Renewable and Sustainable Energy Reviews 153 (2022) 111797

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Renewable and Sustainable Energy Reviews

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Bioenergy production from the organic fraction of municipal solid waste

and sewage sludge using mesophilic anaerobic co-digestion: An
experimental and kinetic modeling study
Afsane Sedighi a, Mohsen Karrabi a, *, Bahar Shahnavaz b, Morteza Mostafavinezhad c
a
Department of Civil Engineering, Faculty of Engineering, Ferdowsi University of Mashhad, Mashhad, Iran
b
Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran
c
Department of Civil Engineering, Faculty of Engineering, Khavaran Institute of Higher Education of Mashhad, Mashhad, Iran

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

Keywords: Following the rapid surge in global demand for energy as well as the rising generation of municipal solid waste
Methane (MSW) worldwide, much more attention has been drawn to sustainable waste management solutions to tackle
Anaerobic both problems. The present study investigated the potential for biogas production from the anaerobic co-
Co-digestion
digestion (ACD) of the organic fraction of MSW (OFMSW) and sewage sludge after physical and chemical pre­
Solid waste
Sewage sludge
treatments at three inoculum-to-substrate (I/S) ratios (ISR: 2, 4, & 6) and three levels of total solids (TS: 3, 5, &
Kinetic 7%). A series of experiments were further carried out in laboratory-scale (1-L) single-stage digesters in batch
mode at the temperature of 35 ◦ C (viz. Under mesophilic conditions) for 50 days. The results demonstrated that
the digestion produced the best outputs at TS = 5% and ISR = 2, whereby it offered the biogas yield of 0.697, the
methane (CH4) production of 0.448 m3/Kg-volatile solid (VS)added, and the total VS (TVS) removal by 65%.
Moreover, it was observed that most reactors with TS = 7% encountered the phase separation problem while
augmenting the TS content from 3 to 5% increased the biogas yield and CH4 production. The kinetic study of the
mesophilic ACD additionally showed that both models of modified Gompertz (GM) equation and logistic function
(LF) provided a reasonably accurate description of biogas production, but the first-order (FO) model failed in this
task. Furthermore, the amount of sludge (namely, ISR) in the digester enlarged and the models showed a better
fit to the experimental data.

incineration or landfilling, to prevent air and groundwater pollution.

1. Introduction
Statistics show that the global generation of municipal solid waste
(MSW) is increasing at an annual rate of 2–3%. Indeed, waste manage­
ment has long been one of the major challenges facing urban manage­
ment in big cities [1]. Among the components constituting MSW, its
organic fraction (OF), which contains food waste, paper products, and
green waste, can be decomposed through the biochemical process,
called anaerobic co-digestion (ACD), inducing biogas as the final prod­
uct [2]. In addition to MSW, cities also produce other types of wastes
such as a large amount of sludge from municipal wastewater treatment
plants, which can impose substantial treatment and disposal costs on
urban management organizations. In recent years, some limitations
have been placed on common sewage-sludge disposal methods, such as
This has accordingly drawn much more attention to ACD as a process for
sludge disposal, which offers features such as the excellent stabilization
of organic matter (OM) and pathogenic microorganisms as well as
biogas production [3].
Considering the finite nature of non-renewable energy sources (most
importantly, fossil fuels), their frequent price fluctuations, and rising
awareness about the environmental impacts of greenhouse gas (GHG)
emissions, energy and environment policy-makers have long searched
for alternative energy sources with high renewability as well as effec­
tiveness in soothing environmental concerns and preventing climate
change [4]. Over recent years, growing attention has been directed to­
ward the energy generation potentials of biomass resources including
MSW and sewage sludge, which are estimated to account for over 20% of
the world’s total energy supply in 2020 [1]. As compared with other

* Corresponding author.
E-mail addresses: afsane.sedighi@alumni.um.ac.ir (A. Sedighi), karrabi@um.ac.ir (M. Karrabi), shahnavaz@um.ac.ir (B. Shahnavaz), morteza.mostafavinezhad@
yahoo.com (M. Mostafavinezhad).

https://doi.org/10.1016/j.rser.2021.111797
Received 7 July 2021; Received in revised form 18 September 2021; Accepted 17 October 2021
Available online 23 October 2021
1364-0321/© 2021 Elsevier Ltd. All rights reserved.
A. Sedighi et al.
Nomeclature
TS Total solid (% of waste)
VS Volatile Solids (% of TS)
TVS Total Volatile Solids
ISR Inoculum to Substrste Ratio (VS/VS)
pH Power of Hydrogen (− )
MSW Municiple Solid Waste
OFMSW Organic Fraction of Municiple Solid Waste
AD Anaerobic digestion
ACD Anaerobic co-digestion
FAs Fatty Acids
CNR Carbon to Nitrogen Ratio (− )
renewable energies, the use of these biomass resources has advantages,
such as (i) universal availability, (ii) no environmental problems
including GHG emissions, (iii) clean energy production, and (iv)
recruitment of simple technologies for conversion purposes [4,5].
Within ACD, anaerobic bacteria decompose OM in the absence of oxygen
(O), producing biogas, such as methane (CH4) and carbon dioxide (CO2).
Thanks to its biochemical nature and complex reactions, the ACD effi­
ciency can be affected by various factors, such as the
inoculum-to-substrate (I/S) ratio (ISR), carbon-to-nitrogen (C/N) ratio
(CNR), moisture content, digestion duration, process temperature, po­
tential hydrogen (pH), alkalinity, and volatile matter percentage [6].
In Izumi et al. (2010), reflecting on biogas production from food
waste and the effect of particle size on its performance, waste crushing
had increased its solubility by 30%, but too much particle size reduction
could lead to the accumulation of volatile fatty acids (FAs), which could
ultimately result in reduced solubility and lower CH4 production [7]. As
well, Parra et al. (2016) investigating the effect of mixing ratio on the
ACD of municipal bio-waste with domestic wastewater sludge had re­
ported that using particles smaller than 2 mm had resulted in 0.128
L/g-volatile solid (VS) (~19%) more CH4 production from reactors
compared with larger particles [8]. Brown et al. (2012) had also
examined the process of biogas production from the co-digestion of food
and backyard wastes with different mixing ratios and had analyzed how
it would be affected by the use of different feedstock-to-inoculum (F/I)
ratios (FIRs) [9].
Chen et al. (2014) had correspondingly studied the application of
food and green waste mixtures at six different ratios for biogas pro­
duction. They had reported that as the food waste percentage in the
feedstock had increased, so did the CH4 yield, but the retention time had
declined once the green waste percentage had augmented. After
selecting the food waste-green waste ratio of 40:60 as the optimal
mixing one, they had also examined the effect of the total solid (TS) level
at the range of 5–25%. During this examination, they had found that CH4
production had been higher in the digestion with lower moisture
(namely, higher TS levels) [10]. Alrawashdeh et al. (2017) had similarly
reported that the co-digestion of untreated sludge and the OFMSW had
resulted in up to 36% higher biogas production and 94% higher CH4
content than the individual activated sludge digestion [11]. Moreover,
Keucken et al. (2018) had established that the ACD of mixed sludge and
OFMSW at a ratio of 1:1, based on the VS levels, had led to stable mi­
crobial activity and rapid adaptation to acetate degradation, thus
resulting in higher CH4 production compared with the individual acti­
vated sludge digestion [12]. As well, Duan et al. (2012), investigating
the anaerobic digestion (AD) of sewage sludge had reported that
increasing the TS level from 10 to 15% had boosted the CH4 yield by
7.4% and then reduced the removal of organic solids by 6% [13].
In order to compare thermophilic and mesophilic AD, Ince et al.
(2017) had further applied maximum organic loads onto two thermo­
philic (55 ◦ C) and mesophilic (35 ◦ C) reactors. They had further found
2
Renewable and Sustainable Energy Reviews 153 (2022) 111797
CRD Randomized Design Completely
FO First Order Model
MG Modified Gompertz Model
LF Logistic Function Model
S (m3/kg-VS) Maximum Biogas Production
Rm Maximum Cumulative Biogas Production rate (m3/kg-VS.
d)
λ Lag phase (d)
k Reaction Rate Constant (d− 1)
t Time (d)
RMSE Root Mean Square Error (m3/kg-VS)
i Data Point
that thermophilic reactors could offer higher digestion performance and
lower digestion duration [14]. However, Guo et al. (2018) had reported
that certain acetoclastic-methanogenic and propionate-butyrate oxida­
tive activities during the AD of food waste had been significantly higher
in mesophilic reactors. Under mesophilic conditions, the particles of
1–2 mm in diameter had also formed crystalline calcium complexes,
which had mitigated the inhibitory effect of long-chain FAs during AD. It
had become clear that both temperature regimes could be suitable,
depending on substrate and retention time. In other words, the ther­
mophilic condition could be more appropriate for the digestion of
high-fiber biomasses with short retention times, while the mesophilic
condition could be preferred for the digestion of biodegradable mate­
rials with a long retention time [15]. Given the raw nature of OMs, the
AD of the wastes that included such materials may thus affect
biochemical reactions, especially at the hydrolysis, acidogenesis, ace­
togenesis, and methanogenesis stages. However, the ACD of certain
wastes (including MSW and sewage sludge) together may improve the
process at various stages and boost biogas production efficiency [16].
In general, widespread access to fossil fuel resources, as well as the
very low costs of energy carriers, has thus far made energy decision-
makers reluctant to develop renewable energies. This has also led to
growing consumption of fossil fuels and consequently environmental
damage. In addition, in countries such as Iran, the food industry is not
sufficiently developed in terms of proper food processing and packaging,
and the culture of food consumption has given rise to the burial of a
large number of biodegradable food materials along with MSW in
landfills. This can undoubtedly increase GHG emissions and contami­
nate groundwater resources [17,18]. In the meantime, conducting
fundamental and applied research on biomass resources and sustainable
energy development can influence government approaches toward new
energy sources. As little research has been so far conducted on the ACD
of OFMSW along with sewage sludge, the effect of two parameters, viz.
TS and ISR, on co-digestion performance was simultaneously investi­
gated in this study. In addition, the ACD kinetics, less examined in
previous works, was explored to simulate biogas production. Of note,
the most widely used kinetic model in AD has been the first-order (FO)
equation, which provides information on the rate constant of hydrolysis.
However, this model fails to assess some very important aspects, espe­
cially operating batch-mode anaerobic reactors. Therefore, predicting
biomass decomposition behavior and its relationship with AD
by-products (mainly, CH4 production) has doubled the need to use other
kinetic models. In this regard, some models have been developed to
demonstrate the important features of AD, including the lag phase of
biogas production as well as the daily biogas yield. For this purpose, the
present study aimed to (i) investigate the potential for biogas production
from the mesophilic ACD of OFMSW together with sewage sludge, (ii)
evaluate the effect of TS at three levels (viz. 3, 5, & 7%), and ISRs (that
is, 2, 4, & 6%) on ACD, and (iii) present the kinetic modeling of biogas
production using models such as FO, logistic function (LF), and modified
A. Sedighi et al.
Gompertz (MG).
2. Materials and methods
In this study, the anaerobic sludge from a municipal wastewater
treatment plant was used as the inoculum and the OFMSW was utilized
as the substrate of ACD. In general, ACD could be performed at one or
multiple stages and under dry or wet conditions, in thermophilic
(50–60 ◦ C) or mesophilic (30–40 ◦ C) conditions, and continuously or in
batch modes [19,20]. In this study, ACD was completed at the
single-stage batch mode under wet and mesophilic (35 ◦ C) conditions.
The raw materials were also subjected to physical pretreatment (i.e.,
crushing to increase contact surface) as well as chemical pretreatment
(namely, adjusting pH and alkalinity) to optimize the process conditions
for maximum biogas production. In this study, the effects of ISR (at 2, 4,
& 6%) and TS levels (at 3, 5, & 7%) on the biogas production were
further investigated in laboratory-scale (1-L) digesters. The CNR was
also tested before loading. The TS level, total VS (TVS) content, pH, and
alkalinity were then measured at the retention times of 5, 15, and 50
days. Of note, the biogas production and the CH4 content were measured
every day.
2.1. Specifications of raw materials
During the ACD in this study, the substrate was OFMSW. To prepare
this substrate, five fresh waste samples were collected from the Waste
Management Organization of Mashhad Municipality (Mashhad, Iran)
and 2 Kg of each one were mixed to reach a completely random 10 Kg
final sample [21]. The ratio of different components in this waste after
separation and weighing is illustrated in Table 1. This waste contained
non-degradable components such as plastics, glasses, and metals, which
had to be separated and discarded. To prepare the waste for ACD, all the
materials were crushed until reaching a homogenous mix of particles
smaller than 2 mm. The crushed materials were then stored in a sealed
plastic container at 4 ◦ C for 24 h before loading into the digesters [22].
The inoculum of the ACD in this study was the sludge from a
municipal wastewater treatment plant. To prepare this inoculum, 150 L
of anaerobic sludge was collected from a municipal wastewater treat­
ment plant located in the city of Torqabeh (Mashhad, Iran) and then
transferred to the Environmental Engineering Laboratory of the Fer­
dowsi University of Mashhad, Mashhad, Iran. To ensure full anaerobi­
cization of the sludge and prevent impurities (i.e., nutrients) to affect the
experiments, the sludge was stored in a sealed plastic container under
anaerobic conditions for 1 month at room temperature (with an average
of 18 ◦ C). The main purpose during this process was to make sure that all
the nutrients were consumed by microorganisms and the sludge could
reach the endogenous phase. After this, the supernatant was removed to
obtain a more concentrated sludge with higher TS levels. The product
was subsequently mixed until perfect homogeneity was reached.
Following this process, 40 L of the prepared sludge was separated and
placed in sealed plastic containers at 4 ◦ C, for 24 h, before digestion
[22].
Table 1
The composition of OFMSW used in the present study.
Components Values (%)
Fruit waste 35
Vegetable waste 20
Protein products 5 bags with a 60-mL syringe [24]. As well, CH4 production analysis was
Cellulose products 5 performed using the method developed in Kaluza et al. (2014) by
Carbohydrates 15
Inseparable and identifiable materials 20
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Renewable and Sustainable Energy Reviews 153 (2022) 111797
2.2. Experiment procedure
Upon preparing both substrate and inoculum, moisture, TS, and TVS
contents, they were measured using the standard methods instructed by
the United States Environmental Protection Agency (EPA)-821-R-01-
015, alkalinity was calculated as instructed in the American Public
Health Association (APHA) 2017, pH was determined via a Hanna
(HI98194-USA) multi-parameter probe, and the CNR was established
through the standard method of APHA (namely, CHNS experiment)
[23]. The experiments were also developed using a completely ran­
domized factorial design for two factors of ISRs (viz. 2, 4, & 6) and TS at
three levels (i.e., 3, 5, 7%) with three replications and three control
samples. The experiments were further conducted in 36 pre-designed
anaerobic digesters. The quantity of each material to be placed in
each digester was also determined based on the target ISR and TS levels
and the effective volume of the digesters (700 mL). It should be noted
that some treatments required sludge concentration, which was done by
the use of a JB-Eliminator vacuum pump (DV-6E-250 – USA), an
Erlenmeyer vacuum flask, a Buchner funnel, and Whatman filter papers.
In addition, the previously collected supernatant was utilized to provide
the required moisture for some treatments. With regard to the low pH of
the raw materials (i.e., 6.2 for the sludge and 6.9 for the OFMSW) and
the possibility of further pH drop during acidogenesis, which could
result in insufficient intrinsic alkalinity (due to insufficient protein
compounds), a chemical pretreatment was performed by adding 7 M
sodium hydroxide (NaOH, Merck – Germany) solution until a pH of 7.5
was reached. For each group of digesters (that is, 9 groups), a control
digester containing sludge with similar characteristics was also consid­
ered. The information obtained from these control digesters was
consequently employed to assess the changes in the biogas production
process.
As mentioned, biogas production and CH4 content were measured on
a daily basis. Given the difficulty of installing a drain valve on each 1-L
reactor and the possibility of errors due to the large volume of samples
relative to the total volume, TS and TVS contents, as well as pH and
alkalinity were measured in two other reactors under the same condi­
tions at the retention times of 5, 15, and 50 days at each stage of the
experiment. The experiment was further carried out using a semi-
automatic system built at the Environmental Engineering Laboratory
of the Ferdowsi University of Mashhad, Mashhad, Iran. The main com­
ponents of this system included a set of reactors, biogas collection and
storage bags, and water baths (as a system for digester temperature
control).
As shown in Fig. 1, 1-L glass digesters (DURAN®– Germany) were
firstly coupled via pneumatic connections to 2-L gas collection bags.
Inside the digesters, washers and silicone adhesives were also used to
prevent biogas leakage from inside and air infiltration from outside in
order to create completely anaerobic conditions. All 1-L reactors were
further loaded to an effective volume of 0.7 L to reach a 1:1 reactor
diameter to height ratio. To generate an anaerobic environment, first,
the air inside the contents of the reactor was vacuumed out by a JB-
Eliminator vacuum pump (DV-6E-250 – USA), and then the remaining
air was taken out by injecting nitrogen gas through a V-shaped (namely,
two-way) fitting installed above the lid of the reactors. To keep the re­
actors at 35 ◦ C, a 200-L insulated water bath with a water circulator was
made in the laboratory environment. The temperature of this water bath
was also controlled by a precise electrical circuit (RTD temperature
sensor, Autonics TC4y thermostat – South Korea, 100mc contactor, two
2000-W industrial elements) with a maximum variation of ±0.5 ◦ C [22].
The produced biogas was stored in a series of bags made from
corrosion-resistant plastic, which were then connected to the digesters.
The volume of biogas produced was further measured by vacuuming the
titration [25]. For this purpose, 5 mL of the produced biogas was
injected into 7 M NaOH solution in an Einhorn’s saccharometer. Given
A. Sedighi et al. Renewable and Sustainable Energy Reviews 153 (2022) 111797

Fig. 1. Schematic diagram of the anaerobic bioreactor, 1) Electric circuit with temperature display, 2) External heating tank, 3) Element (2000w), 4) Submersible
pump, 5) RTD temperature Sensor, 6) Discharge valve, 7) Baffle, 8) Digester, 9) Pneumatic connections, 10) Three way stopcock, 11) Gas storage bag [22].

the rapid dissolution of hydrogen sulfide (H2S) and CO2 (namely, other regression [34], in such a way as to achieve the highest fit to the
gasses in the produced biogas) in a strong base solution, the gas that experimental data [32]. During this regression process, the coefficient of
remained after this process was considered to be completely CH4 [19]. determination (R2) and the root mean square error (RMSE) (Equation
(4)) were further employed as the measure of the model’s ability to
predict cumulative biogas production [35,36].
2.3. Kinetic modeling √̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅̅
√N ( )2
√∑
It should be noted that the ACD of OFMSW and sewage sludge pro­ √
√i=1 Se,i − Sm,i
duces some biogas, which can serve as a renewable energy source. The RMSE = (4)
N
kinetic analysis of this ACD is also essential for predicting its biogas
production behavior [26]. The mathematical modeling in the kinetic wherein Se,i represents the biogas yield measured in the experiment, Sm,i
process is thus crucial for ACD performance optimization, simulation, refers to the biogas yield simulated by the model, and N indicates a
and monitoring [27]. In this study, the kinetics of the biogas production number of measurements (namely, data points). The lowest RMSE value
process was modeled with MG, LF, and FO equations, and the resulting also suggested the highest compliance of the model with the experi­
models were utilized to simulate the cumulative biogas production. Each mental data.
of these models had its own advantages. For example, while the FO
model (Equation (1)) could give more detailed information on the hy­ 3. Results and discussion
drolysis rate constant, the MG model (Equation (2)) could provide more
information on the lag period and the maximum biogas production rate, The results of the experiments performed to determine the moisture
which could be employed to optimize process design parameters content, TS, TVS, alkalinity, pH, and CNR are provided in Table 2. Upon
[28–32]. The LF model (Equation (3)) was further suitable for an initial preparing the raw materials and mixing them based on the target ISRs
exponential increase, leading to stabilization at the highest biogas pro­ and TS levels, a few preliminary tests were performed to measure pH and
duction levels [33]. alkalinity inside the digesters (Table 3), revealing that the alkalinity and
S(t) = S.(1 − exp(− k.t)) (1) pH of the environment would fall outside the optimal range for the ac­
tivity of methanogenic bacteria right from the beginning of the experi­
( (
Rm
)) ment. Therefore, to ensure ACD in all treatments with different ISRs and
S(t) = S.exp − exp .exp(λ − t) + 1 (2) TS levels, it was necessary to adjust the alkalinity and pH of the raw
S
materials, which was fulfilled by conducting an alkaline pretreatment
S with concentrated NaOH to adjust the pH of all treatments to 7.5 and
S(t) = ( ) (3)
4Rm
1 + exp S
(λ − t) + 2
Table 2
where S(t) refers to the cumulative biogas production at time t (m3/kg- The specifications of OFMSW and anaerobic sludge.
VS), S stands for the maximum biogas production (m3/kg-VS), Rm rep­ parameter OFMSW Sewage sludge
resents the maximum cumulative biogas production rate (m3/kg-VS.d), Total Solids (%) 16 ± 0.5 3 ± 0.2
λ shows the lag period before the start of biogas production (d), k de­ Volatile Solids (%) 93 ± 3 75 ± 2
notes the FO reaction rate constant (d− 1), and t suggests the time for pH 6.9 ± 0.1 6.2 ± 0.1
which the process has been in progress (d). For these equations, the Alkalinity (mg/l CaCO3) 1800 ± 85 1250 ± 55
C/N ratio 33.2 ± 2.5 14.9 ± 1.4
kinetic parameters of Rm, λ, and k were estimated via a nonlinear

4
A. Sedighi et al. Renewable and Sustainable Energy Reviews 153 (2022) 111797

Table 3
The specifications of treatments before feeding and at the beginning of the anaerobic process.
ISR 3 Total Solids (%) 7

2 4 6 2 4 6 2 4 6

Before feeding pH 6.4 ± 0.1 6.3 ± 0.1 6.1 ± 0.1 6.7 ± 0.1 6.5 ± 0.1 6.4 ± 0.1 7 ± 0.1 6.7 ± 0.1 6.4 ± 0.1
Alkalinity (mg/l as 1450 ± 65 1350 ± 60 1200 ± 42 1650 ± 75 1600 ± 72 1500 ± 68 2100 ± 1750 ± 80 1600 ± 72
CaCO3) 105
pH 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1 7.5 ± 0.1
Fed-batch Alkalinity (mg/l as 2200 ± 2200 ± 2100 ± 2350 ± 2250 ± 2200 ± 2500 ± 2350 ± 2200 ±
reactors CaCO3) 130 130 120 140 135 130 150 140 130
TVS (%) 77 77 77 77 77 77 77 77 77
TVS (gr) 16 16 16 27 27 27 38 38 38
NaOH added (gr/l) 0.57 0.59 0.71 0.65 0.53 0.54 0.33 0.53 0.45

their alkalinity to 2300 ± 200, as shown in Table 3. Accordingly, the TVS
content of all treatments was 77%.
3.1. CH4 production
Fig. 2 shows the diagram of cumulative CH4 production for different
TS levels and ISRs. At TS = 3%, the highest CH4 production (6847 mL)
occurred at ISR = 6. There was also no significant difference between the
amount of CH4 produced at ISRs = 4 and 2, indicating the effect of the
population of anaerobic bacteria on CH4 production, because the pres­
ence of larger populations of these bacteria could increase the anaerobic
reactions, thereby leading to a faster decline in TVS content and more
biogas production. The TVS reduction rate observed at different ISRs
additionally supported this argument. It is worth noting that both CNR
and VS had a deep impact on ACD. Moreover, C could further increase
CH4 production capacity, and N did the same by augmenting the
metabolism in the materials being digested. Reaching the best results at
ISR = 6 could be also due to the high amount of N compared with C in
municipal wastewater, which resulted in more active reaction, degra­
dation, and digestion of the materials in this wastewater, and ultimately
produced more CH4 [37–39]. To generate more CH4, the raw materials
needed to have a large VS content. While CNR and VS level were both
higher in OFMSW than in sewage sludge, the digesters that contained
more sludge at TS = 3% produced more CH4, because the population of
microorganisms could have a greater impact on biogas production
where there were fewer microorganisms and nutrients at lower

Fig. 2. Cumulative CH4 production and yield at the TS levels of (a) 3, (b) 5, and (c) 7%.

5
A. Sedighi et al.
concentrations (that is, higher moisture contents).
Biogas production and CH4 content could be also influenced by
several other factors including pH and alkalinity. During the experi­
ments in this study, biogas production showed a decreasing trend after
the first day. Given the decrease in pH and alkalinity (Fig. 3), the growth
and activity of acidogenic microorganisms were also much faster than
methanogenic ones, leading to the slight acidification of the environ­
ment. In Borowski (2015), reflecting on the co-digestion of OFMSW and
sewage sludge, it had been further reported that the hydrolysis of the
compounds in the early days and then CO2 production during acido­
genesis could lower pH, and thus reduce biogas production [40]. After
some time, the acidogenesis phase could end and the population of
methanogenic microorganisms could start to adapt and grow; as well,
volatile FAs could be consumed, and pH and alkalinity could start to
increase (due to ammonia [NH3] production), resulting in higher biogas
production. The studies carried out by Fonoll et al. (2015) and Siciliano
et al. (2016) had similarly reported a surge in pH and biogas production
following acid consumption [41,42]. Therefore, pH in the digestion
environment could be assumed as an important indicator of digestion
optimality and stability.
At ISR = 2, biogas production started to drop after the first day and
this trend continued until the thirteenth day, but then started to in­
crease. According to the pH and alkalinity measurements made on the
fifth and fifteenth days, the pH of this treatment also started to elevate
roughly on the thirteenth day. Kangle et al. (2012) had further shown
that the decomposition of OM, especially proteins and N-containing
substances in a digester, could produce NH3, which could amplify the
alkalinity of the environment [43]. In the present study, a 30% rise in
alkalinity on the fifteenth day additionally prevented the accumulation
Fig. 3. pH and alkalinity at the TS levels of (a) 3, (b) 5, and (c) 7%.
6
Renewable and Sustainable Energy Reviews 153 (2022) 111797
of FAs produced during the decomposition phase, leading to higher pH.
The same trend was also observed at ISR = 4; but, at ISR = 6, the initial
pH drop in the hydrolysis and acidogenesis phases was smaller than that
at ISRs = 2 and 4, probably due to the higher concentration of NaOH
injected into this digester. Furthermore, at ISR = 6, the 45% increase in
alkalinity on the fifteenth day indicated the faster onset of the meth­
anogenesis phase and the better activity of microorganisms during this
phase thanks to the presence of better conditions compared with other
digesters, which had led to an escalation in biogas production. Ac­
cording to Zupancic et al. (2012), the optimal pH ranges for acidogenic
and methanogenic microorganisms were respectively 5.2–6.3 and
6.7–7.5 [44]. Therefore, the better results of ISR = 6 and TS = 3% could
be attributed to more suitable pH and alkalinity conditions for the
growth and activity of methanogenic microorganisms (because of the
NaOH concentration injected into the alkaline pretreatment process).
At TS = 5%, the highest CH4 production and the highest VS removal
rates were further related to ISRs = 2, 4, and 6, respectively, which was
the opposite of what occurred at TS = 3% (wherein ISR = 6 had the
highest CH4 production). To explain the better results of TS = 5%
compared with TS = 3%, firstly, it was argued that the higher dilution of
the digesters with TS = 3% had reduced the microbial population and its
required substrate, resulting in lower biogas and CH4 production. Sec­
ondly, the quality of the substrate in terms of CNR and VS level could
play a more decisive role in CH4 yield because of the larger microbial
population at TS = 5%. As well, given the better quality of the substrate
at ISR = 2 compared with ISR = 4 and at ISR = 4 compared with ISR = 6,
the best digesters in terms of biogas and CH4 production and primary
CH4 yield were those with ISR = 2 and then ISR = 4 above those with
ISR = 6 (Table 4). In Ahmadi Pirloo et al. (2017), the ACD of MSW and
A. Sedighi et al.
Table 4
CNR at the onset and end of ACD.
Total Solids (%)
3 5
C/N ISR 2 4 6 2
Fed-batch reactors 20.8 ± 1.2 19.6 ± 1.5 18.5 ± 1.1
End of process 12.9 ± 0.5 10.4 ± 0.4 11.5 ± 0.5
sewage sludge with four mixing ratios (viz. 50:50, 40:60, 20:80, &
10:90) had been also investigated at 37 ◦ C for 30 days, and the optimal
MSW-sewage sludge ratio had been determined by 40:60. Based on this
ratio, the effect of TS levels of 5, 10, 15, and 20% had been also
examined, revealing that CH4 yield was higher at the TS content of 5%.
These researchers had further reported that biogas and CH4 production
had respectively increased by 15 and 18% after pretreating the feedstock
that had TS = 5% with 6% (by the weight of raw material) NaOH [45].
However, the best results in this regard had been obtained at TS = 7% in
the studies by Cesaro et al. (2012) and Patil et al. (2014) [46,47].
It should be noted that the treatment with ISR = 6 reached its CH4
yield at TS = 5 and 3% within a shorter retention time. In Boulanger
et al. (2012), examining the ACD of MSW, it had been also reported that
increasing ISR had boosted CH4 yield and decreased the retention time,
attributed to the reduced risk of inhibition due to pH drop and the
increased population of methanogenic bacteria [48]. In the present
study, the growth in biogas and CH4 production after the initial reduc­
tion started from the fifth day. Considering the rise in pH and alkalinity
between the fifth and fifteenth days, it was concluded that the reactors
left the acidogenesis phase and the methanogenic bacteria started to
consume the produced acids on the fifth day. With the onset of CH4
generation in the reactor, the volatile FAs produced at the previous stage
were metabolized by CH4-producing bacteria and their amount in the
reactor was reduced. According to a study carried out by Panigrahi et al.
(2020), CNR decreased, resulting in higher NH3 production, which led to
an increase in pH as well as the optimization of the CH4 production
process [36]. At ISR = 4 and 6, the increasing trend of biogas and CH4
production also started from the sixth day, which was as a result of the
consumption of the volatile FAs by methanogenic bacteria, considering
the rise in pH between the fifth and the fifteenth days. In Zhang et al.
(2011), upon reaching the methanogenesis phase of the digestion pro­
cess, NH3 concentration in the environment elevated and the pH rose to
even above 8 [49]. In a study conducted by Kumar et al. (2021), it had
been also reported that pH at the range of 6.8–7.2 could result in stable
CH4 production [50]. Given the obtained results and the similarity of the
pH and alkalinity trends at TS = 5% (Fig. 3), the better outputs obtained
from the digester with ISR = 2 compared with other ISRs could be
attributed to the difference between their CNRs.
In the reactors with TS = 7% and ISRs = 2 or 6, a sharp pH drop one
the early days (because of the activity of the microorganisms at the
hydrolysis and acidogenesis phases) caused the pH of the environment
to fall outside the suitable range for the activity of methanogenic mi­
croorganisms (that is, pH from 6 to 8). This led to the reduced activity of
methanogenic microorganisms compared with acidogenic ones and the
accumulation of acid in the digester (Fig. 3), resulting in a substantial
reduction in CH4 production throughout the digestion period. The
reduction was so severe that the digesters could not even match the
production rates of the control ones. In the reactor with TS = 7% and
ISR = 4, however, the pH drop on the early days was not severe enough
to halt the activity of methanogenic microorganisms (that is, it remained
above 6). Over time, the activity of these microorganisms increased
alkalinity by 25%, which also augmented pH. The better control of
conditions (viz. pH and alkalinity) in this digester could be due to the
higher amount of NaOH injected into the alkaline pretreatment stage,
which created more resistance to pH drop on the early days, and
consequently led to the better activity of methanogenic microorganisms
in the following period.
Renewable and Sustainable Energy Reviews 153 (2022) 111797
7
4 6 2 4 6
27.5 ± 1.7 24.8 ± 1.5 20.2 ± 1.1 20.4 ± 1.1 19.2 ± 0.9 18.8 ± 0.8
10.6 ± 0.4 13.2 ± 0.6 9.8 ± 0.4 11.7 ± 0.5 9.3 ± 0.4 10.4 ± 0.4
The observations made during the experiment (an example is dis­
played in Fig. 4) showed that the high concentration of the load in a
small digestion environment without a stirrer in the digesters with TS =
7% and ISRs = 2 or 6 resulted in the separation of its materials into a
solid phase (viz. sediments at the bottom), a liquid phase (namely, semi-
opaque phase in the middle) and a semi-solid phase (floating above the
liquid phase) on the early days of the process. At ISR = 4, this occurred
well after the early days of the process (in the middle). In any case, this
separation reduced the contact surface between the materials in the
reactor and decreased the effective digestion volume.
3.2. Kinetic study
Fig. 5 displays the fit of the FO, MG, and LF models to the experi­
mental data (namely, cumulative biogas production) at TS = 3% for
ISRs = 2, 4, and 6 as well as the control sample (i.e., sludge). These
results revealed that the models had a better fit to the experimental data
as ISR and the amount of sludge in the digester increased. Indeed, the
best fit to the experimental data with the lowest RMSE was associated
with the sludge as the control sample, due to the presence of more
anaerobic microorganisms in the digesters with higher ISR. As the
amount of the sludge relative to the substrate in the digester elevated,
the lag phase of the experimental curve (viz. the roughly horizontal part
of the curve before the twentieth day) drastically shortened so that this
part was removed for the control sample, allowing the models to achieve
a better fit to the experimental data.
The results also established that the worst fit to the experimental
data in all cases belonged to the FO model and the best fit was for the LF
one, which could always perform slightly better than the MG equation in
this respect. It should be noted that here the best model was in the one
with the highest R2 and the lowest RMSE values, indicating its better
performance in explaining general variations in the data [33]. The ki­
netic parameters calculated in the modeling process are given in Table 5.
Accordingly, there was only a small difference between the RMSE of the
MG and LF models, but they were notably lower than the values of the
FO model. The λ value was also at the range of 1–2 days in all cases
except for the control sample. This suggested that the microorganisms
had a lag of 1–2 days before starting to decompose the substrate and
produce the biogas. As presented in Table 5, the reaction rate constant
(k) rose as the ISR and the amount of sludge in the digesters increased.
Since a larger k indicated a higher biodegradation rate, this might prove
the effect of microbial population on the biogas production rate [51].
The highest Rm values given by the MG (24.35 m3/kg-VS.d) and LF
(22.33 m3/kg-VS.d) models were thus related to ISR = 6, which had the
highest cumulative biogas yield.
As shown in Fig. 6, the models exhibit a better fit to the experimental
data at TS = 5% than TS = 3%. At TS = 5%, the MG model accordingly
produced accurate cumulative biogas production estimates for all ISRs
and sludge amounts, and its outputs had the highest consistency with the
experimental data (namely, the lowest RMSE). At this TS level, the es­
timates of the LF model were also highly corresponding to the experi­
mental data, and even the FO model produced somewhat acceptable
biogas production estimates. As presented in Table 6, at TS = 5%, λ was
mostly zero, indicating no lag in biogas production. This could be due to
the presence of more anaerobic sludge in the digester compared with TS
= 3%, which allowed biogas production to take place even before the
substrate was introduced. As with TS = 3%, the reaction rate constant
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A. Sedighi et al. Renewable and Sustainable Energy Reviews 153 (2022) 111797

Fig. 4. The phase separation of digester contents with TS = 7% into three distinct layers.

Fig. 5. Kinetic models and experimental data in TS = 3% for the ISRs of (a) 2, (b) 4, (c) 6, and (d) the sludge.

(k) at TS = 5% also elevated with the increase in the ISR and the amount kinetic models and the experimental data. Accordingly, the models
of sludge loaded to the digester. achieved a better fit to the experimental data as the ISR and the amount
Fig. 7 depicts the biogas production trend at TS = 7%, as given by the of sludge in the digester increased. For TS = 7%, the MG and LF models

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A. Sedighi et al. Renewable and Sustainable Energy Reviews 153 (2022) 111797

Table 5
Parameters estimated by the kinetic models of biogas production in TS = 3%.
First-order Kinetic models Logistic function
parameter
Modified Gompertz

ISR 2 4 6 Sludge 2 4 6 Sludge 2 4 6 Sludge

k 0.067 0.073 0.091 0.078 – – – – – – – –
Rm - – – – 16.51 17.37 24.35 15.67 17 16.43 22.33 12.4
λ - – – – 0.84 1.62 1.09 1.81 2.13 1.94 1.25 0
R2 0.78 0.882 0.912 0.913 0.814 0.937 0.972 0.733 0.92 0.952 0.888 0.798
RMSE 0.1088 0.1027 0.0904 0.0346 0.0596 0.0391 0.0252 0.0246 0.0547 0.0299 0.0253 0.0246

Fig. 6. Kinetic models and experimental data in TS = 5% for the ISRs of (a) 2, (b) 4, (c) 6, and (d) the sludge.

could accurately predict the biogas production trends in all cases except
for ISR = 2, but the predictions of the FO model had a slight inconsis­
tency with the experimental data (as reflected in the RMSE values in
Table 7). At this TS level, the best results in terms of fit (namely, con­
sistency with the experimental data) were related to ISR = 6. According
to Table 7, the λ value at TS = 7% was mostly zero, much like that at TS
= 5%. It seemed that increasing the concentration of the microorgan­
isms in the digester shortened the lag phase of biogas production. At TS
= 7%, the highest Rm values given by the MG (14.21 m3/kg-VS.d) and LF
(11.21 m3/kg-VS.d) models were also related to ISR = 4.
In this study, the FO reaction rate constant (k) for all concentrations
and all sludge-to-waste ratios was at the range of 0.054–0.095 d− 1.
Previous studies on the AD of OMs had further reported a wide range of k
values from 0.027 to 0.49 d− 1 [26,31,35,52,53]. The FO model could be
thus recruited to determine the degradability of OM, which had a direct
impact on the maximum biogas production [51]. The results of this
study also showed no particular pattern in how the hydrolysis constant
had changed following the increase in TS content (at least not at the

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A. Sedighi et al. Renewable and Sustainable Energy Reviews 153 (2022) 111797

Table 6
Parameters estimated by the kinetic models of biogas production in TS = 5%.
parameter First-order Kinetic models Logistic function

Modified Gompertz

ISR 2 4 6 Sludge 2 4 6 Sludge 2 4 6 Sludge

k 0.054 0.092 0.095 0.083 – – – – – – – –
Rm - – – – 19.14 21.88 19.79 11.91 21.89 17.51 15.67 9.73
λ - – – – 0.64 0 0 0 3.87 0 0 0
R2 0.955 0.984 0.953 0.933 0.992 0.98 0.971 0.964 0.953 0.973 0.962 0.966
RMSE 0.0714 0.0358 0.0327 0.0263 0.0216 0.0116 0.0366 0.0102 0.0253 0.0252 0.0263 0.0145

Fig. 7. Kinetic models and experimental data in TS = 7% for the ISRs of (a) 2, (b) 4, (c) 6, and (d) the sludge.

concentrations examined). However, the outputs revealed that the lag
period tended to approach zero as the TS level elevated, which could be
due to the presence of larger amounts of anaerobic sludge. At all TS
levels studied, the RMSE value was also minimized with the growing
trend in the amount of sludge in the reactor, suggesting that the higher
the number of anaerobic microorganisms in the digester, the better the
kinetic models could fit the experimental data. The lower fit of the
models to the experimental data for ISR = 2 at all TS levels also sup­
ported this argument.
In Deepanraj et al. (2017), examining the AD kinetics of food waste,
it had been also reported that the MG model had provided a better
description of the kinetic process than the LF equation [54]. Pramanik
et al. (2019) had further studied the AD kinetics of food waste with FO,
MG, and LF models, and had reported that the MG model could give an
over 99% accurate description of biogas production in this process [31].
Moreover, Zahan et al. (2018) had found similar results for the ACD of
industrial-agricultural wastes [29]. In Donoso et al. (2010), wherein the
MG, LF, and reaction curve-type models had been used to compare the

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A. Sedighi et al. Renewable and Sustainable Energy Reviews 153 (2022) 111797

Table 7
Parameters estimated by the kinetic models of biogas production in TS = 7%.
parameter First-order Kinetic models Logistic function

Modified Gompertz

ISR 2 4 6 Sludge 2 4 6 Sludge 2 4 6 Sludge

k 0.084 0.086 0.075 0.083 – – – – – – – –
Rm - – – – 6.19 14.21 6.57 8.09 3.34 11.21 5.24 6.61
λ - – – – 0 1.72 0 0 0 0 0 0
R2 0.964 0.97 0.918 0.927 0.862 0.864 0.945 0.958 0.929 0.903 0.954 0.959
RMSE 0.0248 0.0280 0.0074 0.0163 0.0106 0.0130 0.0920 0.0071 0.0262 0.0117 0.0100 0.0106

anaerobic degradation of untreated and pre-treated wastewater sludge,
the results had established that the reaction curve-type model had
offered better descriptions of this process than the other two cases [33].
In addition, Behera et al. (2010), Ramos et al. (2012), and Nielfa et al.
(2015) had reported that the MG model was well adapted to experi­
mental data and could describe the development of the AD of food waste
[55–57]. In the present study, the results revealed that, among the MG,
LF, and FO models, the MG one produced the most accurate estimations
of the experimental biogas production data. While the LF model could
give slightly worse but mostly acceptable biogas production estimates,
the FO one did not outperform in this case, especially at low ISRs.
4. Conclusion
The study results revealed that the ACD of biomass materials, espe­
cially those serving as major substrate and inoculum sources, could
substantially improve biogas production efficiency in this process. This
study also investigated the potential for biogas production from the
mesophilic ACD of OFMSW and sewage sludge at three ISRs (viz. 2, 4, &
6) and three TS levels (namely, 3, 5, & 7%). The combination of high
microbial load in the sludge with the good CNR and the nutrient content
of OFMSW also provided favorable conditions for the co-digestion of
these wastes. The experiments in this study were further carried out in
single-stage anaerobic digestion reactors operating in batch mode and
under mesophilic conditions. The results showed that the highest biogas
yields were 0.642 and 0.697 m3/kg-VSadded at TS = 3 and 5%, respec­
tively, and were observed at ISRs = 6 and 2, in that order. The experi­
ments indicated that the injection of more NaOH (within the range used
in this study) into the alkaline pretreatment resulted in better preser­
vation of optimal pH conditions (that is, lower pH drop) inside the
digester during acidogenesis, which ultimately led to higher yields. In
general, biogas production, CH4 production, and primary CH4 yield
increased as TS levels elevated from 3 to 5%. However, the raw mate­
rials tended to form a layered structure separated by density at TS = 7%,
because of the high concentration of the materials in the absence of
stirring, which in turn resulted in slow and incomplete ACD (the highest
biogas yield was 0.481 m3/kg-VSadded at ISR = 4). The CH4 yield of the
anaerobic reactions could be further influenced by a range of factors and
their interactions. A high CNR alone did not necessarily lead to higher
CH4 production, as CNR in the mixture could play an important role at
different digestion stages. The TS level and VS content of the materials
were also of particular significance. If a substrate had a low TS level and
VS content, CH4 production capacity would be inevitably limited
regardless of CNR.
The kinetic modeling of the biogas production process using FO, MG,
and LF models also demonstrated that the amount of sludge fed into the
digesters affected how well these models could fit the experimental data.
Accordingly, as ISRs increased in the reactor, the models were effec­
tively able to predict the biogas production trends. In general, bioenergy
extraction from MSW and sewage sludge, while reducing the destructive
effects of waste disposal in the environment, could be a sustainable so­
lution for waste management and renewable energy use. The present
study was thus of utmost importance from the perspective of pollution
control and low-cost energy production in developing countries.
Credit author statement
Mohsen Karrabi: Supervision, Conceptualization, Writing – review
& editing; Bahar Shahnavaz: Validation, Formal analysis, Writing –
review & editing; Afsane Sedighi: Methodology, Investigation, Writing
– original draft; Morteza Mostafavinezhad: Investigation, Writing –
original draft
Authors’ contributions
MK and BS devised the project, the main conceptual ideas and proof
outline. AS and MM carried out the experiment. AS and MK derived the
kinetic models and analyzed the data. AS and MM wrote the manuscript
in consultation with MK and BS. All authors discussed the results and
commented on the manuscript.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgment
This research study was conducted with the support of the Vice
Chancellor of Research of the Ferdowsi University of Mashhad through
grant number 48955.
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