Bioresource Technology 359 (2022) 127497

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Anaerobic biogas formation from crops’ agricultural residues –

Modeling investigations
Marvin Scherzinger a, *, Martin Kaltschmitt a, Amr Y. Elbanhawy b
a
Hamburg University of Technology (TUHH), Institute of Environmental Technology and Energy Economics (IUE), Eissendorfer Strasse 40, 21073 Hamburg, Germany
b
Ain Shams University (ASU), Energy Technology and Climate Change Laboratory, Faculty of Engineering, Elsarayat Street 1, Cairo / Abbasia, Egypt

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Biogas production was modelled based

on kinetic Gompertz-Models.
• The model can be applied to other sub­
strates using relatively simple batch
studies.
• Indications regarding optimal operation
of small-scale biogas plants are supplied.
• A complete feed analysis of various
biogenic residues from Egypt is
presented.

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

Keywords: The cultivation of field crops necessarily produces large quantities of organic residues, which could be used for
Biogas the production of biogas. However, this is only successfully possible if the operators of such biogas plants have
Modeling sufficient expertise in running these plants, e.g. to avoid an overload of the biogas system. Against this back­
Co-digestion
ground, the anaerobic degradability of various Egyptian agricultural residues is determined by laboratory testing;
Gompertz-Models
Lignocellulose
this includes both a determination of degradation behavior in batch tests and a feed analysis. The residues
studied produce biogas yields ranging from 303 to 496 mLN g-1 VS. Co-digestion experiments demonstrate that
hardly any interaction effects occur during the fermentation process for different mixtures of the investigated
residues. Based on these findings, a model is developed to estimate biogas production using the investigated
agricultural residues in continuous operation and to give recommendations for the optimal mode of operation.

1. Introduction agricultural primary production of field crops. While in some places

large parts of such herbaceous residues are already being put to viable
Wherever agricultural activities take place, significant amounts of use (e.g. as construction material or as animal fodder), in many other
organic residues also accrue; among others, this is especially true for areas there is not yet a broader valorization established (Nour et al.,
straw being the most important organic (waste) side stream from 2021). For this reason, farmers in many regions of this world burn large

* Corresponding author.
E-mail address: marvin.scherzinger@tuhh.de (M. Scherzinger).

https://doi.org/10.1016/j.biortech.2022.127497
Received 17 May 2022; Received in revised form 13 June 2022; Accepted 15 June 2022
Available online 22 June 2022
0960-8524/© 2022 Elsevier Ltd. All rights reserved.
M. Scherzinger et al.
parts of the occurring straw in the field in order to get rid of it without
bearing additional costs; this is especially true for regions where such
practices are not prevented and/or controlled by legislation (El-Dewany
et al., 2018). In this perspective, the most prominent example is the
burning of rice straw. According to current knowledge, the proportion of
rice straw burned in the fields varies from 0.6 to up to 90 % of the total
rice straw produced, depending on the country of production (Singh
et al., 2021). Fortunately, more and more use options are being devel­
oped for the use of these agricultural residues, so that in many countries
(such as China, which is the main rice-growing country) field burning is
steadily declining. Current estimates indicate that in Egypt, for example,
about 25 % of the lignocellulosic residual streams are already being
used, with an upward trend (the main use is as livestock feed) (Nour
et al., 2021). However, economically promising and environmentally
sound utilization possibilities must also be found for the remaining
quantities unused so far. In this way, negative effects, such as unwanted
air pollution and the release of climate-impacting gases associated with
unsustainable processing strategies, can be reduced and possibly pre­
vented in the long term.
Anaerobic digestion of such residues could be one promising utili­
zation option in this regard. During this process, various microorganisms
decompose the organic material resulting in the release of a biogas as the
main product; this can then be used as fuel gas to substitute fossil fuels.
The fermentation residues from biogas production could furthermore be
used as valuable fertilizers to close nutrient cycles. However, such her­
baceous residues from agricultural production have only suboptimal
degradability characteristics. Amongst others, these include an inap­
propriate Carbon to Nitrogen ratio (C / N-ratio), a shortage of trace el­
ements, and a high proportion of substances that are recalcitrant to
degradation (Paul and Dutta, 2018). A very efficient and frequently used
method to compensate for these limitations is the co-digestion of various
substrates with different characteristics, whereby more favorable
degradation conditions can be artificially adjusted (Siddique and Wahid,
2018; Kasinath et al., 2021).
Another significant obstacle preventing the use of such agricultural
residues for biogas production is their predominantly seasonal occur­
rence. Although this can be partially compensated by processes for
preservation (e.g. storage of straw bales), it is often not possible to
guarantee a steady substrate mixture (or substrate quality) throughout
the year, especially on smaller farms without large storage areas.
Furthermore, at such farms in rural areas of developing countries chal­
lenges due to a lack of technical knowledge occur. Especially the lack of
knowledge about the size of biodigesters and the volume of organic
substrates often leads to low biogas production rates and digestate
pollution near the respective biogas plants (Sikora, 2021).
The development of models to predict biogas production volumes for
specific systems has been used in attempts to counteract these knowl­
edge gaps and to provide valuable tools for plant operators to evaluate
the effects of substrate feeding. The models thereby differ greatly in
terms of their level of detail. A very detailed model is e.g. ADM1
including multiple steps describing biochemical as well as physico-
chemical processes (Batstone et al., 2002). The original ADM1 model
has been adapted several times, e.g. to be able to represent the co-
digestion of agricultural residues. However, the complexity of param­
eter determination and model calibration was also highlighted in this
context (Hagos et al., 2017). A much simpler method compared to the
above-mentioned modeling approach is the empirical determination of
kinetic models. Such models are typically based on enzymatic equations
or refer to the canonical algebraic form of equations with dimensionless
coefficients (Pererva et al., 2020). A large number of studies reporting
on kinetic modeling of various substrates exist and a good overview on
several models based on biogas formation batch test data is given in the
literature (Velázquez-Martí and Meneses-Quelal, 2019). Currently,
almost all these kinetic models are derived from laboratory-scale ex­
periments and further use to predict biogas production in continuously
fed systems is not considered (Siddique and Wahid, 2018). However, in
2
Bioresource Technology 359 (2022) 127497
cases where there are no interaction effects between different substrates,
biogas formation from any substrate mixture can be calculated as the
sum of biogas production from its individual components. This has been
demonstrated for a biogas plant that was continuously fed with a sub­
strate mixture of cow slurry, pig slurry, slaughterhouse waste and corn
silage (Zepter et al., 2021).
Against this background, in this paper the results of biogas formation
experiments of typical herbaceous residues from agriculture occurring
in Egypt are investigated. Based on this, co-fermentations of the inves­
tigated substrates are carried out in order to evaluate if interaction ef­
fects occur in the anaerobic digestion process. Based on these
experiments and data from literature, a simple model based on the
degradation kinetics of various agricultural residues within an anaerobic
digestion process is developed and presented. This model is intended to
enable operators of biogas plants to easily calculate the daily input
quantity for a freely selectable fermenter size as well as the resulting
biogas quantity for various substrates or substrate mixtures based on
current knowledge of the framework conditions of anaerobic fermen­
tation (e.g., taking into account restrictions regarding loading rate per
unit volume). Thus, this study aims to provide a tool with which the
kinetic models widely described in the literature can be usefully applied
to predict biogas formation by anaerobic fermentation in both existing
and new biogas plants to be built. The developed model can easily be
extended to include other substrates for which data regarding degra­
dation kinetics are already available.
2. Material and methods
2.1. Investigated substrates
Different agricultural residues from different crops and/or gover­
norates in Egypt are investigated regarding their potential as substrates
for biogas production via anaerobic digestion. The residues (namely rice
straw (Oryza sativa), corn stalks (Zea mays), wheat straw (Triticum aes­
tivum), sugarcane bagasse (Saccharum officinarum), grape pruning (Vitis
vinifera) and orange pruning (Citrus × sinensis L)) were collected from
farms in August 2021 and then sun-dried for a period of three days. In
the case of rice straw and maize straw, one sample each from the gov­
ernorates of Behaira and Sharkia were used. Grape and orange pruning
originated from the district of Gharbia, wheat straw was collected in the
governorate Behaira and sugarcane bagasse came from the governorate
Cairo. For further analyses, the dried samples were sent to Hamburg
University of Technology.
2.2. Determination of characteristic values
2.2.1. General
Prior to the investigations regarding the biogas formation potential,
all samples were analyzed with respect to various characteristic prop­
erties. Both for this and for the subsequent investigations of the biogas
formation potential, a comminution of the samples was necessary. This
comminution was performed using a laboratory cutting mill (MF10, IKA,
Germany) and the target particle size was < 1 mm. In order to obtain
subsamples that were as homogeneous as possible, the ground samples
were further divided using a rotary cone sample divider (Laborette27,
Fritsch, Germany).
2.2.2. Water content and volatile solids
Water content (W) and volatile solids (VS) were determined gravi­
metrically according to DIN EN ISO 18134 (DIN, 2015) and DIN EN
15935 (DIN, 2012) in triplicates, respectively. The drying process for
water content analysis was performed in a standard drying oven (U80,
Memmert, Germany) at 105 ◦ C. For the determination of volatile solids,
a muffle furnace (M104, Thermo Scientific Heraeus, USA) was used. The
ashing took place at a temperature of 550 ◦ C. The respective values for
water content and volatile solids were calculated using Equation (1) and
M. Scherzinger et al. Bioresource Technology 359 (2022) 127497

Equation (2), respectively. Table 1

mw Substrate mixtures in the co-digestion experiments.
W= (1)
(md + mw ) Substrate fractions

( ) Substrate Sugarcane bagasse Grape pruning

ma Mixture 1 (Cairo) (Gharbia)
VS = 1 − (2) (SM1) a) 25 %VS 75 %VS
md
b) 50 %VS 50 %VS
mw is the total mass of analysis sample, md is the mass of analysis c) 75 %VS 25 %VS
sample after drying at 105 ◦ C and ma is the mass of sample after incin­ Substrate Sugarcane bagasse Rice straw
Mixture 2 (Cairo) (Sharkia)
eration at 550 ◦ C.
(SM2) a) 25 %VS 75 %VS
b) 50 %VS 50 %VS
2.2.3. Elemental analysis c) 75 %VS 25 %VS
An elemental analysis quantifying the main elements nitrogen (N), Substrate Corn stalks Wheat straw
carbon (C), hydrogen (H) and sulfur (S) was carried out using a NCHS Mixture 3 (Sharkia) (Behaira)
(SM3) a) 25 %VS 75 %VS
analyzer with internal conductivity detector (vario MACRO cube, Ele­ b) 50 %VS 50 %VS
mentar, Germany). The content of oxygen (O) was determined by the c) 75 %VS 25 %VS
difference method by subtracting the above mentioned elements as well Substrate Rice straw Wheat straw Corn stalks
as the ash content from 100 %. Mixture 4 (Behaira) (Behaira) (Behaira)
(SM4) a) 20 %VS 40 %VS 40 %VS
b) 40 %VS 20 %VS 40 %VS
2.2.4. Weender feed analysis extended by Van Soest method c) 40 %VS 40 %VS 20 %VS
The organic components of the various agricultural residues were
further investigated using the Weender feed analysis extended by Van
Soest method in duplicates (Van Soest, 1967). More precisely, a subdi­ was confirmed by a reference measurement using microcrystalline cel­
vision was made into the following fractions: cellulose, hemicellulose, lulose as substrate (more than 90 % of the theoretically producible
lignin, raw lipids, raw protein, and non-fiber-carbohydrates. The steps biogas (=745 mLN g-1 VS) was formed). In the first test series (i.e. mono-
for determining the individual parameters are briefly described below. digestion), 2.7 gVS substrate was added into each reactor, and only 2.5
gVS in the second series (i.e. co-digestion). This adjustment was neces­
• Cellulose, hemicellulose and lignin. The amounts of the cell wall sary due to minor changes in the inoculum composition; the requirement
components cellulose, hemicellulose and lignin were determined defined within the measurement guideline that the ratio of the organic
according to the consecutive Van Soest dissolution method described components of the substrate to the organic components of the inoculum
in the VDLUFA method manual volume 3 (VDLUFA, 2012). The should be <0.5 was fulfilled in that way. The duration of the studies
method includes three dissolution steps, after each of which the ranged from 35 to 45 days. During that time, the amount of biogas
sample is dried and weighed. In the end, the remainder is incinerated formed was measured by the use of eudiometer tubes connected to the
following the above-mentioned method for volatile solids determi­ sample bottles and filled with a sealing liquid consisting of sodium
nation. The proportions of the substances dissolved out or remaining chloride solution acidified with citric acid. All results were corrected for
in each case can then be determined by calculation (see e.g. biogas production from the inoculum without substrate addition. In
(Scherzinger et al., 2021)). addition, the biogas formed was collected in each case and analyzed for
• Raw lipids. The amount of raw lipids was determined based on the methane content using a portable gas analyzer (Biogas5000, Geotech,
DIN EN ISO 11085 (DIN, 2016). Briefly, around 2 g sample mass was United Kingdom) equipped with a dual-wavelength IR sensor.
subjected to extraction with n-hexane (boiling point 69 ◦ C) in a
Soxleth apparatus for 24 h. The amount of dissolved mass corre­ 2.4. Modeling
sponds to the raw lipid content.
• Raw protein. The raw protein content was calculated by multiplying 2.4.1. General
the amount of nitrogen by the factor 6.25 (a standard protein con­ The modeling approach chosen to simulate biogas production is
tains 16 % nitrogen (N); i.e. the factor 6.25 is always used when there shown in a simplified form in Fig. 1. In the following, the individual
are no other specifications). parts of this modeling concept are discussed and the most important
• Non-fiber-carbohydrates. The amount of non-fiber-carbohydrates equations contained in the model are presented. Wet fermentation is
was calculated by subtracting the above mentioned feed compo­ assumed, with a substrate addition once a day.
nents (i.e. cellulose, hemicellulose, lignin, raw lipids and raw pro­
tein) and the ash content from 100 %. 2.4.2. Choice of substrate mixture
The first part of modeling is related to the choice of a suitable sub­
2.3. Fermentation in batch experiments strate mixture. For this, the previously determined contents of water,
volatile solids as well as the contents of carbon and nitrogen are applied.
The batch tests to investigate the anaerobic degradability of the Equation (3) shows the calculation of the water content of the feed (WF).
agricultural residues were conducted in two runs. First, the biogas for­ Analogous to this procedure, the volatile solid content and the C/N-ratio
mation potential of the individual agricultural residues was investi­ of the substrate mixture is calculated from the sum of the individual
gated, followed by co-digestion studies for selected mixtures of the components multiplied by the respective proportion of the individual
individual substrates. Table 1 provides an overview of the substrate component. With the help of the dry matter content, however, a con­
mixtures used in the second investigation series. version was carried out in these cases in order to relate the results to
All batch experiments were performed in triplicate determinations fresh mass.
according to the VDI guideline 4630 at mesophilic conditions (i.e. 37 ±
WF = (WRM1 × SRM1 ) + ⋯ + (WRMn × SRMn ) (3)
1 ◦ C). As reactors, 500 mL glass bottles were used. As inoculum 400 ± 1
g of digested sludge from a municipal wastewater treatment plant South WF and WRM are the water content of the chosen substrate feed and
of Hamburg was added to each reactor; the sludge was allowed to outgas the raw material, respectively. SRM is the proportion of the respective
for one week prior to substrate addition and all reactors were purged raw material in the substrate mix.
with nitrogen after substrate addition. The suitability of the inoculum The C/N-ratio of the substrate mixture should be between 10 and 30

3
M. Scherzinger et al.
Fig. 1. Modeling approach to simulate biogas production.
for anaerobic degradation according to current knowledge (FNR, 2016).
A corresponding query was implemented in the model to check this. If
the value is not in the optimal range, an adjustment of the substrate
mixture is recommended. Furthermore, a pumpable substrate mixture is
required for the present consideration (wet fermentation). In the model,
it is assumed that this occurs from a water content of 88 % or higher. If
the water content of the substrate mixture is below this threshold value,
additional water to be added (Wadd) is calculated using Equation (4).
Parts of the water can be re-used resp. recycled after the fermentation
process by mixing it with fresh substrate. Therefore, the fermentation
residues are pressed; here it is assumed that pressing is possible up to a
water content of 50 %.
0.88 − WF
Wadd = (4)
(1 − 0.88)
2.4.3. Operating parameters
The model is designed to freely select the fermenter volume (VF).
This also applies to the hydraulic retention time (HRT) of the substrate
mixture used, which should, however, be at least 20 days to ensure
stable operation. A density (ρ) of 1,000 kg m− 3 is assumed for the sub­
strate mixture, which either naturally has a water content above 88 % or
is brought to this water content by adding additional water. The daily
volume flow (V˙) is calculated via Equation (5). Subsequently, the
loading rate per unit volume (BR) is calculated via Equation (6). The
model also checks whether loading rate per unit volume (BR) is in an
appropriate range (between 1.5 and 3.5). If this is not the case, it pro­
vides a recommendation for action by adjusting the hydraulic residence
time. c is the concentration of organic matter in the substrate mixture.
V̇ =
VF
(5)
HRT
V̇ × ρ × c
BR = (6)
VF
2.4.4. Determination of kinetic parameters
The core of the chosen modeling approach is the prediction of biogas
formation from the selected substrate mixture. This is realized based on
kinetic models of the individual substrate components. More precisely,
modified Gompertz models are used (Velázquez-Martí and Meneses-
4
Bioresource Technology 359 (2022) 127497
Quelal, 2019). To obtain the kinetic parameters of these models, ki­
netic model fits are initially performed. The averaged biogas production
data from the batch experiments of the mono-substrates are used and a
nonlinear model fit based on the modified Gompertz equation (Equation
(7)) is performed using the “curve_fit” function from the Python module
“SciPy”. The kinetic parameters for the substrates studied here are
shown in the results (Table 3); P is the maximum biogas production
potential in mLN g-1VS, Rm is the maximum rate of biogas production in
mLN g-1 -1
VS d and λ is the lag phase in d. In addition, the root mean square
error RMSE was calculated to determine the quality of the models (the
smaller the value, the more accurate the model). Since a large number of
such kinetic studies for different substrates are available in literature,
the model can be extended relatively easy for other substrate types. M(t)
is the biogas yield in mLN g-1VS with respect to time t in days and e is the
Euler constant (2.718).
{ ( )}
Rm e
M(t) = P × exp − exp (λ − t) + 1 (7)
P
2.4.5. Calculation of the expected biogas / methane yield
The model can be used to calculate the biogas / methane formation
of a certain substrate mixture over time. Additionally, it is also possible
to implement changes in the substrate mixture at certain points in time.
Therefore, the daily biogas production is formed as the sum of the
specific biogas production rates (M˙(t)) of the individual components of
the substrate mixture multiplied by their respective shares. The specific
biogas production rates of the individual components are calculated
using Equation (8) by taking the difference in biogas formation between
two subsequent time steps.
Ṁ(t) = M(t) − M(t − 1) (8)
A conversion of the daily biogas production is performed by multi­
plication with the measured methane contents of the individual com­
ponents in the substrate mixture. This does not take into account that the
methane content of the formed biogas could change over time.
3. Results and discussion
In the following, the results of the laboratory investigations of the
substrates are presented and discussed. Building on this, the results of
M. Scherzinger et al. Bioresource Technology 359 (2022) 127497

the developed model are then presented using a specific example. Table 2
Main elements (in g kg-1
DM) and C / N – ratio of the investigated agricultural
residues.
3.1. Characteristic values of the investigated agricultural residues C H N S O C / N-ratio

Rice straw (Behaira) 380 45 5.2 < 2.0 379 73

Due to sun drying of agricultural residues, the water content in all Rice straw (Sharkia) 390 46 6.2 < 2.0 352 63
samples ranged from 5.7 ± 0.1 % (for corn stalks from Sharkia and Corn stalks (Behaira) 470 55 6.2 < 2.0 429 76
Behaira) to 7.8 ± 0.2 % (for orange pruning from Gharbia). The water Corn stalks (Sharkia) 440 48 6.9 < 2.0 403 64
Wheat straw (Behaira) 430 49 6.2 2.6 403 69
contents were used to ensure conversion of the results of the analytic-
Sugarcane bagasse (Cairo) 470 58 3.0 < 2.0 456 157
moist samples to dry reference basis. The results of the Weender feed Grape pruning (Gharbia) 480 58 6.1 < 2.0 407 79
analysis extended by the Van Soest method are accordingly shown in Orange pruning (Gharbia) 460 55 6.9 < 2.0 410 67
Fig. 2 in terms of dry matter (the numerical values can be found in the
supplementary material). The total of cell wall components hemicellu­
lose, cellulose, and lignin accounted for the majority of plant biomass for Table 3
all samples studied. Rice straw and sugarcane bagasse samples had the Kinetic parameters of the determined Gompertz-models (RMSE root mean
lowest content of these substances with a total of 70.6 ± 0.3 % for square error) and mean CH4 content of the investigated agricultural residues.
sugarcane bagasse and 70.7 ± 1.4 % and 70.2 ± 0.8 % for rice straw
P Rm λ RMSE CH4 in Mass related
from Behaira and Sharkia, respectively. In the case of the rice straw biogas conversion
samples, this comparatively low total amount of cell wall constituents
mLN mLN d mLN % % of VS
was due to the relatively high ash content of 19.1 ± 0.7 % and 20.6 ± g-1 g-1 -
g-1
VS VS d VS
1.4 % for samples from Behaira and Sharkia, respectively. In the case of 1

sugarcane, a relatively large amount of cell ingredients was found; e.g. Rice straw 450.6 41.4 0.15 16.8 57.3 55.0
the amount of non-fiber-carbohydrates, which mostly are cell in­ (Behaira)
Rice straw 475.9 43.1 0.16 15.6 55.9 59.0
gredients such as simple sugars, starch or pectin, accounted for 25.6 %.
(Sharkia)
During the production of biogas by anaerobic fermentation, various Corn stalks 489.4 41.8 − 0.07 9.1 58.8 58.8
microorganisms can degrade a large part of the organic components of (Behaira)
the biomass. However, due to its complex structure, lignin is an excep­ Corn stalks 392.4 25.5 1.45 8.3 62.3 45.3
(Sharkia)
tion, which cannot be degraded within anaerobic processes. Further­
Wheat straw 470.6 39.1 − 0.39 15.7 58.1 56.9
more, lignin coats the degradable substances and therefore partly (Behaira)
prevents the microorganisms from degrading them. Thus, high lignin Sugarcane 467.7 29.5 − 2.72 21.9 55.8 58.0
contents of the lignocellulosic feedstock hinders an anaerobic fermen­ bagasse
tation. In this case, pretreatment is essential; in most cases, a purely (Cairo)
Grape 298.5 16.4 − 0.52 6.7 60.5 35.2
mechanical shredding of the biomass is realized increasing the surface
pruning
area and thus improving the accessibility of the degradable substances (Gharbia)
(Paul and Dutta, 2018). Here, highest lignin content was found for grape Orange 378.5 11.0 − 5.46 15.3 63.3 43.2
pruning with 19.9 ± 0.2 % and lowest lignin content was found for rice pruning
(Gharbia)
straw form Behaira with 1.5 ± 0.1 %. Typically, severely lignified ma­
terial such as orange pruning is not used for anaerobic fermentation.
However, due to the relatively high proportions of easily degradable Highest amount of carbon was found for grape pruning (480 g kg-1 DM)
substances (e.g. 15.2 % non-fiber-carbohydrates), this material was also while lowest amount of carbon was found for rice straw from Behaira
being further investigated for this purpose. (380 g kg-1
DM). This is well in line with expectations and can be explained
The fat content in all the samples tested was very low, with a by the previously discussed molecular composition. High carbon content
maximum of 2.1 ± 0.1 % for grape pruning. This is typical for such in grape pruning is related to the high amount of lignin, which usually
agricultural residues; in particular, high-fiber biomass such as straw shows the highest carbon content of the investigated molecular sub­
usually has only very low fat contents (Tufail et al., 2018). stances varying from 51 to 67 % (Christ et al., 2017). Lower amounts of
The results of the elemental analysis are presented in Table 2.

Fig. 2. Molecular composition of the investigated agricultural residues.

5
M. Scherzinger et al.
carbon in the rice straw samples can be explained by the high amount of
inorganics.
With regard to anaerobic biogas formation, however, not only the
carbon content is important, but also the C/N-ratio. This ratio should be
between 10 and 30 (chapter 2.4.2). Too high C/N-ratios lead to an
insufficient metabolism in which the available carbon cannot be
completely converted. In the opposite case, excess nitrogen can lead to
the excessive formation of ammonia (NH3) inhibiting the growth of
bacteria even in low concentrations; in the worst case, it could even lead
to the complete collapse of the entire microorganism population (FNR,
2016). None of the agricultural residues studied had a C/N-ratio within
the optimum range. The lowest C/N-ratio of 64 was measured for corn
stalks from Sharkia, while the highest C/N-ratio of 157 was determined
for sugarcane bagasse. This also means that no suitable substrate
mixture can be produced by mixing the residual materials investigated
here; thus, organic residues and waste streams with a clearly higher
nitrogen content, such as sewage sludge or animal manure, could be
used to compensate for the lack of nitrogen (Neshat et al., 2017). In the
anaerobic degradation studies conducted, this was implemented by
using sewage sludge as inoculum.
3.2. Fermentation of mono-substrates
The results of biogas production of the individual agricultural resi­
dues over time and the corresponding Gompertz models are shown in
Fig. 3. The highest total yield of 496 ± 6 mLN g-1
VS was measured for corn
stalks from Behaira. Similar high yields were also obtained from both
rice straw samples, wheat straw, and corn stalks from Behaira. Signifi­
cantly less biogas (405 ± 2 mLN g-1 VS) was formed from corn stalks from
Sharkia. This can be explained by the molecular composition. Compared
to the cornstalk sample from Behaira, the sample from Sharkia have had
lower contents of easily degradable non-fiber carbohydrates and hemi­
cellulose. Additionally, the content of non-degradable lignin was
somewhat higher. So, although it is the same plant and even the same
part of the plant, there are significant differences in the case of corn
Fig. 3. Biogas formation of the investigated agricultural residues over time – Measured values and corresponding kinetic Gompertz-models.
6
Bioresource Technology 359 (2022) 127497
stalks. Therefore, for the most accurate prediction of biogas formation, a
laboratory investigation of the substrates to be used seems essential.
In contrast, both the molecular composition and the biogas forma­
tion potential of the two rice straw samples are very similar. Of the
investigated agricultural residues, grape pruning has the lowest biogas
formation potential (303 ± 7 mLN g-1 VS). This rather low biogas yield is
justified by the strong lignification of this material (see chapter 3.1).
This is also true for orange pruning, which has a biogas potential of 373
± 15 mlN g-1 VS.
In addition to the maximum biogas potential, however, the biogas
production rate is also of decisive importance in the technical imple­
mentation of an anaerobic fermentation plant. In fact, (too) slow biogas
formation rates are among the biggest challenges regarding the eco­
nomic operation of a biogas plant (Shen et al., 2015). Using the Gom­
pertz functions derived from the measured data, the maximum biogas
formation rates can be directly compared. Following this, the highest
maximum biogas formation rate of 43.1 mLN g-1 -1
VS d was found for rice
straw from Sharkia while the lowest maximum biogas formation rate of
11.0 mLN g-1 -1
VS d was found for orange pruning (Table 3). Based on the
biogas formation progressions shown in Fig. 1, no inhibitions of the
process were detected. However, both the two rice straws and corn
stalks from Sharkia exhibit lag phases; i.e., a delayed biogas production
after substrate addition can been observed. The reason for this behavior
might be that such substrates contain only very few amounts of readily
degradable substances. In this case, hydrolytic microorganisms first
decompose certain biomass components before the desired biogas for­
mation can begin. This is typical for a broad variety of lignocellulosic
substrates, where the hydrolysis phase is usually the rate-determining
step during anaerobic fermentation (Shrestha et al., 2017).
For the other substrates, biogas production starts with a phase of
linear or exponential growth and then decreases over time. An exception
to this is the sugarcane bagasse studied; here, the biogas formation de­
creases after about 5 days, before it increases again in a second increase
phase after about 8 days. Such diauxic behavior is rather atypical for
lignocellulosic substrates, but occurs frequently for other substrates such
M. Scherzinger et al.
as e.g. gelatin (Gomes et al., 2021). In the case of sugarcane bagasse, it
could be that all easily degradable components were metabolized within
the first 5 days; the second phase of biogas formation then refers to the
conversion of the more hardly degradable cell wall components hemi­
cellulose and cellulose. Both before substrate addition and after
completion of the experiments, the pH value was measured. While the
pH value before substrate addition was about 7.0, it increased slightly in
all reactors (up to about 7.4), but thus remained always in the neutral
and optimal range for biogas formation. Inhibitions due to changes in
the pH value are therefore considered very unlikely in the investigations
carried out.
In addition to the course of biogas production, the methane content
in the biogas formed was also determined. For this purpose, the biogas
was collected in the eudiometer tubes until enough gas was available for
a measurement (about 300 mL of biogas was needed). Hence, the
number of measurements carried out during the biogas tests depended
on the total amount of biogas formed. These data are then used to
calculate the average contents of methane relative to the total biogas
volume; those values are shown in Table 3. Due to this procedure, no
exact statements can be made regarding the temporal course of methane
formation. However, it can be deduced from the individual measured
values at different times that the methane content of the biogas formed
increases over time. This can be explained by the fact that easily
degradable components such as carbohydrates have a lower methane
potential (i.e. 415 mL CH4 g-1VS) than components that are more difficult
to degrade, such as proteins or lipids (i.e. 496 and 1,014 mL CH4 g-1 VS,
respectively) (Xue et al., 2020).
Based on the amount of biogas formed and the proportions of CH4
and CO2 and their density, respectively (it was assumed that biogas
consists only of these two gases; ρ of CH4 = 0.657 kg m− 3 and ρ of CO2 =
1.997 kg m− 3), the mass-based conversion was calculated. The highest
mass related conversion was obtained for rice straw from Sharkia with
59 % of VS, while the lowest mass related conversion was achieved for
Grape pruning with only 35.2 % of VS (see Table 3). Mass conversion
Fig. 4. Measured and predicted biogas formation of the investigated substrate mixtures (for definition of the substrate mixtures see Table 1).
7
Bioresource Technology 359 (2022) 127497
affects the amount of digestate produced in continuous digestion. The
lower the conversion, the more (solid) biomass is present in the digestate
at the end.
3.3. Fermentation of co-substrates
Fig. 4 shows the results of the co-digestion experiments. In addition,
these results are compared with the predicted values, which were
calculated on basis of the previously determined Gompertz equations
(data from Table 3). Here, it becomes obvious that biogas production
from the different substrate mixtures can be predicted accurately in most
cases using the Gompertz models derived from the monosubstrates. The
difference between the last measured value after 34 days and the pre­
dicted value is usually <5 %; only for SM1 a) and SM3 c) larger de­
viations of 10.8 % and 9.2 % were measured, respectively.
This means that the co-fermentation approach can achieve targeted
changes in both degradation rate and total biogas yield when the kinetic
constants of the substrates to be fermented are known. Furthermore, a
controlled change of the lag phase is possible. This is evident, for
example, from SM3, where a distinct lag phase was observed in the
fermentation of cornstalks from Sharkia. By co-fermenting with a sub­
strate without such a lag phase (i.e. wheat straw), this lag phase could be
almost completely avoided. In this context, it has to be noted that the
possible changes can only take place within a certain range depending
on the properties of the respective substrates.
These findings are in good agreement with insights from other
studies. For example, it was demonstrated that the lag phase occurring
during mono-digestion of raw sludge could be significantly reduced by
the addition of lignocellulosic biomass. This was attributed to the faster
degradation of carbohydrates from the lignocellulosic biomass
compared to the proteins mainly contained in the raw sludge (Zou et al.,
2018).
Moreover, in the present case, no significant interaction effects were
observed during the co-fermentation of the substrates. This is mainly
M. Scherzinger et al.
due to the fermentation procedure according to the VDI guideline 4630,
in which the fermentation takes place under the most optimal conditions
possible. A nutrient deficiency is excluded within this method by the
choice of inoculum and an overload of the fermentation system is
avoided by the relatively small amount of substrate used (the actually
unfavorable C/N-ratios of the substrate mixtures, of which the numeri­
cal values are listed in the supplementary material, can be compensated
by the sewage sludge (inoculum)). This also means that not all possible
benefits of co-digestion can be captured by the studies conducted here.
Co-digestion can, among other things, increase the buffer capacity,
change the C/N-ratio and dissolve toxic substances, all of which can
have a positive effect on the biogas yield (Xu et al., 2018). Furthermore,
non-biogenic factors can also influence the process of biogas formation.
For example, the methane-forming microorganisms require certain trace
elements such as iron, cobalt and nickel. During continuous co-
digestion, an optimal supply of these trace elements can be ensured by
adjusting the substrate mixture (Li et al., 2014). Moreover, during
continuous co-digestion, adaptations of microorganisms to the substrate
mixture could also occur, which could improve biogas formation.
Additionally, substrate-related inhibitions, e.g. due to the formation of
floating or sinking layers, could arise. Therefore, the findings obtained
here should also be verified in continuously fed biogas reactors in order
to obtain results independent from the used inoculum and under more
practice-oriented environments.
3.4. Modeling results
3.4.1. General
Despite the mentioned limitations due to the applied method, the
collected data can be used in a modeling approach based on the iden­
tified kinetic constants. However, during modeling, care must be taken
that the parameters are selected in such a way that the best possible
fermentation is ensured, as otherwise effects may occur that cannot be
represented by the model. To investigate such effects (like e.g. acidifi­
cation of the reactor), a more advanced model would be required.
In the following, the developed model is presented based on a
Fig. 5. Simplified representation of the assumptions and individual substrate mixture-dependent parameters in the presented modeling case.
8
Bioresource Technology 359 (2022) 127497
specific use case.
3.4.2. Choice of fermenter volume, hydraulic retention time and substrate
mixture
In rural Egypt, biogas is mainly produced in so-called “Chinese fixed-
dome household digesters” consisting of a closed cylindrical chamber
with an immovable gas space and both a digestate in- and outlet
(Ioannou-Ttofa et al., 2021). The typical volume of such a simple
digester is up to about 6 m3. Therefore, a small fermenter volume was
also assumed for the example case described here (i.e. 5 m3). In general,
however, the fermenter volume can be freely selected in the model. The
case considered here is illustrated in simplified form in Fig. 5.
In the case shown here, a substrate mixture consisting only of agri­
cultural residues from the governorate Behaira was selected as initial
substrate mixture (i.e. corn stalks, rice straw and wheat straw, ratio
1:1:1 based on fresh mass) and the hydraulic retention time (HRT) was
set to be 40 d. In order to obtain a pumpable substrate mixture, 6.72 kg
of water must be added per kg of substrate under the assumptions made
in chapter 2.4. Of this, 5.87 kg of water can subsequently be recovered
by squeezing. A high water recovery rate is particularly important in
water-scarce regions, of which Egypt is one. For the operation of the
plant with the selected substrate mixture, this means that for each kg of
fresh substrate mass, an additional 0.85 kg of water must be provided.
The total amount of fresh mass to be fed daily is close to 16.2 kg.
Accordingly, an adequate supply of water (i.e. 13.8 kg d–1) is considered
very likely.
The resulting loading rate per unit volume (BR) under these specifi­
cations is 2.66, which is in the optimal range. Although it has already
been demonstrated that, in particular by co-fermenting different sub­
strates, higher loading rates per unit volume (BR) can also be realized
without process disturbance, the relatively conservative range selected
means that, for example, an accumulation of fatty acids can be largely
ruled out (Hegde, 2019). The loading rate per unit volume is in direct
interaction with the hydraulic retention time; the model makes recom­
mendations to lengthen the hydraulic retention time (HRT) if the
loading rate per unit volume (BR) is too high and to shorten it if it is too
M. Scherzinger et al.
low.
The C/N-ratio for the selected substrate mixture is 72.8, which is
significantly higher than the optimal C/N-ratio. However, an optimal
ratio cannot be achieved by mixing the agricultural residues investi­
gated here, since all investigated residues show a too low nitrogen
content. A warning is therefore displayed in the model. It could be that
due to the unfavorable C/N-ratio, lower biogas yields are achieved in
practice than calculated in the model. But, nitrogen is concentrated by
the decomposition of carbon compounds within the digestate. This is
due to the fact that relatively more carbon compounds are converted,
while large proportions of nitrogen compounds pass into the liquid
phase (Dong et al., 2020). Hence, by returning the process liquid to the
fermenter, the C/N-ratio could be lowered through the recirculation of
nitrogen, bringing it closer to the optimum range in theory.
Due to the varying accumulation of agricultural residues in the
course of the year, the influence of a substrate change was also inves­
tigated in the model. Therefore, it was assumed that the substrate
mixture described above would only be used in the initial phase (up to
day 25).
From this point on, a substrate mixture of cornstalks, rice straw and
sugarcane bagasse (ratio 1:1:3 based on fresh mass) is used. This also
changes the amount of water to be added to obtain a pumpable substrate
mixture (i.e. 6.75 kg water need to be added per kg substrate mixture, of
which 5.89 kg can be recovered by squeezing). With this new substrate
mixture, the loading rate per unit volume changes slightly to 2.84 and
still lies in the optimum range. The total amount of fresh mass to be fed
daily is 16.1 kg from that day on.
3.4.3. Feeding management and biogas generation
The developed model is based on a daily one-time feeding of the
biogas plant, with the same amount of substrate being fed each day.
Accordingly, the biogas formation is always related to the substrate
addition on a given day; this is illustrated in Fig. 6. Consequently, the
total biogas production is then the sum of the biogas production of the
respective fed substrate mixture at the respective time. It also follows
from this presumption that under steady state conditions the same
amount of biogas (with the same methane content), is formed every day.
A steady state is formed only after at least one hydraulic residence time
has elapsed, although in practice rather longer periods are needed (i.e. 3
to 5 residence times) for this (VDI, 2016). Accordingly, in the modeling
results shown in Fig. 6, a steady state is not achieved. Rather, a start-up
phase is shown from day 0, which then slowly approximates to a steady
state phase after about 20 days. The figure indicates also that the biogas
production of one specific substrate addition using the first substrate
mixture spans approx. over a time period of 30 days, whereby already
after approx. 20 days only small biogas amounts are still formed.
Fig. 6. Modeling results for the selected use case; the gray curves show the daily biogas production with respect to the individual feeds (left axis), the solid black line
shows the total daily biogas production (right axis) and the dotted black line is the time of substrate change.
9
Bioresource Technology 359 (2022) 127497
Until the substrate change on day 25, the biogas production conse­
quently remains largely constant. Due to the slightly lower biogas for­
mation potential of the new substrate mixture and the changed course of
degradation (e.g. lower maximum daily biogas formation), the biogas
yield then initially decreases before it slightly increases again over time
and settles at a level comparable to that immediately before the sub­
strate change.
This developed model is intended to enable operators of biogas
plants to easily obtain common process data regarding the anaerobic
digestion of various agricultural residues. That this can be achieved via
such an approach has already been demonstrated in another study
(Zepter et al., 2021). In contrast to the aforementioned study, however,
an adjustment of substrate properties (i.e., change of substrate mixture)
was included here. This is considered to be highly relevant for plant
operators due to varying substrate availability throughout the year.
A major open issue is that the developed model has not yet been
validated on real measured data. It is therefore not clear to which extent
the model can provide reliable data. In the optimal range of anaerobic
digestion, it is highly probable that reliable data can be provided (this
was confirmed by the co-digestion experiments). However, under un­
favorable conditions, this is rather not the case as the model cannot
include inhibitory effects. Also, no trace element deficiency was
considered in the model. Due to these clear limitations, the model is only
recommended for the applications mentioned above. Here, it is mainly
characterized by its simplicity, which should facilitate a broad use.
4. Conclusion
A user-oriented model that predicts the most optimal process pa­
rameters as well as the expected biogas production for small-scale biogas
plants was developed. The model is both based on kinetic Gompertz
models and specific substrate properties. For the investigated substrates,
it was demonstrated that under optimum conditions almost no interac­
tion effects occur during co-digestion. The deviation of predicted results
from obtained results from co-fermentation experiments was <5 % in 10
out of 12 cases. Therefore, it can be assumed that the model provides a
good approximation of the actual biogas production also in a quasi-
continuous operation.
CRediT authorship contribution statement
Marvin Scherzinger: Conceptualization, Methodology, Software,
Investigation, Writing – original draft, Writing – review & editing,
Visualization. Martin Kaltschmitt: Supervision, Project administration,
Funding acquisition. Amr Y. Elbanhawy: Resources, Supervision,
Project administration.
M. Scherzinger et al.
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.
Data availability
Data will be made available on request.
Acknowledgements
The results presented in this publication were obtained as part of the
“Co-Agrow+” project (grant number: 01DH21002), which was funded
by the German Federal Ministry of Education and Research.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.biortech.2022.127497.
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