Energy Conversion and Management 117 (2016) 538–547

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Energy Conversion and Management

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Effect of VS organic loads and buckwheat husk on methane production

by anaerobic co-digestion of primary sludge and wheat straw
Mahmoud Elsayed a,d,⇑, Yaves Andres a, Walid Blel b, Ali Gad c, Abdelkader Ahmed d
a
GEPEA UMR CNRS 6144, 4 rue Alfred Kastler, 44307 Nantes cedex 3, Environmental Engineering Department, Ecole Des Mines De Nantes, Nantes University, Nantes, France
b
CNRS, GEPEA, UMR 6144, CRTT, Environmental Engineering Department, Nantes University, France
c
Civil Engineering Department, Faculty of Engineering, Assiut University, 71516 Assiut, Egypt
d
Civil Engineering Department, Faculty of Engineering, Aswan University, 81542 Aswan, Egypt

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

Article history: An environmentally acceptable disposal of sewage sludge and agro-wastes presents an urgent problem
Received 22 January 2016 facing many countries. Anaerobic digestion (AD) is a robust and suitable technique for producing renew-
Received in revised form 22 March 2016 able energy from wastes. This study aims to improve methane production from anaerobic co-digestion of
Accepted 23 March 2016
primary sludge (PS) and wheat straw (WS) depending on their volatile solids (VS) organic load and by
adding a proposed waste material of buckwheat husk (BH) based on their carbon to nitrogen (C/N) ratio.
Mesophilic anaerobic batch tests were carried out in 500-mL digesters. Individual and six mixtures of PS
Keywords:
and WS at different VS organic loads were anaerobically digested to optimize VS load for the greatest gas
Primary sludge
Agro-wastes
production. The highest cumulative methane yield (CMYs) occurred with combined substrates at a VS
Anaerobic co-digestion load of 7.50 gVS/L. In general, the optimized organic loads that gave the highest cumulative biogas yield
Methane production (CBYs) and CMYs were in the range of 6–8 gVS/L. In addition, AD of individual substrates of PS, WS, and
Volatile solids organic load BH and of their mixture at different C/N ratios was investigated regarding to the methane yields. Multi-
Carbon to nitrogen ratio component substrates produced the greatest CMY at a C/N ratio of 10.07. The CMYs was increased by
39.26% when the proposed waste material of buckwheat husk (BH) was added to the different mixtures
of PS and WS compared to the co-digestion of PS and WS. Experimental results were approved using sta-
tistical analysis by ANOVA test at P-value less than 0.05. Purification of methane and biodegradation of VS
were evaluated. The results emphasized positive synergy of anaerobic co-digestion for improving CMY
and best feedstock utilization.
Ó 2016 Elsevier Ltd. All rights reserved.

1. Introduction tally sensitive problem. With some traditional disposal routes

Currently, the management of sewage sludge produced from
wastewater treatment plants is one of the most serious challenges
in biological wastewater treatment. Its amount is increasing
worldwide as the quantity of wastewater being treated rises [15].
Fresh and untreated sludge will have many pathogens, a high pro-
portion of water, high biochemical oxygen demand (BOD), and is
normally putrid and odorous. Treatment and disposal of sewage
sludge accounts for about half, even up to 60%, of the total cost
of wastewater treatment [38]. It is an expensive and environmen-
⇑ Corresponding author at: GEPEA UMR CNRS 6144, Ecole Des Mines De Nantes, 4
rue Alfred Kastler, 44307 Nantes cedex 3, France; Civil Engineering Department,
Faculty of Engineering, Aswan University, 81542 Aswan, Egypt.
E-mail addresses: Mahmoud-Abdelazez.BASTAWEY@mines-nantes.fr,
engzezo1111@yahoo.com (M. Elsayed).
(landfill, land application, and incineration) coming under pressure
and others such as sea disposal having been phased out, the chal-
lenge facing sludge managers is to find cost-effective and innova-
tive solutions whilst responding to environmental, regulatory,
and public pressures.
Simultaneously, a huge quantity of wastes is generated from
agricultural crops all over the world. Amount of agriculture resi-
dues is proportional to the production volume and they are gener-
ally disposed of by incineration or land filling leading to damaging
effects on the environment. Agro-wastes contain valuable
resources which can be recovered for many and very diverse eco-
nomic, social, and environmental purposes. Among agricultural
products commonly found worldwide, wheat is considered one of
the most important. It is grown in large amounts all over the world
and represents the second major edible commodity after rice [4].
As well, buckwheat is an important crop, which is widely grown

http://dx.doi.org/10.1016/j.enconman.2016.03.064
0196-8904/Ó 2016 Elsevier Ltd. All rights reserved.
 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547
in Europe and Asia. It can be used in human nutrition, as feed for
livestock, and as a honey crop. The increasing amount of
agro-wastes from wheat and buckwheat crops encourages
researchers to pay more attention to particular aspects of its man-
agement, especially to recycling and waste-to-energy issues.
A cost-effective disposal of sewage sludge and agricultural resi-
dues in a way that is environmentally acceptable is one of the most
urgent problems facing many countries [28]. Both aerobic and
anaerobic processes can be used for waste stabilization. Anaerobic
digestion (AD) has several advantages such that it consumes a little
or even no energy, stabilizes the feedstock, and the residual cake
can still be used as a soil ameliorant. In addition, it is considered
one of the most suitable techniques for producing renewable
energy from wastes. This technique converts organic matters in
sewage sludge [45] and crop residues [44] into biogas (rich with
methane) by a consortium of microorganisms, which can be used
for heating and producing electricity [39].
Sewage sludge is a valuable feedstock for anaerobic digestion
(AD) as it contains a high percentage of nitrogen, which can pro-
vide a buffering capacity and an improved balance of nutrients
[31]. Although convenient and feasible, it has been recognized that
the individual digestion of sewage sludge did not achieve the most
efficient production of biogas due to its inherent deficiency of car-
bon; i.e., low carbon/nitrogen (C/N) ratio [21]. In contrast, agro-
wastes contain a high percentage of carbon and a low percentage
of nitrogen (high C/N ratio). Co-digestion of sewage sludge and
agro-wastes is considered the best solution for enhancing methane
yields [10], improving the C/N ratio [20], and decreasing the risk of
ammonia inhibition [9]. Different types of agro-wastes can be used
in admixture with sewage sludge, such as wheat straw [36] and
rice straw [13]. A high biogas yield was obtained from the co-
digestion of food wastes and rice husk based on C/N ratio of the
mix [6].
The effect of total solids (TS) concentration on biogas and
methane yields from anaerobic digestion of a single type of wastes
has been investigated in several previous studies. Beevi et al. [3]
investigated the effect of TS concentration on anaerobic digestion
of organic fraction of municipal solid wastes using varying
substrate concentrations. Yavini et al. [42] studied the effect of
TS concentration on biogas yields of agricultural wastes. Tanimu
et al. [32] studied the effect of volatile solids (VS) feeding load on
both biogas and methane production in batch mesophilic
anaerobic digesters treating food wastes. Although, the anaerobic
digestion (AD) is considered an economic and environmentally
friendly technology, it confronts certain limitations regarding high
retention times, restricted methanogenic production, and low
overall organic dry solids degradation efficiency [1]. A reliable
overcoming of these restrictions in a wide range would be of a
great value.
Most of the previous studies were concerned with the effect of
the total solids concentration on biogas production from anaerobic
digestions. There are fewer studies about the effect of volatile
solids (VS) organic load in biochemical methane potential (BMP)
tests for the optimal methane production and the best utilization
of feedstock components. On the other hand, the optimization of
methane production from co-digestion of different feedstock mate-
rials can significantly improve methane yields, anaerobic digestion
performance, and biodegradability of the feedstock. The main
objective of this research is to investigate the possibility of improv-
ing the methane yields from anaerobic co-digestion of nitrogen-
rich primary sludge in combination with carbon-rich wheat straw
based on their VS organic loads and by adding a proposed waste
material of buckwheat husk depending on the carbon to nitrogen
ratio that achieve a better balance of nutrients and positive syner-
gisms in the digestion medium.
539
2. Methodology
2.1. Preparation of the substrates
A primary sludge (PS) substrate was collected from a full-scale
municipal wastewater treatment plant located in Nantes, France.
The plant system follows the activated sludge treatment method.
Primary sewage sludge samples were dried for stabilization and
stored for the later determination of their characteristics and the
following anaerobic digestion processes. A wheat straw (WS) sub-
strate was obtained from a farm located in Nantes and dried at the
room temperature. Yong et al. [43] recommended that the size of
the straw is better to be ranged from 0.30 to 1.0 mm for economic
and energy-saving reasons. The size of WS substrate was reduced
to be less than 1.0 mm using a hammer mill followed by a coffee
grinder. The grinded WS feedstock was kept for determining its
characteristics and feeding anaerobic digesters. A buckwheat husk
(BH) substrate, a waste material that was suggested for improving
methane production, was supplied by the Voltière mills in Gar-
nache, France. It was dried, grinded, and kept for use as the other
feedstock components.
2.2. Inoculum
Samples of fresh cow manure were collected from a small farm
located in Nantes, France to be used as an inoculum in the anaer-
obic digestion processes. To remove the dissolved methane and
residual organic matter contained in the fresh manure, it was
stored in an anaerobic headspace for more than one month. To
readapt microorganisms for mesophilic conditions, the inoculum
was kept at a constant temperature of 37 °C using a water bath
for 15 days before starting the batch tests [7]. The measured value
of TS of the inoculum was found to be 5.59% relative to its dry
weight while its VS amounted to 73.18% of its TS. The inoculum
had a pH value of 7.21.
2.3. Analytical techniques
Volatile solids (VS), total solids (TS), and pH values for the sub-
strates of anaerobic digestion and residues of the digestion pro-
cesses were determined according to APHA Standard Methods
[2]. In addition, total carbon (TC), total nitrogen (TN), total oxygen
(TO), and total hydrogen (TH) were estimated with a thermal con-
ductivity detector using a FLASH EA 1112 Series CHN Analyzer. The
composition of the produced biogas from anaerobic digestion was
determined by a Clarus 500 Gas Chromatograph (Perkin Elmer).
After absorbing carbon dioxide from the produced biogas, the vol-
ume of the produced methane was measured daily by the water
displacement method. Characterization results of the feedstock
Table 1
Characteristics of the feedstock components used in the biochemical methane
potential (BMP) tests.
Characteristics Primary sludge (PS) Wheat straw (WS) Buckwheat husk (BH)
VS (TS %) 82.50 ± 0.10 95.64 ± 0.05 97.60 ± 0.08
TS (dry wt.%) 81.70 ± 0.15 90.82 ± 0.20 85.00 ± 0.36
TC (dry wt.%) 39.90 ± 0.44 47.62 ± 0.58 47.50 ± 0.51
TN (dry wt.%) 6.70 ± 0.25 0.30 ± 0.04 2.30 ± 0.12
TO (dry wt.%) 28.30 ± 0.19 44.10 ± 0.42 43.37 ± 0.30
TH (dry wt.%) 5.40 ± 0.09 5.54 ± 0.17 6.10 ± 0.12
C/N ratio 5.96 158.73 20.65
Notes: VS = volatile solids, TS = total solids, TC = total carbon, TN = total nitrogen,
TO = total oxygen, TH = total hydrogen, and C/N = nitrogen to carbon. The data
represent the means ± SD, n = 4.
540 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547

components that were used in the biochemical methane potential its height in a temperature-controlled water bath (Polystat 23 from
(BMP) tests are given in Table 1. Bio Block Scientific) to maintain a constant incubation temperature
of 37 ± 1 °C for the mesophilic digestion. Each reactor was sealed
2.4. Carbon dioxide separating unit with a poly-vinyl chloride cap that was provided with two accesses
for plastic tubes; one tube was connected to the carbon dioxide
Biogas produced from anaerobic digestion does not consist (CO2) separating unit while the other was plugged with a plastic
entirely of methane as it also contains carbon dioxide (CO2), smal- tubing clamp. After passing through the separating unit, the CO2
ler amount of other gases such as nitrogen and hydrogen sulphide, free biogas left through a plastic tube to displace an equivalent vol-
and traces of volatile compounds. The high ratio of CO2 in the pro- ume of water in the measuring vessel. The volume of the produced
duced biogas reduces its heating value and increases its compres- methane was measured by the water displacement method as
sion and transportation costs. As reported by Horan et al. [10], CO2 shown in Fig. 1. According to Zhang et al. [45], each reactor was
was excluded from the produced biogas by absorbing the later in shaken manually for one minute per day before the gas volume
3 M solution of sodium hydroxide (NaOH). Glass bottles with vol- was measured.
umes of 250 mL were filled with NaOH solution to a volume of The methane gas produced from the inoculum was subtracted
230 mL and sealed with poly-vinyl chloride caps. Each cap had from the sample assays and the produced methane could be calcu-
two circular accesses for plastic tubes, one of these tubes was con- lated from the following equation:
nected to a reactor bottle for the incoming biogas and the other
delivers the gas to the volume measuring device.
V ðfeedstockþinoÞ  V ino
BMP ¼ ð1Þ
VSfeedstock
2.5. Experimental design and set-up
where BMP is the biochemical methane potential in mL of CH4/
gVSadd, V ðfeedstockþinoÞ is the volume of the produced methane from
Biochemical methane potential (BMP) tests were carried out in
the mixed feedstock and inoculum in mL of CH4, V ino is the volume
duplicate under mesophilic (37 ± 1 °C) conditions to get the opti-
of the produced methane from the individual inoculum in mL of
mal volatile solids (VS) organic load and C/N ratio for an effective
CH4, and VSfeedstock is the mass of volatile solids in the feedstock in
methane production. Anaerobic batch reactors with total and effec-
gm of VSadd. The tests continued for 30 days with an effective biogas
tive volumes of 500 and 400 mL, respectively, were used. In the
production and they were ended when the biogas ceased continu-
first group of experiments, six different weights of VS of primary
ously for 15 days, according to the method described by Xie et al.
sludge (PS) and wheat straw (WS) were mixed in anaerobic reac-
[41]. At the end of each experiment, the digestion residue in each
tors referred as B1 to B6 to get different VS organic loads in the
reactor was sampled for measuring the TS, VS, and pH to determine
range from 3.0 to 12.0 gVS/L as presented in Table 2. For compar-
the VS removal rate and biodegradability of the feedstock.
ison, the C/N ratios for all digesters were adjusted to be approxi-
mately the same. Two more anaerobic digesters, C1 and C2, with
an individual feedstock of PS and WS, respectively, were used as 2.6. Statistical analysis
controls.
In the second group of experiments, five different amounts of In the present study, the measured values of cumulative
primary sludge (PS), wheat straw (WS), and buckwheat husk methane yields (CMYs) were performed as the means of two repli-
(BH) were mixed to produce different C/N ratios ranges from cates. The statistical analysis was carried out by STATGRAPHICS
10.07 to 25.25 and digested in five anaerobic reactors referred as Centurion XV software (Virginia, USA) using ANOVA analysis. In
R1 to R5 as given in Table 3. Unmixed substrates of PS, WS, and
BH in three additional anaerobic reactors referred as C1, C2, and
Table 3
C3, respectively, as given in Table 3 were used as controls. In each Different mixtures of substrates of PS, WS, and BH in anaerobic co-digestion process
digester of this group, the total VS organic load of the feedstock at different C/N ratios.
was adjusted to be 7.50 gVS/L.
Reactor number PS WS BH C/N
The digesters of the two groups of experiments were partially (gVS/400 mL) (gVS/400 mL) (gVS/400 mL) ratio
filled with 150 mL of inoculum along with the appropriate
R1 1.55 0.73 0.72 10.07
amounts of the specific feedstock components. A constant ratio
R2 1.07 0.97 0.96 13.06
of 2.0 was kept between the VS of inoculum and the total VS of R3 0.86 1.07 1.07 15.01
substrates in the second experiment to minimize the diffusion R4 0.50 1.25 1.25 20.03
and to avoid acidification or toxicity inhibition as recommended R5 0.27 1.37 1.36 25.25
C1 3.00 0.00 0.00 5.96
by Rico et al. [26]. Deionized water was supplied to each reactor
C2 0.00 3.00 0.00 158.73
to reach the working volume of 400 mL. Initial pH value for all C3 0.00 0.00 3.00 20.65
digesters was adjusted to a value of 7 ± 0.1 using 6 M sodium
Notes: PS = primary sludge, WS = wheat straw, BH = Buckwheat husk, and C/
hydroxide (NaOH). The headspace of each reactor was flushed with
N = carbon to nitrogen. The reactors R1 to R5 were fed by mixtures of PS, WS, and
nitrogen (N2) gas for three minutes to produce an absolute anaer- BH whereas C1, C2, and C3 were fed by a single substrate of PS, WS, and BH,
obic condition. After that, each reactor was immersed up to half of respectively.

Table 2
Different mixtures of substrates of PS and WS in anaerobic co-digestion process at various volatile solids (VS) organic loads.

Reactor number B1 B1 B3 B4 B5 B6 C1 C2
PSadded (gVS/400 mL) 0.45 0.90 1.10 1.20 1.50 1.80 3.00 0.00
WSadded (gVS/400 mL) 0.75 1.50 1.90 2.00 2.50 3.00 0.00 3.00
Mixture load (gVS/L) 3.00 6.00 7.50 8.00 10.00 12.00 7.50 7.50
C/N ratio 15.19 15.27 15.59 15.26 15.24 15.24 5.96 158.73

Notes: PS = primary sludge, WS = wheat straw, and C/N = carbon to nitrogen. The reactors B1 to B6 were fed by mixtures of PS and WS while C1 and C2 were fed by an
individual substrate of PS and WS, respectively.
 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547
Fig. 1. Schematic diagram of the biochemical methane potential (BMP) tests set-up.
this series of experiments, the P-value of the F-test is less than 0.05.
The differences between the values of the mean cumulative
methane produced at the different levels of VS organic loads and
C/N ratios were analyzed at a confidence interval of 95%.
3. Results and discussion
3.1. Characterization of feedstock components
In anaerobic digestion processes, it is important to characterize
the feedstock components to ensure its balance in terms of volatile
solids (VS), total solids (TS), total carbon (TC), and total nitrogen
(TN) contents. Four measurements were performed for each
parameter and the arithmetic mean was estimated. In addition, a
statistical analysis was performed and standard deviations (SD)
were calculated. As shown in Table 1, it is clear that wheat straw
(WS) and buckwheat husk (BH) are rich in their carbon contents
with ratios of 47.62% and 47.50% relative to their dry weights,
respectively. However, they have low nitrogen contents with per-
centages of 0.3% and 2.30% for WS and BH relative to their dry
weights, respectively. In contrast, primary sludge (PS) is character-
ized by a relatively low carbon content of 39.90% and a relatively
high nitrogen content of 6.70% from its dry weight. In light of these
results, WS and BH are found to have higher C/N ratios of 158.73
and 20.65, respectively, compared with the low ratio of 5.96 in
the case of PS. This great variance in C/N values between PS and
agro-wastes foretells that their mixture may increase the methane
production as compared to the individual substrates by enhancing
C/N ratio.
3.2. Cumulative methane yields at different volatile solid organic loads
For anaerobic digestion of multi-component substrates, there
are few studies regarding to the effect of volatile solid (VS) organic
loads on the methane (CH4) production, although they play an
541
important role. The cumulative methane yields (CMYs) from anaer-
obic co-digestions of PS and WS through continuous 30 days of
experiments under mesophilic conditions at different VS organic
loads are presented in Fig. 2. The CMYs from anaerobic co-
digestions of PS and WS at VS organic loads of 12.0, 10.0, 8.0, 7.5,
6.0, and 3 gVS/L showed a higher methane yield of 1.81, 1.89,
1.97, 2.30, 2.06 and 1.45 times more, respectively, than digesting
PS alone. Similarly, the organic loads of 12.0, 10.0, 8.0, 7.5, and
6.0 gVS/L produced CMYs greater than digesting WS separately.
However, the CMY at the organic load of 3.0 gVS/L showed a
decrease of 16.35% compared to the single digestion of WS. This
low CH4 production may be come from the small amounts of the
added feedstock components at this load.
As shown in Fig. 2, it is clear that as the VS organic load deviated
from the optimum value (7.50 gVS/L), the CMY became less. More-
over, through 30 days of experiments, the highest CMYs (345.5 mL/
gVSadd) from anaerobic co-digestion of PS and WS occurred at an
optimum organic load of 7.50 gVS/L. While, the minimum CMYs
were observed at organic loads of 3 and 12 gVS/L. This may be
occurred as a result of many reasons; firstly, the optimum organic
load (7.50 gVS/L) provides suitable conditions for microorganisms
to grow and proliferate consequently transfer the highest carbon
content of the feedstock to CH4 through the methanization process.
Secondly, the anaerobic reactor does not operate effectively with
low organic loads because the microorganisms will show a low
metabolic activity and low amounts of gas will be produced [25].
Thirdly, biogas production decreased with the increase in organic
loads after optimum concentration due to the high production of
volatile fatty acids (VFAs) which cannot be consumed as rapidly
as they produced and result in toxic conditions in the reactor [5].
The maximum CMY obtained in this study (345.5 mL/gVSadd) is
higher than some of the yields reported by others such as Wang
et al. [36] from the co-digestion of dairy, chicken manure, and
wheat straw (234.7 mL/gVS), Sahito et al. [29] from the co-
digestion of crop residue with buffalo dung (322.0 CH4/gVSadd),
and Wei et al. [37] from the co-digestion of barley straw with
542
400
Cumulative methane yields (mL/gVSadd)
350
300
250
200
150
100
50
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
Fig. 2. Transient changes of the cumulative methane yields from the anaerobic co-digestions of PS and WS substrates at different VS organic loads.
two animal manures (233.4 mL/gVS). However, it is lower than the
CMY that was reported by Tanimu et al. [32] from the co-digestion
of treating food wastes (535.0 mL/gVS). These incompatibilities can
be relayed to many reasons; Firstly, this study used a feedstock mix
of PS and WS with a C/N ratio of around 15.0 (Table 2) while Tan-
imu et al. [32] digested food wastes at a C/N ratio of 30.0. Secondly,
Tanimu et al. [32] used only four feeding loads in the range from
0.5 to 5.5 gVS/L, while in this study six VS organic loads that were
varied from 3.0 to 12.0 gVS/L. Thirdly, the difference in methane
yield can be attributed to the variation in substrate composition.
Indeed, agro-wastes biomass is mainly composed of cellulose,
hemicelluloses, and anaerobically non-digestible lignin [12], while
the main composition of food wastes are carbohydrate and protein
[34]. Finally, the VS removal ratio (96%) which was found by Tan-
imu et al. [32] is higher than the ratio obtained in this study
(71.77%) due to the low degradability of lignin in agro-wastes com-
position. Accordingly, the type of wastes, biomass constituents,
and the C/N ratio control the level of biochemical activity that will
occur in the anaerobic digester.
The statistical analysis for two replicates of the measured CMYs
was performed using STATGRAPHICS Centurion XV software (Vir-
ginia, USA) using ANOVA analysis. In this approach, the data are
purely experimental and the studied system was considered as a
black box.
At a P-value of the F-test less than 0.05, a statistically significant
difference was found between the cumulative methane yields from
one volatile solids organic load level to another at 95.0% confidence
level. The VS organic load of 7.50 gVS/L is statistically considered as
the most optimized concentration which induces high methane
production.
3.3. Methane contents in the produced biogas at different VS organic
loads
The produced biogas from anaerobic digestions of wastes con-
tains around 50–80% methane and 20–40% carbon dioxide [8]
besides small amounts of nitrogen and hydrogen sulphide gases
and traces of volatile compounds. For economical and transporta-
tion requirements, it is preferred to reduce CO2 from the produced
biogas and increase its CH4 content before using for heat and elec-
tricity or as a fuel for power generation [17]. The volumetric
methane contents in the purified biogas at specific times of 5, 10,
15, 20, 25, and 30 days through one month of BMP tests for anaer-
obic co-digestions of PS and WS at different volatile solids (VS)
organic loads were plotted as shown in Fig. 3. From the figure,
the average CH4 contents in the purified biogas at all investigated
organic loads for the multi-component substrates of PS and WS
M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547
12 gVS/L 10 gVS/L
8 gVS/L 7.5 gVS/L
6 gVS/L 3 gVS/L
PS WS
Inoculum
Experimental time (days)
were found to be higher than those resulted from the single
digestion.
Table 4 presents the volumetric cumulative biogas yields
(CBYs), average volumetric CH4 contents in the purified biogas, vol-
umetric cumulative methane yields (CMYs), average removed per-
centage of the volatile solids (VS), and the end pH values at
different VS organic loads for anaerobic co-digestions of PS and
WS through BMP tests. From the table, the highest average CH4
contents of the purified biogas through BMP tests (96.51% and
95.90%) were observed at organic loads of 6.0 and 7.50 gVS/L,
respectively, while the lowest was 93.80% and occurred at an
organic load of 3.0 gVS/L. The average monthly CH4 contents in
the purified biogas for the used VS organic loads were relatively
high and ranged from 93.80% to 96.51%. In addition, the average
monthly CH4 contents from co-digestion of PS and WS at all tried
VS organic loads are higher than its contents from their individual
digestion as presented in Table 4. In addition, the highest monthly
removal ratio of VS (71.77%) occurred with the optimum organic
load (7.50 gVS/L) that reflects the maximum utilization of the feed-
ing substrates. The pH values at the end of BMP tests were found to
be still within the acceptable range of anaerobic microorganisms.
3.4. Effect of adding a proposed waste material of BH on anaerobic co-
digestions of PS and WS depending on their C/N ratio
3.4.1. Methane production from the single digestion of PS, WS, and BH
Fig. 4 presents the results of the daily methane production from
anaerobic co-digestions of primary sludge (PS), wheat straw (WS),
and buckwheat husk (BH) under mesophilic conditions though one
month of BMP tests at different C/N ratios. From the figure, the
peaks of the daily CH4 yields from the individual digestion of the
used PS, WS, and BH were found to be 17.70, 26.80, and
16.90 mL/gVSadd, respectively and occurred after 15, 6, and 14 days
from the beginning of BMP tests. The peak values of the daily CH4
yields of the agro-wastes (BH and WS substrates) occurred earlier
than PS, which may be due to its small particle size and high car-
bon content that make the digestion process more easy and fast.
The cumulative methane yields (CMYs) from anaerobic co-
digestions of primary sludge (PS), wheat straw (WS), and buck-
wheat husk (BH) though one month of BMP tests are shown in
Fig. 5. From the figure, it is clear that the CMYs recorded through
one month of anaerobic single digestion of WS and BH were
260.0 and 215.1 mL/gVSadd, respectively, and they were higher
than that was obtained from digesting PS (150.4 mL/gVSadd). The
results of this study comply with other researches that WS has
been proven to be the best feedstock for anaerobic single digestion
[36]. Sahito et al. [29] studied anaerobic digestion of WS separately
 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547 543

100

98

Volumetric methane content (%)

96

94

92

90

88 12 gVS/L 10 gVS/L 8 gVS/L 7.5 gVS/L

86 6 gVS/L 3 gVS/L PS WS

84
5 10 15 20 25 30
Experimental time (days)

Fig. 3. The average methane content in the purified biogas from anaerobic co-digestions of PS and WS at different VS organic loads.

Table 4
The measured CBYs, average methane content, CMYs, volatile solids (VS) removal, and pH values for the anaerobic co-digestions of PS and WS at different VS organic loads.

Parameter CBYs (mL/g VSadd) Average methane content (%) CMYs (mL/g VSadd) Volatile solids removal (%) pH Value
3.0 gVS/L 231.88 ± 12.0 93.80 217.50 ± 11.3 68.44 7.33
6.0 gVS/L 321.42 ± 10.2 96.51 310.20 ± 9.8 69.87 7.28
7.5 gVS/L 360.22 ± 18.4 95.90 345.50 ± 17.6 71.77 7.22
8.0 gVS/L 313.84 ± 12.0 94.24 295.80 ± 11.3 65.47 7.32
10.0 gVS/L 299.28 ± 16.2 94.96 284.20 ± 15.4 55.24 7.52
12.0 gVS/L 290.22 ± 13.0 93.86 272.40 ± 12.2 64.22 7.45
PS 162.06 ± 15.5 92.83 150.40 ± 14.5 58.42 7.66
WS 279.76 ± 6.8 92.92 260.00 ± 6.3 67.87 7.52

Notes: CBYs = cumulative biogas yields, CMYs = cumulative methane yields, PS = primary sludge, and WS = wheat straw. The data of CBYs and CMYs represent the means ± SD,
n = 2.

and got a CMY of 322.0 mL/gVSadd that is higher than the collected 3.4.2. Methane production from the co-digestion of PS, WS, and BH
volume of this study. However, Wang et al. [36] digested WS sep- For an effective methane production, it will be useful to co-
arately and reported a CMY of 121.2 mL/gVSadd that is lower than digest nitrogen-rich primary sludge (PS) from wastewater treat-
the recorded volume of this study (260.0 mL/gVSadd). On the other ment plants in combination with carbon-rich agriculture residues
hand, the CMY observed from anaerobic single digestion of PS to mediate potential effects of ammonia inhibition and overcome
(150.40 mL/gVSadd) is lower than the range of 239.0–267.0 mL/ nutrient deficiencies in terms of carbon. In this study, an optimiza-
gVSadd that was achieved by Komatsu et al. [14]. tion of the methane production from anaerobic co-digestion of PS

40
C/N=10.07

35 C/N=13.06
Daily methane production (mL/g VS)

C/N=15.01
30
C/N=20.03

25 C/N=25.25

BH
20
WS

15 PS

10

0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
Experimental time (days)

Fig. 4. Variations of the volumetric daily methane production from anaerobic co-digestions of PS, WS, and BH at different C/N ratios.
544
500
450
Cumulative methane yields (mL CH4/g VS)
400
350
300
250
200
150
100
50
0
1 3
Fig. 5. Transient changes of the cumulative methane yields from anaerobic co-digestions of PS, WS, and BH at different C/N ratios.
and WS by adding the buckwheat husk (BH) as a proposed waste
material based on their carbon to nitrogen ratio was performed.
The variations of the daily CH4 production from anaerobic co-
digestions of primary sludge (PS), wheat straw (WS), and buck-
wheat husk (BH) under mesophilic conditions though one month
are illustrated in Fig. 4. The peaks of the measured daily CH4 yields
from anaerobic co-digestion of PS, WS, and BH at C/N ratios of
10.07, 13.06, 15.01, 20.03, and 25.25 were found to be 34.6, 31.1,
34.6, 32.3, and 34.2 mL/gVSadd, respectively. These peak values
are around two folds the peaks of the single digestion of PS, WS,
and BH.
The transient changes of the measured cumulative methane
yields (CMYs) from anaerobic individual and co-digestions of pri-
mary sludge (PS), wheat straw (WS), and buckwheat husk (BH)
under mesophilic conditions though one month of BMP tests were
plotted in Fig. 5. The CMYs from anaerobic co-digestion of PS, WS,
and BH at C/N ratios of 10.07, 13.06, 15.01, 20.03, and 25.25 were
481.1, 436.7, 435.2, 410.1, and 337.9 mL/gVSadd, respectively.
These results reveal an increase of 3.20, 2.90, 2.90, 2.73, and 2.25
times more compared with digesting PS separately (150.4 mL/
gVSadd), an increase of 1.85, 1.68, 1.67, 1.58, and 1.30 times more
compared with the individual digestion of WS (260.0 mL/gVSadd),
and higher CMYs of 2.24, 2.03, 2.02, 1.91, and 1.57 times more
compared with the single digestion of BH (215.1 mL/gVSadd). Co-
digestion of PS, WS, and BH depending on their C/N ratios can sig-
nificantly improve CH4 yields and consequently biodegradability of
the feedstock. Mixing different substrates in anaerobic digestion
depending on their C/N ratios greatly improves biogas production
[36] and enhances anaerobic digestion performance [47]. Rizk
et al. [27] reported that the methane production was improved
for a mixture of sewage sludge with fruit and vegetable wastes
based on their C/N ratio. Wu et al. [40] studied the co-digestion
of swine manure with three crop residues and indicated that all
crop residues increased biogas and methane production at all C/
N ratios. Zhang et al. [44] showed that co-digestion of goat manure
with corn stalks or rice straw enhanced biogas production at all C/
N ratios. These previous studies support the results obtained in this
work and this enhancement can be related to the balanced nutri-
ents in the feedstock that improve the bacterial growth and the
CH4 production.
Analysis of variance using ANOVA test on the results of the
cumulative methane yields (CMYs) for co-digestion tests showed
a P-value of the F-test that is less than 0.05 and there is a statisti-
M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547
C/N=10.07
C/N=13.06
C/N=15.01
C/N=20.03
C/N=25.25
BH
WS
PS
Inoculum
5 7 9 11 13 15 17 19 21 23 25 27 29
Experimental time (days)
cally significant difference between the mean cumulative methane
from one level of C/N ratio to another at 95.0% confidence level
with an optimal C/N ratio of 10.07 for the methane production.
In turn, anaerobic co-digestion of the proposed waste material
of buckwheat husk (BH) with PS and WS increased the methane
yields by 39.26% compared to the anaerobic co-digestion of PS
and WS. The benefits of co-digesting primary sludge (PS) with
plant materials represent in that PS can provide a buffering capac-
ity and a wide range of nutrients, while added plant materials with
high carbon content can improve C/N ratio of the mixture, thus
potentially improving the methane yields. Kim et al. [13] investi-
gated co-digestion of rice straw and sewage sludge; the results
showed a higher biogas production by adding 41 g rice
straw/150 mL sewage sludge. Wang et al. [35] observed a higher
methane production from co-digestion of swine manure with
wheat straw. Sahito et al. [29] have got the maximum biogas pro-
duction from the co-digestion of buffalo dung with wheat straw.
As shown in Fig. 5, the produced CMYs through one month of
anaerobic co-digestion of PS, WS, and BH increased gradually by
decreasing the C/N ratio and reach the peak value (481.1 mL/
gVSadd) at a C/N ratio of 10.07. This optimum ratio (10.07) fits the
ideal range of 9.0–30.0 for the anaerobic digestion of wastes, which
was recommended by Horan et al. [10]. Two reasons may explain
the highest CMY that occurred at the C/N ratio of 10.07. Firstly,
there was a higher content of PS compared to the other C/N ratios
as presented in Table 3. Biogas is more easily produced from PS,
which is readily biodegradable and consists of more easily digested
carbohydrates and fats [18]. Secondly, this optimum C/N ratio con-
tains a lower percentage of agricultural wastes (WS and BH) than
the other ratios as presented in Table 3, consequently less lignin.
Lignin is considered the least degradable material of all crop resi-
dues in the anaerobic digestion process [40]. In this study, the max-
imum CMY was 481.1 mL/gVSadd, which showed a high volume
compared with the value of 234.7 mL/gVSadd that was collected
by Wang et al. [35] for co-digestion of dairy and chicken manure
and wheat straw, more than 311.0 mL/gVSadd that was found by
Komatsu et al. [14] for co-digestion of sewage sludge and rice straw,
and greater than 322 mL/gVSadd that was obtained by Sahito et al.
[29] for co-digestion of crop residue with buffalo dung.
3.4.3. Methane contents in the produced biogas at different C/N ratios
The produced biogas from the anaerobic digestion can be used
directly to generate energy, but the high percentage of CO2 reduces
 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547
its heating value, increases its transport costs and limits its eco-
nomic feasibility [46]. Purification of the produced biogas can be
fulfilled by absorbing CO2 and therefore increasing its methane
content. In this study, the produced biogas was purified from CO2
by passing it through 6 M NaOH solution. Fig. 6 illustrates the vol-
umetric methane content in the produced biogas that was pro-
duced from anaerobic co-digestions of PS, WS, and BH based on
different C/N ratios at different times through the BMP tests. The
highest volumetric methane contents in the purified biogas were
recorded at C/N ratios of 15.01 and 10.07 while the lowest value
was observed at a C/N ratio of 25.25. This may be a result of the
balanced nutrients in the feedstock at C/N ratios of 15.01 and
10.07, which enhanced bacterial growth, methanization process,
and methane yields. Fig. 7 presents average monthly volumetric
methane and carbon dioxide contents in the produced biogas from
anaerobic co-digestions of PS, WS, and BH before and after the
purification processes. The range of the average methane content
increased from 58.91% to 63.05% before the purification process
to 92.46–95.30% after the purification process. In contrast, the
CO2 content decreased sharply after the purification process and
reached to zero at some C/N ratios. The main difference in compo-
sition between biogas and natural gas relates to the carbon dioxide
content. Carbon dioxide is one of the main components of biogas,
while natural gas contains very low amounts. By separating carbon
dioxide from the biogas in the purification process, the energy con-
tent of the purified biogas becomes comparable to natural gas that
can be used in different applications [22].
3.4.3.1. Potential use of the purified biogas. Biogas can be used out-
side the laboratory in a number of applications including fuel for
natural gas vehicles. The main environmental advantages are that
fossil fuels like petrol and diesel can be replaced and the reduction
of green house gas emissions can be as much as 100% [22]. Biogas
has to be purified to natural gas quality in order to be used in nor-
mal vehicles, designed to use natural gas [24]. Sweden is today the
only country in the world with a national standard for biogas as
vehicle fuel. This standard essentially states that the methane con-
tent must be higher than 95% [11]. Depending on this standard, the
potential use of the cleaned biogas of this study as a vehicle fuel
can be implemented for organic loads of 6 (96.51%) and 7.50
(95.90%) gVS/L in the first experiment and at C/N ratio of 15.01
(95.30%) in the second experiment. The methane content below
95% of the produced biogas can be injected into the natural gas grid
fuel, used in industrial processes, and to provide clean fuel for
cooking and lighting [23].
100
98
96
Methane content (%)
94
92
90
88
C/N=10.07
86
C/N=20.03
84
BH
82
80
5 10 15
Expermental time (Day)
Fig. 6. The average methane content from anaerobic co-digestions of PS, WS and BH based on their different C/N ratios after the purification process.
545
3.4.4. VS and pH characteristics of the digested feedstock
At the end of each of the biochemical methane potential (BMP)
test, samples were taken from the digestion residual to measure
their volatile solids (VS) and pH value. From the measured values
of VS at the beginning and at the end of the BMP test, the volatile
solids removal ratio through anaerobic digestion could be calcu-
lated. As shown in Table 5, the volatile solids (VS) removal ratios
in the single digestion of WS and BH were higher than the removal
ratio of digesting PS separately. This can be relayed to the small
particle size of WS and BH, which can be easily digested in the
anaerobic digestion process in addition to the special cell structure
of PS. The highest VS removal ratio for the anaerobic co-digestion
of PS, WS, and BH was found to be 73.45% and recorded at C/N ratio
of 10.07 as shown in Table 5. On the other side, the lowest VS
removal ratio was observed at C/N ratio of 25.25 with a value of
59.55%. These results strengthen and assure the findings of this
study regarding to the highest and lowest value of cumulative
methane yields (CMYs) that were occurred at C/N ratios of 10.07
and 25.25, respectively. As much as methanization process suc-
cesses, feedstock components are well utilized to produce methane
and volatile solids degrade more.
Krishania et al. [16] reported that pH is an important parameter
affecting anaerobic digestion processes and growth of microorgan-
isms, thus it must be maintained at certain stable ranges. Through
anaerobic digestion processes, different pH levels are required for
the two processes of acidification (pH above 5.0) and methanogen-
esis (a minimum pH value of 6.2). Methanogenic bacteria can pro-
duce biogas and methane effectively at a pH value from 6.5 to 8.0
[30]. In this study, the initial pH level for all reactors was adjusted
to a value of 7 ± 0.1 using 6 M NaOH solution. As illustrated in
Table 5, the BMP tests maintained a stable pH values as at the
end and they were found to be in the range of 7.18–7.47, therefore
anaerobic co-digestion of PS, WS, and BH retained the buffer capac-
ity of the anaerobic digestion system.
3.5. Effective retention time for biochemical methane production
As shown in Figs. 1 and 5, the flow rate of the produced
methane from the two groups of experiments decreased gradually
after 20 days from the beginning of biochemical methane potential
(BMP) tests. Through the first 20 days of BMP tests, methane yields
for both the first and second experimental groups were in the
range of 85–90% and 90–95%, respectively, relative to the total
methane produced within the first 30 days of BMP tests. The diges-
ter retention time presents a key process design constraint as it
C/N=13.06 C/N=15.01
C/N=25.25 PS
WS
20 25 30
546 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547

70 (A)
CH4 CO2
60

Methane and CO2 contents (%)

50

40

30

20

10

0
C/N=10.07 C/N=13.06 C/N=15.01 C/N=20.03 C/N=25.25 PS BH WS
The C/N ratio and feedstock component
100 (B)
CH4 CO2
90
Methane and CO2 contents (%)

80

70

60

50

40

30

20

10

0
C/N=10.07 C/N=13.06 C/N=15.01 C/N=20.03 C/N=25.25 PS BH WS
The C/N ratio and feedstock component

Fig. 7. The average methane and CO2 contents in the produced biogas from anaerobic co-digestions of PS, WS, and BH based on their different C/N ratios before (A) and after
(B) the purification processes.

Table 5
The measured CBYs, average methane content, CMYs, volatile solids (VS) removal, and pH values for the anaerobic co-digestions of PS, WS, and BH at different C/N ratios.

C/N ratio CBYs (mL/g VSadd) Average methane content (%) CMYs (mL/g VSadd) Volatile solids removal (%) pH Value
10.07 510.55 ± 6.0 94.24 481.15 ± 5.7 73.45 7.24
13.06 464.94 ± 8.2 93.93 436.72 ± 7.7 70.67 7.38
15.01 456.68 ± 6.8 95.30 435.21 ± 6.5 63.48 7.18
20.03 443.58 ± 6.6 92.46 410.14 ± 6.1 60.54 7.35
25.25 365.21 ± 8.9 92.52 337.89 ± 8.2 59.55 7.40
PS 162.06 ± 15.5 92.83 150.40 ± 14.5 58.42 7.66
WS 279.76 ± 6.8 92.92 260.00 ± 6.3 67.87 7.52
BH 231.51 ± 9.3 93.00 215.09 ± 8.6 60.47 7.44

Notes: CBYs = cumulative biogas yields, CMYs = cumulative methane yields, PS = primary sludge, WS = wheat straw, and BH = buckwheat husk. The data of CBYs and CMYs
represent the means ± SD, n = 2.

affects the growth of microorganisms and their reproduction in the sible to decrease the detention time to 10–15 days, which reduces
reactor [19]. In addition, it is economically to operate anaerobic the investment costs.
digesters at the maximum rate of gas production. In this study,
the first 20-days of anaerobic digestion of PS with agro-wastes
under mesophilic conditions presents the most effective period 4. Conclusions
for the biochemical methane production. Previous studies support
the results obtained in this work. Sahito et al. [29] studied the In this study, two experiments were performed; in the first, dif-
anaerobic co-digestion of buffalo dung with crop residue. The ferent mixtures of PS and WS were prepared based on their VS
results showed that 20 days retention time can be advised for organic loads to obtain the best VS organic load for greater
the used crop residue as the percent of methane production methane yields. The maximum cumulative methane yield (CMY)
through this period was ranging from 85% to 91% from the total and highest VS removal rate from the anaerobic co-digestion of
produced methane. Timmerman et al. [33] showed that it was pos- PS and WS in mesophilic batch experiments were recorded at
 M. Elsayed et al. / Energy Conversion and Management 117 (2016) 538–547
7.50 gVS/L and they were higher than those from digesting PS and
WS separately, whereas the minimum CMY was observed at an
organic load of 3.0 gVS/L. In the second run, the methane yield
from anaerobic co-digestion of buckwheat husk (BH) as a proposed
waste material with PS and WS could be increased by 39.26% com-
pared with that from anaerobic co-digestion of PS and WS.
Methane production and VS removal rate at the tested C/N ratios
were higher than the results from the single digestion of PS, WS,
and BH. The maximum methane production and highest VS
removal rate were recorded at a C/N ratio of 10.07. The average
methane content of the produced biogas after purification was
raised to be in the range from 92.46% to 95.30% with an increase
of 56.95% and 51.13% compared to that without purification. The
results showed that the co-digestion of nitrogen-rich PS in combi-
nation with carbon-rich WS at the optimal VS organic load with the
addition of the proposed buckwheat husk waste material can
enhance methane production and performance of the reactor.
Acknowledgements
This work was supported by both GEPEA UMR CNRS 6144
(Ecole Des Mines De Nantes, France) and the Ministry of Higher
Education and Scientific Research in Egypt. The authors thank
‘‘CARENE/Saint-Nazaire agglomeration” (WWTP of Saint-Nazaire,
France) for their kind collaboration during this project.
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