.
Note: (1) “—” = Information not provided. (2) All values are in mg/L unless otherwise mentioned.
Water 2019, 11, 372 12 of 27
Therefore, several researchers have investigated the feasibility of installing a hydrolysis unit
prior to UASB to reduce the loading of CODP on the reactor. The following section includes a review
of studies carried out on a two-phase UASB with pre-hydrolysis units.
Among the main components of primary sludge (e.g., carbohydrates, lipids and proteins),
carbohydrates are known to be easily and rapidly converted to simple sugars via hydrolysis
and subsequently fermented to volatile fatty acids (VFA). Protein is hydrolyzed to amino acids
and further degraded to VFA either through anaerobic oxidation linked to hydrogen production or
via fermentation. The former is dependent on the presence of hydrogen-scavengers while the latter is
independent of the methanogenic activity in the reactor. Among the lipids, triglycerides are hydrolyzed
to long chain fatty acids and further oxidized via β-oxidation to acetate or propionate [106].
In a primary fermentation unit, COD removal of about 25% was observed in almost all cases,
even though with HRTs of 2.8 and 3.3 h, suspended solid removal was about 70% [101]. This imbalance
between suspended solid and COD removal is due to the solubilization of a segment of the particulate
organic matter in the wastewater. The reduction in COD, due to the lowering of CODP concentration,
is offset by an increase in soluble COD (CODS ) concentrations. No production of dissolved COD
was observed with an HRT of 1.1 h and it increased gradually with an HRT between 2.1 and 2.8 h.
A maximum produced dissolved COD of around 30 mg/L was observed for HRT between 2.1 and 2.8 h.
At HRT > 2.8 h, the concentration of the filtered COD decreased, due to the action of methanogenic
microorganisms. At a HRT of 4.3 h, dissolved COD concentration was on the order of 19 mg/L [101].
The hydrolysis of lipids and carbohydrates increases with increasing SRT, whereas protein hydrolysis
only occurs under methanogenic conditions [106]. This study employed five completely mixed stirred tank
reactors (CSTRs) with an effective volume of 5 L and operated them to maintain SRTs of 3, 5, 8, 10, and 15
days. The process temperature was controlled at 25 ± 1 ◦ C by recirculation of temperature-controlled
water through the double walls of the reactors. The reactors were inoculated with diluted digested
primary sludge (20 gTS/L), i.e., settled solids of domestic sewage from a wastewater treatment plant.
The feed included diluted primary sludge from the same wastewater treatment plant. A certain volume
of digested sludge was withdrawn from the reactor and an equal volume of primary sludge was pumped
into each reactor. The reactors operated at 3 and 5 d SRTs were fed two times a day to avoid shock
loading [91]. The study revealed a decrease in protein hydrolysis and acidification under acidogenic
conditions. The low values obtained for protein hydrolysis and acidification have been partially explained
by the relatively high ammonium-nitrogen concentration in the influent, which suggests that easy
degradable protein was already hydrolyzed before the sludge was used in the CSTRs. Also, low pH
and high lipid concentrations could affect the hydrolysis and precipitation of ammonium as struvite
may contribute to the decrease of the ammonium-nitrogen levels, which might have contributed to
a lower calculated hydrolysis [91]. Also, the hydrolysis rate of entrapped organics has been reported to
be significantly affected by temperature, that is, 58% of entrapped particulate organics liquefy at 25 ◦ C,
which decreases to 33% at 13 ◦ C [78]. Maximum solubilization occurs only when CH4 production is
effectively suppressed. Little, if any, methanogenesis will develop in the hydrolytic reactor, because the pH
is depressed by acid fermentation. Only a part of the entrapped matter will be hydrolyzed and excess
sludge will have to be discharged from the reactor at a relatively high frequency. This means that
the sludge age will be too low for the slow growing methanogens. The effluent from the hydrolytic reactor
will be mainly dissolved compounds, so that it can be conveniently treated in second phase reactor [63].
The results of pre-hydrolysis of domestic wastewater in bench and pilot-scale solubilization reactors
under different working conditions are presented in Table 4. Studies on the hydrolysis of particulates
in sewage under anaerobic conditions have, so far, been carried out on bench scale reactors [76]. In all
the cases, hydrolysis is accompanied by reduction in COD (Table 4). The range of COD reductions is
about 25–59%. The reduction of CODS , in most cases, indicates a conversion of soluble fractions into
CH4 gas. This indicates that methanogenesis has taken place, in addition to hydrolysis. The reasons for
methanogenesis in hydrolysis reactor or pretreatment unit may be due to good amount of seed sludge
or a low food to microorganism ratio (F/M < 1) and other microenvironment parameters. Efforts are
required to check methanogenesis in hydrolysis unit.
Water 2019, 11, 372 14 of 27
Table 4. Hydrolysis performance on bench-scale and pilot-scale solubilization reactors for the treatment of domestic wastewater.
Hydrolytic Up-flow
Urban Wastewater 2.2–4.5 525–710 257–344 186–268 153–232 6–29 - The methanogenic activity of the inoculum used was
Digester 0.69–0.89 [107]
(20) (4.42) (33%–47%) (13%–30%) (55%–68%) (52%–63%) (25–107) 0.18 g CODCH4 /g VSS·d at 20 ◦ C
(2 L)
Prefermenter - SRT of 5 day and 10 day were tested and the best
Domestic Sewage 1.3 399 126 305 5.1
Reactor — 0.90 results were obtained in a covered pre-fermenter [109]
(17–20.5) (7.36) (25%) (9.5%) (61%) (14)
(3.3 L) with a 5 d SRT.
Table 4. Cont.
in both aerobic and anaerobic parts of the system. In the anaerobic unit, the first order kinetic
constant is 0.31 h−1 in summer temperatures (~19 ◦ C) and 0.20 h−1 in winter temperatures (~12.5 ◦ C),
whereas in aerobic units, the values are higher (~2.0 h−1 ) [114]. Table 5 gives the characteristic values
of maximum growth rate constants of biomass and first-order hydrolysis constants used in METHANE
model [115,116]. The hydrolysis step of complex organic matter has been identified as the rate-limiting
in anaerobic digestion [115,117]. Acetogenesis or methanogenesis might be the rate-limiting stages in
complex waste. For complex waste, stimulation of hydrolysis (mechanically, chemically or biologically)
could lead to a further inhibition of acetogenesis or methanogenesis stages (these stages could be rate
limiting for complex waste), which ultimately affects hydrolysis as well [117].
Table 5. The characteristic values of maximum growth rate constants (day 1) of biomass
and first-order hydrolysis constants used in METHANE model. [115].
Process Source of
Feed, Temperature
Hydrolysis Acidogenesis Acetogenesis Methanogenesis Experimental Data
5.6 (B1),
Cellulose, 35 ◦ C 0.1 0.56 0.56 (H) [100]
4.1 (B2)
5.0 (B1),
Sewage Sludge, 5 ◦ C 0.25 0.8 0.5 (A), 2.0 (H) [118]
5.0 (B2)
Note: B1 = acetate-producing acidogens, B2 = propionate-producing acidogens, A = acetate-utilizing methanogens,
H = hydrogen-utilizing methanogens.
The International Water Association (IWA) developed a structured generic model for anaerobic
digestion titled “Anaerobic Digestion Model No 1 (ADM1)”, which included multiple steps to
describe the biochemical as well as physico-chemical processes during anaerobic digestion [119].
The biochemical processes were divided into: (a) disintegration of dead biomass to particulate
constituents, (b) enzymatic extracellular hydrolysis of these particulates to their soluble monomers,
(c) intercellular degradation of soluble materials (resulting in biomass growth and subsequent
decay) [119]. Meanwhile, the physico-chemical processes were not biologically mediated and included
ion association/dissociation, and gas-liquid transfer [119]. This model assumed a disintegration step
of complex particulate waste to carbohydrate, protein, and lipid particulate substrate and particulate
and soluble inert material to facilitate modeling of waste-activated sludge digestion. The disintegration
step was included to precede more complex hydrolytic steps and this step makes the complex
polymeric material more bioavailable for subsequent hydrolysis. This disintegration step includes
an array of processes such as lysis, non-enzymatic decay, phase separation, and physical breakdown
(e.g., shearing). ADM1 is a powerful tool for predicting steady-state behavior of anaerobic digesters
treating sewage sludges [120]. Determination of the fraction of the biodegradable versus the total solids
in the feed sludge is critical while using the ADM1 model. A first-order disintegration process was
employed to describe the complex composite particulate waste breakdown to particulate substrates
of carbohydrate, protein, and lipid along with inert materials. When the disintegration step was
considered, before the hydrolytic steps in ADM1, it was found to be the rate-limiting step compared to
the hydrolysis step and hence it affected the final modeling results.
The use of the ADM1, with the first-order rate constant assigned for disintegration (Kdis )
and hydrolysis (Khyd ) as 0.5 day−1 and 10 day−1 , respectively, reflected the steady-state full-scale
anaerobic digestion data collected from two different sized wastewater treatment plants with
satisfactory level of accuracy [120]. The ADM1 is used extensively for modeling anaerobic digestion of
solid wastes and has been studied and modified to reflect different physico-chemical and biochemical
processes in the anaerobic digestion process [121–124]. The ADM1 models the disintegration
and hydrolysis steps in anaerobic digestion following a first-order kinetics and assuming that their rates
do not depend on disintegration/hydrolytic biomass concentration. However, for complex substrates,
the first-order kinetics can be modified to account for slowly degradable material [117,125]. It has
been demonstrated that when hydrolysis is coupled with the growth of hydrolytic microorganisms,
then the model prediction is enhanced at high or fluctuant organic loading [117]. Ramirez et al. [125]
Water 2019, 11, 372 18 of 27
developed a slightly modified ADM1 model to simulate thermophilic anaerobic digestion of thermally
pretreated waste activated sludge by using the Contois model for disintegration and hydrolysis steps
instead of first-order kinetics and the Hill function to model for ammonia inhibition of aceticlastic
methanogens instead of a non-competitive function.
The modified ADM1 was calibrated and validated using batch experimental data sets and the model
parameters involve three disintegration biochemical parameters, nine hydrolytic biochemical parameters
and four stoichiometric parameter values. The model was capable of predicting the data measured
under different pretreatment conditions and was able to explain the dynamics of acetate accumulation in
batch experiments [125]. ADM1 was also calibrated and validated (using 360 days operation data)
to satisfactorily model the dynamic performance of a full-scale anaerobic digester treating mixed
(primary and secondary) sludge in a wastewater treatment plant under mesophilic condition [126].
The calibration of the ADM1 parameters requires a deep understanding of the interaction between each
process, functional microorganism and environmental conditions. Thus, only the most important
parameters should be calibrated to improve the compatibility between measured data and model outputs.
A minimum set of parameters (disintegration rate of particulates, fractionation of particulate organics
and kinetics of acetate, propionate and hydrogen utilizers), which were reported as the most sensitive
parameters in the simulation results, were manually calibrated [126].
in one-stage UASB systems. On the other hand, the hydrolysis stage of the two-phase process indicated
CODT removal efficiencies of 11–59% (Table 4). Table 6 demonstrates the overall CODT and CODS
removal performance of a two-phase system. The overall CODT removal efficiency of a two-phase
process has been reported to be between 51–82%. It is to be noted that CODT removal efficiency depends
on issues like temperature and HRT. For similar temperatures, the two-phase process exhibited better
COD removal than the one-stage UASB process [32]. Removal efficiency of CODT increases considerably
with the decrease of the HRT, as a treatment efficiency of 76% was achieved at HRT = 10 h (8 h for Phase I
and 2 h for Phase II), while the treatment efficiency was 82% at HRT = 6 h (4 h for Phase I and 2 h for
Phase II) [127]. The two-step AF→AH system, operated at an HRT of 4 + 8 h and a temperature of 13 ◦ C,
provided high removal efficiencies for all fractions of COD. The CODT removal efficiency was as high
as 71%, similar to that in one-step UASBRs in tropical countries. Application of the AF→AH system
results in high values for hydrolysis, acidification, and methanogenesis. No significant difference was
observed at different HRTs. At the imposed HRTs of 2 + 4 to 4 + 8 hours in the AF→AH system, 60–74% of
the removed CODT was converted to CH4 [54]. These ranges are significantly higher than those reported
by Uemura and Harada (2000) [78], who observed that 35% of the removed CODT was converted to CH4
while treating domestic sewage in a one-stage UASBR (at HRT = 4.7 h and temperature = 13 ◦ C).
the performance of UASB, but also on subsequent post-treatment and sludge disposal. Considering
the literature on (i) hydrolysis, (ii) the performance of the full scale UASB process, and (iii) bench-scale
two-phase treatment of sewage, a need to investigate hydrolysis of CODP without losing carbon
as CH4 was observed. Therefore, efforts are required to limit methanogenesis in the hydrolysis unit.
To achieve this, it is suggested to startup the hydrolytic reactor and to recover CH4 from the reactor. It is
also suggested that the F/M ratio in the system should be varied and pH value also monitored to reach
an optimum condition where methanogens will not be encouraged in the first-phase hydrolytic reactor.
Few researchers have investigated optimum mixing strategies for increasing efficiency
of hydrolysis and subsequent acidification stage of AD. Ma et al. [130] performed the characterization
of dissolved organic matter and key microorganisms to understand the effect of mixing for Sewage
Sludge (SS) treatment. They reported that a mixing speed of 90 rpm provides the highest efficiency
for hydrolysis and acidification phase (HAP). However, the efficacy of the mixing intensity is yet to be
revealed properly for the maximization between methanogenesis and HAP.
Temperature is another parameter that can play a crucial role in the acidification process in AD.
Ambient temperature can have a deteriorating effect on the hydrolysis thereby significant decrease
in acidification as well. Studies showed that operating AD under thermophilic condition (55 ◦ C)
can improve short chain fatty acid formation. Liu et al. [131] investigated HAP enhancement for
elevated temperatures such as mesophilic (35 ◦ C), thermophilic (55 ◦ C) and extreme thermophilic
(70 ◦ C) conditions. They reported that the optimum temperature for acidification is the thermophilic
zone, which shows 115% and 12% more efficiency than mesophilic, and extreme thermophilic zone
respectively. Later, Zhang et al. [132] found similar results of increased acidification of about 15.7% due
to higher temperature (55 ◦ C) in semi-continuous reactors compared with 35 ◦ C operating temperature.
Although the AD process has a potential to attract world-wide attention due to the production
of net positive energy and other valuable byproducts such as VFA, organic fertilizer,
and bioproducts [133], there is very limited research on full-scale study for the understanding of one of
the key aspects of VFA production during anaerobic digestion process. However, some researchers have
worked on the full-scale applications to produce VFA using chemical routes [134], but production
of VFA through biological pathways from SS, municipal sludge and industrial organic wastes has
yet to be investigated at commercial scale. Recently, one paper was published by Liu et al. [135]
describing full-scale operation to produce VFAs. They reported the hydrolysis rate in pretreatment
and the VFA yield in fermentation to be 68.7% and 261.32 mg COD/g VSS, respectively, for sewage
sludge. However, for more comprehensive understanding, further research is required to explore
the influence of HAP/VFAs productions and their downstream applications, such as enhancing biogas
production and/or for the biological nutrient-removal process in WWTPs [135].
The application of culture-independent methods to anaerobic digesters, together with crucial
complementary techniques such as imaging, isotope labeling, and chemical analyses, has provided us
with understanding of microbial community composition and the function of dominant populations [136].
Metagenomics, the sequencing of bulk DNA extracted from samples, provides direct access to the metabolic
potential of a microbial community [137]. Improvements in sequence throughput and bioinformatics
tools have contributed to a more widespread application of metagenomics to study natural and engineered
systems [136]. Application of metagenomics to understand anaerobic digestion process have enabled
better perspective on the representative microbial communities, the shift in communities during
the entire digestion process, and the relationship between reactor performance and microbial community
shifts [137–139]. Throughout the entire single-stage anaerobic digestion of high-strength food wastewater,
the abundance of phylum Chloroflexi decreased significantly, and the methanogenic microorganisms
shifted from aceticlastic to hydrogenotrophic methanogens with high increase in the proportion of
syntrophic bacterial communities [138]. Application of metagenomics has opened up a new direction
to appraise the complex interconnected processes performed by microbial communities, and to
understand how microbial community dynamics, interactions and functionality influence digester
efficiency and stability [136–140]. Use of culture-independent metagenomics might add useful information
Water 2019, 11, 372 21 of 27
Author Contributions: Conceptualization R.R.; Writing-Original Draft Preparation R.R. and M.R.C.; Methodology,
Investigation and Resources N.A. and M.R.C.; Supervision, Writing—Review & Editing and funding acquisition
B.G. and M.S.R.
Funding: This research was funded by Natural Sciences and Engineering Research Council (NSERC) of Canada
[Grant No. 505734-16].
Acknowledgments: The authors gratefully acknowledge the Natural Sciences and Engineering Research Council
(NSERC) of Canada for providing funding supports for this project.
Conflicts of Interest: The authors declare no conflict of interest.
References
1. Aiyuk, S.; Forrez, I.; Lieven, D.K.; van Haandel, A.; Verstraete, W. Anaerobic and complementary treatment of
domestic sewage in regions with hot climates—A review. Bioresour. Technol. 2006, 97, 2225–2241. [CrossRef]
[PubMed]
2. Chan, Y.J.; Chong, M.F.; Law, C.L.; Hassell, D.G. A review on anaerobic-aerobic treatment of industrial
and municipal wastewater. Chem. Eng. J. 2009, 155, 1–18. [CrossRef]
3. Zeeman, G.; Lettinga, G. The role of anaerobic digestion of domestic sewage in closing the water and nutrient
cycle at community level. Water Sci. Technol. 1999, 39, 187–194. [CrossRef]
4. Mahmoud, N.; Zeeman, G.; Gijzen, H.; Lettinga, G. Anaerobic sewage treatment in a one-stage UASB reactor
and a combined UASB-Digester system. Water Res. 2004, 38, 2347–2357. [CrossRef] [PubMed]
5. Seghezzo, L.; Zeeman, G.; Van Lier, J.B.; Hamelers, H.V.M.; Lettinga, G. A review: The anaerobic treatment
of sewage in UASB and EGSB reactors. In Bioresource Technology; Elsevier: Amsterdam, The Netherlands,
1998; Volume 65, pp. 175–190. ISBN 0960-8524.
6. Chong, S.; Sen, T.K.; Kayaalp, A.; Ang, H.M. The performance enhancements of upflow anaerobic sludge
blanket (UASB) reactors for domestic sludge treatment—A State-of-the-art review. Water Res. 2012,
46, 3434–3470. [CrossRef] [PubMed]
7. Ekstrand, E.M.; Karlsson, M.; Truong, X.B.; Björn, A.; Karlsson, A.; Svensson, B.H.; Ejlertsson, J. High-rate
anaerobic co-digestion of kraft mill fibre sludge and activated sludge by CSTRs with sludge recirculation.
Waste Manag. 2016, 56, 166–172. [CrossRef] [PubMed]
8. Tiwari, M.K.; Guha, S.; Harendranath, C.S.; Tripathi, S. Influence of extrinsic factors on granulation in UASB
reactor. Appl. Microbiol. Biotechnol. 2006, 71, 145–154. [CrossRef] [PubMed]
9. Schellinkhout, A. UASB technology for sewage treatment: Experience with a full scale plant and its
applicability in Egypt. Water Sci. Technol. 1993, 27, 173–180. [CrossRef]
10. Lettinga, G.; De Man, A.; Van der Last, A.R.M.; Wiegant, W.; Van Knippenberg, K.; Frijns, J.; Van Buuren, J.C.L.
Anaerobic treatment of domestic sewage and wastewater. Water Sci. Technol. 1993, 27, 67–73. [CrossRef]
11. Seghezzo, L. Anaerobic Treatment of Domestic Wastewater in Subtropical Regions; Wageningen University:
Wageningen, The Netherlands, 2004; Volume 58, ISBN 90-8504-029-9.
12. Gomec, C.Y. High-rate anaerobic treatment of domestic wastewater at ambient operating temperatures:
A review on benefits and drawbacks. J. Environ. Sci. Health Part. A Toxic/Hazard. Subst. Environ. Eng. 2010,
45, 1169–1184. [CrossRef]
13. Yu, H.Q.; Fang, H.H.P.; Tay, J.H. Enhanced sludge granulation in upflow anaerobic sludge blanket (UASB)
reactors by aluminum chloride. Chemosphere 2001, 44, 31–36. [CrossRef]
14. Yu, H.Q.; Tay, J.H.; Fang, H.H.P. The roles of calcium in sludge granulation during UASB reactor start-up.
Water Res. 2001, 35, 1052–1060. [CrossRef]
15. Yu, H.Q.; Fang, H.H.; Tay, J.H. Effects of Fe2+ on sludge granulation in upflow anaerobic sludge blanket
reactors. Water Sci. Technol. 2000, 41, 199–205. [CrossRef]
16. Sondhi, A.; Guha, S.; Harendranath, C.S.; Singh, A. Effect of aluminum (Al3+ ) on granulation in upflow
anaerobic sludge blanket reactor treating low-strength synthetic wastewater. Water Environ. Res. 2010,
82, 715–724. [CrossRef] [PubMed]
Water 2019, 11, 372 22 of 27
17. Tiwari, M.K.; Guha, S.; Harendranath, C.S.; Tripathi, S. Enhanced granulation by natural ionic polymer
additives in UASB reactor treating low-strength wastewater. Water Res. 2005, 39, 3801–3810. [CrossRef]
[PubMed]
18. Tiwari, M.K.; Guha, S.; Harendranath, C.S. Enhanced granulation in UASB reactor treating low-strength
wastewater by natural polymers. Water Sci. Technol. 2004, 50, 235–240. [CrossRef]
19. Wang, Y.; Show, K.Y.; Tay, J.H.; Sim, K.H. Effects of cationic polymer on start-up and granulation in upflow
anaerobic sludge blanket reactors. J. Chem. Technol. Biotechnol. 2004, 79, 219–228. [CrossRef]
20. Show, K.Y.; Wang, Y.; Foong, S.F.; Tay, J.H. Accelerated start-up and enhanced granulation in upflow
anaerobic sludge blanket reactors. Water Res. 2004, 38, 2292–2303. [CrossRef]
21. Mungray, A.K.; Patel, K. Coliforms removal in two UASB + ASP based systems. Int. Biodeterior. Biodegrad.
2011, 65, 23–28. [CrossRef]
22. Cao, Y.S.; Ang, C.M. Coupled UASB-activated sludge process for COD and nitrogen removals in municipal
sewage treatment in warm climate. Water Sci. Technol. 2009, 60, 2829–2839. [CrossRef]
23. Moawad, A.; Mahmoud, U.F.; El-Khateeb, M.A.; El-Molla, E. Coupling of sequencing batch reactor and UASB
reactor for domestic wastewater treatment. Desalination 2009, 242, 325–335. [CrossRef]
24. Guimaraes, P.; Melo, H.N.S.; Cavalcanti, P.F.F.; van Haandel, A.C. Anaerobic-Aerobic Sewage Treatment
Using the Combination UASB-SBR Activated Sludge. J. Environ. Sci. Health 2003, 38, 2633–2641. [CrossRef]
25. Aiyuk, S.; Amoako, J.; Raskin, L.; Van Haandel, A.; Verstraete, W. Removal of carbon and nutrients
from domestic wastewater using a low investment, integrated treatment concept. Water Res. 2004,
38, 3031–3042. [CrossRef] [PubMed]
26. Kalogo, Y.; Verstraete, W. Development of anaerobic sludge bed (ASB) reactor technologies for domestic
wastewater treatment: Motives and perspectives. World J. Microbiol. Biotechnol. 1999, 15, 523–534. [CrossRef]
27. Leitão, R.C.; Van Haandel, A.C.; Zeeman, G.; Lettinga, G. The effects of operational and environmental
variations on anaerobic wastewater treatment systems: A review. Bioresour. Technol. 2006, 97, 1105–1118.
[CrossRef] [PubMed]
28. Halalsheh, M.; Koppes, J.; Den Elzen, J.; Zeeman, G.; Fayyad, M.; Lettinga, G. Effect of SRT and temperature
on biological conversions and the related scum-forming potential. Water Res. 2005, 39, 2475–2482. [CrossRef]
[PubMed]
29. Lew, B.; Tarre, S.; Belavski, M.; Green, M. UASB reactor for domestic wastewater treatment at low
temperatures: A comparison between a classical UASB and hybrid UASB-filter reactor. Water Sci. Technol.
2004, 49, 295–301. [CrossRef]
30. Lew, B.; Lustig, I.; Beliavski, M.; Tarre, S.; Green, M. An integrated UASB-sludge digester system for raw
domestic wastewater treatment in temperate climates. Bioresour. Technol. 2011, 102, 4921–4924. [CrossRef]
31. Agrawal, L.K.; Harada, H.; Okui, H. Treatment of dilute wastewater in a UASB reactor at a moderate
temperature: Performance aspects. J. Ferment. Bioeng. 1997, 83, 179–184. [CrossRef]
32. Wang, K. Integrated Anaerobic and Aerobic Treatment of Sewage; Wageningen Agricultural University:
Wageningen, The Netherlands, 1994; Volume 90, ISBN 90-5485-232-1.
33. Corstanje, R. The Anaerobic Digestion of Waste Activated Sludge in a Anaerobic Upflow Solids Removal
Reactor Coupled to an Upflow Sludge Digester. Master’s Thesis, Wageningen Agricultural University,
Wageningen, The Netherlands, 1996.
34. Lettinga, G.; Rebac, S.; Zeeman, G. Challenge of psychrophilic anaerobic wastewater treatment. Trends Biotechnol.
2001, 19, 363–370. [CrossRef]
35. Kujawa-Roeleveld, K.; Zeeman, G. Anaerobic treatment in decentralised and source-separation-based
sanitation concepts. Rev. Environ. Sci. Biotechnol. 2006, 5, 115–139. [CrossRef]
36. Schink, B. Anaerobic digestion: concepts, limits and perspectives. Water Sci. Technol. 2002, 45, 1–8. [CrossRef]
37. Bajpai, P. Anaerobic Technology in Pulp and Paper Industry; Springer: Berlin/Heidelberg, Germany, 2017;
ISBN 978-981-10-4129-7.
38. Dos Santos, A.B.; Cervantes, F.J.; van Lier, J.B. Review paper on current technologies for decolourisation
of textile wastewaters: Perspectives for anaerobic biotechnology. Bioresour. Technol. 2007, 98, 2369–2385.
[CrossRef] [PubMed]
39. Mao, C.; Feng, Y.; Wang, X.; Ren, G. Review on research achievements of biogas from anaerobic digestion.
Renew. Sustain. Energy Rev. 2015, 45, 540–555. [CrossRef]
Water 2019, 11, 372 23 of 27
40. Angenent, L.T.; Karim, K.; Al-Dahhan, M.H.; Wrenn, B.A.; Domíguez-Espinosa, R. Production of bioenergy
and biochemicals from industrial and agricultural wastewater. Trends Biotechnol. 2004, 22, 477–485. [CrossRef]
41. Gallert, C.; Winter, J. Bacterial Metabolism in Wastewater Treatment Systems. In Environmental Biotechnology
Concepts and Applications; Jördening, H.-J., Winter, J., Eds.; WILEY-VCH Verlag GmbH & Co.: Weinheim,
Germany, 2005; pp. 1–41. ISBN 3-527-30585-8.
42. Khanal, S.K. Anaerobic Biotechnology for Bioenergy Production: Principles and Applications; Wiley-Blackwell,
John Wiley & Sons, Ltd.: West Sussex, UK, 2008.
43. Hutñan, M.; Drtil, M.; Derco, J.; Mrafková, L. Methanogenic and Nonmethanogenic Activity of Granulated
Sludge in Anaerobic Baffled Reactor. Chem. Pap. 1999, 53, 374–378.
44. Young, J.C.; McCarty, P.L. The anaerobic filter for waste treatment. J. Water Pollut. Control Fed. 1969, 41.
[CrossRef]
45. Lettinga, G.; van Velsen, A.F.M.; Hobma, S.W.; de Zeeuw, W.; Klapwijk, A. Use of the upflow sludge blanket
(USB) reactor concept for biological wastewater treatment, especially for anaerobic treatment. Biotechnol.
Bioeng. 1980, 22, 699–734. [CrossRef]
46. Switzenbaum, M.S.; Jewell, W.J. Anaerobic attached-film expanded-bed reactor treatment. J. Water Pollut.
Control Fed. 1980, 52, 1953–1965.
47. Murray, W.D.; Berg, L.V.D. Effect of Support Material on the Development of Microbial Fixed Films
Converting Acetic Acid to Methane. J. Appl. Bacteriol. 1981, 51, 257–265. [CrossRef]
48. Barber, W.P.; Stuckey, D.C. The use of the anaerobic baffled reactor (ABR) for wastewater treatment: A review.
Water Res. 1999, 33, 1559–1578. [CrossRef]
49. De Man, A.W.A.; van der Last, A.R.M.; Lettinga, G. The use of EGSB and UASB anaerobic systems or
low strength soluble and complex wastewaters at temperatures ranging from 8 to 30 ◦ C. In Proceedings of
the 5th International Symposium on Anaerobic Digestion; Hall, E.R., Hobson, P.N., Eds.; Monduzzi Bologna:
Milano, Italy, 1988; pp. 197–208.
50. Bogte, J.J.; Breure, A.M.; Van Andel, J.G.; Lettinga, G. Anaerobic treatment of domestic wastewater in small
scale UASB reactors. Water Sci. Technol. 1993, 27, 75–82. [CrossRef]
51. Van Lier, J.B. Thermophilic Anaerobic Wastewater Treatment: Temperature Aspects and Process Stability;
Wageningen University: Wageningen, The Netherlands, 1995; ISBN 90-5485-436-7.
52. Mortezaei, Y.; Amani, T.; Elyasi, S. High-rate anaerobic digestion of yogurt wastewater in a hybrid EGSB
and fixed-bed reactor: Optimizing through response surface methodology. Process Saf. Environ. Prot. 2018,
113, 255–263. [CrossRef]
53. Mockaitis, G.; Pantoja, J.L.R.; Rodrigues, J.A.D.; Foresti, E.; Zaiat, M. Continuous anaerobic bioreactor with
a fixed-structure bed (ABFSB) for wastewater treatment with low solids and low applied organic loading
content. Bioprocess Biosyst. Eng. 2014, 37, 1361–1368. [CrossRef] [PubMed]
54. Elmitwalli, T.A.; Oahn, K.L.T.; Zeeman, G.; Lettinga, G. Treatment of domestic sewage in a two-step anaerobic
filter/anaerobic hybrid system at low temperature. Water Res. 2002, 36, 2225–2232. [CrossRef]
55. Bodkhe, S.Y. A modified anaerobic baffled reactor for municipal wastewater treatment. J. Environ. Manag.
2009, 90, 2488–2493. [CrossRef] [PubMed]
56. Hickey, R.F.; Wu, W.M.; Veiga, M.C.; Jones, R. Start-up, operation, monitoring and control of high-rate
anaerobic treatment systems. Water Sci. Technol. 1991, 24, 207–255. [CrossRef]
57. Van der Last, A.R.M.; Lettinga, G. Anaerobic treatment of domestic sewage under moderate climatic (Dutch)
conditions using upflow reactors at increased superficial velocities. Water Sci. Technol. 1992, 25, 167–178.
[CrossRef]
58. Lettinga, G.; Hulshoff Pol, L. UASB-Process design for various types of wastewates. Water Sci. Technol. 1991,
24, 87–107. [CrossRef]
59. Van Haandel, A.; Kato, M.T.; Cavalcanti, P.F.F.; Florencio, L. Anaerobic Reactor Design Concepts for
the Treatment of Domestic Wastewater. Rev. Environ. Sci. Biotechnol. 2006, 5, 21–38. [CrossRef]
60. Sabry, T. Application of the UASB inoculated with flocculent and granular sludge in treating sewage
at different hydraulic shock loads. Bioresour. Technol. 2008, 99, 4073–4077. [CrossRef] [PubMed]
61. Elmitwalli, T.A. Anaerobic Treatment of Domestic Sewage at Low Temperature; Wageningen University:
Wageningen, The Netherlands, 2000; ISBN 90-5808-277-6.
62. Verstraete, W.; Vandevivere, P. New and broader applications of anaerobic digestion. Crit. Rev. Environ. Sci. Technol.
1999, 29, 151–173.
Water 2019, 11, 372 24 of 27
63. Van Haandel, A.C.; Lettinga, G. Anaerobic Sewage Treatment: A Practical Guide for Regions with a Hot Climate;
John Wiley and Sons Ltd.: Chichester, UK, 1994; ISBN 0471951218.
64. Rebac, S. Psychrophilic Anaerobic Treatment of Low Strength Wastewater; Wageningen Agricultural University:
Wageningen, The Netherlands, 1998; ISBN 90-5485-943-1.
65. Seghezzo, L.; Cuevas, C.M.; Trupiano, A.P.; Guerra, R.G.; González, S.M.; Zeeman, G.; Lettinga, G. Stability
and activity of anaerobic sludge from UASB reactors treating sewage in subtropical regions. Water Sci. Technol.
2006, 54, 223–229. [CrossRef]
66. Lettinga, G. Anaerobic digestion and wastewater treatment systems. Antonie Van Leeuwenhoek 1995, 67, 3–28.
[CrossRef] [PubMed]
67. Lettinga, G.; Roersma, R.; Grin, P.; de Zeeuw, W.; Pol, H.L.; van Velsen, L.; Hobma, S.; Zeeman, G.
Anaerobic treatment of sewage and low strength wastewaters. In Proceedings of the 2nd International Symposium
on Anaerobic Digestion, Travemünde, Germany, 6–11 September 1981; pp. 271–291.
68. Technologien, N.; Wirtschaftsberatung, B. Anaerobic Treatment of Municipal Wastewater in UASB—Reactors;
TBW GmbH: Frankfurt, Germany, 2001.
69. Kooijmans, J.L.; Lettinga, G.; Passa, G.R. The UASB-process for domestic wastewater treatment in developing
countries. J. Inst. Water Eng. Sci. 1985, 39, 437–451.
70. Vieira, S.M.M.; Souza, M.E. Development of Technology for the Use of the Uasb Reactor in Domestic Sewage
Treatment. Water Sci. Technol. 1987, 18, 109–121. [CrossRef]
71. Schellinkhout, A.; Collazos, C.J. Full-scale application of the UASB technology for sewage treatment.
Water Sci. Technol. 1992, 25, 159–166. [CrossRef]
72. Draaijer, H.; Maas, J.A.W.; Schaapman, J.E.; Khan, A. Performance of the 5 MLD UASB Reactor for Sewage
Treatment at Kanpur, India. Water Sci. Technol. 1992, 25, 123–133. [CrossRef]
73. Khalil, N.; Sinha, R.; Raghava, A.K.; Mittal, A.K. UASB Technology for Sewage Treatment in India:
Experience, Economic Evaluation and its Potential in Other Developing Countries. In Proceedings of
the Twelfth International Water Technology Conference 2008, Alexandria, Egypt, 27–30 March 2008.
74. Pandey, N.; Dubey, S.K. Up-flow Anaerobic Sludge Bed (UASB) based sewage treatment plant (STP)
at Mirzapur: A Review. Int. Res. J. Environ. Sci. 2014, 3, 67–71.
75. Heffernan, B.; Van Lier, J.B.; Van Der Lubbe, J. Performance review of large scale up-flow anaerobic sludge
blanket sewage treatment plants. Water Sci. Technol. 2011, 63, 100–107. [CrossRef]
76. Prashanth, S. Treatment of Sewage Using UASB Process; Indian Institute of Technology: Roorkee, India, 2003.
77. Mahmoud, N.J.A. Anaerobic Pre-Treatment of Sewage under Low Temperature (15 ◦ C) Conditions in an Integrated
UASB-Digester System; Wageningen University: Wageningen, The Netherlands, 2002.
78. Uemura, S.; Harada, H. Treatment of sewage by a UASB reactor under moderate to low temperature
conditions. Bioresour. Technol. 2000, 72, 275–282. [CrossRef]
79. Prashanth, S.; Kumar, P.; Mehrotra, I. Anaerobic Degradability: Effect of Particulate COD. J. Environ. Eng.
2006, 132, 488–496. [CrossRef]
80. Singh, K.S.; Viraraghavan, T. Start-up and operation of UASB reactors at 20 C for municipal wastewater
treatment. J. Ferment. Bioeng. 1998, 85, 609–614. [CrossRef]
81. Ruiz, I.; Soto, M.; Veiga, M.C.; Ligero, P.; Vega, A.; Blazquez, R. Performance of and biomass characterisation
in a UASB reactor treating domestic waste water at ambient temperature. Water SA 1998, 24, 215–222.
82. Bodik, I.; Herdova, B.; Drtil, M. Anaerobic treatment of the municipal wastewater under psychrophilic
conditions. Bioprocess Eng. 2000, 22, 385–390. [CrossRef]
83. Gomec, C.; Letsiou, I.; Ozturk, I.; Eroglu, V.; Wilderer, P. Identification of Archaeal population in the granular
sludge of an UASB reactor treating sewage at low temperatures. J. Environ. Sci. Health Part A Toxic/Hazard.
Subst. Environ. Eng. 2008, 43, 1504–1510. [CrossRef]
84. Lettinga, G.; Roersma, R.; Grin, P. Anaerobic treatment of raw domestic sewage at ambient temperatures
using a granular bed UASB reactor. Biotechnol. Bioeng. 1983, 27, 1701–1723. [CrossRef]
85. De Man, A.W.A.; Grin, P.C.; Roersma, R.E.; Grolle, K.C.F.; Lettinga, G. Anaerobic treatment of municipal
wastewater at low temperatures. In Proceedings of the Aquatech’86 Conference—Anaerobic Treatment,
Amsterdam, The Netherlands, 15–19 September1986; pp. 453–466.
86. Singh, K.S.; Harada, H.; Viraraghavan, T. Low-strength wastewater treatment by a UASB reactor. Bioresour. Technol.
1996, 55, 187–194. [CrossRef]
Water 2019, 11, 372 25 of 27
87. Leitao, R.C.; Silva-Filho, J.A.; Sanders, W.; van Haandel, A.C.; Zeeman, G.; Lettinga, G. The effect
of operational conditions on the performance of UASB reactors for domestic wastewater treatment.
Water Sci. Technol. 2005, 52, 299–305. [CrossRef]
88. Mahmoud, N. High strength sewage treatment in a UASB reactor and an integrated UASB-digester system.
Bioresour. Technol. 2008, 99, 7531–7538. [CrossRef]
89. Halalsheh, M.M.; Abu Rumman, Z.M.; Field, J.A. Anaerobic wastewater treatment of concentrated sewage
using a two-stage upflow anaerobic sludge blanket-anaerobic filter system. J. Environ. Sci. Health Part.
A Toxic/Hazard. Subst. Environ. Eng. 2010, 45, 383–388. [CrossRef]
90. Lettinga, G.; DeMan, A.; Grin, P.; Hulshoff Pol, L.W. Anaerobic wastewater treatment as an appropriate
technology for developing countries. Trib. Cebedeau 1987, 40, 21–32.
91. Vieira, S.M.M.; Garcia, A.D., Jr. Results and Recommendations for Design and Utilization. Water Sci. Technol.
1992, 25, 143–157. [CrossRef]
92. Vieira, S.M.M.; Carvalho, J.L.; Barijan, F.P.O.; Rech, C.M. Application of the UASB technology for sewage
treatment in a small community at Sumare, Sao Paulo State. Water Sci. Technol. 1994, 30, 203–210. [CrossRef]
93. Monroy, O.; Fama, G.; Meraz, M.; Montoya, L.; Macarie, H. Anaerobic digestion for wastewater treatment in
Mexico: State of the technology. Water Res. 2000, 34, 1803–1816. [CrossRef]
94. Halalsheh, M.; Sawajneh, Z.; Zu’bi, M.; Zeeman, G.; Lier, J.; Fayyad, M.; Lettinga, G. Treatment of strong
domestic sewage in a 96 m3 UASB reactor operated at ambient temperatures: two-stage versus single-stage
reactor. Bioresour. Technol. 2005, 96, 577–585. [CrossRef]
95. Walia, R.; Kumar, P.; Mehrotra, I. Performance of UASB based sewage treatment plant in India: Polishing by
diffusers an alternative. Water Sci. Technol. 2011, 63, 680–688. [CrossRef]
96. Fan, L.T.; Erickson, L.E.; Baltes, J.C.; Shah, P.S. Analysis and Optimization of Two-stage Digestion.
Water Pollut. Control Fed. 2000, 45, 35–52.
97. Ghosh, S. Pilot-scale demonstration of two-phase anaerobic digestion of activated sludge. Water Sci. Technol.
1991, 23, 1179–1188. [CrossRef]
98. Beccari, M.; Bonemazzi, F.; Majone, M.; Riccardi, C. Interaction between acidogenesis and methanogenesis in
the anaerobic treatment of olive oil mill effluents. Water Res. 1996, 30, 183–189. [CrossRef]
99. Zeeman, G.; Sanders, W.T.M.; Wang, K.Y.; Lettinga, G. Anaerobic treatment of complex wastewater and waste
activated sludge—Application of an upflow anaerobic solid removal (UASR) reactor for the removal
and pre-hydrolysis of suspended COD. Water Sci. Technol. 1997, 35, 121–128. [CrossRef]
100. Noike, T.; Endo, G.; Chang, J.-E.; Yaguchi, J.-I.; Matsumoto, J.-I. Characteristics of carbohydrate degradation
and the rate-limiting step in anaerobic digestion. Biotechnol. Bioeng. 1985, 27, 1482–1489. [CrossRef]
[PubMed]
101. GonCalves, R.F.; Charlier, A.C.; Sammut, F. Primary fermentation of soluble and particulate organic matter
for wastewater treatment. Water Sci. Technol. 1994, 30, 53–62. [CrossRef]
102. Ligero, P.; De Vega, A.; Soto, M. Influence of HRT (hydraulic retention time) and SRT (solid retention time)
on the hydrolytic pre-treatment of urban wastewater. Water Sci. Technol. 2001, 44, 7–14. [CrossRef] [PubMed]
103. Lin, H.-Y.; Ouyang, C.-F. Up-flow Anaerobic Sludge Digestion in a Phase Separation System.
Water Sci. Technol. 1993, 28, 133–138. [CrossRef]
104. Pascual, A.; de la Varga, D.; Arias, C.A.; Van Oirschot, D.; Kilian, R.; Álvarez, J.A.; Soto, M. Hydrolytic anaerobic
reactor and aerated constructed wetland systems for municipal wastewater treatment—HIGHWET project.
Environ. Technol. 2017, 38, 209–219. [CrossRef]
105. Álvarez, J.A.; Zapico, C.A.; Gómez, M.; Presas, J.; Soto, M. Anaerobic hydrolysis of a municipal wastewater
in a pilot-scale digester. Water Sci. Technol. 2003, 47, 223–230. [CrossRef]
106. Miron, Y.; Zeeman, G.; Van Lier, J.B.; Lettinga, G. The role of sludge retention time in the hydrolysis
and acidification of lipids, carbohydrates and proteins during digestion of primary sludge in CSTR systems.
Water Res. 2000, 34, 1705–1713. [CrossRef]
107. Ligero, P.; Vega, A.; Soto, M. Pretreatment of urban wastewaters in a hydrolytic upflow digester. Water SA
2001, 27, 399–404. [CrossRef]
108. Ligero, P.; Soto, M. Sludge granulation during anaerobic treatment of pre-hydrolysed domestic wastewater.
Water SA 2002, 28, 307–311. [CrossRef]
109. Barajas, M.G.; Escalas, A.; Mujeriego, R. Fermentation of a low VFA wastewater in an activated primary
tank. Water SA 2002, 28, 89–98. [CrossRef]
Water 2019, 11, 372 26 of 27
110. Speece, R.E. Anaerobic biotechnology for industrial wastewater treatment. Environ. Sci. Technol. 1983,
17, 416–427. [CrossRef] [PubMed]
111. Bal, A.S.; Dhagat, N.N. Upflow anaerobic sludge blanket Reactor—A review. Indian J. Environ. Health 2001,
43, 1–82. [PubMed]
112. McInerney, M.J. Anaerobic hydrolysis and fermentation of fats and proteins. In Biology of Anaerobic
Microorganisms; Zehnder, A.J.B., Ed.; Wiley: New York, NY, USA, 1988; pp. 373–415. ISBN 0471882267.
113. Henze, M.; Harremoes, P.; Cour Jansen, J.I.; Arvin, E. Wastewater Treatment Biological and Chemical Processes,
2nd ed.; Springer-Verlag: Berlin/Heidelberg, Germany, 1997; ISBN 978-3-662-22607-0.
114. Castillo, A.; Llabres, P.; Mata-Alvarez, J. A kinetic study of a combined anaerobic-aerobic system for treatment
of domestic sewage. Water Res. 1999, 33, 1742–1747. [CrossRef]
115. Vavilin, V.A.; Rytov, S.V.; Lokshina, L.Y. A description of hydrolysis kinetics in anaerobic degradation of
particulate organic matter. Bioresour. Technol. 1996, 56, 229–237. [CrossRef]
116. Vavilin, V.A.; Rytov, S.V.; Lokshina, L.Y. Two-phase model of hydrolysis kinetics and its applications to
anaerobic degradation of particulate organic matter. Appl. Biochem. Biotechnol. 1997, 63–65, 45–57. [CrossRef]
[PubMed]
117. Vavilin, V.A.; Fernandez, B.; Palatsi, J.; Flotats, X. Hydrolysis kinetics in anaerobic degradation of particulate
organic material: An overview. Waste Manag. 2008, 28, 939–951. [CrossRef]
118. Siegrist, H.; Renggli, D.; Gujer, W. Mathematical modeling of anaerobic mesophilic sewage sludge treatment.
In Proceedings of the International Symposium on Anaerobic Digestion of Solid Waste, Venice, Italy,
14–17 April 1992; pp. 51–64.
119. Batstone, D.J.; Keller, J.; Angelidaki, I.; Kalyuzhnyi, S.V.; Pavlostathis, S.G.; Rozzi, A.; Sanders, W.T.M.;
Siegrist, H.; Vavilin, V.A. The IWA Anaerobic Digestion Model No 1 (ADM1). Water Sci. Technol. 2002,
45, 65–73. [CrossRef]
120. Shang, Y.; Johnson, B.R.; Sieger, R. Application of the IWA Anaerobic Digestion Model (ADM1) for simulating
full-scale anaerobic sewage sludge digestion. Water Sci. Technol. 2005, 52, 487–492. [CrossRef]
121. Maharaj, B.C.; Mattei, M.R.; Frunzo, L.; van Hullebusch, E.D.; Esposito, G. ADM1 based mathematical
model of trace element precipitation/dissolution in anaerobic digestion processes. Bioresour. Technol. 2018,
267, 666–676. [CrossRef]
122. Uhlenhut, F.; Schlüter, K.; Gallert, C. Wet biowaste digestion: ADM1 model improvement by implementation
of known genera and activity of propionate oxidizing bacteria. Water Res. 2018, 129, 384–393. [CrossRef]
[PubMed]
123. Spyridonidis, A.; Skamagkis, T.; Lambropoulos, L.; Stamatelatou, K. Modeling of anaerobic digestion of
slaughterhouse wastes after thermal treatment using ADM1. J. Environ. Manag. 2018, 224, 49–57. [CrossRef]
[PubMed]
124. Charnier, C.; Latrille, E.; Jimenez, J.; Torrijos, M.; Sousbie, P.; Miroux, J.; Steyer, J.P. Fast ADM1 implementation for
the optimization of feeding strategy using near infrared spectroscopy. Water Res. 2017, 122, 27–35. [CrossRef]
125. Ramirez, I.; Mottet, A.; Carrère, H.; Déléris, S.; Vedrenne, F.; Steyer, J.P. Modified ADM1 disintegration/hydrolysis
structures for modeling batch thermophilic anaerobic digestion of thermally pretreated waste activated sludge.
Water Res. 2009, 43, 3479–3492. [CrossRef] [PubMed]
126. Ozgun, H. Anaerobic Digestion Model No. 1 (ADM1) for mathematical modeling of full-scale sludge digester
performance in a municipal wastewater treatment plant. Biodegradation 2018, 4. [CrossRef] [PubMed]
127. Sayed, S.K.I.; Fergala, M.A.A. Two-stage UASB concept for treatment of domestic sewage including sludge
stabilization process. Water Sci. Technol. 1995, 32, 55–63. [CrossRef]
128. Wang, K.; van der Last, A.R.M.; Lettinga, G. The hydrolysis upflow sludge bed (HUSB) and the expanded
granular sludge blanket (EGSB) reactors process for sewage treatment. In Proceedings of the 8th International
Conference on Anaerobic Digestion, Sendai, Japan, 1997; pp. 301–304.
129. Foresti, E.; Zaiat, M.; Vallero, M. Anaerobic processes as the core technology for sustainable domestic
wastewater treatment: Consolidated applications, new trends, perspectives, and challenges. Rev. Environ.
Sci. Biotechnol. 2006, 5, 3–19. [CrossRef]
130. Ma, S.; Ma, H.J.; Hu, H.D.; Ren, H.Q. Effect of mixing intensity on hydrolysis and acidification of
sewage sludge in two-stage anaerobic digestion: Characteristics of dissolved organic matter and the key
microorganisms. Water Res. 2019, 148, 359–367. [CrossRef] [PubMed]
Water 2019, 11, 372 27 of 27
131. Liu, X.; Dong, B.; Dai, X. Hydrolysis and acidification of dewatered sludge under mesophilic, thermophilic
and extreme thermophilic conditions: Effect of pH. Bioresour. Technol. 2013, 148, 461–466. [CrossRef]
132. Zhang, D.; Jiang, H.; Chang, J.; Sun, J.; Tu, W.; Wang, H. Effect of thermal hydrolysis pretreatment on
volatile fatty acids production in sludge acidification and subsequent polyhydroxyalkanoates production.
Bioresour. Technol. 2019, 279, 92–100. [CrossRef]
133. Stazi, V.; Tomei, M.C. Enhancing anaerobic treatment of domestic wastewater: State of the art, innovative
technologies and future perspectives. Sci. Total Environ. 2018, 635, 78–91. [CrossRef]
134. Chen, Y.; Jiang, X.; Xiao, K.; Shen, N.; Zeng, R.J.; Zhou, Y. Enhanced volatile fatty acids (VFAs) production
in a thermophilic fermenter with stepwise pH increase—Investigation on dissolved organic matter
transformation and microbial community shift. Water Res. 2017, 112, 261–268. [CrossRef] [PubMed]
135. Liu, H.; Han, P.; Liu, H.; Zhou, G.; Fu, B.; Zheng, Z. Full-scale production of VFAs from sewage sludge
by anaerobic alkaline fermentation to improve biological nutrients removal in domestic wastewater.
Bioresour. Technol. 2018, 260, 105–114. [CrossRef]
136. Vanwonterghem, I.; Jensen, P.D.; Ho, D.P.; Batstone, D.J.; Tyson, G.W. Linking microbial community structure,
interactions and function in anaerobic digesters using new molecular techniques. Curr. Opin. Biotechnol.
2014, 27, 55–64. [CrossRef]
137. Vanwonterghem, I.; Jensen, P.D.; Rabaey, K.; Tyson, G.W. Genome-centric resolution of microbial diversity,
metabolism and interactions in anaerobic digestion. Environ. Microbiol. 2016, 18, 3144–3158. [CrossRef]
138. Jang, H.M.; Kim, J.H.; Ha, J.H.; Park, J.M. Bacterial and methanogenic archaeal communities during
the single-stage anaerobic digestion of high-strength food wastewater. Bioresour. Technol. 2014, 165, 174–182.
[CrossRef] [PubMed]
139. Narihiro, T.; Sekiguchi, Y. Microbial communities in anaerobic digestion processes for waste and wastewater
treatment: A microbiological update. Curr. Opin. Biotechnol. 2007, 18, 273–278. [CrossRef] [PubMed]
140. Yang, Y.; Yu, K.; Xia, Y.; Lau, F.T.K.; Tang, D.T.W.; Fung, W.C.; Fang, H.H.P.; Zhang, T. Metagenomic
analysis of sludge from full-scale anaerobic digesters operated in municipal wastewater treatment plants.
Appl. Microbiol. Biotechnol. 2014, 98, 5709–5718. [CrossRef]
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