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Journal Name, Year, Volume 1

Biogas production from agricultural residues: A review

Mohsen Taherdanak, Hamid Zilouei*

Department of Chemical Engineering, Isfahan University of Technology, Isfahan 84156-83111, Iran

Abstract: Nowadays, the energy security as well as the environmental concerns has made great attempts to find
economically and environmentally friendly energy resources. Biogas production via anaerobic digestion has been shown as a
sustainable, renewable and carbon neutral energy source which can reduce the global fossil fuels dependency. Agricultural
activities generate huge amounts of organic residues annually worldwide. These are including crop residues, straw, stalks,
branches, leaves, waste from pruning, bagasse, residue from cotton ginning, oil cakes and animal manure. Traditional
methods such as burning of these residues cause adverse effects on the environment such as greenhouse gas emission and
pollution of air and water, while they could be sufficiency converted into valuable products through anaerobic digestion
process. In order to enhance the efficiency of anaerobic digestion of the agricultural residues, a sufficient pretreatment may be
necessary to make the feedstock ready for microbial decomposition. Besides, the optimization of microbial activity of
anaerobic digestion via manipulation and control of operational parameters is necessary and important. This paper presents a
comprehensive review on the latest studies investigating the potential of agricultural wastes as feedstock for biogas. The
review includes the investigations on the biomethane potential of agricultural residues as well as the investigated
pretreatments and operational conditions for the improvement of anaerobic digestion of agricultural residues.

Keywords: Anaerobic digestion, agricultural waste, biogas, pretreatment, renewable energy.

1. INTRODUCTION which could be sufficiently produced from biomass via

Fossil fuels, the world’s primary energy source, have
faced the world with sever environmental problems such as
the global warming and the air/water/land pollution [1]. The
greenhouse effect of carbon dioxide which is released from
the fossil fuel combustion leads to the global warming. The
energy production by fossil fuel combustion releases the
carbon monoxide, nitrogen oxides, sulfur oxides and
carbohydrates into the atmosphere which infects the air. The
environment (water, air and land) is polluted through
extraction, transportation and application of fossil fuels [2,
3]. Energy supply from the wind/hydroelectric power,
solar/geothermal energy, and biomass is considered as the
best solution for the aforementioned problems [3].
Biomass, the stored form of the solar energy on the earth,
comes from a variety of sources including the wood from the
forest and woodlands, forestry plantation and residues,
agricultural and agro-industrial residues, animal wastes,
municipal and industrial wastes, food processing wastes and
sewage. The energy generation from the biomass obviates
not only the mentioned problems, but also the challenges
with the waste disposal and management [2, 4].
Bioethanol, biomethanol, butanol, acetone, biodiesel,
biomethane, and biohydrogen are the most known biofuels
*Address correspondence to this author at the Department of chemical
engineering, Isfahan university of technology, Isfahan 84156-83111, Iran;
Tel/Fax: +98 3133915632, +98 3133912677; E-mails: h.zilouei@ce.iut.ac.ir
biological processes [1, 2].
Biogas, a biofuel with a high energy value, could be
produced via anaerobic digestion (AD) of a wide variety of
biomass known around the world, and it is mainly composed
of methane (50-75%), carbon dioxide (25-50%) and
hydrogen sulfide (0-3%) [1, 3]. Concisely, through the AD
process, the biomass is first hydrolyzed by hydrolytic
bacteria followed by conversion to the volatile fatty acids by
acidogenesis bacteria. In the next step, the acetate, hydrogen
and carbon dioxide are produced and in the last step, these
are converted to methane and carbon dioxide by
methanogenesis [1].
The aim of this paper is to review the potential of
biomethane generation from agricultural residues. In the first
section the most popular and traditional agricultural residues
around the world are reviewed. Then, different treatment
processes in order to enhance the efficiency of the digestion
which is necessary for the special structure type of
agricultural residues is presented. At the end, the effective
operational parameters on the AD process which have been
investigated in different researches are reviewed.
2. AGRICULTURAL RESIDUES AS FEEDSTOCK
The annual global production of agricultural residues is
nearly 140 billion tonnes which their energy potential is
estimated to be equivalent to 50 billion tons of oil [3, 5]. The
agricultural residues including the residues from field

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2 Journal Name, 2014, Vol. 0, No. 0 Principle Author et al.

agricultural crops (straw, stalks, branches, leaves, waste from seed) in 2013 was nearly 33 million tonnes [5]. According to
pruning, etc.) and biomass from the by-products of the the estimated residues to seed ratio of 0.8-1.6 [8], the
processing of agricultural products (bagasse, residue from generation of wheat straw in 2013 was approximately 26-53
cotton ginning, oil cakes, etc.) can potentially be used for million tonnes, which points to the high potential of wheat
biogas production. However, the efficiency of the AD straw for biogas production. The dominants for cellulose,
process depends on both the substrate characteristics and the hemicellulose, lignin and protein contents of the wheat straw
operational conditions. The following describes the are 22-37%, 19-22%, 18-38%, and 3-4%, respectively
characteristics of the potential agricultural residues in more (Table 1). Because of the high cellulose, hemicellulose and
details. protein contents, the wheat straw is known as an important
substrate for microbial cultures, either in the rumen or in
2.1. Rice straw
digesters. Different studies have investigated the AD of rice
Rice straw is one of the most abundant agricultural straw which the biomethane potential of the raw wheat straw
residues around the world [2]. Its global production (in the was obtained in the ranges of 180 and 270 mL/g-VS (Table
form of seeds) in 2013 was approximately 18 million tonnes 2).
[5]. It is estimated that 1.35 tons of rice straw is remained as
2.3. Barley straw
field residues from one ton of harvested rice [3]. The huge
rice residue generation makes it attractive for both developed Barley, one of the major cereal crops, is primarily grown
and under developed countries to use it as feedstock for for its grain, but it also generates valuable straw left in the
bioenergy production [6]. Rice straw is mainly consists of field after the grain harvesting. The global production of
cellulose (30-35%), hemicellulose (17-30 %), and lignin (6- barely in 2013 was nearly 8 million tonnes, and its residue to
30 %) (Table 1). Different studies have investigated the seed ratio is reported in the range of 0.8-1.3 [5, 8]. Based on
biomethane production from the rice straw using different Table 1, the cellulose, hemicellulose, lignin, and protein
inoculums and the biomethane potential of the raw rice straw contents of the barley straw are reported 38%, 22%, 17%,
was obtained between 70-200 mL/g-VS (Table 2). and 3%, respectively. The fine nutritional species of barley
straw makes it a suitable feedstock for anaerobic digestion
2.2. Wheat straw either in cattle rumen or in biogas digesters. Studies have
Wheat straw, the above-ground plant which remains after demonstrated that the biomethane potential of the barley
the harvesting of wheat grain, including leaves and stalks, is straw would be nearly 240 mL/g-VS (Table 2).
an abundant and accessible agricultural residue around the
world [7]. The global production of wheat (in the form of
Table 1. Characteristics of agricultural residues reported in literatures.
Chemical composition
Feedstock Ref.
Cellulose (%) Hemicellulose (%) Lignin (%) Protein (%) C/N ratio
Bagasse stillage residue 28.9 8.3 22.0 8.9 ― [9]
Barley straw 38 21.9 17.3 2.8 50-100 [10]
Corn stover 33.7– 42.3 19.1– 29.8 7.1– 15.2 4.13– 4.4 63.2– 79.7 [11-16]
Cotton stalk 38.3– 47.1 19.5– 24.1 21.6– 22.0 ― 64.4 [17, 18]
Ensiled sorghum forage 47.5 27.4 7.0 8.9 29.0 [19]
Palm oil empty fruit bunches 32.4 8.7 24.6 ― ― [20]
Rice straw 3.0− 35.0 16.7− 29.8 6.5− 32.0 0.87− 3.2 47.0− 69.0 [6, 21, 22]
Sorghum 31.1–35.8 17.2–20.1 22.3–37.6 7.1–9.7 36–48 [23, 24]
Sugarcane bagasse 25−47 20−35 15−35 4..3 62.5– 63.5 [25-28]
Sunflower stalk 32.6-34.0 20-21 13.3-30 5.2 ― [29, 30]
Wheat straw 22.1− 36.7 18.9− 21.8 18.1− 37.9 3.4− 4.0 41.5− 69.8 [31-33]

2.4. Corn stover utilization of this vast area of lignocellulosic feedstock [12,
35]. In such utilizations, not only the waste is not completely
The stalk and leaves of maize plant in a field left after the
disposed, but also the air and land pollutions are occurred
harvesting of cereal grain is known as corn stover. It is a
[34]. The corn stover is mainly composed of cellulose (33-
very common agricultural residue in areas with great
47%), hemicellulose (19-29%), lignin (9-22%), and protein
amounts of corn production [12, 34]. The global production
(4%) (Table 1). In different studies, the corn stover was used
of maize (seeds) in 2013 was approximately seven million
as the substrate for AD which the methane potential from the
tonnes [5]. The residue to seeds ratio for maize (0.9-1.2) raw corn stover was between 244 and 247 mL/g-VS (Table
indicates the huge amount of the left residues from corn
2).
processing [8]. During the last decade, the maize production
was increased by nearly 40%, and accordingly the produced 2.5. Sorghum stalk
corn stover residues was increased by the same proportion
Sweet sorghum is one of the crops with high sugar
[5]. Use for animal feed, or burning for cooking and house
content and biomass yield, which is used for food, animal
energy generation in rural fields are some of the primarily
fodder and biofuel production [36]. The global production of
Title of the Article
sorghum (seeds) in 2013 was approximately 800,000 tonnes
[5]. The sorghum stalk is rich in soluble sugars and insoluble
carbohydrates which could be utilized as feedstock for
biogas production [36]. The cellulose, hemicellulose, lignin
and protein contents of the sorghum straw lie in the ranges of
31-36%, 17-20%, 22-26%, and 7-10%, respectively (Table
1). The biomethane potential of the raw sorghum stalk used
as feedstock in AD has been reported 75.3 mL/g-VS [36].
2.6. Sugarcane bagasse
Sugarcane bagasse, the fibrous matter remained after the
extraction of juice from sugarcane stalks, is widely used in
the manufacture of pulp and building material as well as the
biofuel production [37]. The global production of sugarcane
in 2013 was nearly two billion tonnes [5]. It is estimated that
the remained sugarcane bagasse from each tonnes of
sugarcane crushed in nearly 0.3 tonnes of wet sugarcane
bagasse, which indicates the huge amount of the left residues
from the sugarcane processing [3]. The cellulose,
hemicellulose, lignin, and protein contents of the sugarcane
bagasse are reported in the ranges of 24-47%, 20-35%, 15-
35%, and 4-6%, respectively (Table 1). The methane yield
of 226.42 ml/g-VS of the raw sugarcane bagasse has been
reported from its anaerobic digestion [37].
2.7. Cotton residues
Cotton residues including stalk, leaves and cottonseed
hull could be utilized as livestock feed, composting, paper
production and biofuel production [38]. The world’s cotton
production in 2013 was approximately 4.5 million tonnes
[5]. The studies have shown that the cellulose, hemicellulose
and lignin contents of the cotton stalk are in the ranges of
38.3-47.1%, 19.5-24.1%, and 21.6-22%, respectively (Table
1). The biogas production from cotton residues via AD was
performed in batch reactors and the biomethane potentials of
cottonseed, cotton stalk and cotton oil cake were obtained
86, 65, and 78 mL/g-substrate, respectively [38].
2.8. Sunflower stalk
Sunflower, the fourth oil-seed source worldwide, is a
quickly gaining popularity as a feedstock crop with more
than 25 million hectares cultivated land surface. According
to [5], the global production of sunflower (seeds) in 2013
was nearly 700,000 tonnes, which its residue to seed ratio is
2.2-3.2 [8]. After the seed harvesting, the stalks are left in
the fields. Usually, the left biomass is burned leading to the
associated disposal costs and environmental problems [29].
The left residues are estimated to be three to seven tons of
dry biomass per hectare, including the heads and the stalks.
Therefore, this inexpensive, renewable, and use-less biomass
could be utilized for biofuel (especially biogas) production
[3, 39]. According to Table 2, the cellulose, hemicellulose,
and lignin contents of the sunflower stalk are in the ranges of
32.6-34.0%, 20-21%, and 13.3-30%, respectively. The
biomethane potential of 92 mL/g-VS has been reported via
mesophilic AD of the raw sunflower stalk as feedstock [30].
2.9. Sunflower oil cake
The world’s sunflower seed production in 2013 was
approximately 44 million tonnes [5]. Sunflower oil
cake/meal is the solid residues remained after oil extraction
from the sunflower seeds. Its composition widely varies
depending on the growing condition, seed’s quality, and the
method of extraction [40]. The protein, fiber and ash content
Journal Name, 2014, Vol. 0, No. 0 3
of the sunflower oil cake are nearly 34%, 13%, and 7%,
respectively [41]. The biomethane potential of 143 mL/g-VS
has been achieved from the AD of sunflower oil cake under
mesophilic condition (Table 2).
2.10. Palm oil residues
The oil production from palm has grown tremendously in
the last decade. The growing palm oil production indicates to
the growing palm oil residues including such as palm oil mill
effluent (POME), palm oil from fresh fruit bunch (FFB),
empty fruit bunch (EFB), palm kernel shell (PKS) [20, 42].
These lignocellulosic matters could be used as feedstock for
biofuel (especially biogas) production. In a previous study,
the EFB was used as feedstock for biogas production via AD
process. The analysis revealed that the cellulose,
hemicellulose, and lignin contents of raw EFB were nearly
32%, 8%, and 25%, respectively, and the biomethane
potential of the raw EFB was obtained nearly 200 mL/g-VS
[20].
3. PRETREATMENT OF AGRICULTURAL
RESIDUES
Agricultural residues, composed of mainly lignocellulosic
and starchy feedstocks, have different resistances against
biodecomposition through anaerobic digestion process [43].
The lignocellulosic biomass is mainly composed of
cellulose, hemicellulose and lignin, which the protection of
the celluloses and hemicelluloses by lignin (hard cover)
makes it difficult for microorganisms and enzymes to diffuse
and convert the celluloses and hemicelluloses into valuable
products. Therefore, the feedstock should be treated prior to
its utilization as substrate for biomethane production [44].
The pretreatment would separate the cellulose, hemicellulose
and lignin, and alter the structure of the lignocellulosic
biomass for biodegradation. During the few past decades,
different pretreatment techniques involving physical,
chemical, and biological pretreatments have been employed
to improve the efficiency of biogas production from the
agricultural residues [45]. The following reviews some of the
most applied pretreatment methods for enhancement of the
efficiency of AD on agricultural residues.
3.1. Physical pretreatments
3.1.1. Comminution
Comminution, the reduction of the feedstock’s particle size,
is usually applied to change the ultrastructure of the
lignocellulosic feedstock, reduce the crystallinity index of
cellulose, and increase the accessible surface area of
feedstock which facilitates the microbial attack followed by
enhance efficiency of AD [3, 44]. Comminution of biomass
could be performed by using milling or grinding machines,
such as vibro, ball, hammer, knife, two-roll, colloid, attrition
mills and extruders [46]. The selection of the comminution
machine depends on the moisture content of the feedstock.
For example, use of vibro and ball could be utilized to
comminution of both dry and wet materials, while knife,
hammer, two-roll, and attrition could be utilized when the
moisture content of the feedstock is less than 15%, and the
extruders and colloid mills are sufficient for the
comminution of materials with the moisture content higher
than 15% [45, 46]. Some of the achieved improvements in
methane yield by using comminution pretreatments are
presented in Table 2.
4 Journal Name, 2014, Vol. 0, No. 0 Principle Author et al.

3.1.2. Liquid hot water (hydrothermolysis) microorganisms and enzymes. The enhanced sugar
extraction, lower inhibitor production, pentose recovery, and
Liquid hot water (LHW), the hydrothermal pretreatment
higher cellulose degradability are the advantage of LHW
which does not require addition of chemicals, is usually
pretreatment in comparison with the acid-catalyzed stream
utilized in the pulp industry for lignocellulosic feedstock
explosion and dilute acid pretreatments [44]. The LHW has
pretreatment. Through this pretreatment, both pressure and been applied to improve the biogas production from barely
temperature are elevated to keep the water in the liquid phase straw under mesophilic conditions which the methane yield
[44]. During the pretreatment, the feedstock is cooked under
was improved by 40% (Table 2).
high temperature and pressure, and the water can penetrate
into its cell structure followed by cellulose hydrating, 3.1.3. Steam explosion (autohydrolysis)
hemicellulose solubilizing, and partly lignin removal [44,
Steam explosion (autohydrolysis) is one of the most common
47]. The LHW pretreatment increases the accessible surface
area of cellulose and enhance the cellulose degradation by pretreatments applied for agricultural feedstocks [48]. In
steam explosion pretreatment, the feedstock is heated using
Table 2. Enhanced effects of the pretreatments on biomethane production form agricultural residues reported in literature.
Methane
Feedstock Pretreatment AD’s condition Reactor type Ref.
Yield (mL/g-VS) Enhancement (%)
Barley straw Mechanical (particle size of 0.5 cm) Mesophilic Batch 370 54 [49]
Barley straw Mechanical (particle size of 2 cm) Mesophilic Batch 339 41 [49]
Barley straw Mechanical (particle size of 5 cm) Mesophilic Batch 286 19 [49]
Barley straw Thermal (120 °C) Mesophilic Batch 338 41 [49]
Barley straw Thermal (90 °C) Mesophilic Batch 340 42 [49]
Cassava residues Biological Thermophilic Batch 260 97 [50]
Grass Mechanical Mesophilic Batch 200 9 [51]
Ley crop silage Mechanical Mesophilic Batch 180 59 [52]
Maize stalk Mechanical (particle size of 0.2 cm) Mesophilic Batch 272 11 [49]
Maize stalk Mechanical (particle size of 2 cm) Mesophilic Batch 254 3 [49]
Maize stalk Thermal (120 °C) Mesophilic Batch 267 9 [49]
Mango leaves Ionic liquid Mesophilic Batch 125 140 [53]
Palm oil empty fruit bunches Acidic-thermal Thermophilic Batch 280 47 [20]
Palm oil empty fruit bunches Alkaline-thermal Thermophilic Batch 404 100 [20]
Paper tube residuals Steam Explosion and Chemical Thermophilic Batch 493 107 [54]
Rice straw Ionic liquid Mesophilic Batch 233 224 [53]
Rice straw Mechanical (particle size of 5 cm) Mesophilic Batch 203 3 [49]
Rice straw Thermal (120 °C) Mesophilic Batch 261 33 [49]
Rice straw Thermal (90 °C) Mesophilic Batch 207 5 [49]
Sorghum Ensilage-alkaline-thermal Mesophilic Batch 355 32 [55]
Straw Mechanical Mesophilic Batch 370 11 [51]
Sugarcane bagasse Acidic-thermal Mesophilic Batch 434 92 [37]
Sunflower oil cake Acidic-thermal Mesophilic Batch 302 55 [56]
Sunflower oil cake Ultrasonic Mesophilic Batch 220 54 [57]
Sunflower stalk Alkaline-thermal Mesophilic Batch 259 35 [30]
Sunflower stalk Ca(OH)2-thermal Mesophilic Batch 241 26 [30]
Sunflower stalk FeCl3-thermal Mesophilic Batch 248 29 [30]
Sunflower stalk H2O2-thermal Mesophilic Batch 256 33 [30]
Sunflower stalk HCl-thermal Mesophilic Batch 233 21 [30]
Sunflower stalk Thermal (1h, 170°C) Mesophilic Batch 219 14 [30]
Sunflower stalk Thermal (24h, 55°C) Mesophilic Batch 198 3 [30]
Sweet sorghum Organosolv Mesophilic Batch 278 270 [36]
Wheat plant Acidic-thermal Thermophilic Batch 322 13 [4]
Wheat plant Alkaline-thermal Mesophilic Batch 404 55 [1]
Wheat plant Alkaline-thermal Thermophilic Batch 411 44 [7]
Wheat straw Alkaline-thermal Mesophilic Batch 340 67 [55]
Wheat straw Mechanical (particle size of 0.2 cm) Mesophilic Batch 334 84 [49]
Wheat straw Mechanical (particle size of 5 cm) Mesophilic Batch 285 57 [49]
Wheat straw Microwave Mesophilic Batch 344 28 [58]
Wheat straw Thermal (120 °C) Mesophilic Batch 299 64 [49]
Wheat straw Thermal (90 °C) Mesophilic Batch 295 62 [49]

high-pressure saturated steam (160-260 °C and 0.69-4.83 3.2. Chemical pretreatments

MPa) for a short period of time (several seconds-few
minutes) and then the pressure is quickly reduced to
terminate the reactions [44, 59]. Through this pretreatment,
the hemicelluloses are hydrolyzed to their components
(simple sugars) and the lignin is partly removed, making the
feedstock more degradable for microorganisms and enzymes.
Usually the addition of acids such as acetic acid can improve
the hydrolysis of hemicelluloses [59, 60]. The steam
explosion has been applied to improve the mesophilic AD of
paper tube residue which the methane yield was enhanced by
107% (Table 2).
3.2.1. Alkaline pretreatment
Alkaline pretreatment refers to the utilization of alkaline
solutions such as NaOH, KOH, Ca(OH) 2, NH3.H2O, to
increase the accessibility of microorganisms and enzymes to
celluloses as well as remove the hemicellulose and lignin
[45]. Alkaline pretreatment breaks the crosslinks and leads to
the swelling of the feedstock’s structure, decreases the
crystallinity index of the cellulose, increases the porosity and
the internal surface of the feedstock, and destructs the lignin
structure and removes it from the feedstock [44]. As the
Title of the Article
temperature of the pretreatment and the concentration of the
used base is increased, the time duration of the pretreatment
is decreased and vice versa [1, 45]. Alkaline pretreatment
using NaOH was applied to enhance the biogas production
from wheat plant under mesophilic conditions, and the yield
of methane was increased by 55% compared to untreated
feedstock (Table 2).
3.2.2. Acid pretreatment
During the acid pretreatment, both inorganic and organic
acids, including sulfuric acid, phosphoric acid, hydrochloric
acid, maleic acid, acetic acid, and nitric acid are employed to
decompose the celluloses and hemicelluloses into their
components [4]. The acid pretreatment could be performed
either by dilute acid at high temperatures (e.g. 0.1-4% and
230 °C) or by concentrated acid (e.g. 30-70%, 40 °C) [44].
Although concentrated acid have a higher impact on
cellulose decomposition, it is highly toxic, corrosive, and
dangerous. Moreover, it needs special reactor and equipment
such as glass-lined reactor. Dilute-acid hydrolysis is one of
the most commonly applied methods for enhancement of the
biogas production from agricultural feedstocks [3].
Depending on the pretreatment conditions, it can hydrolyze
the hemicellulose into its component sugars (e.g. xylose,
arabinose and galactose) up to 100%. The high lignin
disruption could be achieved by this pretreatment. However,
it is not so effective on dissolving the lignin. Dilute acid
pretreatment is well-known for the increase in the cellulose
accessibility to microorganisms and enzymes [44, 61]. Dilute
sulfuric acid pretreatment has been applied to enhance the
efficiency of mesophilic AD of sunflower oil cake which the
methane yield was enhanced by 55% (Table 2).
3.2.3. Organosolv pretreatment
During the organosolv pretreatment, an organic/aqueous
organic solvent (e.g. alcohols, phenols, glycols, ketones,
organic acids, esters, and ethers) is applied to disrupt the
structure of the lignin and make the celluloses and
hemicelluloses available for microbial degradation [3, 45].
During the process, feedstock is heated in the organic solvent
and water, and the lignin and a part of cellulose are dissolved
in the liquid phase and the reactive cellulose remains in the
solid phase. The temperature of the process could be
decreased while the lignin removal could be enhanced by
addition of the certain catalysts (e.g. oxalic, salicylic, and
acetylsalicylic acid) in the process [45]. In order to increase
the efficiency of mesophilic AD of sweet sorghum, the
organosolv pretreatment was applied which enhanced the
methane yield by 270% (Table 2).
3.2.4. Ionic liquid pretreatment
During the pretreatment using ionic liquids (e.g. 1-n-butyl-3
methylimidazolium chloride (BMIMCl), N-
methylmorpholine-N-oxide mono-hydrate (NMMO), 3-
methyl-N-bytylpyridinium chloride (MBPCl), 1-allyl-3-
methylimidazolium chloride, and benzyldimethyl
(tetradecyl) ammonium chloride), the oxygen and hydrogen
atoms of cellulose hydroxyl groups interact with the ionic
liquid. This interaction breaks the hydrogen bonds between
molecular chains of the cellulose and leads to dissolution of
cellulose [44, 62, 63]. Then, the dissolved cellulose could be
precipitate by addition of anti-solvents (e.g. ethanol,
methanol, acetone, or water) to the system. Ionic liquids,
known as the efficient and “green” solvents of cellulose,
Journal Name, 2014, Vol. 0, No. 0 5
could efficiently dissolve the cellulose under mild conditions
(90-130 °C and ambient pressure). In addition to the low
energy consumption, they could be recovered even up to
100% [44, 53]. In comparison with the other organic
solvents of cellulose, they have high reaction rates, broad
selection of anion and cation combinations, high
electrochemical and thermal stability, low viscosity, low
toxicity, low hydrophobicity, low volatility accompanied,
minimum negative impacts on the environment accompanied
by the non-flammability [44, 64]. As tabulated in Table 2,
the application of ionic liquids enhanced the yield of AD of
mango leaves and rice straw by 140% and 224%,
respectively.
3.3. Biological pretreatments
The biological pretreatments (e.g. pretreatment by microbial
consortium, fungal pretreatment, enzymatic pretreatment,
and even ensiling) utilize the microorganisms to alter the
feedstock’s structure [44, 45]. The major objectives of these
pretreatments are to maximize the lignin removal and
minimized the loss of carbohydrates [44]. In comparison
with the physical and chemical pretreatments, the biological
pretreatments are performed with much lower energy input
without using chemicals. The feedstock is pretreated at mild
conditions (ambient temperature and pressure), and
therefore, the nutritional composition of the feedstock is
disrobed accompanied by less inhibitor production for
microbial digestion. However, because of the long duration
of these pretreatments, often they are not commercially used
[35, 44]. In a previous study, the cassava residue was used as
feedstock for biomethane production via AD in batch
reactors. The feedstock was first biologically pretreated
which its methane yield was enhanced by 97% (Table 2).
4. INFLUENCE OF OPERATIONAL PARAMETERS
The activity of microorganisms and their metabolism in AD
process are affected by several different parameters [1]. The
sudden changes in the system’s operational conditions (e.g.
temperature, pH or even the substrate concentration) may
lead to shut down the microbial metabolism and therefore
gas production. Usually it takes 2-3 weeks to adapt the
microorganisms to the new environmental conditions [65].
Moreover, since the AD is a four-step process containing
different kinds of microorganisms in each step, the required
environmental conditions can vary from one step to another
[13, 61]. The followings describe the impact of the
temperature, pH, organic loading rate (OLR), Hydraulic
retention time (HRT)/Solid retention time (SRT), C/N ratio,
and the metal ions on the efficiency of AD process,
accompanied by the review on the related literatures (Table
3).
4.1. Temperature
Generally, the anaerobic bacteria are classified according to
their growing temperature: psychrophilic (4-25 °C),
mesophilic (25-40 °C), thermophilic (50-60 °C), and
hyperthermophilic (>65 °C) [65]. For each group, the
activity of enzymes/co-enzymes as well as the microbial
metabolic pathway is dependent on the temperature of
system. For instance, through the AD process, the
endothermic reactions such as the conversion of propionate
to acetate, hydrogen and carbon dioxide are facilitated at
higher temperatures, while the exothermic ones such as the
conversion of acetate and hydrogen to methane are disturbed
6 Journal Name, 2014, Vol. 0, No. 0
and vice versa [66]. In general, the optimum temperature
range for the hydrolysis/acidogenesis step is 25-35 °C, while
that of the methanogenesis step is 32-42 °C (mesophilic)/50-
58 °C (thermophilic) [2, 3, 67]. Appels et al. [66], pointed
out that the stability of the AD process could be disturbed as
the temperature of the system fluctuates in a rate higher than
0.6 °C/day. In comparison with the mesophilic/thermophilic
bacteria, the psychrophilic bacteria are often used in smaller-
scale digesters which have lower AD efficiency. The lower
temperatures lead to the lower phosphorus assimilation and
lower organic nitrogen degradation [68]. Likewise, the
thermophilic digesters have higher loading rate and higher
pathogen elimination than the mesophilic digesters. But, they
are generally more sensitive to toxicants and higher
operational costs than the mesophilic ones [65]. A previous
study reported that the biogas production under thermophilic
conditions was at least two folds higher than that of
mesophilic condition [67]. Komemoto et al. [69],
demonstrated that the food waste was solubilized in a higher
rate under mesophilic conditions than thermophilic ones.
Moreover, it has been demonstrated that the mesophilic
anaerobic digestion of high-ammonia substrates was more
stable than that of thermophilic one [65]. However, some
studies have shown that the inhibitory effect of ammonia on
the AD process under thermophilic conditions is less than
under mesophilic conditions [67]. It has been demonstrated
that the rising temperature of the AD process can increase
the volatile fatty acids (VFAs) production at a rate higher
than their consumption by methanogens and therefore, the
accumulation of the VFAs in the system can disturb the
methanogen’s activity [65].
4.2. pH
The AD process could be classified into two stages
according to its pH requirements. The
hydrolysis/acidogenesis step which needs pH between 5.2 to
6.3, and the acetogenesis/methanogenesis step which its
required pH lies within 6.7 and 7.5 [65]. Therefore, in order
to prepare the required optimum condition of each step, it is
preferred to perform the AD in two consequent reactors [70].
In general, the neutral pH favors all types of
microorganism’s activity and therefore, it is important to
maintain the pH of the system in the neutral range [65].
Increasing the organic loading leads to increase the
production of intermediates mainly acetic acid, butyric acid,
and propionic acid which cause pH drop of the system
followed by failure of the synthrophic methanogenesis.
Therefore, in order to preserve the stability of the system’s
optimum pH, the total concentration of VFAs must be kept
below 2 g/L [65]. Moreover, the degradation of proteins can
lead to the ammonia accumulation in the system which could
increase the pH [71]. In order to provide the buffering
capacity of the system, some coupled acid-base
combinations such as ammonia-ammonium, bicarbonate-
carbon dioxide, and hydrogen phosphate-dihydrogen
phosphate could be supplemented to the system [65, 72].
4.3. Organic loading rate
The stability and performance of the anaerobic digestion are
highly dependent on the OLR, the rate of the addition of
feedstock to the system (kg-VS/m 3.d for solids and kg-
COD/m3.d for liquids) [73-75]. It usually affects the
temperature, pH and the efficiency of the biological
processes [75]. A previous study demonstrated that the OLR
Principle Author et al.
can even change the bacterial community; the Firmicutes
were observed at lower OLR, while the dominant bacteria
were Actinobacteria, Bacteroidetes, Deferribacteres and
Gamma-proteobacteria at higher OLR [76]. Usually, at the
system start-up period, a low OLR is preferred and after the
stability of the system, higher OLRs could be performed
[77]. At the lower OLRs, the system is not so disturbed and
can operate well, but its operation is not economically
acceptable. However, very high OLRs can increase the rate
of the VFA production more than the rate of VFA
consumption by the methanogens. The accumulation of the
VFAs in the system can reduce the pH of the system
followed by gas production failure [77-79]. The OLR is
often optimized according to the reactor specifications and
higher OLRs can eventually reduce the size and the capital
cost of the reactor [80].
4.4. Hydraulic/Solid retention time
The HRT and SRT are referred to the time that the
liquid/solid phase spends in the digester [65]. The HRT/SRT
is dependent on the substrate’s physical characteristics, solid
concentration and the temperature of the system [80]. It
introduces the required time for the microbial growth and the
substrate conversion [71]. When the substrate has a high
resistance against the biodegradation, the HRT is more
significant than the SRT and vice versa [65, 81, 82]. Lower
capital costs and more efficient processes are expected at
shorter HRTs/SRTs [65]. While system operates at shorter
HRTs/SRTs, the OLR must be low enough to convert the
substrate into biogas in a more efficient manner [65, 83].
Therefore, the optimum value of HRT/SRT must be
determined in accordance with the equilibrium between the
HRT/SRT and the OLR [65].
4.5. C/N ratio
The microbial growth is deeply affected by the availability of
the carbon and nitrogen sources in the system [6, 84]. The
shortage of the nitrogen source in the system can disturb the
microbial utilization of the carbon source followed by
disturbing the efficiency of the AD process [6, 85]. Through
the AD, the released nitrogen from the nitrogenous materials
is converted into ammonia which is in equilibrium with
ammonium. The change in the operational conditions of the
system (e.g. rising temperature and pH) can shift the
equilibrium toward the ammonia formation followed by its
accumulation in the system, which leads to the proton
imbalance in the system and efficiency reduction [65, 86]. It
is reported that the required range of C/N ratio for the
hydrolysis/acidogenesis step is 10-45, while that of the
methanogenesis step is 20-30 [2, 65]. Generally, the
optimum C/N ratio depends on the inoculum and the
substrate characteristics and the operational conditions [71,
80].
4.6. Metallic ions
Through the AD process, lots of the biochemical reactions
are catalyzed by enzymes and lots of them include the
metallic particles (metalloenzymes) [2]. Therefore, metallic
ions such as Na, K, Ca, Mg, Al, Fe, Ni, Cu, Zn, Co, and Ag
ions are necessary for the enzyme expression and the
enzyme’s activity [67]. Although the availability of the
metallic ions in the system can enhance the efficiency of AD
process, at high concentrations they can inhibit the process
by disturbing the enzyme’s structure and activity. This
Title of the Article Journal Name, 2014, Vol. 0, No. 0 7

inhibitory effect depends on both total concentration of the reported that the optimum sodium concentration for the AD
metallic ions and the form of the metallic ions [2]. Moreover, process would be 350 mg/L, and the potassium at
the pH and the redox potential of the system can influence concentrations lower than 400 mg/L could increase the AD
the inhibitory effect of the metallic ions [67]. Moreover, the efficiency [67].
pH and the redox potential of the system can influence the
inhibitory effect of the metallic ions [2, 87]. It has been Yu et al. [88] demonstrated that the supplementation of
calcium at concentrations of 350-400 mg/L can increase the
Table 3. Effective parameters investigated in AD of agricultural residues.
Studied parameter Optimum methane yield (mL/g-
Feedstock Inoculum Reactor type Ref.
Range optimum VS)

Temperature

Cow manure Cow manure Batch 5.0-30 30.0 196.8 [89]

De-oiled Jatropha Sewage sludge Batch 25-65 45 178.4 [90]
cattle and
Olive debittering and washing Effluent Batch 35-55 55 350 [91]
pig manures
Pig manure Cow manure Batch 5.0-30 30 211.4 [89]
30.0- a
Rice straw Anaerobic sludge Batch 35.6 346 [6]
45.0
Rice straw Anaerobic sludge Batch 25-55 35 239.7 [92]
Sewage sludge and sugar beet pulp
Primary sludge Batch 35-55 55 544.4 [93]
lixiviation
Semi-
Vinasses Digested sludge 35-55 55 1230b [94]
continues
c
Water hyacinth pig slurry Batch 25-65 55 124.5 [95]
pH
Dairy waste sludge Batch 4.0-9.5 7.0 2000b [96]
Kitchen waste Cow manure Batch 6.0-8.0 7.5 8579b [97]
Sewage sludge and sugar beet pulp
Primary sludge Batch 4.8-7.8 7.7 544.4 [98]
lixiviation
Swine manure and
Anaerobic sludge Batch 6.0-8.0 7.0 146.0 [99]
maize stalks
Semi-
Vinasses Digested sludge 4.5-6.5 6.5 2775b [94]
continues
Vinasses Rumen fluid Batch 6.0-8.0 7.0 3.81d [100]
OLR
25.0-
Corn husk Anaerobic sludge Continues 43.6 42.9e [101]
45.0
Semi- 1.05-
Maize Anaerobic sludge 5.25 770 [102]
continues 6.30
Anaerobic f
Maize silage leachate Batch 5.0-17.5 17.5 8.2 [74]
effluent
e
Scraped manure Anaerobic sludge continues 0.3-1.3 0.3 88.3 [75]
HRT
chicken dung Animal manure Batch 10-40 25 5.6g [103]
Semi- b
Vinasses Digested sludge 1-5 5 2775 [94]
continues

C/N ratio

Food waste and rice husk Cow dung Batch 20-35 20 584 [104]
Food, fruit, vegetable and paper wastes Mixed culture Batch 27-32 27 327 [84]
15.0-
Rice straw Anaerobic sludge Batch 29.6 353 [6]
34.0
a b c d e f g 3
biogas yield, mL, mL/g-substrate, mL/g-COD, mL/d, L/L.d, m /ton

efficacy of the AD process. Another study reported that CONCLUSION

supplementation of the calcium even at concentration of 7
g/L had no inhibitory effect of the process [67]. It has been The increasing global demand for energy could be
answered by recycling the bioenergy from the huge annual
demonstrated that the supplementation of iron ions to the
amount of the agricultural residues. Either mesophilic or
system at concentrations of 150 and 350 mg/L, could
increase the efficiency of the acidogenesis step of the AD thermophilic AD is a reliable technology for methane
production from agricultural residues. In order to improve
process which accordingly could increase the efficiency of
the AD efficiency, an efficient pretreatment is necessary to
the AD process [105]. Similar studies reported that the
alter the structure of the agricultural residues (lignocellulosic
addition of copper, zinc, magnesium, and nickel at
biomass) for microbial hydrolysis. The type of the
concentrations of 400.0, 250.0, 23.6 and 0.1 mg/L, have the
pretreatment must be selected according to the composition
optimum positive effect on the acidogenesis step’s efficiency
[106]. and the structure of the residue. To keep the efficiency of the
8 Journal Name, 2014, Vol. 0, No. 0
AD process at its high level, the operational conditions such
as temperature and pH must be controlled at their optimum
levels. The OLR, SRT, and HRT are in equilibrium with the
temperature and the pH of the system, and the changes in
their value would change the temperature and the pH of the
system. The C/N ratio of the substrate must be matched with
the microbial requirements and in some cases a combination
of two or more different feedstocks can provide the required
C/N ratio. Moreover, the addition of metallic ions to the
system can improve the enzyme expression as well as the
microorganism’s activity which enhance the AD efficiency.
ACKNOWLEDGEMENT
The authors would like to thank Mrs. Freshteh Shams for
her invaluable contribution through this work, especially in
the pretreatment section.
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