Fuel 107 (2013) 209216

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Fuel
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Biomethane production from maize and liquid cow manure Effect of share
of maize, post-methanation potential and digestate characteristics
Mari Seppl a,, Ville Pyykknen a,1, Ari Visnen b, Jukka Rintala a,2
a
b

University of Jyvskyl, Department of Biological and Environmental Science, P.O. Box 35, FI-40014 University of Jyvskyl, Finland
University of Jyvskyl, Department of Chemistry, P.O. Box 35, FI-40014 University of Jyvskyl, Finland

h i g h l i g h t s
" Co-digestion of cow manure and maize was studied in different share of maize.
" The highest methane yield was achieved, when share of maize was 40% VS of feed.
" Post-methanation potential was low, when methane yield from reactor was high.
" Trace elements content decreased in digestate when the share of maize increased.

a r t i c l e

i n f o

Article history:
Received 23 April 2012
Received in revised form 14 December 2012
Accepted 18 December 2012
Available online 8 January 2013
Keywords:
Anaerobic digestion
Trace elements
Manure
Methane production
Maize

a b s t r a c t
This study investigates the co-digestion of liquid cow manure and maize with different share of maize in
continuously stirred tank reactors (CSTRs). The objective was to determine the methane yield of reactor
and the post-methanation potential of the digestate from different reactor trials. The highest specic
methane yield (259 Nl CH4/kg volatile solids (VSs)) was obtained when the share of maize in the feedstock was 40% (VS) and the second highest specic methane yield was when the proportion of maize
was 60% (VS) (234 Nl CH4/kg VS). The post-methanation potential of the digestate was determined in
batch assays. The minimum value (maize 40%, 75 1 Nl CH4/kg VSfeed) occurred when the methane yield
in the reactor was at a maximum and when the methane yield was at a minimum (maize 67%,
153 46 Nl CH4/kg VS) the potential at a maximum (140 Nl CH4/kg VSfeed). Plant nutrients and trace elements were determined in the feedstock and the digestate. As maize contained fewer nutrients than
manure the concentration of the nutrients and trace elements in the digestate decreased when the share
of maize in the feedstock increased. Thus it seems that even though the CSTR co-digesting maize and
manure can be operated with high organic loading rate (OLR) and short hydraulic retention time (HRT)
a signicant part of the methane yields of the feedstock may be lost, if the post-methane potential is
not considered. Based on the yield results of the reactor trials a biogas plant (270 kW) could produce
energy (methane) almost 2400 MW h, when the reactor operates at OLR 2.5kg VS/m3/d and feedstock
consists of 60% VS of maize and 40% VS of liquid cow manure.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
Biogas production is one of the most promising ways to produce
renewable energy from energy crops and other organic materials.
The benets of anaerobic digestion include the production of two
usable products: biogas and digestate. The biogas can be used in
heat and power production or upgraded to biomethane and used
a vehicle fuel or injected into the gas grid. Agricultural wastes like
Corresponding author. Tel.: +358 40 8053904; fax: +358 14 617 239.
E-mail address: mari.p.seppala@jyu. (M. Seppl).
Present address: MTT Agrifood Research Finland, Animal Production Research, FI71750 Maaninka, Finland.
2
Present address: Tampere University of Technology, Department of Chemistry
and Bioengineering, P.O. Box 541, FI-33101 Tampere, Finland.
1

0016-2361/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.fuel.2012.12.069

cow and pig manures are commonly available and are used feedstock in the biogas production. Manures have a low biogas yield
potential and that is why co-digestion with crop material or organic wastes is useful. Manures provide good buffering capacity and a
wide range of nutrients, while the addition of energy crops increases the energy yield of the process. The anaerobic digestion
is also recycling the nutrients as it captures nutrients from agricultural wastes and energy crops and recycles them to the land in the
form of organic fertilizer, which is a valuable fertilizer for crops.
Maize (Zea mays) is considered a chill-sensitive species with a
relative high temperature required for germination, development,
and dry matter accumulation. Improvements in temperature tolerance would assist maize cultivation in northern Europe. Maize cultivation for biogas production has recently gained interest also in

210

M. Seppl et al. / Fuel 107 (2013) 209216

Table 1
The characteristics of inoculum, liquid cow manure, and maize, variety valdes, standard deviation.

Inoculum
Maizea
Cow manure Ia
Cow manure II

TS (%ww)

VS (%ww)

Ntot (g/l)

NH4N (g/l)

SCOD (g/l)

5.0
16.4
5.7
5.0

3.9
15.2
4.7
4.3

2.5
15b
2.3
1.5

0.15b
0.76
0.45

12.0
134b
13.1
8.6

Specic methane yield

Nl CH4/kg VS

Nl CH4/kg TS

Nl CH4/kg ww

81
360 8
212 18

104
334 7
172 16

4
55 1
10 2

Cow manure I was used in feedstock during days 166, and cow manure II during days 67136.
a
Methane production of inoculum subtracted.
b
mg/g TS.

Finland. In previous studies maize cultivation in southern Finland

have been promising with yields of 16 t total solids (TSs)/ha and
60009000 Nm3 CH4/ha for biogas production [1]. In comparison
average grass cultivation yields have been on average 10 t TS/ha
and energy yields of 12003600 Nm3 CH4/ha [2]. Despite this signicant difference in yield, the total energy balance of the supply
and production chain (LCA) should be considered to make a full
comparison.
Nowadays, maize breeders have developed new biogas maize
varieties that are late maturing, and the TS yields per hectare of
these are over 30 t TS/ha in Central Europe [3]. Silage maize is a
key component of ruminant diets in Central Europe due to its high
yield and energy content and that is why maize has characterized
by good digestibility for biogas production. The digestibility and
nutritional value of the maize (cell wall concentration, composition
and degradability) has been studied, improved and developed for
energy use in biogas production [4,5]. Maize has been the most
popular feedstock for biogas plants in Germany and Austria, because it produces the highest net energy yield per hectare in comparison with other energy crops like sunower, grasses and whole
crop grain silage [6,7]. Mono-digestion of energy crops (i.e. maize
or grass) have been of interest, still the co-digestion of energy
crops and manure is the most stable way to produce methane.
Biogas production has moved towards more and more economically viable activity, which relies on maximizing prot for the sale
of renewable energy. The efcient and reliable process technology
and energy production per unit forces to use a high organic loading
rate (OLR) and short hydraulic retention time (HRT). There has
been little research into the co-digestion of feedstock with a high
share of the energy crops. One research has been made with energy
crops, when share of crop has been 4060% of volatile solids (VSs)
of the feedstock [8]. Co-digestion with maize and manure has been
carried out differing shares of maize and using various HRT and
OLR [9,10] but the post-methanation potential of the digestate
was not measured in any of these studies with maize [810]. The
post-methanation potential is of importance because it describes
the methane potential of digestate and even some energy yield
would be lost from the methane yield of the digestate. In order
to maximize the yield of produced methane, necessary nutrients
must be present in the reactor, which allow using higher OLR, lower HRT and to get higher energy production. To achieve high rates
of methane production, trace nutrients are needed so that microorganisms are supplied with essential nutrients [11,12]. A lack of
trace nutrient have been demonstrated research papers and it is
generally problem in biogas plants which use only energy
crops or some other single substrate material, such as organic
waste [11].
The objective of the study was to investigate the methane yield
of co-digestion of maize and liquid cow manure with increasing
share of maize in the feedstock. The aim was also to measure the
post-methanation potential of the digestate from different share
of maize in the feedstock. The trace elements of the feedstock
and digestate were determined also.

2. Material and methods

2.1. Origin of materials
The maize, variety Valdez (middle late ripening, FAO 290), used
in this study was cultivated at Agrifood Research Finland (MTT) in
Piikki in southern Finland. The cultivation data can be found from
Seppl et al. [1]. After harvesting the maize material was cut using
chopper (SD 180 E, Wolf Garten, Germany) to a particle size of
approximately 3 cm. Subsequently, maize material was then frozen and stored at 20 C in plastic bags in portions equivalent to
three to four days worth of feedstock for the reactors described below. After thawing the maize particles were further reduced using
scissors to particle size about 0.5 cm to assist syringe feeding. Analyses of the maize (Table 1) were performed on melt maize material
after size reduction (to 0.5 cm). The feed mixtures for reactor studies were prepared daily.
Liquid cow manure was obtained from a dairy farm, Kalmari, in
Laukaa Central Finland. The shipment of manure (Cow manure I
and II) was obtained twice from the farm during the reactor runs
and was stored at 4 C. Inoculum was obtained from a mesophilic
farm digester (Kalmari, in Laukaa) processing dairy manure, energy
crops and industrial by-products from a candy factory.

2.2. Experimental set-up

The reactor experiments were carried out in three parallel continuously stirred tank reactors (CSTRs). The reactors (referred as
R1, R2 and R3) were constructed of glass, each with a total volume
of 5 l and a liquid volume of 4 l, stirred continuously at 300 rpm
and incubated at 35 1 C. The reactors were inoculated on day 0
with 4 l of inoculum and when the methane concentration of the
produced biogas rose to 50% feeding of the feedstock was started.
R1 was fed with manure and R2 and R3 with the mixture of manure and maize, using different share of maize and manure (Table 2).
The reactors were fed with a syringe once a day, 5 d per week and
the equivalent volume of the digestate was removed from the reactor. In the reactor R3 the liquid fraction of the digestate was recirculated back to the reactor along with the daily feed, consequently
that the HRT will get the long enough. The liquid fraction was obtained by centrifuging (10 min, 350 rpm) the removed digestate.
The biogas was collected in aluminium gas bags via silicone tubes.
The gas in the tubes was sampled through glass septa.
The assays used to measure specic methane yields were performed in triplicate 1 l glass bottles at 35 1 C. Firstly, 300 ml of
inoculum was added to each bottle followed by the addition of
substrate in a VSsubstrate/VSinoculumratio of 1 and using methods
described by Seppl et al. [2]. The batch assays were incubated
for 80100 d.
The post-methanation potentials of the digestates were measured in batch experiments in triplicate 120 ml serum vials incubated at 35 1 C. The digestate (40 g) was added to the vials

Table 2
Operational conditions, feedstock and digestate characteristic, and methane production in CSTRs standard deviation.
Substrate
Cow manure

Cow manure and maize (low feed)

Cow manure and maize (high feed)

R1
I

R2
I

II

III

R3
I

II

III

(% VS)
(% ww)

0
0

20
8

30
11

40
17

50
18

60
26

67
35

(% ww)
(kg VS/m3 /d)
(d)

2
23

2
27

2
28

2
30

29
2
25

15
2.5
25

6
3
25

(d)
d/HRT

042
1.8

042
1.6

4391
1.5

98140
1.4

042
1.7

4383
1.6

8491, 98126
1.4

TS (%)
VS (%)
SCOD (g/l)
NH4N (g/l)
Ntot (g/l)

5.7
4.7
13.6
0.72
2.3

6.4
5.4
14.5
0.67
2.4

6.2
5.4
11.7
0.45
1.6

6.7
5.9
11.7
0.33
1.7

5.7
5
14.8
0.63
2.2

6.9
6.3
13.9
0.41
1.8

8.3
7.5
14.9
0.32
1.9

TS (%)
VS (%)
SCOD (g/l)
NH4N (g/l)
Ntot (g/l)
NO3N (g/l)
pH
VFAtot (mg/l)
Soluble P (g/l)
Ptot (g/l)
Ktot (g/l)

4.8 0.2
3.7 0.3
10.0 1.0
0.87 0.04
2.5 0.1
0
7.5 0.1
60 104
0.11
0.51
3.38

5.0 0.4
3.9 0.4
10.4 0.4
0.81 0.05
2.5 0.1
0
7.5 0.1
69 119
0.16
0.51
3.47

4.5 0.1
3.4 0.1
12.2 1.0
0.84 0.03
2.3 0.1
0
7.4 0.1
1918 568
0.15
0.46
3.22

4.0 0.1
3.1 0.1
10.3 1.0
0.66 0.03
1.9 0.1
0
7.4 0.1
55 22
0.18
0.37
3.05

4.7 0.0
3.7 0.1
10.5 0.8
0.79 0.04
2.4 0.0
0
7.47 0.1
112 165
0.13
0.44
3.42

4.7 0.2
3.7 0.2
9.1 0.7
0.65 0.05
2.2 0.1
0
7.4 0.1
1079 129
0.15
0.38
3.55

4.9 0.1
4.0 0.1
12.7 1.0
0.53 0.05
1.9 0.1
0
6.9 0.3
4854 798
0.21
0.35
3.4

TS removal (%)
VS removal (%)
CH4 content (%)

16
21
57 2

22
27
52 1

28
36
44 6

41
48
52 2

18
26
51 3

32
41
48 2

41
47
41 9

(Nl/kg VS)

212

241

256

271

286

301

311

(Nl/kg VS)
(Nl/kg ww)

193 2
9.0 0.1
91

198 7
10.7 0.4
82

194 84
10.5 4.5
76

259 6
15.3 0.3
95

221 14
11.0 0.7
77

234 16
14.6 1.0
78

153 46
11.5 3.5
49

Reactor
Feeding regime (FR)
Share of maize

Share of recycled digestate

OLR
HRT
Duration

Feedstock

M. Seppl et al. / Fuel 107 (2013) 209216

Digestate

Calculated specic CH4 yield

of feedstock
Specic CH4 yield of reactor

% Of total CH4 potential

in substrates

211

212

M. Seppl et al. / Fuel 107 (2013) 209216

and they were sealed with butyl rubber stoppers and aluminium
crimps. The contents of the vials were ushed with nitrogen gas
for 3 min to remove residual oxygen. The post-methanation potentials of the digestates were measured from R2 and R3, when the
share of maize in the feedstock was 30%, 40%, 60%, and 67%. The
post-methanation assays were incubated for a total of 125136 d.

2.3. Analysis and calculations

The methane content of the biogas was measured using a gas
chromatograph (Perkin Elmer Arnel Clarus 500, Alumina column
30 m  0.53 mm) with ame-ionization detectors. Operating conditions were: oven temperature 100 C, detector 225 C and injection
port 250 C. Argon was used as the carrier gas (14 ml/min). The
volume of the biogas produced was measured by use of a water
displacement. Volatile fatty acids (VFAs) were measured chromatographically (Perkin Elmer Autosystem XL, HP-INNOWax-column, 30 m  0.32 mm) with ame ionization detector. The
operating conditions were: oven temperature 100160 C (25 C/
min), injection port and detector 225 C. Helium was used as the
carrier gas.
The TS and VS were determined according to Finnish standard
methods [13] and pH was measured with a Metrohm 774 pHmeter. Total nitrogen (Ntot) and ammonium nitrogen (NH4N)
were determined according to the Tecator application note
(Perstorp Analytical/Tecator AB, 1995) with a Kjeltec system
1002 distilling unit (Tecator AB). The soluble chemical oxygen
demand (SCOD) and NH4N from crop samples were analysed
after extraction and ltered with GF50-glass bre lter papers
(Schleicher and Schuell). SCOD was analysed according to SFS
5504 [14].
Water soluble phosphorus (P) was measured (from the digestates) to take 30 g of non-dried slurry and shake with 150 ml of
water for 1 h. The sample was extracted with lter paper (approximately 8 lm pore size) [15]. P content of the extract was measured after peroxodisulphatedigestion [16,17] with a Lachat
autoanalyser. NitrateN was measured from 1:5 water extract
(EN 13652) with a Lachat autoanalyser. Total phosphorus (Ptot)
and total potassium (Ktot) were determined from 0.51.0 g of dried
matter (105 C) with dry ashing (450 C) and dissolved with 100 ml
of 0.2 M HCl. P was determined colourimetrically with the modied ammoniumvanadatemolybdate method [18].
The digestate and feedstock samples for trace element analysis
were rst dried (24 h, 105 C), and then milled in a rotor mill (Pulverisette 14, Fritsch, Germany) and stored at 22 C until analyzed.
The concentration of trace elements was measured using a Perkin
Elmer (Norwalk, CT, USA) Optima 4300 DV ICP-OES using the following default parameters of the instrument: nebulizer ow 0.5
1.0 l/min, auxiliary gas ow 0.2 l/min, plasma gas ow 15 l/min
and plasma power of 1300 or 1400 W [19].
HRT and OLR in the reactor experiments were calculated for the
ve feeding days per week while for practical (weekly) values the
two unfed days during weekends should be considered. Values for
the gas production, feedstock and digestate characteristics are presented either as weekly averages or as averages over the last
2 weeks feeding period. The specic methane yield is given in normal litres per kg VS (Nl CH4/kg VS) i.e. the volume of methane is
based on normal conditions: 273 K and 101.3 kPa. The specic
methane yields from methane potential assays were calculated
as Nl CH4/kg VS added, Nl CH4/kg TS added and Nl CH4/kg wet
weight (WW) added with CH4 content of inoculum subtracted.
The methane yields of post-methanation potential (Nl CH4/
kg VSfeed, Nl CH4/kg TSfeed) assays has been calculated per VS and
TS per feed of CSTR reactor.

3. Results and discussion

3.1. Specic methane yield of maize
In the batch assays the specic methane yields of the maize was
360 8 Nl CH4/kg VS (Table 1), which is in the same range as previously reported for various maize species. In earlier studies the specic methane yield of early, middle and late ripening varieties was
found to be between 350 and 400 Nl CH4/kg VS in Denmark [20]
and in Germany the specic methane yields of silage maize hybrids
(energy maize prototypes) were 282419 Nl CH4/kg VS [21]. The TS
yield of the studied maize was 16.7 t TS/ha and methane yield per
hectare was 5600 m3/ha [1], while the TS yield of maize species
(FAO 300400) have been 2429 t TS/ha, corresponding 7000
9000 Nm3 CH4/ha in Slovenia [22]. The specic methane yield of
the cow manure was 212 18 Nl CH4/kg VS (Table 1), which is in
the upper range of previously reported values, 131230 Nl CH4/
kg VS [8,9,23]. The different specic methane yields of manures
have been a consequence of different feeding intensities and forage
composition (grass/maize silage) for dairy cows [23].
3.2. Co-digestion of maize and liquid cow manure
To evaluate the methane yields and process performance of the
co-digestion of liquid cow manure and maize three parallel CSTR
reactors were operated simultaneously for 136 d. Two reactors
(R2 and R3) were at rst fed with the feedstock containing 20%
(R2) and 50% (R3) of VS maize and one reactor (R1) was run as a
control and fed with manure only. OLR in all reactors was
2 kg VS/m3/d. After 42 d, control reactor (R1) was stopped while
in R2 the share of maize was rst increased to 30% and subsequently to 40% on day 98 of feedstock VS while maintaining OLR
of 2 kg VS/m3/d, HRT 2730 d. In R3 the share of maize was increased from 50% to 60% of feedstock VS and OLR was 2.5 kg VS/
m3/d. On day 84 the share of maize was increased at 67% of the
feedstock VS and the OLR to 3 kg VS/m3/d and HRT was 25 d. There
was a break for feeding the reactors in the days 9297.
The share of maize in the feedstock increased the methane yield
in the reactors. With stable process performance the highest specic methane yield (259 Nl CH4/kg VS) was obtained when the
share of maize in the feedstock VS was 40% (R2) and the second
highest specic methane yield was when the proportion of maize
in the feedstock VS was 60% (234 Nl CH4/kg VS) (Fig. 1, Table 2).
The specic methane yield of manure (R1) was 193 2 Nl CH4/
kg VS and the lowest methane yield was obtained in R3 when
the proportion of maize in the feedstock was 67% (153 46 Nl CH4/
kg VS). The highest specic methane yield (share of maize 40%)
was 95% of the calculated specic methane yield of the feedstock
determined in the batch assays (Table 2). The lowest specic methane yield was due to apparent overloading of the R3, when the
share of maize was 67% of the feedstock VS. In that case, also the
methane content of biogas decreased to 41 9% CH4 and further
to 21% CH4.
The HRT of this study was 23d in cow manure (R1) and 2530 d
in two maize reactors (R2 and R3). Thus the present study as well
as other recent studies [6,8] suggests that mesophilic biogas reactors can be operated with a HRT of 30 d, which is shorter than that
used in full-scale biogas plants. The typical HRT of biogas plants
which treat energy crops together with manure is between 60
and 90 d [24]. The OLR of the reactors in this study was 2
3 kg VS/m3/d and share of maize used in the feedstock was similar
to or lower than that found in previous studies of the same nature
(Table 3) [9,10,25]. The highest single OLR (6 g VS/l/d) has been reported in laboratory scale in CSTR with maize share 50% and gave a
methane yield of 210 l CH4/kg VS. This OLR however was not optimal because of a decrease in methane yield [10]. Furthermore, in

213

M. Seppl et al. / Fuel 107 (2013) 209216

Specific methane yield (Nl CH4/kgVS)

300
R2

250
200
150
100
R3

50
R1

R2

R3

0
0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

days (d)
Fig. 1. Specic methane yields as weekly averages in digestion of manure alone (R1) and co-digestion of cow manure with maize 20%, 30% and 40% of feedstock VS (R2) and
50%, 60% and 67% of feedstock VS (R3). The vertical dashed line indicates the change of the share of maize in the feedstock.

Table 3
The co-digestion of the animal manure and plant material in CSTRs operated within the mesophilic temperature range as reported in literature.
Feedstock (ratio on VS basis)

Reactor volume (l)

OLR (kg VS/m3/d)

HRT (d)

VS removal (%)

Specic CH4 yield (Nl CH4/kg VS)

CH4 (%)

Reference

Liquid cow manure, maize (80:20)

Liquid cow manure, maize (70:30)
Liquid cow manure, maize (60:40)
Liquid cow manure, maize (50:50)
Liquid cow manure, maize (40:60)
Liquid cow manure, maize (33:67)
Pig manure, corn stover (75:25)
Cattle slurry, maize (100:0)
Cattle slurry, maize (67:33)
Cattle slurry, maize (50:50)
Cattle slurry, maize (40:60)
Cattle slurry, maize (50:50)
Cattle slurry, maize (50:50)
Cattle slurry, maize (50:50)
Cattle slurry, maize (50:50)

5
5
5
5
5
5
30
5
5
5
5
5
5
5
5

2
2
2
2
2.5
3
3.84
2
3
4
5
3
4
5
6

27
28
30
25
25
25
16
33
30
28
26
29
22
18
15

27
36
48
26
41
47
46
n.r.
n.r.
n.r.
n.r.
n.r.
n.r.
n.r.
n.r.

198
194
259
221
234
153
210
171
263
304
300
240
220
210
210

52
44
52
51
48
41
67
n.r.
n.r.
n.r.
n.r.
n.r.
n.r.
n.r.
n.r.

This
This
This
This
This
This
[28]
[9]
[9]
[9]
[9]
[10]
[10]
[10]
[10]

study
study
study
study
study
study

n.r. = not reported.

Table 4
The post-methanation potential of the digestate at 35 C calculated per feedstock (TS or VS) of CSTR.
Share of maize (OLR)

30%
40%
60%
67%

(2 kg VS/m3/d)
(2 kg VS/m3/d)
(2.5 kg VS/m3/d)
(3 kg VS/m3/d)

TS (%)

6.2
6.7
6.9
8.3

VS (%)

5.4
5.9
6.3
7.5

full scale digester the OLR was doubled from 2.11 to 4.25 kg VS/m3/
d and stable operation was still achieved at OLR 5.5 kg VS/m3/d
with maize and pig manure [25]. In this study the overloading of
reactor (R3) occurred when the share of maize in the feedstock
was increased to 67% and OLR was 3 kg VS/m3 d. In co-digestion
of crop silage and cow manure the overloading occurred the OLR
7.78 g VS/l/d and the share of crop silage in the feedstock was
81% VS [6], while the process performed well at OLR of 5.15 g VS/
l/d and the cow manure VS:crop silage VS was 29:71 [6]. In the
present study the HRT was 25 d while in the study by Comino it
was ca 42 d [6]. In this study the HRT was too short for efcient
degradation, as the amount of non-degraded matter in the digestate increased leading to an increase in the post-methanation potential (Table 4).
The post-methanation potential of the digestate was determined for digestates when the share of maize was 30%, 40%, 60%

Post-methanation potential
Nl CH4/kg/VSfeed

Nl CH4/kg/TSfeed

Nl CH4/kg/ww

99
75
101
140

86
66
92
127

5
4
6
11

and 67% of feedstock (Table 4). The post-methanation potential

was the lowest (maize 40%, 75 1 Nl CH4/kg VSfeed) when the
methane production in the reactor was the highest
(259 6 Nl CH4/kg VS) and when the methane production in the
reactor was low (maize 67%, 153 46 Nl CH4/kg VS) the postmethanation potential was high (140 Nl CH4/kg VSfeed). In this
study the post-methanation potential increased (from 75 to
140 Nl CH4/kg VSfeed) when the OLR increased from 2 to 3 kg VS/
m3/d. Also in previous study the post-methanation potential has
increased when the OLR have been risen in the reactor [8] and
when the share of energy crops in the feedstock increased [26].
The measured methane yields of the reactor experiments and
post-methanation experiments were about 1020% higher
than the calculated methane yields from the feedstock. The
post-methanation potentials were 2348% of the total methane
yields (methane yields of the reactor plus post-methanation exper-

M. Seppl et al. / Fuel 107 (2013) 209216

SCOD (g/l)

214

15
14
13
12
11
10
9
8
7

R1

R2

R3
R2

R3

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

110

120

130

140

150

110

120

130

140

150

110

120

130

140

150

6000

VFAtot (mg/l)

5000
R3

4000
3000
2000
R2

1000
0
0

10

20

30

40

50

60

70

80

90

100

NH4 -N (g /l)

1.2
1.0

R2

0.8
0.6
R3

0.4
0

10

20

30

40

50

60

70

80

90

100

6.0

TS (%)

5.5

R3

5.0
4.5
4.0
R2

3.5
0

10

20

30

40

50

60

70

80

90

100

Time (d)
Fig. 2. The characteristics (SCOD, VFA, NH4-N, TS% of digestate) of the digestates in digestion of the manure alone (R1) and the co-digestion of the cow manure with maize
20%, 30% and 40% of the feedstock VS (R2) and 50%, 60% and 67% of the feedstock VS (R3). The vertical dashed line indicates the change of the share of maize in the feedstock.

iments). Thus it seems that even though the CSTR co-digesting

maize and manure can be operated with higher OLRs and shorter
HRTs a signicant part of the methane potentials of the feedstock
may be lost, and in the worst case even emitted to the atmosphere.
The relatively high post-methanation potentials also suggest that
degradation of the feedstock was apparently not limited by lack
of nutrients but rather with the retention time and the microbial
population in the reactor. For energy crops digestion with manure,
two-stage digester systems are preferred because of high gas yield
and reduced post-methanation potential. In two stage systems the
rst reactor is high-loaded main reactor and the second reactor is
low-loaded, which treats the digestate from the rst reactor [7,27].
Also the storage tank should be covered to recovery the methane.
The studies [9,10,28] presented in Table 3 did not report the
post-methanation potentials of the digestates.
The recirculation of the digestate back into the reactor (R3) did
not increase the specic methane yield. The recycling of the digestate has been shown to improve the gas production marginally,

because the microbes have been returned to the digester, providing

an additional microbial population [27,29]. Increasing the recirculation of the digestate (only 6% ww of feedstock) the overloading of
R3 might be prevented. Also too short HRT (only 25 d) of the reactor might be one reason for overloading. In the previous studies
with maize the HRT has been 26 d when the share of maize in
the feedstock was 60% VS [9]. Cornell [10] has also used a short
HRT only 1529 d when the reactors were fed OLR 36 g VS/l/d
(50% of VS maize and 50% of VS cattle slurry), but then the methane
yields were only 210240 Nl CH4/kg VS. The HRT was 3343 days
when crop silage was fed as 5171% of the feedstock VS [6]. In both
studies there was no indication of overloading or process
inhibition.
The pH in the reactors was about 7.5 except in the R3 during
overloading (maize 67%) when it dropped to 6.9. SCOD ranged from
9.1 to 12.7 g/l while VFAtot concentrations were generally less than
1100 mg/l except with maize 30% (R2) when the VFAtot concentration was higher (1918 mg/l) as there were some feeding problems

215

M. Seppl et al. / Fuel 107 (2013) 209216

Table 5
The trace element (plant nutrient and heavy metal) content of the maize, liquid cow manure, inoculum and digestate from CSTR in this study and biogas plant in Austria (maize
silage and digestate 100% maize) [30].
Plant nutrients

S
(mg/kg TS)

Mg
(mg/kg TS)

K
(mg/kg TS)

Na
(mg/kg TS)

Maize
Maize silagea
Inoculum
Cow manure I
Cow manure II

534
990
3728
3706
2855

1428
1250
6164
5595
4181

19327
8000
47360
40753
33207

90
50
2751
1616
2496

2089
3590
8322
7346
6046

3659
3840
3597
3195
3336
3169
2915
3610

7200
7488
6704
5875
6254
5427
4375
4120

50673
57327
50600
49273
62617
49370
43377
28020

2413
2680
2599
2705
2764
2009
1722
680

9663
9809
9347
8621
7994
7562
7041
14170

Reactor

Share of
maize

1
2
2
2
3
3
3
Digestate 100%
maizeb

0
20
30
40
50
60
67

P
(mg/kg TS)

Heavy metals

Cd
0.82.7c
(mg/kg TS)

Cu
1.24.3c
(mg/kg TS)

Cr
1.13.7c
(mg/kg TS)

Maize
Maize silagea
Inoculum
Cow manure I
Cow manure II

0.0
n.a.
0.0
0.0
0.0

4.3
4.9
20.8
18.1
14.7

0.0
<0.05
2.7
2.5
1.3

0.0
0.0
0.0
0.0
0.0
0.0
0.0
n.a.

17.5
18.1
20.0
22.3
17.2
16.8
14.2
23.9

4.1
3.6
1.3
1.7
2.8
1.7
1.7
1.9

Reactor

Share of
maize

1
2
2
2
3
3
3
Digestate
100% maizeb

0
20
30
40
50
60
67

Si
(mg/
kg TS)

Ca
(mg/kg TS)

C
(g/
kg TS)

N
(g/
kg TS)

C:N rate

108
80
526
345
430

515
n.a.
3071
1925
960

2991
1800
10597
9675
7225

442
n.a.
413
441
438

16
n.a.
22
20
16

28
n.a.
19
22
27

555
562
618
773
463
536
554
1640

4840
5719
2007
2083
6687
2274
1778
n.a.

12543
12867
11723
10670
11600
9779
8987
9140

416
412
407
403
409
410
430
n.a.

23
24
23
24
25
23
23
n.a.

18
17
18
17
16
18
19
n.a.

Fe
(mg/kg TS)

Ni
26.6c
(mg/kg TS)

Zn
310c
(mg/kg TS)

Pb
1.86c
(mg/kg TS)

As
1.75.5c
(mg/kg TS)

0.0
0.7
2.8
2.6
2.1

21.5
22.4
92.5
85.2
65.1

0.0
n.a.
0.0
0.0
0.0

0.0
n.a.
0.0
0.0
0.0

3.0
3.0
4.7
16.6
2.3
2.0
0.0
3.1

109.1
93.4
90.2
90.7
85.4
80.8
72.5
150.0

0.0
0.0
0.0
0.0
0.0
0.0
0.0
n.a.

0.0
0.0
0.0
0.0
0.0
0.0
0.0
n.a.

0.0 = Under detection limit.

n.a = Not analyzed.
a
Trace element content of maize silage [30].
b
Trace element content of digestate from biogas plant in Austria, feedstock only maize silage [30].
c
Detection limit denition limit.

with syringe during operating the reactor (Fig. 2). Also during the
overloading the VFAtot concentration was high, 4854 798 mg/l,
when 67% feedstock VS was maize (Fig. 2, Table 2), consisting
mainly of acetate (3950 mg/l) and propionate (980 mg/l). The
VFAtot concentrations were about 1% of SCOD concentrations in
reactors, except when the share of maize was 30%, 60% and 67%.
In that case the VFA contributed for 16%, 12% and 38% of the SCOD,
respectively. During the trials in the three reactors, the ammonium
concentration of the digestate varied from 0.53 to 0.87 g/l (Fig. 2),
Ntot from 1.9 to 2.5 g/l indicating decreasing trends during the trials due to the lower nitrogen content of maize as compared to
manure and inoculum (Table 1, Fig. 2). TS and VS removal were calculated for all operational conditions in the reactors. TS removal
was 16% and VS removal 21% of liquid cow manure reactor (R1).
When the share of maize was increased (share of maize 30% in
the feedstock) the TS and VS removals increased being 28% and
36%, respectively and 41% (TS) and 48% (VS), when the share of
maize was 40% in the feedstock (Table 2). The TS and VS removals
were at low level, which is due to short HRT. This can be noticed
also in the relatively high post-methanation potentials (Table 4).
Based on the results of these reactor experiments a biogas plant
(270 kW) could produce about 240,000 Nm3 of methane annually if
the reactor (liquid volume 1000 m3) operates with OLR 2.5 kg VS/

m3/d and the feedstock consists of 60% VS of maize and 40% VS

of the liquid cow manure. The biogas plant would use maize at
3600 tww/a and liquid cow manure 7800 tww/a as a feedstock and
cultivation area of maize would equate to (in boreal conditions)
39 ha (TS yield 15 t TS/ha, VS 15.2%). The energy yield (energy content as methane) of the biogas plant would be about 2400 MWh. If
the produced biogas is upgraded to biomethane to vehicle fuel,
about 190 passengers cars (consumption 6.2 m3 CH4/100 km,
20,000 km/a) could operate on biomethane. The digestate from
the biogas plant could be used as fertilization in the maize cultivation and the methane emissions to atmosphere, inherent from liquid cow manure could be avoided.
3.3. Trace elements
The trace elements were determined in the feedstock and in the
digestate at the end of the each feeding regime. As maize contained
fewer trace elements than the manure the concentration of the
trace elements in the digestate decreased when the share of maize
in the feedstock increased (Table 5). In an earlier study the maize
silage contained fewer plant nutrients and heavy metals than the
digestate (Table 5) [30]. The concentrations of S, Mg, K, Na, P, Si,
and Ca in the digestate were the lowest in the share of maize

216

M. Seppl et al. / Fuel 107 (2013) 209216

67% VS (R3) (Table 5). In previous studies the deciency of the

trace elements was clear from the relatively low OLR, when the
maize silage has been the sole substrate. The lack of the trace elements does not occur immediately but only after a certain operation time [11]. The lack of the trace elements has been reported
in mono-digestion of grass-clover [31], but especially in case of
the mono-digestion of maize the lack of the trace elements is reported to be a challenge with many biogas plants [11]. Adding
the trace elements to the reactor should attain the long-term process stability, high methane production and also operation at higher OLR [11]. The co-digestion of maize with liquid manure helps to
maintain the trace elements concentration in a sufcient level of
the methane production as manures typically contain higher nutrient and trace elements concentration [32]. On the other hand, as
crops contains different concentrations of trace elements, which
depend on the concentration of soil nutrients and regional variation of soil [33], the need of the trace nutrients in biogas plants
should be assessed case dependently.
In previous studies the most limiting trace elements in biogas
production have been reported to be cobalt (Co), molybdenum
(Mo), selenium (Se) and nickel (Ni) [11,34]. The Co, Mo and Se were
not analyzed in the digestate in this study, but Ni concentration of
the digestate were 216.6 mg/kg TS (Table 5) corresponding to
0.090.66 g/m3. Ni addition stimulated both the biogas production
and the methane content of the biogas in batch study, where cattle
dung was used as a substrate [35]. The optimum concentration of
Ni for batch cultures of methanogens was reported to range between
0.012 and 5 g/m3 [36]. Also in previous studies the appropriate addition of Ni and Co of CSTR experiment (model substrate maize silage)
gives an opportunity to increase the OLR in the reactor [32].
4. Conclusions
 The highest methane yield 259 Nl CH4/kg VS was achieved
when the share of maize was 40% of the feedstock VS and the
OLR was 2 kg VS/m3/d.
 If the biogas process has operated with a short HRT and high
OLR, the post-methanation potential has to be taken into
account. When planning biogas plants have to be ensured that
the process runs at stable conditions and that the produced
methane can be used efciently.
 Maize contains fewer trace elements than liquid cow manure.
Anaerobic digestion of maize and manure operates effectively
without the addition of trace element.

Acknowledgements
This study was supported by grants from the University of
Jyvskyl and the Fortum Foundation. The authors wish to thank
the Kalmari biogas farm for providing the liquid cow manure and
inoculum, and Agrifood Research Finland (MTT) in Piikki for providing the maize material. Furthermore, Mervi Koistinen is kindly
acknowledged for her help in the laboratory.
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