Journal of Cleaner Production 289 (2021) 125775

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Journal of Cleaner Production

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

Solid-state anaerobic co-digestion of food waste and cardboard in a

pilot-scale auto-fed continuous stirred tank reactor system
Sameena Begum a, b, Tanmoy Das a, Gangagni Rao Anupoju b, Nicky Eshtiaghi a, *
a
Chemical and Environmental Engineering, School of Engineering, RMIT University, 124 La Trobe St, Melbourne, VIC, 3000, Australia
b
Bioengineering and Environmental Sciences Division, EEFF Department, CSIR-Indian Institute of Chemical Technology, Tarnaka, Hyderabad, 500007, India

a r t i c l e i n f o a b s t r a c t

Article history: Increasing volume of Food waste (FW) and cardboard (CB) which are the two major fractions of
Received 29 January 2020 municipal solid waste increases environmental concerns. This study evaluated the performance of a
Received in revised form pilot-scale auto-fed continuous stirred tank reactor (CSTR) system for the solid-state anaerobic co-
11 September 2020
digestion (ACoD) of acetic acid rich food waste (FW) and cardboard (CB). The objective to co-digest
Accepted 29 December 2020
Available online 2 January 2021
acids rich FW and CB originates from the fact that acetic acid can act as a pre-treatment agent. Can
the frequency of feeding and feeding pattern assists the system to withstand an organic loading rate >4 g
Handling editor: Prof. Jiri Jaromir Kleme
s volatile solids (VS)/(L.d) without addition of any pH boosting chemicals? Four mixing ratios of 100:0,
80:20, 60:40, 50:50 (v/v %) of FW and CB on biogas yield and process stability at an OLR > 4 g VS/(L.d) at a
Keywords: hydraulic residence time (HRT) of 40 d was investigated. Results revealed that the reactor R1 operated
Food waste with FW:CB in the ratio of 100:0 achieved 76% biodegradability (the ratio of experimental to theoretical
Cardboard methane yield) while it was 39%, 32% and 22% for FW:CB ratios of 80:20, 60:40 and 50:50, respectively.
Operational strategy This study concludes that exposing the CB to acetic acid-rich FW assisted the disintegration and depo-
Auto-fed continuous stirred tank reactor
lymerization of CB without any pre-treatment step. Micro-feeding the reactor on an hourly basis assisted
Anaerobic co-digestion
to overcome the susceptible failure of the system. The outcomes of this study provides new insight into
Biodegradability
the potential of implementing decentralized AD units for the treatment of FW and CB at source, which
not only diverts waste from landfills but creates a revenue model through bioenergy generation and
digestate sales.
© 2021 Elsevier Ltd. All rights reserved.

1. Introduction
Food waste (FW) is one of the largest components of solid waste
attained increasing attention worldwide due to growing concern
about its emission of greenhouse gases and leachate generation.
One third of the food produced for human consumption is wasted
every year, producing an estimated 1.3 billion tonnes of FW
(Ishangulyyev et al., 2019). In majority of the under developed
countries, FW is diverted to open landfills rather than being utilized
through alternative and energy-efficient routes of valorization
(Garcia-Garcia et al., 2019). Cardboard (CB) is another major frac-
tion of municipal solid waste. About three tons of wood is required
to produce one ton of CB (General Kinematics, 2019). Recycling CB is
the best option to produce new CB which offers dual environmental
and economic benefits. It is also known that not all CB is recyclable
* Corresponding author.
E-mail address: nicky.eshtiaghi@rmit.edu.au (N. Eshtiaghi).
for instance, waxed CB such as pizza boxes, wet and soiled CB are
not recyclable and they are diverted to the landfills. About 40% of CB
is sent to landfills every year which creates a similar environmental
hazard like FW (Department of the Environment and Energy, 2019).
Anaerobic digestion (AD) is one of the most efficient technolo-
gies for waste treatment and stabilization as it enables the recovery
of valuable chemicals (bioethanol, volatile fatty acids (VFA)) and
energy (hydrogen, biogas), promoting a sustainable circular bio-
economy (Kuruti et al., 2015; Begum et al., 2016; Begum et al.,
2018). Increasing interest in AD of FW had resulted in extensive
laboratory-scale investigations (as reviewed in Ren et al., 2018;
Wang et al., 2018) while mono-digestion of CB is scarcely reported
because of its poor biodegradability due to lignin despite contain-
ing significant organic matter in the form of cellulose and hemi-
cellulose (Li et al., 2019). To the best of our knowledge no full-scale
plant has been installed for the mono-digestion of CB whereas
there are numerous full-scale demonstrations for FW (De Clercq
et al., 2019; Yang et al., 2019). The full-scale FW digesters often
tend to fail, if operated at higher organic loading rates (OLR) due to

https://doi.org/10.1016/j.jclepro.2020.125775
0959-6526/© 2021 Elsevier Ltd. All rights reserved.
S. Begum, T. Das, G.R. Anupoju et al.
rapid acidification. Most studies to date have focused on the gen-
eration of biogas from FW at a maximum OLR of 4 g VS/(L.d) (VS is
volatile solids) using external pH-boosting chemicals to maintain
the pH in the range of 6.8e7.2, as this is a favorable range for the
methanogenic phase of AD to occur. It was reported that an OLR
>4 g VS/(L.d) causes digesters to fail due to overloading (Meng et al.,
2014; Kuruti et al., 2017).
Anaerobic co-digestion (ACoD) of FW and CB is gaining interest
to overcome the limitations associated with mono-digestion
(Guilford et al., 2019). Co-digestion of FW with other co-
substrates such as sewage sludge, cattle manure, yard waste and
domestic waste water was proven to be economically feasible to
overcome the accumulation of VFAs and high solids which leads to
inefficient performance or process failure due to sudden pH drop
(Panigrahi et al., 2020; Zhang et al., 2013). A recent study reported
on co-digestion of FW and CB in batch trials by Asato et al. (2019)
investigated the impact of mixing ratios, total solids and food to
inoculum (F/I) ratio on biogas or methane yield. They stated that
greater fractions of FW resulted in higher methane production rates
and yields, while greater fractions of CB led to longer lag phases. In
another study, Capson-Tojo et al. (2017) studied the impact of F/I
ratios of 0.25, 1 and 4 gVS/gVS in dry ACoD of FW and CB and found
that the reactors produced methane only at F/I of 0.25. An impor-
tant issue for research is that the performance of laboratory scale
studies may not match the performance at full scale, possibly the
semi-continous and continuous studies would. Therefore, pilot-
scale demonstrations on ACoD of FW and CB is needed.
Laboratory scale investigations reported on co-digestion of FW
and CB incorporating physical (size reduction), mechanical (ultra-
sound, microwave etc.) and/or chemical (acid/alkali) pre-treatment
step demonstrated enhanced process performance but the eco-
nomic viability is lower (Zeynali et al., 2017). Addition of buffering
chemicals and nutrients in the pre-treatment step would incur
extra operational costs for a full-scale AD plant operated with any
substrate particularly FW and CB. Previous studies have demon-
strated that AD is very sensitive to disturbances in pH, temperature
and OLR as it is a highly dynamic and multi-step process resulting
from a continuous interaction between substrate, microorganisms/
enzymes and operating conditions which creates a delicate equi-
librium mainly dependent on substrate characteristics (Fisgativa
et al., 2016; Kuruti et al., 2017). The main controls of AD opera-
tion are OLR, hydraulic residence time (HRT), temperature, mixing
and the feeding pattern. When translating the results from batch to
continuous, the frequency of feeding and the feeding pattern is
often overlooked (Svensson et al., 2018). For instance, one time
feeding in done in batch reactors, one time a day in continuously
stirred tank reactors (CSTR) while the full-scale AD plants are fed
continuously throughout the day at regular intervals. Feeding
patterns and its frequency are the two significant parameters that
influences the overall performance of the digesters (Ziels et al.,
2017). A case study investigated by Svenson et al. (2018) proved
that frequently fed digesters produced 20% more methane from FW
and had lower effluent concentrations of long chain fatty acids
compared to one time fed reactor. Therefore, to assess the techno-
economic performance analysis of a full-scale AD plant based on
laboratory scale outcomes, similar feedstock, feeding pattern and
frequency of feeding to the reactors should be taken into consid-
eration. A new operational approach to treat FW and CB containing
high solids >20% without any buffering chemicals and/or pre-
treatment step, is required to fulfil essential conditions such as
robustness and high adaptive capacity of the AD system. Stable
operation at high OLR, low HRT and good economic performance
are necessary for a process to be implemented at full scale
(Cavinato et al., 2013). The optimum HRT required for AD of sub-
strates such as animal manures, FW and agricultural waste falls
2
Journal of Cleaner Production 289 (2021) 125775
between 24 and 90 d (Kuruti et al., 2017; Irena.org, 2019).
This study demonstrated the solid-state ACoD of acetic acid rich
FW and CB in a pilot-scale auto-fed CSTR system. To find the
optimal FW:CB mixing ratio, four different mixing ratios of 100:0,
80:20, 60:40 and 50:50 (v/v %) with a corresponding total solids
(TS) content of 19%, 21%, 23% and 24% were investigated. This study
attempted to answer two major research questions on reactor
operation. Firstly, can an anaerobic digester operated with acetic
acid rich FW in mono or co-digestion without addition of any pH
boosting chemical at an OLR greater than 4 g VS/(L.d) as it is the
threshold value for safe operation? Secondly, can the feeding
pattern and the frequency of feeding influence the overall stability
of the CSTR system? The study investigated the ACoD of FW and CB
at a full HRT of 40 d at a corresponding OLR values in the range of
4.7e5.7 g VS/(L.d). The CSTR system was initially operated at 90 d
HRT, followed by 80 d and 60 d at an interval of 10 d with OLR below
4 g VS/(L.d) to allow acclamatization of micro-organisms with the
substrate. The CSTR system was operated at a complete HRT of 40 d
(OLR > 4 g VS/(L.d)) to find the optimal ratio of CB that can be co-
digested with FW and overall process stability. The outcome of
this study was used for a techno-economic performance analysis of
a full-scale AD plant at one-tonne capacity.
2. Materials and Methods
2.1. Substrate and inoculum collection
25 L of coarsely ground acetic acid rich FW slurry and inoculum
were collected from a mesophilic anaerobic digester operated with
FW at the Aurora sewage treatment plant (Preston, Australia), in
airtight polypropylene containers. The mesophilic inoculum was
stored in an incubator at 37
C for two weeks to allow degassing,
and the FW slurry was stored in the refrigerator at 4
C until the
start of experiments. The co-substrate used for experimentation
was corrugated CB collected from recycle bins in RMIT University.
2.2. Analytical methods
Total solids and volatile solids (VS) were measured according to
APHA Methods 2540 B and 2540 E respectively (APHA, 1998). The
chemical oxygen demand (COD) (total and soluble), ammonia ni-
trogen (NH3eN) and volatile acids (VA) were determined by
colorimetric techniques using a HACH (Model: DR/5000 U) spec-
trophotometer according to APHA Methods 8000, 10031 and 8196,
respectively (Zahan et al., 2018). The samples were centrifuged
(Eppendorf 5702, Germany) at 10,000 rpm for 5 min and then
filtered through 0.45 mm filter paper (mixed cellulose-ester mem-
brane filter, Advantec, Japan) to measure the soluble organic con-
stituents. The density of the CB and FW slurry was calculated from
its mass and volume. The slurry’s pH was measured using a cali-
brated pH meter (Thermo Orion, Model 550 A), and its elemental
composition was determined using a LECO (TruMac) analyzer.
2.3. Preparation of feed mixture (FW and CB) and characterization
The corrugated CB sheets were cut into pieces of 3e4 cm in
length using scissors. The density of the FW was measured and
found to be 1060 kg/m3 while the density of CB was 72 kg/m3. The
feed mixture of FW and CB was prepared in ratios of 100:0, 80:20,
60:40 and 50:50 on the basis of volume. For example, to prepare a
mixture of FW:CB 50:50, the equivalent weight of 50 mL CB
(0.0036 g) was added to 50 mL of FW slurry and soaked for 1 h. The
soaked feed mixture was ground into a paste using a heavy-duty
blender. Characteristics of the raw FW slurry, feed mixture sam-
ples, CB and inoculum are presented in Table 1. The pH of the acetic
S. Begum, T. Das, G.R. Anupoju et al. Journal of Cleaner Production 289 (2021) 125775

Table 1
Physico-chemical characteristics of inoculum, CB and feed mixtures.

Parameter Units Inoculum CB FW:CB (100:0) FW:CB (80:20) FW:CB (60:40) FW:CB (50:50)

pH () 7.4 ± 0.2 ND 3.6 ± 0.1 3.7 ± 0.1 3.7 ± 0.1 3.8 ± 0.1
TS % 4.2 ± 0.1 93 ± 0.2 19 ± 0.2 21 ± 0.2 23 ± 0.2 24 ± 0.2
VS % 2.4 ± 0.1 85 ± 0.1 18 ± 0.1 20 ± 0.1 21 ± 0.1 22 ± 0.1
tCOD g/L 37.6 ± 0.2 ND 624 ± 0.2 550 ± 0.2 518 ± 0.2 520 ± 0.2
sCOD g/L 11.6 ± 0.1 ND 68 ± 0.1 70 ± 0.1 71 ± 0.1 72 ± 0.1
NH3eN g/L 2.5 ± 0.1 ND 1.3 ± 0.1 0.4 ± 0.1 0.8 ± 0.1 0.7 ± 0.1
VA g/L 0.3 ± 0.02 ND 42 ± 0.1 43 ± 0.1 43.5 ± 0.1 43 ± 0.1
Carbon (C) % ND 49 51 ND ND ND
Hydrogen (H) % ND 5.4 10 ND ND ND
Nitrogen (N) % ND 0.3 0.7 ND ND ND
Sulfur (S) % ND 0.3 0.2 ND ND ND
Oxygen (O) % ND 0.2 0.4 ND ND ND

ND-Not determined.

acid rich FW slurry was lower i.e. < 4.0 and the carbon to nitrogen
(C/N) ratio of FW and CB was high i.e. 73 and 163 respectively,
which are not favorable for the AD process, but the operational
strategy significantly reduced the impact of these limitations.
2.4. Equipment description
The equipment used in the present study was the patented
auto-fed CSTR system (Lobster-max-I model) with four reactors
namely R1, R2, R3 and R4 as shown in Fig. 1. The reactors or
anaerobic digesters are cylindrical vessels made of stainless steel
with a reactor volume of 5.2 L and an effective working volume of
5 L and feed syringe volume of 1.5 L each. Each reactor is equipped
with its own beam to automatically feed the digester at the set feed
rate and time. The feed from the feeder syringe is delivered to the
reactor from a feeder port at the bottom using a unique patented
feeding system that moves vertically upwards. A non-return
membrane value is fitted between the digester and the feeder sy-
ringes to prevent the movement of digestate into the feeder section.
Mixing in the reactors is performed by a gas-tight mixer shaft
connected to a 24 V DC motor rotating at 45 rpm. The equipment
includes feed, reactor and digestate valves. The temperature of each
reactor is controlled by an electric heating jacket wrapped around
the external reactor wall. A touchscreen programmable logic
controller (PLC) is used to program the feeder mechanism and for
data logging and monitoring. Gas generation is measured using a
liquid displacement gas flow meter module, which is a single
perspex block with six perspex tumbling buckets of active volume
22e24 mL each. Each reactor is also equipped with 1 L bottles for
the collection of digestate.
2.5. Startup and operational strategy
The feeders of each of the reactors were initially filled using the
top-up feed syringe, keeping the reactor and feed valve open to
prevent a gas lock. The FW:CB ratios in the feed mixture were
100:0, 80:20, 60:40 and 50:50 for R1eR4 respectively. The feeder
syringes were topped up with fresh feed periodically using the top-
up syringe, which pushes the piston downwards automatically.
Subsequently, the reactors were inoculated with 5 L of inoculum
until it started to flow into the digestate bottles by keeping the
reactor-feed isolation valve closed and the digestate valve open.

Fig. 1. The Lobster-max-I auto-fed CSTR digester system.

3
S. Begum, T. Das, G.R. Anupoju et al. Journal of Cleaner Production 289 (2021) 125775

The tubing from the reactors was connected to the gas flow meter
module to estimate the biogas generation potential of the FW and
CB mixture in each reactor.
After filling the feeder syringes and inoculating the reactors, the
input parameters (dynamic parameters) were set in the PLC
controller as summarized in Table 2. The temperature in the re-
actors was maintained at 37 ± 0.1
C and the experiments were run
over 70 d. The reactors were operated at a stepwise decreasing HRT
of 90 d, 80 d and 60 d (for a duration of 10 d each to allow sufficient
time for the inoculum to acclimatize with feed mixture) and a full
HRT of 40 d at an OLR between 4.7 and 5.7 g VS/(L.d). The beam
speed of the reactors was calibrated early in the experiment and
maintained until the end (Table 2). The number of feed cycles per
day, feed cycle period and the feed delivery rate were constant
throughout the experiment. The reactors were fed every hour i.e. 24
feed cycles per day. As the HRT decreased from 90 d to 40 d, OLR,
feed volume per day, feed volume and feed time per cycle increased
accordingly, as presented in Table 2. In spite of the unfavourable pH
of the feed (<4.0), the solid-state ACoD of FW and CB mixture was
feasible with process stability because of frequent micro-feeding
mechanism on an hourly basis.
3. Results and discussion
3.1. Performance analysis of solid-state ACoD of FW and CB in an
auto-fed CSTR system
3.1.1. Impact of HRT on daily and cumulative biogas yield
The impact of HRT on daily and cumulative biogas yield over the
digestion time during mesophilic ACoD of FW and CB in different
mixing ratios in reactors R1, R2, R3 and R4 is shown graphically in
Fig. 2. Fig. 2A shows that the daily biogas yield increased sharply
from 0.1 to 2.0 L/gVSadded during the first 10 d of start-up (HRT of
90 d) in R1, whereas in R2, R3 and R4 the increase in biogas yield
was 0.05e0.6 L/gVSadded. This could be attributed to the volume of
inoculum (abundance of methanogens) available initially compared
to the volume of the input substrate in all the reactors. Subse-
quently, a sharp decrease in biogas yield of 42.5% (from 2.0 to 1.15 L/
gVSadded) was observed over days 11e20 (HRT of 80 d) in R1, while
in R2 it decreased by 10% (from 0.30 to 0.27 L/gVSadded) and in R3
and R4 it increased by 79% (0.58e1.04 L/gVSadded) and 18.4%
(0.38e0.45 L/gVSadded) respectively. Fig. 2A also shows that during
days 21e30 (HRT of 60 d) the biogas yield increased by 28.6%
(1.15e1.48 L/gVSadded) and 185% (0.27e0.77 L/gVSadded) in R1 and
R2 respectively, while in R3 and R4 it decreased by 41.3%
(1.04e0.61 L/gVSadded) and 28.8% (0.45e0.32 L/gVSadded) corre-
spondingly. At the final and full HRT of 40 d, average biogas yields of
0.47, 0.31, 0.07 and 0.18 L/gVSadded were observed from R1, R2, R3
and R4 respectively, with the system attaining stability. These ob-
servations indicated that the biogas yield decreased as the HRT
decreased in reactors R1 and R2, and in contrast, in reactors R3 and
R4 the biogas yield increased as the HRT decreased. This implies
that an increase in the CB:FW ratio improved the buffering capacity
and enhanced the synergistic methanogenic mechanism.
Fig. 2B reveals that the cumulative biogas yield in R1 was 0.74 L/
gVSadded which is significantly higher than the yield achieved from
R2, R3 and R4 (0.32, 0.30 and 0.21 L/gVSadded respectively). It can be
clearly stated that the biodegradability of the FW:CB in the ratio of
100:0 in R1 was higher than the biodegradability of FW:CB in the
ratio of 80:20, 60:40 and 50:50 in R2, R3 and R4 correspondingly. It
is known that VFA is the precursor for methanogens for the pro-
duction of biogas through methanogenesis (Begum et al., 2020). In
reactor R1 the feed contained only FW without any CB, therefore
the VFA available in FW was utilized by methanogens to produce
more biogas, while in reactors R2, R3 and R4 the VFA was being
absorbed by CB for the degradation of cellulose. The average cu-
mulative biogas yield in R1 and R3 sharply increased as HRT
decreased from 90 d to 60 d, while in R2 and R4 the difference in
average cumulative biogas yield was smaller (Fig. 2B). At the
beginning of 40 d HRT, the average cumulative biogas yield
decreased in reactors R1, R3 and R4, while in R2 it increased
slightly. It is interesting to note from Fig. 2B that the reactor R2 with
FW:CB 80:20 attained stability with stable biogas yield in the range
of 0.3e0.35 L/gVSadded within the first 10 d of operation (HRT of
90 d) while in reactor R4 the system attained stability in the first 15
days but the biogas yield relatively decreased by 40% (0. 35 to

Table 2
The fixed and dynamic paramaters showing the operating conditions of auto-fed CSTR system.

Reactor/Operational parameters R1 (FW:CB, 100:0) R2 (FW:CB, 80:20) R3 (FW:CB, 60:40) R4 (FW:CB, 50:50)

Fixed parameters
Beam speed (mm/h) 108 111 111 111
Feed cycle period (min) 60 60 60 60
Feed cycles per day 24 24 24 24
Feed delivery rate (mL/min) 12.75 12.75 12.75 12.75
Dynamic parameters
HRT (d) 90 90 90 90
OLR (g VS/(L.d) 2 2.3 2.4 2.5
Feed volume per day (mL) 57.7 57.7 57.7 57.7
Feed volume per cycle (mL) 2.4 2.4 2.4 2.4
Feed time per cycle (s) 12.2 12.2 12.2 12.2
HRT (d) 80 80 80 80
OLR (g VS/(L.d) 2.3 2.6 2.7 2.9
Feed volume per day (mL) 65 65 65 65
Feed volume per cycle (mL) 2.7 2.7 2.7 2.7
Feed time per cycle (s) 12.73 12.73 12.73 12.73
HRT (d) 60 60 60 60
OLR (g VS/L/d) 3.1 3.5 3.6 3.8
Feed volume per day (mL) 86.6 86.6 86.6 86.6
Feed volume per cycle (mL) 3.6 3.6 3.6 3.6
Feed time per cycle (s) 16.98 16.98 16.98 16.98
HRT (d) 40 40 40 40
OLR (g VS/(L.d) 4.7 5.2 5.5 5.7
Feed volume per day (mL) 130 130 130 130
Feed volume per cycle (mL) 5.4 5.4 5.4 5.4
Feed time per cycle (s) 25.47 25.47 25.47 25.47

4
S. Begum, T. Das, G.R. Anupoju et al. Journal of Cleaner Production 289 (2021) 125775

Fig. 2. Impact of HRT on biogas generation profile of anaerobic co-digestion of high-solids FW and CB at four mixing ratios (A) Daily biogas yield; (B) Cumulative biogas yield.

0.21 L/gVSadded). In reactors R1 and R3, the biogas yield sharply respectively. This could be attributed to the frequent micro-feeding
increased till 30 days of operation (i.e. from HRT of 90 de60 d) strategy of the substrate (which was at low pH (<4.0)) in the re-
followed by a decrease at a complete HRT of 40 d. This signifies that actors on an hourly basis which facilitated the complete utilization
the reactors R1 and R3 took longer time to attain stability. of the organic matter by the methanogens, in addition to the OLR
remaining below the threshold of 4 g VS/(L.d). This could be
credited to the feeding pattern, that is, the feed volume per cycle
3.1.2. Impact of OLR on cumulative biogas yield
(mL) at HRTs of 90 d, 80 d and 60 d, which was 2.37 mL, 2.7 mL, and
The increase in OLR corresponding to the decrease in HRT from
3.6 mL respectively per hour in all the reactors (Table 2). No buff-
90 d to 40 d and its impact on cumulative biogas yield is presented
ering chemicals were added to the substrate (which was at low
in Fig. 3. In reactors R1 and R3, the cumulative biogas yield
pH < 4.0) to increase the pH to neutral before feeding, because the
increased sharply from 0.1 to 1.1 L/gVSadded and 0.07e0.57 L/
ratio of volume of inoculum in the reactor to volume of feed fed to
gVSadded, respectively, as the OLR increased from 2 to 3.1 gVS/(L.d)
the reactors per hour was high, which balanced the pH in the
in R1 and 2.3 to 3.5 gVS/(L.d) in R3 respectively on the basis of HRT.
reactor by creating self-buffering in the system. This suggests that
In reactors R2 and R4, gradual increase in cumulative biogas yield of
the buffering capacity in the reactors could be retained with
0.05e0.38 L/gVSadded and 0.05e0.35 L/gVSadded were observed as
smaller amounts of feed being fed to the reactors frequently.
the OLR increased from 2.4 to 3.6 gVS/(L.d) and 2.5 to 3.8 gVS/(L.d),
5
S. Begum, T. Das, G.R. Anupoju et al.
Fig. 3. Impact of increasing OLR as a result of decreasing HRT on cumulative biogas yield during solid-state anaerobic co-digestion of FW and CB.
Decrease in cumulative biogas yield from 1.1 to 0.74 L/gVSadded,
0.57 to 0.29 L/gVSadded and 0.35 to 0.29 L/gVSadded were observed in
reactors R1, R3 and R4 after the beginning of 40 d HRT; i.e. increase
in OLRs from 3.1 to 4.7 gVS/(L.d) in R1; 3.6 to 5.5 gVS/(L.d) in R3;
and 3.8 to 5.7 gVS/(L.d) in R4 correspondingly. In reactor R2, the
decrease in biogas yield from 0.38 to 0.32 L/gVSadded as the OLR
increased from 3.5 to 5.2 gVS/(L.d) was lower compared to those in
other reactors (Fig. 3). This could be due to the impact of the FW:CB
mixing ratio (80:20) that was adopted by the methanogens, even
though the feed volume per cycle increased to 5.4 mL as the OLR
increased. This could be the reason reactor R2 attained stability in
biogas production with in the first 10 days of operation. As the OLR
increased in reactors R1, R3 and R4, the feed volume per cycle (mL)
also increased from 3.6 mL to 5.4 mL per hour (Table 2), which was
reflected in the increased amount of CB fed to the reactors in R3 and
R4 that resulted in the decrease in biogas yield. However, in reactor
R1 the decrease might have been due to organic shock loading.
Although a decrease in biogas yield from 1.1 to 0.74 L/gVSadded was
observed in R1, it was still significant as the reactor was operated at
an OLR >4 g VS/(L.d) which is a threshold value for safe operation
with FW, however an HRT of 40 d might have led to attain stability
even at higher OLR. In reactors R3 and R4, lower biogas yield was
observed at the full HRT of 40 d because the CB:FW ratios were
40:60 and 50:50 respectively, which resulted in lower biodegrad-
ability of CB at reduced HRT. The overall performance of the CSTR
system showed that the reactor with FW:CB of 80:20 is better in
terms of attaining quick process stability and stable biogas yield
during ACoD without the need of any pre-treatment or buffering
6
Journal of Cleaner Production 289 (2021) 125775
chemicals in comparison to other mixing ratios of FW:CB. Never-
theless, the biogas yield from monodigestion of FW (i.e. FW:CB
100:0) was superior among all the ratios.
3.1.3. Impact of volatile acids (VA) present in FW on CB degradation
at OLRs of 2e5.7 g VS/(L.d)
It is well documented that the ACoD process performance of
lignocellulosic biomass, such as wheat straw, rice straw and CB,
improves after pre-treatments such as acid, alkali, ultrasonication
and/or combinations of these (Zeynali et al., 2017). This is because
the pre-treatment disintegrates the structure of the biomass and
cell lysis occurs, which releases the intracellular organic matter
(monomers such as sugars, amino acids and glycerol) and makes
these more bioavailable thereby improving the reaction rates and
efficiency of the digestion process. In particular, acid (H2SO4) and/or
alkaline (NaOH) pre-treatment of lignocellulosic biomass improves
the biogas yield compared to biomass without any pre-treatment
(Mosier, 2005). In the present study, the VA in the form of acetic
acid (CH3COOH) which is a weak acid favored the efficient solubi-
lization of CB by disintegrating its structure during 90 d HRT,
although it is a weak acid in FW. Acetic acid is also a major
degradation product of newsprint paper, among other low molec-
ular weight carboxylic acids such as formic, lactic, malonic, malic
and succinic acids (Gibson and Watt, 2010). C3 and C4 alkyl
substituted cyclohexanols and C4 to C12 aliphatic alcohols, among
numerous volatile organic compounds, have also been identified as
main degradation products in cotton and chemical wood pulp
books. Dupont and Tetreault (2000) observed immediate
S. Begum, T. Das, G.R. Anupoju et al.
significant depolymerization of cellulosic material after it was
exposed to acetic acid in the concentration range of 20e200 mg/m3
(277e2,777 mg/kg) but not after exposure to 3 mg/m3 (41 mg/kg).
Lignocellulose has a recalcitrant and crystalline structure that re-
sists bacterial attack and biodegradation. Subjecting CB to acetic
acid-rich FW increased the biodegradability of lignocelluloses by
decreasing its crystallinity and eliminating its network structure.
Therefore, a major finding of this study is that acetic acid-rich FW
can be utilized to degrade CB during ACoD of FW and CB which
eliminated the use of external pre-treament chemicals.
The concentration of acetic acid in FW that interacted with the
amount of CB fed to the reactors R2, R3 and R4 and its impact on
biogas yield against OLR and HRT, is summarized in Table 3. To
make a comparison of biogas yield obtained in CSTRs as a function
of OLR irrespective of the FW:CB mixing ratios, biogas yield of
reactor R1 is also presented in Table 3. Dupont and Tetreault (2000)
found that the degradation of cellulose by exposure to acetic acid is
dependent on the concentration of the acid and also on the aging or
exposure time. Table 3 shows that the increase in the CB ratio in FW
from 20% in R2 to 40% in R3 and 50% in R4 decreased the concen-
tration of acetic acid from 3.1 to 1.5 to 1.2 gVA/gCB, respectively.
Moreover, in reactor R2, the average cumulative biogas yield
increased from 0.24 to 0.35 (L/gVSadded) as the HRT reduced from
90 d to 40 d (Table 3). In reactors R3 and R4 the ratio of CB in FW
was higher, resulting in lower interaction of VA with CB and
reduced degradation, leading to lower biogas yield than 20% CB in
FW. Dupont and Tetreault (2000) also reported that the depoly-
merization of cellulose by exposure to acetic acid is more likely to
occur over the long term, and the same was observed in R3 and R4.
From HRT of 90 de60 d, the average cumulative biogas yield
increased from 0.31 to 0.56 (L/gVSadded) and 0.27 to 0.35 (L/gVSad-
ded) in reactors R3 and R4, while at 40 d, the biogas yield decreased.
This implies that co-digestion of 40% and 50% CB in FW needs
longer HRT. In reactor R1, as the OLR increased from 2 to 4.7 gVS/
(L.d), the biogas yield decreased from 0.92 to 0.74 L/gVSadded. The
decrease in biogas could be attributed to organic shock loading at
an OLR 4.7 gVS/(L.d) but the system still attained stability at higher
OLR with notable biogas yield.
3.2. Impact of increasing CB content on VS removal efficiencies and
digestate pH during ACoD of FW and CB
The digestate samples collected from the digestate bottles of
reactors R1, R2, R3 and R4 after every five days were analyzed for VS
and pH. The VS removal efficiencies and pH values of the digestate
are shown in Fig. 4. The VS removal efficiencies were between 75
and 90% during the first 20 d (i.e., HRT of 90 and 80 d). At an HRT of
60 d, the VS removal efficiencies declined by 10% i.e. 65e80%. This
reduction in VS removal efficiency could be due to the decrease in
HRT and an increase in OLR. This observation elucidates the exis-
tence of an inverse correlation between organic matter removal
and OLR. The VS removal efficiencies further decreased at an HRT of
40 d, but the system reached steady state with an average removal
Table 3
Impact of VA concentration in FW on the hydrolysis of CB (g VA/g CB) and biogas yield in the reactors R2 (FW:CB 80:20), R3 (FW:CB 60:40) and R4 (FW:CB 50:50) at OLRs of
2.3e5.7 g VS/L/d compared to rector R1 (FW:CB 100:0) at OLR of 2e4.7 g VS/L/d.
R1 (FW:CB 100:0) R2 (FW:CB 80:20) R3 (FW:CB 60:40)
HRT OLR (gVS/L/ Biogas yield (L/ OLR (gVS/L/ gVA/ Biogas yield (L/ OLR (gVS/L/
(d) d) gVS) d) gCB gVS) d)
90 2 0.92 2.3 3.0 0.24 2.4
80 2.3 1.0 2.6 3.0 0.32 2.7
60 3.1 1.1 3.5 3.0 0.34 3.6
40 4.7 0.74 5.2 3.0 0.35 5.5
7
Journal of Cleaner Production 289 (2021) 125775
in the range of 45e65% in all the four reactors. This decline in VS
removal efficiencies is mainly because of the high solids present in
the substrate (i.e. > 19% TS, which is regarded as solid-state or dry
AD). It is interesting to note that the digestate pH in all the reactors
was between 6.8 and 7.5 throughout the operation, which suggests
that efficient digestion had occurred (Fig. 4). The current study
investigated the performance of FW and CB ACoD at an HRT of 40 d
to understand its techno-economic feasibility at full-scale.
Although the co-substrate CB is not as easily biodegradable as
FW, the reactor start-up approach and micro-feeding strategy
improved the overall performance of FW and CB co-digestions
substantially and the system attained steady state without the
need for external buffering. As noted earlier, depolymerization of
cellulose by exposure to acetic acid is more likely to occur over the
long term. Overall, to enhance the performance of FW and CB
anaerobic co-digestion, a pre-treatment step is necessary.
3.3. Mass balance
The CSTR system was continuously operated for 70 d, with
decreasing HRT from 90 d to 80 d followed by 60 d and 40 d. The TS
in the total solid of substrate which is the sum of VS and fixed solids
(FS). The mass balance in terms of input TS, VS and FS fed to the
reactors and the output in terms of biogas generation, FS and un-
digested VS is shown in Fig. 5. During the digestion time of 70 d, the
reactors R1eR4 were fed with TS of 1386 kg, 1530 kg, 1679 kg and
1750 kg, of which the total VS was 1310 kg, 1460 kg, 1535 kg and
1600 kg respectively. FS is the non-biodegradable inert component
of the digestate. Biogas generation of 974 L, 551 L, 452 L and 333 L
over 70 d of digestion time was observed from the total VS fed to
R1eR4 respectively. The undigested fraction of VS and FS consti-
tutes the TS in the digestate. The biogas generation and VS removal
in R1 were higher than in R2, followed by R3 and R4. The major
reason was the composition of the substrate, because the CB de-
grades more slowly than FW.
3.4. Theoretical v/s experimental methane yield of solid-state ACoD
of FW and CB
The theoretical methane yield (TMY) from FW, CB, and the
mixture of FW and CB in different ratios was estimated from the
elemental composition of FW and CB (Table 1) using Eq. (1)
(Achinas and Euverink, 2016).
22:4*½2a þ 8b  4c  3d e
8  4
TMY ¼
12:017*a þ 1:0079*b þ 15:999*c þ 14:0067*d þ 32:065*e
(1)
Where a, b, c, d and e are the masses of the elements C, H, O, N and S
respectively divided by their molar masses in the numerator. The
TMY calculated using Eq. (1) does not determine the biodegrad-
ability and non-biodegradability of the substrate, therefore, a
comparison of the calculated TMY of the substrate and the
R4 (FW:CB 50:50)
gVA/ Biogas yield (L/ OLR (gVS/L/ gVA/ Biogas yield (L/
gCB gVS) d) gCB gVS)
1.5 0.31 2.5 1.2 0.26
1.5 0.49 2.9 1.2 0.30
1.5 0.56 3.8 1.2 0.35
1.5 0.39 5.7 1.2 0.21
S. Begum, T. Das, G.R. Anupoju et al.
Fig. 4. Impact of FW:CB mixing on volatile solids removal efficiency and digestate pH of ACoD of CB and FW over 70 d digestion time.
experimental methane yield (EMY) was essential to find the actual
biodegradability of the feedstock. The methane (CH4) content in the
present study was assumed to be 65%, which is a standard for FW
(Irena.org, 2019; Isola et al., 2018; Kuo and Dow, 2017; Metcalf and
Eddy, 1991). The TMY of FW:CB in the ratio of 100:0 and 0:100 was
estimated considering the elemental composition of the substrates
individually, while the TMY for mixture was estimated based on the
number of g of VSadded (i.e. gVSadded of FW þ gVSadded of CB). A
prediction of TMY was made based on the cumulative gVSadded
from the mixture. The biodegradability (%) was estimated by the
ratio of EMY to TMY, as shown in Eq. (2).
EMY
Biodegradability ð%Þ ¼ *100%
TMY
The TMY, EMY and the biodegradability from the reactors are
summarized in Table 4, which shows that the EMY in each reactor is
lower than the TMY. An important finding of this study is that the
biodegradability of FW in reactor R1 was 76%, while in reactors R2,
R3 and R4 it was 39%, 32%, and 22% respectively. This could be
attributed to the fact that lignocellulosic substrates like CB degrade
slowly due to their recalcitrant characteristic and the presence of
lignin compared to sugar-containing substrates like FW. In addi-
tion, the TS concentration of all the feed mixtures was higher than
18%, so poor accessibility of microbes to organic matter due to
reduced mobility inside the digester could explain the lower
biodegradability of CB. Lower fluidity due to higher rheological
properties of digester media in 40% CB (R3) and 50% CB (R4) in FW
resulted in lower biodegradability than in R1 and R2 (Miryahyaei
et al., 2019). Therefore, the biodegradability of FW and CB is
lower than that of FW alone, because there will always be complex
biochemical reactions produced by the co-digestion of the different
substrates.
Journal of Cleaner Production 289 (2021) 125775
3.5. A techno-economic analysis of solid-state ACoD of FW and CB
at full-scale
The profitable operation of an AD unit relies on low capital and
operational costs. Most of the studies in literature investigated the
biomethane potential of FW and CB as lab-scale batch experiments
that do not represent the output that could be achieved from a full-
scale AD unit. In the present study, a pilot-scale CSTR system that
could represent a full-scale AD unit was operated with FW and
mixture of FW and CB in different mixing ratios for 70 d. Table 5
presents the estimates of techno-economic analysis of a full-scale
AD unit based on the performance of the pilot-scale CSTR system.
It is experimentally proven in the present study that the AD of
(2)
FW alone results in better output than the co-digestion of FW and
CB. The performance output from ACoD of FW and CB illustrated
that R2 with FW:CB of 80:20 outperformed compared to R3 (FW:CB
60:40) and R4 (FW:CB 50:50), and the same is expected at full-scale
(Table 5). The HRT and power required for the operation of the AD
unit was kept constant in all cases. The water and power require-
ment per day were considered from the literature (Begum et al.,
2017a, b; Kuruti et al., 2017). The operational cost of the AD unit,
which includes manpower, water and power, was estimated be-
tween 10,080 $/y in all the cases as the water, power and manpower
required is assumed to be constant in all the cases (Table 5). It can
be illustrated from Table 5 that the biogas generation from R1 (i.e.
1000 kg/d of FW) is 133 m3/day whereas for R2, R3 and R4, it is
52.5 m3/d, 40 m3/d and 27 m3/d, respectively. The poor perfor-
mance of R3 and R4 interms of biogas generation can be attributed
to the increase of CB ratio. Previous research on 1000 kg FW AD
plants demonstrated a maximum biogas generation between 80
and 150 m3/day (Kuruti et al., 2017) which corroborates with the
present study i.e. 133 m3/day (Table 5). No previous researchers
8
S. Begum, T. Das, G.R. Anupoju et al. Journal of Cleaner Production 289 (2021) 125775

waste disposal rather than earning profits, the predicted techno-

economics are highly promising. The complete economic feasi-
bility of these plants can be assessed when the capital cost of these
plants is taken into consideration which is not accounted in this
study. This study suggests that the installation of decentralized AD
units of this kind at the source of waste generation would eliminate
the costs of transportation of waste to centralized landfill and
produce indirect economic benefits through reduction of environ-
mental contamination and associated harm to public health.

3.6. Further research and recommendations

This study concludes that there remains further scope for

research on solid-state ACoD of acetic acid rich FW and CB partic-
ularly with regard to pre-treatment. The present study found that
the biogas yield decreased at an HRT of 40 d with an OLR >4 g VS/
(L.d), even after the system attained stability. The biodegradability
of the ACoD of FW and CB mixture ranged from 22 to 39%, which
could be further enhanced by the introduction of physical, chemical
and/or biological pre-treatment steps, depending on the composi-
tion of waste and the operating conditions. In addition to the pre-
treatment, further studies on rheological properties of FW and CB
with high solids is necessary because the accessibility of organic
matter to the microbes can be influenced by the rheology of the
reactor contents. The cost of pre-treatment and the value of the
extra methane yield must be taken into account in the overall
economic feasibility analysis of an AD unit particularly for FW:CB
ratios of 60:40 and 50:50. The investment costs for pre-treatment
of recalcitrant substrates like CB are high due to the necessary
process engineering steps. Longer HRTs may improve plant effi-
ciency but requires more capital investment due to large reactor
volumes because the land located near waste generation sources is
inevitably expensive. Research into the detailed techno-economic
feasibility analysis of a full-scale AD plant with a pre-treatment
step and rheological properties has not been undertaken to date
Fig. 5. Overall mass balance in reactor inlets and outlets (TS: Total solid, VS: Volatile which is highly recommended.
solid, FS: fixed solids).
4. Conclusion
Table 4
Comparison of the theoretical and experimental methane yield of FW and CB. This study evaluated the performance of a solid-state ACoD of
acetic acid rich FW and CB in a pilot-scale auto-fed CSTR system at a
Reactor name TMY (L CH4/g EMY (L CH4/g Biodegradability
complete HRT of 40 d. ACoD of FW and CB in the ratios of 100:0,
VSadded) VSadded) (%)
80:20, 60:40 and 50:50 with TS concentration between 19% and
FW:CB (0:100) 0.45 e e 24%.
R1 (FW:CB 0.63 0.48 76%
100:0)
This study concluded that the solid-state ACoD of FW and CB
R2 (FW:CB 0.61 0.24 39% without any pre-treatment or addition of buffering chemicals is
80:20) feasible if the micro-feeding strategy and long HRT (for the long-
R3 (FW:CB 0.6 0.19 32% term exposure of CB to acetic acid-rich FW) are implemented.
60:40)
Micro-feeding strategy and frequent feeding pattern on an hourly
R4 (FW:CB 0.6 0.13 22%
50:50) basis assisted in overcoming the acidification phenomena of FW and
attaining stability even at OLR >4 gVS/(L.d). The lower biogas yield
for reactors with CB content more than 40% is due to poor biode-
gradability of CB due to lower interaction of acetic acid (CH3COOH)
have reported results for full-scale co-digestion of FW and CB, while
in FW with CB. The organic matter removal efficiencies (for 50% CB
this study has made an attempt to predict the performance at full-
addition to no CB in food waste) declined from 75 e 90% to 45e65%
scale.
as the HRT was shortened from 90 d to 40 d. The techno-economic
As can be seen from Table 5 that the difference a positive profits
feasibility analysis indicated the system is profitable even without
that can be achieved from R1 and R2, while for R3 and R4 there is no
any pre-treatment for the reactors with foodwaste only or food-
profit. The lower revenue and no net profit in the case of R3 and R4
waste with 20% CB. To make the digestion of more than 40% CB in
can be attributed to lower degradation of CB. Notwithstanding the
food waste profitable, a pre-treatment method need be envisaged. If
fact that the lower revenues can be attributed to the usage of biogas
the safe waste disposal is the primary concern rather than earning
for electricity generation, rather than its usage for heat applications
profits, the installation of full-scale plants of FW and CB can be an
such as cooking fuel. This is because the cost of power is relatively
ideal solution. Installation of AD units at sources of waste generation
lower than the cooking fuel. Nevertheless, if the installation of this
would also eliminate the cost that would otherwise be incurred for
kind of full-scale plants are considered as a solution for the safe
transportation of waste to a centralized landfill location.
9
S. Begum, T. Das, G.R. Anupoju et al. Journal of Cleaner Production 289 (2021) 125775

Table 5
A techno-economic analysis of ACoD of FW and CB at different mixing ratios in a micro-feeding CSTR.

R1 (FW:CB 100:0) R2 (FW:CB 80:20) R3 (FW:CB 60:40) R4 (FW:CB 50:50)

Technical details of the AD unit

Quantity of substrate (kg/d) 1000 800 kg FW and 14.4 kg CB 600 kg FW and 29 kg CB 500 kg FW and 36 kg CB
Organic matter per ton of substrate (kg VS/d) 180 164 139 126
Volume of digester (m3) 38.3 38.5 38.2 38.6
HRT (d) 40 40 40 40
Water requirement (L/d) 250 250 250 250
Power required for operation (kWh/d) 15 15 15 15
Biogas generation (m3/d) 133 52.5 40 27
Electricity generation from biogas (kWh/d) 266 105 80 54
Digestate generation (L/d) 892 732 570 495
Operational cost
Manpower required for (1 person for 1 h)/y @ $25/h ($/d) 25 25 25 25
Cost of water ($/d) 1.0 1.0 1.0 1.0
Cost of power required ($/d) 4.2 4.2 4.2 4.2
Total operating cost ($/d) 30 30 30 30
No. of working days 336 336 336 336
Total operating cost ($/y) 10,080 10,080 10,080 10,080
Revenue from the plant
Revenue from electricity generation ($/d) 75 29.4 22.4 15
Revenue from digestate sales ($/d) 9 7.3 6 5
Total revenue from the plant ($/d) 84 37 28.4 20
Total revenue from the AD unit ($/y) 28,224 12,432 9,542 6,720
Profit (revenue minus operating cost) ($/y) 18,144 2,352 () 538 () 3,360

1 kWh ¼ 28 cents (Canstarblue.com.au, 2020); Water ¼ $0.0036/L (Team Poly Water Tanks | Water Solutions for Li et al., 2019); 1 m3 of biogas ¼ 2 kWh (Aqper.com, 2020);
Digestate ¼ $0.01/L (Kuruti et al., 2017); $ ¼ AUD (Australian Dollar).

CRediT authorship contribution statement
Sameena Begum: Methodology, Formal analysis, Investigation,
Writing - original draft, Visualization. Tanmoy Das: Investigation.
Gangagni Rao Anupoju: Supervision. Nicky Eshtiaghi: Supervi-
sion, Writing - review & editing, Conceptualization, Project
administration.
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.
Acknowledgement
The authors acknowledge RMIT University, Melbourne, Australia
and the Council of Scientific and Industrial Research e Indian
Institute of Chemical Technology for financial support and a
scholarship for Sameena Begum. The authors also acknowledge the
Aurora sewage treatment plant (Preston, Australia) for providing
food waste and inoculum, and Mr Shoutian Li for his help in reactor
operation.
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