Biotechnology and Bioprocess Engineering 16: 1044-1052 (2011)

DOI 10.1007/s12257-011-0117-4

RESEARCH PAPER

Anaerobic Co-digestion of Swine Manure with Energy Crop Residues

Mara Jos Cuetos, Camino Fernndez, Xiomar Gmez, and Antonio Morn

Received: 22 March 2011 / Revised: 2 June 2011 / Accepted: 10 June 2011

The Korean Society for Biotechnology and Bioengineering and Springer 2011

Abstract Anaerobic co-digestion involves the treatment

of different substrates with the aim of improving the
production of biogas and the stability of the process. In this
research, co-digestion of swine manure (SM) and energy
crop residues (ECRs) was studied. The mixtures evaluated
contained SM combined with maize (Mz), rapeseed (Rs) or
sunflower (Sf) residues. Batch and semi-continuous experiments were performed to determine methane (CH4) yields
and the behavior of reactors while co-digesting agricultural
wastes. Three different proportions of ECRs were tested in
batch experiments for co-digestion with SM: 25, 50, and
75% volatile solids (VS). On the basis of the results obtained from batch tests, a mixture with a 50% ECR content
was selected for the second stage of the study. Mesophilic
reactors with a 3 L working volume were used for semicontinuous experiments. The hydraulic retention time (HRT)
was set at 30 days and the reactors were kept under these
operational conditions over four HRTs. The addition of
ECR to the co-digestion system resulted in a major increase
in the amount of biogas produced daily. The highest biogas
yield was obtained when co-digesting Rs (3.5 L/day),
although no improvement was observed in specific gas
production from the addition of the co-substrate.
Keywords: anaerobic digestion, rapeseed, maize, swine
manure, sunflower

1. Introduction
The production of renewable energy, utilization of by-

Mara Jos Cuetos, Camino Fernndez*, Xiomar Gmez, Antonio Morn

Institute of Natural Resources (IRENA), University of Leon, Leon 24071,
Spain
Tel: +34-987-291-841; Fax: +34-987-291-839
E-mail: cferrd@unileon.es

products as fertilizers, a reduction in agricultural wastes,

and the prevention of environmental pollution are several
of the characteristics that have promoted the industrial
application of anaerobic digestion for the treatment of
wastes. Anaerobic digestion is currently a well-established
technology in Europe with large-scale systems developed
primarily in countries such as Germany and Denmark [13]. Nevertheless, studies need to be initiated to improve
current technologies and encourage the use of biogas [4-7].
Anaerobic co-digestion is defined as a treatment that combines different types of wastes with the aim of increasing
biogas yields. This improvement is achieved by balancing
the nutrient content and reducing the negative effects on
the digestion process of toxic compounds. Moreover, it
leads to a more efficient use of equipment and cost-sharing
by processing multiple wastes in a single facility [8].
The production of swine in the European Union (EU) is
a major agricultural industry. In 2009 there were 152
million head of swine in the EU, with 25 million produced
in Spain [9]. It is estimated that 46 million m3 of swine
manure (SM) are produced annually in that country [10].
SM is a plentiful source of organic matter that may be
used as feedstock in anaerobic digesters. It has a high
buffering capacity, which may protect the digestion process
against possible failures due to any build-up of volatile
fatty acids (VFAs) and any consequential drop in the pH of
the system. SM also contains a wide variety of nutrients
necessary for bacterial growth. However, another of its
characteristics is its high nitrogen content, which may
represent a digestion inhibition risk as the result of high
levels of ammonia within the digester when manure is
digested individually [11,12].
Methanogens are the organisms least tolerant and most
likely to cease growing due to ammonia inhibition [13].
Free ammonia concentration has been suggested as the
principal factor responsible for inhibition problems. The
degree of inhibition depends mainly upon total ammonia

Anaerobic Co-digestion of Swine Manure with Energy Crop Residues

concentration, pH, and temperature. Different threshold

values for free ammonia concentrations have been reported
by several authors, ranging from 200 mg of ammonia
nitrogen (N-NH3)/L [14] for unadapted populations, to
higher values (700 ~ 1,100 mg N-NH3/L) for microorganisms that have previously adapted to their presence
[12,15]. For this reason, manure should preferably be codigested with wastes that have a high carbon content, so as
to improve the carbon/nitrogen (C/N) ratio [16,17].
It has been estimated that 104,731 tons/year of energy
crops are produced in Spain [18]. Agricultural wastes may
be present in large quantities when no market is available.
In such cases, wastes may remain in the fields after
harvesting operations. These wastes may be suitable for
anaerobic co-digestion with manure, as a result of their
considerable carbon content. However, the major drawback
when this type of materials is being digested is the lignin
content, which may hinder anaerobic degradation. Various
pre-treatments have been applied in attempts to improve
the biodegradability of agricultural wastes [19]. A reduction in particle size by mechanical pre-treatment leads to an
increase in the specific surface available and thus improves
gas production in the digester and reduces the technical
digestion time [20].
The aim of the research being reported here was to study
the performance of reactors co-digesting SM with three
types of energy crop residues (ECRs) from: maize (Mz),
rapeseed (Rs) and sunflowers (Sf). For this purpose, the
influence of the co-substrates on biogas yield under batch
conditions was first evaluated. Thereafter, the performance
of reactors under semi-continuous operation was studied
using a mixture of SM and ECR as substrate.

1045

with a working volume of 250 mL. Solutions were kept in

a thermostatic water bath maintained at a mesophilic
temperature (35 1C) and continuously stirred at 250 rpm
by magnetic bars. Batch experiments were performed to
determine the biochemical methane (CH4) potential from
individual substrates and mixtures of crop residues with
SM. The duration of experiments was determined by the
point at which biogas production stopped completely. All
experiments were performed in duplicate and blank reactors
were also used, to determine the background gas productivity of the inoculum. In each reactor, substrate and
inoculum were introduced and when necessary, tap water
was added to attain a total volume of 250 mL. No nutrient
solution was added to biochemical CH4 potential tests. The
mass of ECRs added to digestion tests was 1.8 g of volatile
solids (VS), which was the maximum quantity that could
be added to digesters without causing problems with stirring or agglomeration. The inoculum used was diluted to a
concentration of 5.0 g/L VS, with 125 mL of prepared
inoculum added for digestion tests and 85 mL for codigestion tests. Co-digestion experiments were performed
with different ratios of manure to ECR. SM and ECR
mixtures were prepared with proportions of 25, 50, and
75% VS. These percentages represent the VS content of
the ECR in relation to the total quantity of VS in the
mixture.
Gas and liquid samples were taken twice a week to
measure the composition of the biogas, the pH, and the
concentration of VFAs. Gas production was measured daily using a liquid displacement device. The solution used
was saturated to 75% with sodium chloride and acidified to
pH 2. Adjustments to the pH were made by the addition of
an alkaline solution during the first few days of the experiments whenever the pH value fell below 7 units.

2. Materials and Methods

2.1. Substrates and inoculum
Three ECRs were used in this study: Mz, Rs and Sf. These
residues were collected during the harvest period (September-October) from farms located in Castilla y Len (Spain).
Specimens were chopped up with a Viking (GB370) agricultural chopper and then further milled using a laboratory
grinder to reduce their particle size to less than 3 mm. SM
was obtained from a livestock farm located in Alcoba de la
Ribera (Len, Spain). The manure was stored at 4C until
required for use. The anaerobic sludge used as inoculum
was obtained from the wastewater treatment plant of the
city of Leon (Spain). Details of the operating conditions for
this digester are available elsewhere [21].
2.2. Batch experiments
Batch digestion tests were carried out in Erlenmeyer flasks

2.3. Semi-continuously operated reactors

Four completely stirred tank reactors were used to evaluate
the digestion of SM and mixtures with ECRs. The reactors
were made of methacrylate, being cylindrical in shape and
having a heating jacket through which water was circulated. They were stirred continuously with two triple-blade
propellers to prevent particulate material from floating. The
reactors were labelled in accordance with the substrates fed
into them. Thus, R_SM referred to the reactor treating SM,
and similarly for the various mixtures with ECRs. A mixture with a 50% content of ECR was selected for the second
stage of the study on the basis of the results obtained from
batch tests in the previous experimental stage.
Reactors were initially started up with an adaptation
period intended to acclimatize anaerobic microorganisms
to conditions with inhibitory concentrations of ammonia
[22]. The reactors were operated with a hydraulic retention

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Biotechnology and Bioprocess Engineering 16: 1044-1052 (2011)

time (HRT) of 50 days over a 30-day period. These conditions led to the application of an organic loading rate
(OLR) of 0.7 kg VS/m3/day for R_SM, while the codigesting systems had an OLR of 1.4 kg VS/m3/day. Thereafter, the HRT was decreased to 30 day, equivalent to an
OLR of 1.2 kg VS/m3/day for R_SM, and 2.3 kg VS/m3/
day for co-digesting reactors. These operating conditions
were evaluated for four consecutive HRTs.
The daily production of biogas was measured using a
reversible liquid displacement device provided with a wet
tip counter. Gas and liquid samples were taken twice a
week to monitor biogas composition, pH, chemical oxigen
demand (COD), total solids (TS), VS, ammonia, and VFA
concentrations.

Table 1. Chemical characteristics of inoculum and substrates

2.4. Analytical techniques

Kjeldahl nitrogen, TS, VS, COD, alkalinity, ammonium,
and pH were determined in accordance with American
Public Health Association (APHA) standard methods [23].
Cellulose, hemi-cellulose, and lignin content were estimated by an analysis in duplicate of neutral detergent fiber,
acid detergent fiber, and crude fiber [24] in ground samples
using an ANKOM 200 Fiber Analyzer. Total organic
carbon was determined by following the Walkey-Black
method [25], involving oxidation with 1 N potassium
dichromate (K2Cr2O7) and 96% sulphuric acid (H2SO4)
solutions for 30 min. The organic carbon content was
calculated by back-titration with a solution of 0.5 N
Fe(NH4-SO4)26H2O and organic carbon was determined
on the basis of an organic matter content to organic carbon
ratio of 1.7241. The organic carbon was subsequently
divided by the total nitrogen to obtain the C/N ratio.
Biogas composition was analysed using a gas chromatograph (Varian CP 3800 GC) equipped with a thermal
conductivity detector. A column 4 m long, packed with
HayeSepQ80/100, followed by a molecular sieve column
1m long, was used to separate CH4, CO2, N2, H2, and O2.
The carrier gas was helium and the columns were operated
at 331 kPa at a temperature of 50C. VFAs were analysed
using a gas chromatograph (Varian CP 3800 GC) equipped
with a Nukol capillary column (30 m 0.25 mm 0.25
m) from Supelco (Bellefonte, PA, USA) and a flame ionization detector. The carrier gas was helium. The temperature of the injector was 250C and the temperature of the
oven was initially set at 150C for 3 min and thereafter
increased to 180C.

3. Results and Discussion

3.1. Batch digestion experiments
The characteristics of the ECRs and SM used are given in

Characteristics
C/N
TS
VS
TKN(g/kg TS)
NH4+ (mg/L)
Cellulose (%)
Hemicellulose (%)
Lignin (%)

I
6.2
20.1a
13.6a
46.7
650

SM

Mz

1,010.2 64.7
1,055a 672b
1,035a 644b
1,032.2
8.2
1,282

23.6

29.7

7.4

Rs

Sf

60.1
397b
363b
8.6

40.5
21.6
16.6

52.6
888b
771b
9.7

23.7
15.2
12.0

Values expressed in g/L.

Values expressed in g/kg.

Table 1. The ECRs showed a high C/N ratio, which indicates that these substrates may be suitable for co-digestion
with SM. However, the lignin content of these substrates
was also high. The presence of lignin may hinder the
break-down of organic matter, thus resulting in a fraction
of the substrate possibly remaining unavailable for microbial break-down.
Fig. 1A shows the results from batch digestion tests. SM
and Mz gave the highest CH4 yields from the batch digestion of individual substrates. The lower values obtained
for Rs and Sf may be associated with the higher lignin
content of these substrates. The anaerobic microflora probably could not access the organic material encapsulated
within lignin structures.
With regard to the shapes of curves obtained from digestion tests of individual substrates, an extended lag phase
was observed when digesting SM. This result may indicate
unfavourable conditions for anaerobic microflora. However,
once the organisms acclimatized, the digestion proceeded
at a high rate. A similar trend was reported by Lobato et al.
[26] when evaluating the digestion of SM under batch
conditions at a mesophilic temperature. These authors
obtained values for the two manures that they tested of 276
10 and 512 19 mL CH4/g VS added. Under similar
conditions Chae et al. [27] reported a yield of 403 mL
CH4/g VS added. The results obtained in the work being
reported here were in accordance with those findings, with
a value of 357 34 mL CH4/g VS added.
The CH4 potential of manure comes from the digestion
of the organic components in the feces and in the straw
used as bedding material, which are mainly carbohydrates,
proteins, and lipids. In theory, the CH4 yield from carbohydrates (415 mL/g VS) is lower than that from proteins
(496 mL/g VS) or lipids (1,014 mL/g VS) [28]. In view of
this, although no analysis of the protein and lipid content
was performed during the experiments, it can be assumed
that this substrate was composed mainly of carbohydrates
and proteins.
In contrast, when particulate substrates are being di-

Anaerobic Co-digestion of Swine Manure with Energy Crop Residues

1047

Fig. 1. (A) Methane (CH4) yield of individual substrates. Symbols: (--), maize (Mz); (--), rapeseed (Rs); (- -), sunflower (Sf); (- -),
swine manure (SM). CH4 yield of co-digestion mixtures of SM with (B) Mz, (C) Sf, and (D) Rs. (--), mixtures were prepared with a
content of volatile solids (VS) of 25% of energy crop residue (ECR); (- -), 50% VS of ECR; and (--), 75% VS of ECR.

gested, the process is generally limited by the hydrolysis

phase [29,30] and it may then be described by first order
kinetics [31]. This is the case for the ECR curves obtained
from these digestion tests, with the exception of Sf, as may
be seen in Fig. 1A. The shape of Rs and Mz curves match
those for the category of slowly degradable substrates, as
defined by Labatut et al. [32]. However, in the case of Sf
the shape of the curve indicates a high break-down rate for
the material readily accessible to micro-organisms, but the
lignin content of this substrate is likely to have hindered
the breaking down of a considerable amount of the organic
matter, thus leading to low CH4 yields.
CH4 yield curves for mixtures studied are also shown in
Figs. 1B, 1C, and 1D. A reduction in the lag phase was
observed when co-substrate was added. The shape of the
curves can in all cases be seen as sigmoidal. This characteristic shape has also been reported in batch digestion tests
by several authors [33,34]. The lowest CH4 yield was
obtained from co-digestion with Sf in a proportion of
75:25. As previously commented, this substrate was
characterized by high lignin content and thus a low CH4
yield was to be expected. The addition of SM to Sf may
have resulted in a synergistic effect, since the outcomes of
the digestion tests with mixtures having 25:75 and 50:50
proportions were similar. This behavior was also observed
with the co-digestion of the other two ECRs, the mixtures

thus having similar values for CH4 yield with different

proportions of ECR.
Although co-digestion with Rs and Mz led to similar
values for CH4 yield, differences were observed in the
pattern of break-down. The addition of the co-substrates
produced preferential breaking-down, which could be
observed through changes in the rate of CH4 production.
The addition of agricultural residues as co-substrates has
been reported to increase the production of biogas significantly [35]. Differences between our results and a previous report may be explained by the diminutive particle size
used by Wu et al. [35]. The crop residues (corn stalks, oat
straw and wheat straw) were first cut into small sections
and then ground into fine particles, under 40 mesh size
(0.422 mm) before being added to digesters, thus improving the degradability of the co-substrate particles. However,
the present results are in accordance with those reported in
the literature when considering batch conditions. Fujita et
al. [36] studied the co-digestion of corn and manure in
mesophilic conditions (reporting a value of 267 L CH4/Kg
VS added) and thermophilic conditions (reporting a value
of 305 L CH4/Kg VS added). Varying values for the codigestion of manure with wheat straw can be found in the
literature, with reports of 322 L CH4/Kg VS added [37]
and 148 L CH4/Kg VS added [28].
Fig. 2 shows the details of VFA concentrations for

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Biotechnology and Bioprocess Engineering 16: 1044-1052 (2011)

Fig. 2. Volatile fatty acid (VFA) profiles obtained from the digestion of Sf (A), Mz (B), and SM (C). Symbols: (- -), acetic acid; (- -),
propionic acid; and ( ), butyric acid.

digestion and Fig. 3 shows co-digestion tests. In the case of

the digestion of Rs, no detectable amount was found; i.e.,
the values were below the detection limit. This substrate
was also characterized by its high lignin content, as was Sf.
From Fig. 2 it may also be observed that low values were
measured during the digestion of this substrate. In the case
of Mz, VFA concentrations were slightly higher, but only
during the early days of the experiment. These results were
influenced by the limitation imposed by the hydrolysis
stage. This was not the case for SM. Higher concentrations
of these acids were detected at the beginning of the experiment. As the digestion proceeded VFAs were continuously
detected over a 25-day period. On the basis of these results,

the VFA concentrations reported in co-digestion tests (Fig.

3) may be attributed principally to the acidification of SM.
All co-digestion tests presented lower values for VFAs, as
the amount of SM in the mixtures was reduced in accordance with the ratios being tested.
Acetic acid was the main VFA present in all the batch
tests, with the exception of the digestion of Rs, where
VFAs were below the detection limit. Values for the VFA
concentrations measured in the co-digestion tests at a ratio
of 75:25 were much higher than those obtained from the
digestion of SM on its own. The extended lag phase
observed for CH4 production when SM is being digested
may be attributed to nutrient imbalance rather than to a

Fig. 3. VFA profiles obtained during batch experiments from co-digestion of mixtures of SM with Rs (A1) 25% VS of ECR, (A2) 50%
VS of ECR, (A3) 75% VS of ECR, Sf (B1) 25% VS of ECR, (B2) 50% VS of ECR, (B3) 75% VS of ECR, and Mz (C1) 25% VS of
ECR, (C2) 50% VS of ECR, (C3) 75% VS of ECR. Symbols: (--), acetic acid; (- -), propionic acid; and ( ), butyric acid.

Anaerobic Co-digestion of Swine Manure with Energy Crop Residues

1049

build-up of VFAs.
On the basis of the results obtained from batch tests,
mixtures containing 50% of ECR were chosen for performing evaluations under semi-continuous operation. This
decision was based on the minor modifications observed in
CH4 yield obtained at this ratio as compared to the yield
from the digestion of SM alone, and also to avoid possible
solid build-up problems which may occur under continuous operation.
3.2. Semi-continuous operation
Daily biogas production is shown in Fig. 4A. During the
adaptation period (days 1 ~ 30), reactors were characterized by a limited output of biogas, which was in accordance
with the low OLR applied. Setting the operational conditions at a HRT of 30 days resulted in a gradual increase in
the production of biogas for all reactors evaluated. Codigestion systems presented a higher production of biogas
in comparison to that of R_SM. Fig. 4B shows the specific
gas production (SGP) for reactors. No improvement in the
SGP was observed from the addition of the co-substrates.
As such, the synergistic effects which may have been present under batch conditions were not registered under
semi-continuous operation. The higher production of
biogas obtained from co-digesting systems was explained
by the increase in OLR arising from the ECRs.
Biogas production was steady in all systems from day 60
onwards, with the exception of R_Rs. Over this period the
composition of the gas showed minor variations. However,
co-digestion with Rs was characterized by an upward trend
in the production of biogas from day 90 onwards. This
behavior is likely to be due to an accumulation of VS

Fig. 4. (A) Daily biogas production and (B) specific gas

production (SGP) for reactor treating swine manure (R_SM) (--)
and co-digesting systems: R_Rs ( ), R_Sf ( ), and R_Mz
( ).

within the reactor being transformed at a later stage, thus

making a larger amount of material available for microbial
break-down.
Table 2 shows the average values for parameters recorded during days 70 ~ 150. Co-digestion with Rs presented
the highest figure for average daily biogas production. As
stated previously, this high value was explained by the
accumulation of solids within the reactor, detected during

Table 2. Main operational parameters of digesters studied R_Rs, R_Mz, R_Sf, and R_SM under semi-continuous operation (HRT of 30
days)
Parameters*
Biogas (L/day)
SGP biogas (m3/kg/VSfeed)
CH4 yield (m3/kg/VSfeed)
CH4 (%)
CO2 (%)
TS (g/L)
VS (g/L)
VS removal (%)
TS (g/L)end
VS (g/L)end
COD (g/L)
pH
Alkalinity (g/L)
NH4+-N (mg/L)
NH3-N (mg/L)

R_SM
1.6 0.1
0.46 0.03
0.33 0.03
71.5 1.2
28.5 1.2
39.0 3.9
22.2 2.9
36.6 8.4
35.0 0.4
20.0 0.3
28.6 4.9
8.0 0.1
17.5 0.4
4,580 254.2
418.9 55.5

*Means and standard deviations values obtained from days 70 to150.

R_Rs
3.5 0.3
0.51 0.04
0.34 0.03
65.9 3.2
34.1 3.2
60.1 9.1
41.8 9.1
40.3 13.0
136.0 4.1
113.0 4.5
61.2 3.7
7.9 0.1
18.1 1.0
.4438 281.0
332.8 52.4

R_Sf
2.7 0.2
0.39 0.03
0.26 0.02
65.0 2.9
35.0 2.9
60.5 3.6
38.7 2.5
44.7 3.6
87.0 3,2
55.0 2,2
52.7 2.6
7.9 0.1
17.3 0.4
4,368 331.5
319.5 28.7

R_Mz
3.2 0.1
0.46 0.02
0.30 0.01
63.1 2.0
36.9 2.0
54.8 1.4
33.4 1.9
52.3 2.7
91.0 5.0
70.0 4.8
41.2 2.5
7.9 0.1
17.0 0.5
4,440 259.1
346.6 32.4

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Biotechnology and Bioprocess Engineering 16: 1044-1052 (2011)

the experiment due to the formation of a floating layer of

ECR particles. Such an accumulation of particles has previously been reported, with descriptions of the formation of
a crust layer in the upper part of the liquid during experiments involving the co-digestion of energy crops and crop
residues with cow manure [38]. At the end of the experiments, the reactors were dismantled and their contents
were thoroughly homogenized. The amounts of TS and VS
measured at the end of the experiment are also presented
in Table 2. The homogenized slurry presented a much
higher concentration of solids than what was periodically
measured in the effluent obtained from sampling ports. In
this way, the hypothesis of a late breaking-down phase in
system R_Rs was corroborated, since the ECR stayed in
the reactor for a period longer than the 30 days established.
Although all co-digestion systems presented some accumulation of solids, Mz and Sf residues seemed to be broken
down completely during the time they remained in the
reactor and the particles that accumulated did not increase
biogas production at a later stage.
The figures for CH4 yield shown in Table 2 were similar
to results reported in the literature with respect to experiments in which manure was co-digested with agricultural

wastes. Fujita et al. [36] obtained a value of 0.21 m3/kg VS

fed with manure and corn, while Fischer et al. [39] noted
values in the range of 0.22 ~ 0.24 m3/kg VS fed for
mixtures of manure and wheat straw. Lehtomki et al. [38]
tested a mixture with 30% of straw in the feedstock combined with cow manure and achieved a CH4 yield of 0.21
m3/kg VS fed. Comino et al. [40], during trials with a cow
manure and crop silage mixture, recorded CH4 yields
between 237 and 249 L CH4/kg VS.
The reactors showed a continuous increase in the ammonium concentration as the HRT was reduced from 50 to 30
days. Steady values for ammonium were attained once the
reactors completed the second HRT at 30 days. Free ammonia concentrations were maintained within the normal
ranges for anaerobic digestion of wastes with a high nitrogen content, as stated by several authors [14,41].
The evolution of VFAs is presented in Fig. 5. VFAs were
below the detection limit during the adaptation period. The
increase in the OLR resulted in a rapid build-up of VFAs
during the first HRT cycle at 30 days. The reactors presented high concentrations of acetic acid. This period was
also characterized by an increasing production of biogas
from all reactors. Thus, this upward trend may be explain-

Fig. 5. VFA concentration of digesters studied A) R_Rs, B) R_Mz, C) R_Sf, and D) R_SM. Symbols: (--), acetic acid; (- -), propionic
acid; and ( ), butyric acid.

Anaerobic Co-digestion of Swine Manure with Energy Crop Residues

ed by the availability of acid intermediaries which were

used for further biogas production. Although the steep
increase in the OLR produced an imbalance in microbial
consortia, microorganisms were capable of adapting to new
conditions. As a result, VFAs decreased continuously during
the second HRT evaluated, and finally the predominant
acid types showed minor variations during the third and
fourth HRT periods.

1051

7.
8.

9.

4. Conclusion

10.

Co-digestion of SM with three different types of ECR

derived from Rs, Mz, and Sf was studied under batch and
semi-continuous conditions. SM and Mz gave the highest
CH4 yields in batch digestion tests of individual substrates.
The lower amounts obtained from the other two substrates
may be associated with their higher lignin content.
During semi-continuous operation, no improvement in
the SGP was observed from the addition of the co-substrates. Synergistic effects which may have been present
under batch conditions were not noted during semi-continuous operation. An accumulation of solids within the
reactor was observed in all reactors tested. This accumulation led to an upward trend in biogas production for the
co-digesting system with Rs residue; however, this was not
the case for the Mz and Sf co-digesting reactors.

11.
12.
13.
14.
15.
16.
17.

Acknowledgements
This work was supported financially by project PSS
12000-2008-57/PROBIOGAS of the Spanish Ministry of
Science and Innovation and by the European Regional
Development Fund [ERDF].

18.
19.
20.
21.

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