Industrial Crops & Products 131 (2019) 307–314

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Industrial Crops & Products

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

Small scale biogas production with animal excrement and agricultural T

residues
⁎
A.I. Parralejoa, L. Royanoa, J. Gonzáleza, J.F. Gonzálezb,
a
Department of Field Crops: Biomass and Bioproducts, Centre for Scientific and Technological Research of Extremadura (CICYTEX), Consejería of Economy and
Infraestructure, Junta de Extremadura, Guadajira, 06187, Badajoz, Spain
b
Escuela de Ingenierías Industriales, Avda. de Elvas, s/n 06006, Badajoz, Spain

A R T I C LE I N FO A B S T R A C T

Keywords: Anaerobic digestion is a very versatile technology producing biogas, which can be used for heating and elec-
Agricultural residues tricity production or upgraded and used for vehicle fuel or gas-grid injection. The purpose of using anaerobic
Manure digestion is usually related to waste management (agricultural residues, animal manure and other organic
Anaerobic process waste). Animal manure (AM) of Iberian pig, calf and lamb were studied. The methane production was analyzed
Biogas
in co-digestion of the aforementioned animal manure, and finally they were used as substrates in the co-digestion
Kinetic study
with silages of tomato pulp (T), grape byproduct (G) and olive agro-food byproduct (O). The results obtained in
the different experiments showed that anaerobic co-digestion allow to establish the mixture of animal manure
and agricultural residues more suitable to optimize the production of biogas and stabilize the anaerobic digestion
process. A higher proportion of animal manure in the co-digested mixture leads to higher methane yield. Kinetic
parameters results correspond to similar parameters published in literature, they are located in 0.04-0.07 d−1 for
methane production rate constant from first order model fit, 0.58–2.85 NLCH4 kg VS−1 d−1 for maximum
methane production rate and 0.004–18.96 d for lag phase from modified Gompertz model fit, and 0.03-0.07 d−1
for methane production rate constant from Cone model fit and 2.07–3.07 for the model dimensionless Cone
constant (n).

1. Introduction conventional non-renewable resources has favoured its domestic ap-

A sustainable bioeconomy is essential to the reduction of emissions
in the European Energy sector. Bioenergy, currently the EU´s largest
renewable energy source, is expected to remain a key component of the
energy mix in 2030. The transformation towards sustainable, healthy,
nutrition-sensitive, resource-efficient, resilient, circular and inclusive
food and farming systems needs to accelerate. This includes turning
organic waste, residues and food discards into valuable and safe bio-
based products, for instance by deploying small-scale biorefineries,
helping farmers, foresters and fishermen to diversify their revenue
sources and better manage market risks, all while achieving the goals of
the Circular Economy (European Commission, 2018).
Biogas resulting from the anaerobic digestion of solid/liquid wastes
like cattle and pig excreta, kitchen waste, wastewater sludge, agro-in-
dustrial wastes, lignocellulosic biomass has been one of the most fa-
vourable bioenergy sources (El Achkar et al., 2018; Hirano and
Matsumoto, 2018; de la Lama et al., 2017; Neshat et al., 2017; Zhou
et al., 2016; Cheng et al., 2011). The biogas popularity over
plications, especially as cooking fuel in developing countries, and also
large scale plants for power generation (Santarelli et al., 2017;
Papurello et al., 2016; Sawatdeenarunat et al., 2015).
Biogas facilities range from micro-scale digesters in developing
countries, small-scale digesters used on farms and communities to large
scale digesters encompassing centralized systems found in regions and
cities. The use of animal manures and slurries as a feedstock for small-
scale plants, but also in large-scale plants for co-digestion with other
feedstocks or as a sole feedstock is common practice in some countries.
Anaerobic digestion not only reduces GHG emissions by substituting
fossil energy, but also avoids GHG emissions from open storage of
manure and slurries (Neshat et al., 2017). Various residues of the food
industries or fruit processing can be used as biogas feedstock in co-
digestion with animal manure and slurry. The interest for the use of
lignocellulosic wastes is mainly based on their chemical compositions,
which have high carbohydrate content, in the form of cellulose and
hemicelluloses (∼70%), that can be used for bioenergy production
(Gupta and Tuohy, 2013). The high carbohydrate content of these

⁎
Corresponding author.
E-mail address: jfelixgg@unex.es (J.F. González).

https://doi.org/10.1016/j.indcrop.2019.01.059
Received 30 November 2018; Received in revised form 19 January 2019; Accepted 28 January 2019
0926-6690/ © 2019 Elsevier B.V. All rights reserved.
A.I. Parralejo et al. Industrial Crops & Products 131 (2019) 307–314

wastes makes an excellent substrate for methane production through Table 1

solid-state anaerobic digestion (SS-AD) (Xumeng et al., 2016). Some Design of tests for the substrates studied.
alternatives to maximize SS-AD process efficiency involve the use of Assay G/T/O (g VS) AM (g VS)
inocula capable of promoting higher conversion rates of lignocellulosic
wastes polysaccharides into biogas (Sawatdeenarunat et al., 2015). I CM LM PM
Sousa Lima et al. (2018) have published the effect of different thermal
AE – 25 87 79 4
pretreatments and inoculum selection on the biomethanation rate of AM:G 1 66 9 29 32 2
sugarcane bagasse. They observe that use of a combined inoculum, AM:G 2 44 13 39 43 2
made up of upflow anaerobic sludge blanket (UASB) sludge and fresh AM:G 3 87 6 18 20 1
bovine manure (FBM), was more efficient in maximizing methane AM:T 1 61 9 29 32 2
AM:T 2 41 13 39 43 2
production during anaerobic digestion of sugarcane bagasse (SB) than
AM:T 3 87 6 18 20 1
biomass pretreatment processes. AM:O 1 54 9 29 32 2
Literature reports results from anaerobic co-digestion of agro-food AM:O 2 36 13 39 43 2
byproduct with livestock manure or wastewater (El Achkar et al., 2018; AM:O 3 72 10 18 20 1
Guerini-Filho et al., 2018; Patowary and Baruah 2018; Sousa Lima
AM: Animal Manure: I: Inoculum, VM: Veal Manure, LM: Lamb Manure, PM:
et al., 2018; de la Lama et al., 2017; Riggio et al., 2015; Aylin-Alagöz
Pig Manure.
et al., 2015) introducing into the biodigester two or three co-substrates.
G: Grape byproduct.
The potential methane yield obtained in these works ranges 125–460 T: Tomato pulp.
NL CH4 kgVS−1. However, it is not found research works employing O: Olive agro-food byproduct.
livestock manure of diverse animals in the same biodigester, and fur-
thermore the agro-food byproduct. straw. The straw proportion was low but an important amount was
Kinetic models are usually used in anaerobic digestion processes to found in the silages because it has an extremely low density. Grape
check initial hypothesis, to evaluate the experimental results and to byproduct consisted of biomass waste from winery industry from
control and predict the process performance, optimizing the plant de- Extremadura region (Southwestern Spain).
sign (Isik and Sponza, 2005).
For the development of cost-effective technologies, data on optimal
2.2. Test equipment and assays design
mixture ratios, specific methane yield and the kinetic of degradation
process are needed. Increased methane yield and biodegradation pro-
Assays have been carried out using 6 litres cylindrical waterproof
cess rate will result in more methane produced per unit fed mass and
reactors built in stainless steel with 4.5 litres load of substrates. A water
will reduce the reactor size required, making the process economically
jacket surrounding each digester allowed temperature remains constant
more attractive. In this work, the main research goal has been to de-
of about 38 °C (mesophilic range), checked by a thermostat. Mechanical
velop an optimal mixture of Iberian animal manures and agro-food
agitation in the biodigesters is controlled by an independent regulator
byproduct silages to get a future energy recovery for farms of Iberian
allowing optimal contact between substrates. Assays realized with an-
animals employing the anaerobic co-digestion processes. The novelty of
imal manure have been repeated three times. A control assay is a test
this work is to co-digest cattle manures from farms with different lig-
carried out in the same conditions that the assays studied but inoculum
nocellulosic residues. Moreover, it allows to obtain initial conditions of
diluted in water is the only substrate introduced in the digester. The
Total Solids, Volatile Solids and C/N ratio to get an adequate proportion
methane production obtained in this assay will be used to know the
of substrates to develop an optimum anaerobic co-digestion. The co-
methane volume generated by inoculum in the studied assays.
digestion process and the synergetic effects in different wastes mixtures
Anaerobic co-digestion of animal manure was developed for 45 days
were characterized by determining methane yield, effects of substrate
and assays researched with agro-food byproduct were maintained for
proportion in the mixture on the typical parameters in this process and
65 days. Mixture of substrates studied is detailed for each assay in
kinetic development using anaerobic batch digestion assays.
Table 1. The proportions of substrates in the mixture employed in the
assays was calculated based on four requirements: total volume of
2. Material and methods
mixture of 4.5 litres, total Volatile Solids higher than 120 g, Total Solids
lower than 10% and C/N ratio higher than 10. The proportion of Vo-
2.1. Analyzed raw materials: Iberian animal manure and agro-food
latile Solid of calf manure (CM), lamb manure (LM) and pig manure
byproduct silages
(PM) for assay AM were 45%, 41% and 2%, respectively. This pro-
portion was maintained for anaerobic co-digestion assays modifying the
Animal manure samples evaluated in this work have been taken
relation agro-food byproduct/manure in these cases.
from two Iberian feedlots of Extremadura region: a pig farm and a cattle
and sheep cooperative. Thus three different animal samples have been
characterized to study the anaerobic co-digestion process. Calf and 2.3. Analytical methods
lamb manure samples have been chopped carefully using a scissor (an
important straw content was included) to a size of 2 cm (mechanical Substrates characteristics were analyzed according to the Standard
treatment) and stored at -4 °C. Inoculum was employed in the devel- Methods (APHA, 2012). Total Solids (TS) content have been determined
oped assays to increase the methanogenic microorganisms. The in- by drying the sample to constant weight in an oven (JP Selecta Di-
oculum consists of a degraded mixture (for 5 years) of pig manure and githeat, USA) at 105 °C for 48 h (2540 B method). The total Volatile
prickly pear turned into paste. This substrates mixture allows have in- Solids (VS) content has been obtained by heating the dried TS to con-
oculum in a short time due to its fast degradations. stant weight at 550 °C for 2 h in a muffle furnace (Hobersal
The agro-food byproducts employed in this study were silage pre- 12PR300CCH, Spain) at inert atmosphere (2540 E method). pH and
viously used to feed animals. For this reason tomato and olive by- redox potential have been measured with their corresponding elec-
products needed a part of straw in the silaged mixture. Silages were trodes with a pH meter (Crison Basic 20, Spain), alkalinity of the
prepared by mixturing byproducts with straw (except grape byproduct) medium through 2320 method, Chemical Oxygen Demand (COD) ac-
and storing them in plastic bags in the absence of light and oxygen two cording to 410.4 method (EPA, 1983), N-NH4 through E4500-NH3 B
months at least. The tomato byproduct employed was tomato pulp and volumetric titration method and the Volatile Fatty Acids (VFA) ac-
straw, and the olive byproduct was composed by olive pomace and cording to Buchauer (Buchauer, 1997) by titration methods. The initial

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A.I. Parralejo et al.
C/N ratio in substrates has been determined using an elemental ana-
lyser True-Spec CHN Leco 4084 (USA), according to the standard UNE-
EN 16,948 for analysis of biomass. C, N, H (UNE-EN 16,948, 2015). Gas
composition and total gas volume have been automatically in site
monitored during the experiments with a gas analyser Awite System of
Analysis Process serie 9 (Bioenergie GmbH, Germany). This analyser is
composed of two IR sensors to take methane and carbon dioxide mea-
surements, and three electrochemical sensors that supply values of
hydrogen, sulfhidric acid and oxygen content in the biogas produced.
Gas counters (Ritter model MGC-1 V3.2 PMMA, Germany) have been
used to measure produced biogas stored in tedlar bags. Dry gas volume
has been corrected to standard conditions (0 °C, 101.325 kPa). More
important elements in the digestate have been detected by spectroscopy
technique using an ICP-OES Varian 715 ES (Australia). Previously,
samples have been digested in a laboratory microwave Millestone Start
D (Italy).
2.4. Kinetic study
A kinetics fit adapted to methane volume produced for a dis-
continuous process was used. Three kinetic models were used in this
work: first order model (Pagés et al., 2011), and modified Gompertz
and Cone models (Sousa Lima et al., 2018).
The first-order kinetic model assumes that the rate-limiting step of
the anaerobic digestion is the substrate hydrolysis. This model set the
results according to the expression [1].
G = Gm [1 − e−k 0 t ] (1)
−1
where G (L CH4 kg VS ) is the accumulated methane production after
a time t (d), Gm is the maximum methane yield; ko (d−1) is the methane
production rate constant.
The modified Gompertz model is an empirical model that allows the
determination of important parameters of anaerobic digestion such lag
phase or specific methane production. The expression defined by this
model is:
μ e
G = Gm exp ⎧−exp ⎡ m (λ − t ) + 1⎤ ⎫
⎨
⎩ ⎢
⎣ Gm ⎥
⎦⎬⎭ (2)
−1 −1
Where λ (d) is the lag phase; μm (L CH4 kg VS d ) is the maximum
methane production rate and e is the Euler´s number.
The Cone model as the previous model allows the determination of
the specific methane production rate and the behaviour of methane
production by the shape constant (n) which indicates the presence or
absence of lag phase. The expression is the following:
Gm
G=
1 + (kt )−n (3)
−1
where k (d ) is the methane production rate constant and n is the
Cone model constant (dimensionless).
3. Results and discussion
3.1. Characterization of raw materials
Substrates investigated in this work have been submitted to a phy-
sical-chemical analysis to design the anaerobic digestion assays. The
Table 2 shows the results obtained for each substrate.
The substrate called animal manure supplies alkalinity to the di-
gestion medium to compensate the pH low values of substrates silages.
Alkalinity values higher than 2500 mg L−1 ensure the stability of the
process (Fannin, 1987). Also they provide a reduction medium (nega-
tive values in their redox potential) to let an adequate performance of
reaction mechanisms responsible for the generation of methane. The
COD of calf and lamb manure is favorable for the anaerobic digestion
process because is quite elevated. However, the animal manure usually
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Industrial Crops & Products 131 (2019) 307–314
has high values of ammonia nitrogen as it can be observed in the
Table 2 (1640 mg L−1, 4000 mg L−1 and 1840 mg L−1 for calf, lamb
and pig manure, respectively). Ammonia nitrogen is considered an in-
hibitor of the process, according to some authors this parameter ranges
between 2000 mg L−1 (Flotats et al., 1999) and 3000 mg L−1 (Van
Velsen, 1979; Parkin and Owen, 1986). So in this study, a productive
process to generate methane requires a co-digestion process of animal
manure with agro-food subproduct silages with the objective to equi-
librate this parameter.
3.2. Final parameters in anaerobic co-digestion assays
Anaerobic co-digestion has been carried out for animal manure
according to the Table 1 to get the right values of TS and C/N ratio.
Later the agro-food byproducts have been submitted to the anaerobic
co-digestion process with animal manure in the proportion evaluated in
the process realized at the beginning. The most representative para-
meters of anaerobic co-digestion process developed in this work are
detailed in the Table 3.
It can be observed a high final ammonia nitrogen value for assays
carried out with animal manure and it has been got down in the
anaerobic co-digestion assays employing agro-food byproduct silages.
The alkalinity values obtained decreased when the used proportion of
animal manure is lower (AE:G 3, AE:T 3 and AE:O 3), it can be the
reason that the methane yield were the lowest. According to Mao et al.
(Mao et al., 2019) values of alkalinity/VFA ratio over 2.5 are regarded
as optimal for anaerobic digestion. In this study for AE:O 3 assay al-
kalinity/VFA ratio corresponding is 1.99 so an unstable process was
happened. Also the high C/N ratio (higher than right values for anae-
robic digestion process, 15–30 (Speece, 1987), 20–30 (Zhou et al.,
2016)) in assays previously mentioned can contribute to obtain low
methane yields. So the lower values of methane yield belong to the
assays with low proportion of animal manure and the most elevate yield
corresponding to an higher amount of animal manure than agro-food
byproduct. However, the behaviour of olive agro-food byproduct is
different because the most elevated methane yield is found in the equal
proportion with agro-food byproduct and animal manure. In the olive
waste assays probably the organic matter is too elevated in the used
proportion for AE:O 1 assay. The COD and C/N ratio is higher in the
mentioned co-substrate than the grape byproduct or tomato pulp as it is
shown in Table 2.
3.3. Agro-food byproduct influence on biogas and methane production
The biogas production versus time for the three proportions assayed
in this work of animal manure and agro-food byproduct, 1:1, 2:1 and
1:2 are showed in the Fig. 1. The corresponding evolution of the cu-
mulative methane yield for these tests are given in the Fig. 2.
It can be observed that the biogas generation is slower and lower as
the agro-food byproduct content is increased in the load of biodigester
(Compare tests AM:G 1, AM:T 1 and AM:O 1 with AM:G 3, AM:T 3 and
AM:O 3). A higher animal manure content in the mixture proportion
acecelerate the biogas production reducing the generation time, how-
ever, a higher agro-food byproducts content in the mixture proportion
draws out the biogas production and besides reduces its generation.
This effect can be also seen in the curves of the cummulative methane
yield (Fig. 2), and corroborated with los data showed in Table 3.
Therefore the best results were reached with the proportion 2:1, animal
manure:agro-food byproducts, as much for the biogas production as the
cummulative methane yield. Some authors (El Achkar et al., 2016) have
researched the anaerobic digestion of milled pulp and milled seeds of
grape obtaining methane yields of 343 N L kg VS−1 and 214 N L kg
VS−1, respectively. These values are smaller than results obtained in
the AM:G 2 assay (348 N L kg VS−1) because an anaerobic co-digestion
is more productive than anaerobic digestion. Also the agro-food by-
product grape byproduct studied in this work is reffered to pulp, seeds
A.I. Parralejo et al. Industrial Crops & Products 131 (2019) 307–314

Table 2
Raw material characterization for the different runs.
Animal manure / Agro-food byproduct silages

Parameter/Substrate I CM LM PM T O G

Total Solid (TS), % 1.83 30.66 35.69 0.80 30.73 33.06 30.66
Volatile Solid (VS)1,% 46.27 85.99 81.22 51.89 68.32 60.20 85.99
COD, mg L−1 18000 156000 496000 21000 257400 787500 410000
Volatile Fatty Acid, mg L−1 171 – – 2018 – – –
Alkalinity, mg L−1 9648 – – 6362 – – –
Ammonia nitrogen, mg L−1 5440 1640 4000 1840 226 395 < 30
pH 7.85 8.00 8.84 7.50 3.64 4.56 4.29
Redox potential, mV −441 −227 −82 −413 184 51.1 35
Ratio C/N2 10.92 19.05 21.79 13.00 33.41 42.13 37.28
Ca, ppm 642 35527 42726 207 1040 1901 3044
Fe, ppm 21 1628 1118 8 218 106 195
K, ppm 1946 34638 42257 1851 4058 3960 7594
Mg, ppm 333 11268 9496 77 755 303 622
Na, ppm 426 16077 20944 568 221 18 199
P, ppm 166 12559 12225 57 714 273 675
Cd, ppm n.d. n.d. <2 <2 <2 n.d. <2
Cr, ppm n.d. 13 9 n.d. <2 <2 n.d.
Cu, ppm <5 104 109 <5 <5 <5 <5
Ni, ppm <5 24 27 n.d. n.d. <5 <5
Zn, ppm 52 155 303 3 6 1 n.d.
Al, ppm 13 1616 1008 11 357 88 223

n.d.: not detected.

1
over TS.
2
over dry matter (in pig manure over wet matter).

Table 3
Analyses for anaerobic co-digestion assays.
Parameter AM Silages anaerobic co-digestion

AM:G 1 AM:T 1 AM:O 1 AM:G 2 AM:T 2 AM:O 2 AM:G 3 AM:T 3 AM:O 3

VS reduction, % 53 26 59 55 55 43 47 38 63 65
DQO reduction, % 41 36 33 50 48 41 54 17 64 50
Final ammonia nitrogen, mg L−1 4547 1050 960 640 1110 1200 990 690 680 340
Volatile Fatty Acid, mg L−1 968 661 461 573 703 618 1143 491 607 1568
Alkalinity, mg CaCO3 L−1 13897 6115 5845 4908 6764 6184 5707 3834 3676 3128
Ratio C/N 13.33 10.09 9.82 15.10 9.84 7.99 10.72 28.20 25.62 27.46
Methane yield, NL kg VS−1 260 295 365 398 348 404 325 164 289 124
Ca, ppm 2856 2243 1381 755 1761 1992 1128 820 560 556
Fe, ppm 106 75 43 26 46 111 30 17 32 21
K, ppm 1569 488 2991 1414 5594 3600 1723 891 896 995
Mg, ppm 1086 697 649 272 631 668 445 169 224 186
Na, ppm 2060 984 949 95 1129 1186 271 n.d. n.d. n.d.
P, ppm 614 625 335 211 429 652 370 162 173 140
Cd, ppm 54 n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d.
Cr, ppm 20 <2 n.d. <2 n.d. <2 n.d. n.d. <2 <2
Cu, ppm 19 14 <5 <5 <5 <5 <5 <5 <5 <5
Ni, ppm 4 <5 <5 <5 <5 <5 n.d. n.d. <5 <5
Zn, ppm 26 62 9 25 5 15 31 7 18 17
Al, ppm 73 57 47 19 38 88 17 <5 26

AM: Animal Manure.

G: Grape byproduct.
T: Tomato pulp.
O: Olive agro-food byproduct.
n.d.: not detected.

and small branches. The biogas evolution for tomato pulp co-substrate
represented in the Fig. 1 shows a clear trend to keep the production in
the time. The co-substrate mentioned produces the highest obtained
methane yield values (365 N L kg VS−1, 404 N L kg VS−1and 289 N L kg
VS−1 for AM:T 1, AM:T 2 and AM:T 3, respectively). According to re-
searched results (Bacenetti et al., 2015), the anaerobic digestion of
tomato waste and skins, and seeds separately obtains biogas produc-
tions of 506.8 L kg VS-1 and 358.5 L kg VS-1, respectively, it means an
intermediate value should be obtained in this research because the
tomate pulp co-substrate is composed by waste, skins, seeds and straw
(365 N L kg VS-1 for AM:T 1). Studies for olive agro-food byproduct
(Aylin-Alagöz et al., 2015) have obtained methane yield of 320 N L kg
VS-1 in anaerobic co-digestion with waste sludge pre-treated. This value
is very similar to the assay carried out in this investigation (AM:O 2),
325 N L kg VS-1. For AM:O 1 methane yield obtained is exceeded to
theoretical value (398 N L kg VS-1). Other authors have reached me-
thane yield of 450 N L kg VS-1 in semi-continuous anaerobic digestion of
thermally pretreated two-phase olive pomace (de la Lama et al., 2017).

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A.I. Parralejo et al.
Fig. 1. Biogas production of anaerobic co-digestion assays in different proportions of animal manure and agro-food byproduct silages.
3.4. Interactions of parameters studied
The most important variables analyzed to know the development of
the process are represented in Figs. 3–5 for each agro-food byproduct
and the substrates proportion researched. Fig. 3 shows the effects of the
substrate and substrates proportion on the reduction of VS and COD.
One can see that the VS reduction in the medium is higher as the agro-
food byproducts is increased in the biodigester load, which it is not
agree with the biogas and methane production, as it has been men-
tioned above, however the COD reduction is higher for the test with the
highest animal manure proportion, AM:S 2. According to the influence
of the substrate on the VS and COD reduction, one can observe the
sequence O > T > G, indicating a major facility of the olive pomace
for the degradation.
Fig. 4 shows a sharp relationship of ammonia nitrogen and alkali-
nity with the higher proportion of animal manure in the mixture. The
highest methane production is found for tomato pulp co-substrate and
for AM:S 1 and AM:S 2 substrate proportion. The influence of the
substrate on these two parameters is G > T > O, which it doesn´t
correspond with the initial contents of the substrates otherwise with the
degradation suffered by animal manure.
Fig. 5 shows the effects of the substrate and substrates proportion on
the C/N ratio obtained in the medium and methane yield obtained. In
this case, clearly it is brought to light that the highest proportion of
Fig. 2. Cumulative methane yield evolution of anaerobic co-digestion assays in different proportions of animal manure and agro-food byproduct silages.
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Industrial Crops & Products 131 (2019) 307–314
animal manure in the substrates mixture leads to the highest methane
yield and consequently the lowest C/N ratio. The synergetic effect more
important in the substrates assayed in this work is given between an-
imal manure and tomato pulp, due to the high methane yield reached
for this agro-food byproduct.
3.5. Kinetic study
Results obtained from First order, Modified Gompertz and Cone
models fits are shown in the Table 4. First order model doesn´t fit well
to the methane volume produced in AM:G 3 and AM:O 3 assays and
Cone model doesn´t fit well to methane volumen obtained in AM:G 3
assay. The fastest biogas productions belong to assays with high animal
manure content (AM:G 2 and AM:T 2) except for olive agro-food by-
product assay. It is observed for parameters obtained in the three ki-
netic models. The parameter that indicates the lag phase (λ) of Gom-
pertz model is higher in results corresponding to the slowest assays
(AM:G 1, AM:T 1, AM:G 3, AM:T 3 and AM:O 3). For these assays, the n
parameter of Cone model is also higher, indicating that they are the
slowest, given that n is related to the presence or absence of the lag
phase. Analyzing the results presented in Table 4, the assays which lead
to higher maximum methane production rates (μm) and higher methane
production rate constants, k (d−1), are AM:O 1, AM:G 2 and AM:T 2
that correspond to the higher proportions of animal manure in the
A.I. Parralejo et al.
Fig. 3. Effects of several substrate mixture proportion and different substrates on VS and COD reduction. S:Substrate analyzed (G, T or O).
mixtures. Some authors (Thanikal et al., 2015) show degradation ki-
netic constant values higher than the obtained in this work, 0.08 d-1 and
0.10 d-1 for tomato and grape, respectively. A drawback to not observe
the positive effect of the animal manure proportion in all the assays was
the presence of straw in important proportions in the agro-food by-
product silages, used to feed the animals, as it is known the straw is a
difficult material to degrade. However, the literature evidences a wide
variability in the kinetic parameters such as results detailed in the
Table 4. Obviously, high values are obtained for anarobic co-digestion
process in semicontinuous regime (Li et al., 2015), nevertheless the
dairy and rabbit manure show a methane production rate constant si-
milar to parameter obtained for AM assay. Other works developed by
other workers (Sousa Lima et al., 2018, Herrmann et al., 2016 and
Njuguna et al., 2016) obtain kinetic parameters for different vegetable
wastes and animal excrements quite similar to those obtained in this
work. The best kinetic adjustment for methane results is obtained with
the Modified Gompertz model, as it can be observed in the Table 4,
Fig. 4. Effects of several substrate mixture proportion and different substrates on evaluated final ammonia nitrogen and alkalinity. S:Substrate analyzed (G, T or O).
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Industrial Crops & Products 131 (2019) 307–314
since the adjustment correlation coefficients obtained are higher.
3.6. Digestate as fertilizer evaluation
According to the RD 999/2017 /Real Decreto 999/2017) about
fertilizer products in Spain, it is stablished a classification of fertilizers
obtained from waste treated by anaerobic digestion processes. The
maximum level allowed for Cd, Cr, Cu and Ni heavy metals are showed
in the Table 5, and a classification of digestate obtained to use as fer-
tilizer has been included.
Digestates obtained in this study are set in the class A for digestates
below anaerobic co-digestion assays employing agro-food byproduct
silages. However, the digestate from animal excrements exclusively
exceed the limit value in the element Cd, the rest of heavy metals
concentrations belong to class B.
A.I. Parralejo et al. Industrial Crops & Products 131 (2019) 307–314

Fig. 5. Effects of several substrate mixture proportion and different substrates on C/N ratio and methane yield. S:Substrate analyzed (G, T or O).

4. Conclusions respectively). However, the behaviour of olive agro-food byproduct is

The anaerobic co-digestion assays developed with animal manure
and agro-food byproduct silages got down the elevate ammonia ni-
trogen content of animal manure digested. When assays employing
agro-food byproduct silages are compared to each other, the lower
values of methane yield belong to the assays with low proportion of
animal manure and the most elevate yield corresponding to the assays
with a proportion of animal manure higher than agro-food byproduct
(348 N L kg VS-1 and 404 N L kg VS-1 for AM:G 2 and AM:T 2,
different because the most elevate methane yield is found in the pro-
portion with a high content of animal manure (398 N L kg VS-1),
probably due to the COD is too elevate in the used proportion for AM:O
2 assay. It is observed a sharp relationship of ammonia nitrogen and
alkalinity parameters with higher proportion of animal manure.
Slower biogas and methane generations have been obtained in as-
says with highest agro-food byproduct content (AM:G 3, AM:T 3 and
AM:O3). A kinetic fit adapted to methane volume produced by first
order, modified Gompertz and Cone models was carried out for assays

Table 4
Parameters estimated according to the models for methane production from anaerobic co-digestion assays with different proportion of agro-food byproduct silages
with animal manure and bibliographic theoretical values of kinetic parameters.
Assay Kinetic model (model parameters)

First order Modified Gompertz Cone

k1, d−1 R2 μm, NL CH4 kg VS−1d−1 λ, d R2 Kc, d−1 n R2

AM 0.07 0.91 2.85 6.27 0.99 0.07 2.67 0.94

AM:G 1 0.04 0.84 1.27 15.54 0.97 0.03 2.51 0.83
AM:T 1 0.04 0.90 1.37 11.02 0.99 0.04 2.65 0.91
AM:O 1 0.07 0.96 1.83 3.39 0.98 0.07 2.60 0.97
AM:G 2 0.06 0.96 1.48 3.95 0.94 0.05 3.07 0.95
AM:T 2 0.06 0.96 1.72 5.45 0.99 0.06 2.65 0.94
AM:O 2 0.05 0.99 1.06 0.004 0.97 0.03 2.07 0.99
AM:G 3 0.02 0.51 0.58 18.96 0.82 0.36 0.05 0.30
AM:T 3 0.05 0.87 1.34 12.38 0.98 0.04 2.84 0.90
AM:O 3 0.06 0.79 1.55 11.96 0.98 0.02 2.61 0.43
Sugarcane bagasse (Inoculum from UASB reactor)a 0.01 0.98 1.50 5.61 0.98 0.03 1.63 0.99
Sugarcane bagasse (Inoculum from UASB-BR)a 0.02 0.97 2.10 5.72 0.98 0.04 1.98 0.99
Sugarcane bagasse (Inoculum from UASB-FBM)a 0.05 0.80 3.05 0.00 0.71 0.05 0.75 0.90
Pig manureb(OLR: 8 g VS L−1) 0.12 0.98 – – – 0.18 1.60 0.99
Dairy manureb(OLR: 8 g VS L−1) 0.07 0.99 – – – 0.10 1.36 0.99
Chicken manureb (OLR: 8 g VS L−1) 0.12 0.92 – – – 0.17 0.95 0.99
Rabbit manureb (OLR: 8 g VS L−1) 0.07 0.99 – – – 0.09 1.21 0.99
Beet silagec 0.09 – – 3.45 – – – –
Pig waste and grass clippingsd – – 1.61 0.70 0.94 – – –

a
Sousa et al. (2018).
b
Li et al. (2015).
c
Herrmann et al. (2016).
d
Njuguna et al. (2016).

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A.I. Parralejo et al. Industrial Crops & Products 131 (2019) 307–314

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Funds): Desarrollo y validación de nuevas tecnologías para la de C, H y N. Método instrumental.
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from agro-industrial activities: kinetic evaluation and synergetic effects. Bioresour.
Secretary of Science, Technology and Innovation of Junta of Technol. 102, 10834–10840.
Extremadura and European Regional Development fund of the EU. Also, Papurello, D., Silvestri, S., Tomasi, L., Belcari, I., Biasioli, F., Santarelli, M., 2016.
it was finnanced with the Project PCJ1006 (2011-2015) “Planta in- Biowaste for SOFCs. Energy Procedia 101 (November 2016), 424–431.
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Real Decreto 999/2017, de 24 de noviembre, por el que se modifica el Real Decreto 506/
sectors between researcher groups and industries” of Junta of
2013, de 28 de junio, sobre productos fertilizantes. BOE nº 296.
Extremadura (Spain). The authors are also grateful to the programme Riggio, V., Comino, E., Rosso, M., 2015. Energy production from anaerobic co-digestion
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