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Agriculture, Ecosystems and Environment

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

Chemical properties of anaerobic digestates affecting C and N dynamics in

amended soils
José Antonio Alburquerque ∗ , Carlos de la Fuente, María Pilar Bernal
Department of Soil and Water Conservation and Organic Waste Management, Centro de Edafología y Biología Aplicada del Segura, CSIC, P.O. Box 164, 30100 Murcia, Spain

a r t i c l e i n f o a b s t r a c t

Article history: The optimisation of digestate recycling as fertilisers, based on both environmental and agricultural crite-
Received 11 October 2010 ria, requires an evaluation of the effects on C and N dynamics in soil. In the present paper, six digestates
Received in revised form 1 March 2011 from several anaerobic co-digestion experiments, using pig or cattle slurry as the main substrate, were
Accepted 14 March 2011
evaluated in short-term incubations in soil.
Available online xxx
Digestate properties such as dissolved organic-C (DOC), biochemical oxygen demand (BOD) and
digestate organic-C mineralised in the soil during the first 7 days represented properly the digestate
Keywords:
biodegradability. These, together with their ratios with respect to the total nitrogen (TN) concentration
Anaerobic digestion
Animal slurries
in the digestate, were reliable parameters with respect to defining the C and N dynamics in the soil
Organic matter mineralisation and hence the N-fertiliser potential of the digested materials. Therefore, highly biodegradable digested
Biodegradability materials, represented in the present study by digestates from cattle slurry–glycerine mixtures were
Nitrogen immobilisation not suitable for agricultural use as they caused a high CO2 –C production and led to N-immobilisation
and/or denitrification after their application to soil. Contrastingly, for less biodegradable digested mate-
rials (BOD5 d < 6.0 g O2 L−1 fresh weight, DOC < 5.5 g L−1 fresh weight and DOC/TN < 1.5), less CO2 –C was
evolved and ammonium was rapidly nitrified in soil—being an available N source for crops.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction Digestates also present advantages in comparison with untreated

Nowadays, there is increasing interest in Europe in the imple-
mentation of anaerobic digestion in productive sectors such as
livestock and agroindustry, where vast amounts of biodegradable
wastes (animal manure and slurries, agricultural and food industry
wastes, etc.) must be adequately managed, due to the demand for
renewable energy. Co-digestion can enhance the energy produc-
tion from animal manure and slurries, as co-digestible materials
such as slaughterhouse wastes, glycerine and energy crops or
silage can increase the amount of biodegradable organic matter,
dilute potential toxic compounds, improve the nutrient balance
and favour synergistic effects of microorganisms, thereby raising
biogas production (Álvarez et al., 2010; Holm-Nielsen et al., 2009;
Pesta, 2007). However, the sustainability of biogas production
also depends on an appropriate end-use of the digested material
(digestate)—which should be treated, disposed of or re-used in a
proper way, avoiding any negative environmental impact.
The use of digestates as organic fertilisers in agricultural sys-
tems seems the best option for their recycling since they contain
considerable amounts of residual organic-C and plant nutrients.
∗ Corresponding author. Tel.: +34 968 396200; fax: +34 968 396213.
E-mail addresses: jalburquerque@cebas.csic.es, jalburquerquemendez@yahoo.es
(J.A. Alburquerque).
waste, such as greater microbial stability and hygiene and a higher
amount of N present as ammonium (Al Seadi, 2002; Holm-Nielsen
et al., 2009). So, land spreading of digestate can lead to benefits
if integrated into good agricultural practices, by controlling the N
application rate and heavy metal load, and by securing digestate
hygiene (Al Seadi, 2002; Fuchs et al., 2008; Holm-Nielsen et al.,
2009). Nevertheless, the biodegradability of these materials, which
determines organic matter (OM) mineralisation and thus nutrient
turnover in soil, is not well characterised.
The complete exhaustion of the most labile organic fraction
during the anaerobic process, in order to obtain digestates with
a high stability degree, is not easy to achieve at the industrial level,
the main objective of the anaerobic co-digestion being the pro-
duction of a high rate of biogas, rich in methane. This process is
conditioned mainly by the composition of the raw materials and
the development of the anaerobic process, leading in some cases
to the production of unstable digested materials which may exert
negative impacts on the plant–soil system (Abdullahi et al., 2008;
Salminen et al., 2001). In this context, aerobic respiration indices
based on oxygen uptake are considered the most suitable param-
eters for assessing the biodegradability of organic amendments
(Barrena et al., 2006; Copperband et al., 2003). In wastewaters
and animal slurries the biochemical oxygen demand (BOD) test
is a reliable and commonly used method to determine readily
biodegradable OM (APHA, 2005; Brookman, 1997; Williams, 1983).

0167-8809/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
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Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007
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Although there is no threshold value established as a stability and
quality criterion for the agricultural use of waste materials based
on BOD data, it is possible with some caution to compare it with
the limits established in the solid waste field based on oxygen
uptake rate and/or cumulative oxygen consumption indices. Dif-
ferent methodologies for characterising the biological stability of
organic materials have been used, such as SOUR (specific oxygen
uptake rate as the maximum rate of oxygen consumption), OD20 h
(cumulative oxygen demand for the first 20 h of the test; Lasaridi
and Stentiford, 1998) and DRI24 h (dynamic respiration index, which
is the average oxygen uptake rate at 24 h of maximum biological
activity; Adani et al., 2004).
Also, little information is available about the degree of stabil-
ity of digestates and their C and N dynamics in amended soils
(Sánchez et al., 2008; Tambone et al., 2009). The information gained
from decomposition studies in digestate-treated soil (OM mineral-
isation, N mineralisation–immobilisation, etc.) may be useful for
assessing N availability, and for optimising the digestate applica-
tion rate to agricultural soils. Thus, defining the main digestate
properties affecting such C and N dynamics in soil can help achieve
the sustainable use of digestates as fertilisers in soil–plant system,
which will have both agricultural and environmental benefits.
The present paper evaluates the dynamics of C-mineralisation
and inorganic-N in soil amended with six digestates produced from
representative anaerobic co-digestion processes in Spain. These
effects were investigated in aerobic incubation experiments, an
appropriate tool to evaluate the feasibility of the use of organic
amendments in agricultural soils (Qiu et al., 2008; Trinsoutrot
et al., 2000). Based on these considerations, the main objective of
this paper is to identify the most relevant parameters related to
digestate composition for assessing the maximum benefits of these
materials as fertilisers.
2. Materials and methods
2.1. Incubation experiments in soil
Six digestates were collected as representative samples from
anaerobic co-digestion experiments based on cattle or pig slurry
mixed with agro-industrial wastes (co-digestion mixtures): cat-
tle slurry + 4% glycerine (CG4), cattle slurry + 6% glycerine (CG6),
cattle slurry + 4.3% cattle manure + 11.6% maize–oat silage (CMS),
cattle slurry + 5% orange peel waste (CO), pig slurry + 1.0% sludge
from a slaughterhouse wastewater treatment plant + 6.5% biodiesel
wastewaters (PSB), and pig slurry + 0.6% pasteurised slaughter-
house waste (PS). Digestates were sampled directly after anaerobic
digestion (without post-treatments), stored at a temperature <4 ◦ C
and processed quickly to prevent any chemical or biological alter-
ation. Their main characteristics and the specifications of anaerobic
co-digestion performance are shown in Table 1. CMS and PSB sam-
ples came from industrial-scale co-digestion processes, while the
rest of the samples came from laboratory-scale experiments, run
mainly to optimise the production of biogas through anaerobic
co-digestion.
From an agricultural soil at La Alberca (Murcia, Spain), soil was
taken from the top 20 cm, air-dried and sieved to 2 mm before use.
Its main characteristics were: 24% CaCO3 , pH 7.5 and electrical
conductivity (EC) 1.72 dS m−1 (both saturated paste, with water),
24.3 g kg−1 OM, 14.1 g kg−1 total organic-C (TOC) and 1.85 g kg−1
total nitrogen (TN), with 14.8% clay, 22.3% silt and 62.9% sand.
The digested materials were mixed thoroughly with the soil in
a proportion of 4 g of fresh digestate per 100 g of dry soil (equiva-
lent to a field application of 96 m3 ha−1 ). This application rate was
selected in order to avoid excessively low inputs of organic-C to
the soil with some digestate samples, which could limit the accu-
racy of the C-mineralisation study, while keeping the N addition
(140–380 kg N ha−1 ) realistic for the requirements of agricultural
crops (MARM, 2010).
The digestate-soil mixtures were incubated in darkness under
aerobic conditions, at 26 ± 1 ◦ C for 56 days. Each treatment was
run in triplicate, and soil without digestate was used as the control.
Soil moisture was maintained at 60% of the water-holding capac-
ity during incubation, with deionised water. To follow N dynamics,
a set of destructive samples of the digestate-soil mixtures were
placed in 50-mL tubes without drainage holes; then, they were
closed with parafilm – which allows gas exchange – to retain soil
moisture and avoid anaerobic conditions (de la Fuente et al., 2010).
Periodically, three replicates per treatment were removed from the
incubator (at 0, 2, 7, 14, 28, 42 and 56 days) for analysis of inorganic-
N (NH4 –N and NO3 –N). Since the digestate samples were mixed
homogeneously with the soil at the time of application and there
was no airflow at the soil surface during incubation, N-loss through
volatilisation was negligible—as demonstrated by de la Fuente et al.
(2010). Net organic-N mineralisation was evaluated by the accu-

Table 1
Main characteristics of the digestate samples (mean value ±standard deviation, data expressed on a fresh weight basis).

Parameter CG4 CG6 CMS CO PSB PS

BOD24 h (g L−1 ) 7.5 ± 1.5 35.0 ± 3.0 1.6 ± 0.2 0.5 ± 0.1 0.7 ± 0.1 1.1 ± 0.1
BOD5 d (g L−1 ) 37.5 ± 3.5 52.5 ± 3.5 5.9 ± 0.7 1.3 ± 0.1 2.2 ± 0.2 2.3 ± 0.2
pH 5.64 ± 0.01 7.35 ± 0.03 7.50 ± 0.01 7.86 ± 0.01 8.20 ± 0.02 7.95 ± 0.03
EC (dS m−1 ) 14.5 ± 0.3 11.7 ± 0.2 25.7 ± 0.8 8.7 ± 0.2 30.3 ± 0.9 21.1 ± 0.1
DM (g L−l ) 38.3 ± 0.5 72.9 ± 5.8 90.1 ± 0.2 24.4 ± 0.3 19.5 ± 0.1 21.0 ± 2.4
−l
OM (g L ) 26.4 ± 0.1 56.4 ± 0.9 66.4 ± 0.2 18.0 ± 0.1 8.5 ± 0.1 11.4 ± 0.2
−l
TOC (g L ) 17.8 ± 0.1 42.8 ± 0.1 33.7 ± 0.5 9.4 ± 0.1 5.9 ± 0.1 5.8 ± 0.1
DOC (g L−l ) 10.6 ± 0.3 27.6 ± 0.9 5.4 ± 0.1 1.2 ± 0.1 2.4 ± 0.1 1.2 ± 0.1
TN (g L−l ) 1.88 ± 0.02 2.32 ± 0.07 3.97 ± 0.02 1.44 ± 0.01 3.96 ± 0.03 2.89 ± 0.02
NH4 –N (g L−l ) 0.97 ± 0.03 0.89 ± 0.01 2.43 ± 0.01 0.76 ± 0.01 3.46 ± 0.04 2.21 ± 0.01
TOC/TN ratio 9.5 ± 0.1 18.5 ± 0.5 8.5 ± 0.1 6.6 ± 0.1 1.5 ± 0.1 2.0 ± 0.1
Anaerobic co-digestion performance
Operation Discont Discont Cont Discont Cont Cont
Scale L L I L I L
◦
Temperature ( C) 35 35 38.5 38 37 35
HRT (days) 40 40 25 28 21 20

BOD24 h and BOD5 d : 24-h and 5-d biochemical oxygen demand, respectively. EC: electrical conductivity, DM: dry matter, OM: total organic matter, TOC: total organic-C, DOC:
dissolved organic-C and TN: total nitrogen. Discont: discontinuous operation, Cont: continuous operation, L: laboratory-scale (2–6 L digester), I: industrial-scale (3000 m3
digester) and HRT: hydraulic residence time CG4: cattle slurry + 4% glycerine, CG6: cattle slurry + 6% glycerine, CMS: cattle slurry + 4.3% cattle manure + 11.6% maize–oat
silage, CO: cattle slurry + 5% orange peel waste, PSB: pig slurry + 1.0% sludge from a slaughterhouse wastewater treatment plant + 6.5% biodiesel wastewaters, and PS: pig
slurry + 0.6% pasteurised slaughterhouse waste.

Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007
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mulation of inorganic-N in the soil, since small changes in organic 2.3. Statistical analyses
pools are usually difficult to detect and gaseous losses were consid-
ered negligible under these experimental conditions (de la Fuente Basic statistical analyses of data, correlation coefficients and
et al., 2010). Net N-mineralisation (net N-min) in each treatment regression equations were calculated using the SPSS 18.0 pro-
(in digestate amended and non-amended soil) was calculated by gramme for Windows. The normal distribution of the data was
subtracting the soil inorganic-N content at day 0 from the amount checked by the Shapiro–Wilk’s test; when data failed this test, they
present in soil after 56 days of incubation, and expressed as ␮g were adjusted to a normal distribution through log-transformation.
inorganic-N g−1 dry soil. The N-mineralisation from the digestates
(Nm ) was calculated as:
3. Results and discussion
Nm (%) = 100 × [(inorg − N56 d − inorg-N0 d )soil + digestate − (inorg -
N56 d − inorg-N0 d )soil ]/added TN.while nitrification (nitrate conver-
3.1. Digestate microbial stability
sion) was determined as the percentage of the added-N from the
digestate that had been converted into nitrate after 56 days.
In the present study, all digestate samples showed an initial
phase of maximum respirometric activity within the first 24 h of
NC (%) = 100 × [(NO3 -N56 d − NO3 -N0 d )soil+digestate testing. As a result, the BOD24 h accounted for 20, 67, 27, 39, 30 and
− (NO3 -N56 d − NO3 -N0 d )soil ]/added TN. 47% of the BOD5 d for CG4, CG6, CMS, CO, PSB and PS, respectively
(Table 1). This indicated the presence of an easily biodegradable
organic fraction probably including decaying microbial biomass
The dynamics of C-mineralisation was determined in a sepa-
from the digestate. When the BOD24 h data were expressed as the
rate set of incubations lasting 56 days, using 500-mL, hermetically
average oxygen uptake rate (mg O2 g−1 OM h−1 ), the following
closed glass vessels. A small vial with 10 mL of 0.1 M NaOH was
results were obtained: 1.0, 1.2, 3.2, 3.9, 11.8 and 25.9 for CMS, CO,
placed inside each vessel to trap the CO2 evolved during the incuba-
PSB, PS, CG4 and CG6, respectively. Our results exceed the limit
tion, and empty vessels were used as blanks. These were opened for
value established by Adani et al. (2004) of 0.5 mg O2 g−1 OM h−1 as
several minutes when the NaOH vials were replaced, to maintain
the average oxygen uptake over a 24-h period of the most intense
adequate aerobic conditions. The CO2 was measured periodically
biological activity (DRI24 h ) for highly stable materials (mature
(after 2, 4, 7, 14, 28, 42 and 56 days), by titration of the NaOH
compost); only the CMS digestate behaved like a material of
solution with 0.1 M HCl in an excess of BaCl2 to precipitate car-
medium stability (limit of 1.0 mg O2 g−1 OM h−1 ). By using the less
bonates. The mineralisation of the organic-C from the digestates
restrictive classification proposed by Ponsá et al. (2010) and based
(Cm ) was calculated as the difference between the CO2 –C evolved
on DRI24 h data, the CMS, CO and PSB showed a low biodegradabil-
in the amended soils and that produced in the control (unamended)
ity (<2 mg O2 g−1 DM h−1 ): 0.8, 0.9 and 1.4 mg O2 g−1 DM h−1 ,
soil, and was expressed as a percentage of the TOC added with
respectively. The PS had a moderate biodegradability
the digestates. The data for the C-mineralisation from the diges-
(2–5 mg O2 g−1 DM h−1 ) of 2.1 mg O2 g−1 DM h−1 and the CG
tates were fitted to kinetic functions by the non-linear least-square
samples showed a high biodegradability (>5 mg O2 g−1 DM h−1 ):
technique (Marquardt–Levenberg algorithm), using the Sigma-Plot
8.2 and 20.0 mg O2 g−1 DM h−1 for CG4 and CG6, respectively.
computer programme (SPSS Inc.). The statistical significance of the
These results are in agreement with those of Orzi et al. (2010)
curve-fitting, residual mean square (RMS) and F-values were also
and Tambone et al. (2009), who compared stability data from
calculated.
digested and non-digested materials based on cumulative oxygen
demand after 20 h using the SOUR-test (OD20 h ). They noted clear
decreases in the OD20 h from 235 to 264 mg O2 g−1 DM in the
2.2. Analytical methods
non-digested mixtures to 30–95 mg O2 g−1 DM after anaerobic
digestion, the latter values being comparable to those shown by
The following parameters were determined in the digestate
stabilised materials. When these values were compared to the
samples: EC and pH (directly, after sample homogenisation); mois-
BOD24 h data obtained in this work (Table 2), the CG samples
ture content, after drying to constant weight at 105 ◦ C; the volatile
showed values of oxygen demand similar to those reported for
solids, which reflect the OM content, by loss on ignition at 500 ◦ C for
non-digested mixtures, while the rest of the digestates had lower
24 h. The TOC and TN were measured by automatic microanalysis
values, within the range proposed for stabilised materials. These
(EuroVector elemental analyser, Milan, Italy) of freeze-dried sam-
results can be attributed to the high content in CG digestates of OM
ples and the dissolved organic-C (DOC) using an automatic analyser
easily degradable by microorganisms (DOC, Table 1), which greatly
for liquid samples (TOC-V CSN Analyzer, Shimadzu) after sample
affected the degree of stability of the digestate. The DOC accounted
filtration (0.45 ␮m pore-diameter). Ammonium was extracted by
for 60, 64, 16, 13, 41 and 21% of the TOC for CG4, CG6, CMS, CO, PSB
steam-distillation of fresh samples alkalised with MgO, trapped
in boric acid and titrated with HCl. The 5-day biochemical oxy-
gen demand (BOD5 d ) was measured with a respirometric Oxitop® Table 2
IS 6 (WTW, Germany) based on pressure measurement, which is Biochemical oxygen demand (BOD) values, expressed as mg O2 g−1 DM, obtained
after 24 h and 2, 3, 4 and 5 days of testing.
automatically transformed into mg O2 L−1 . In the Oxitop® system,
cumulative oxygen consumption measurements were made each Digestate BOD24 h BOD2 d BOD3 d BOD4 d BOD5 d
day during a 5-day period. CG4 195.8 456.9 587.4 718.0 979.1
The soil TOC and TN were determined with an automatic CG6 479.9 514.2 651.3 685.6 719.8
microanalyser. The CaCO3 content was measured with a Bernard CMS 18.0 33.3 45.8 55.5 66.0
calcimeter. As incubation progressed, a two-step sequential extrac- CO 20.5 32.8 41.0 45.1 53.2
PSB 33.4 54.0 82.3 100.3 110.5
tion procedure was carried out for inorganic-N determination: PS 50.0 73.8 85.7 97.6 107.1
ultrapure water (1:5 w/v) for NO3 -N and 2 M KCl (1:5 w/v) for
CG4: cattle slurry + 4% glycerine, CG6: cattle slurry + 6% glycerine, CMS: cattle
NH4 –N. The NO3 –N was measured using a nitrate-ion selective
slurry + 4.3% cattle manure + 11.6% maize–oat silage, CO: cattle slurry + 5% orange
electrode (USEPA, 2007), while NH4 –N was determined by a colori- peel waste, PSB: pig slurry + 1.0% sludge from a slaughterhouse wastewater treat-
metric method based on Berthelot’s reaction (Sommer et al., 1992). ment plant + 6.5% biodiesel wastewaters, and PS: pig slurry + 0.6% pasteurised
All values refer to soil dried at 105 ◦ C for 24 h. slaughterhouse waste.

Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007

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(a)
2250
Soil
Cumulative CO2-C evolved (µg C g-1)
2000 Soil+ CG4
Soil+ CG6
Soil+ CMS
1750
Soil+ CO
Soil+ PSB
1500 Soil+ PS
1250
1000
750
500
250
0
0 7 14
Incubation time (days)
Fig. 1. Cumulative CO2 –C evolved from soil during incubation (mean value ±standard deviation; where absent, bars fall within symbols) (a) and cumulative mineralised-C
from digestate samples added to soil: symbols are experimental data (mean value ±standard deviation, n = 3) and lines represent the curve-fitting (b). CG4: cattle slurry + 4%
glycerine, CG6: cattle slurry + 6% glycerine, CMS: cattle slurry + 4.3% cattle manure + 11.6% maize–oat silage, CO: cattle slurry + 5% orange peel waste, PSB: pig slurry + 1.0%
sludge from a slaughterhouse wastewater treatment plant + 6.5% biodiesel wastewaters, and PS: pig slurry + 0.6% pasteurised slaughterhouse waste.
and PS, respectively. Therefore, the higher such percentage was,
the lower microbial stability exhibited.
Although a significant correlation between the BOD24 h and
BOD5 d data was obtained (P < 0.01), indicating that both param-
eters can be used to characterise the microbial stability of the
digested materials, the 5-day measurement can give more reliable
information since the tested materials usually maintained a high
respirometric activity beyond the first 24 h of testing. The cumula-
tive data followed a linear tendency over time in all samples after
the first 24 h of BOD testing, reaching steady rates without showing
clear decreases indicating exhaustion of the easily biodegradable
OM fraction in the digestate (Table 2).
The BOD5 d values varied considerably among the digestate sam-
ples (Table 1). Considering that BOD5 d values for slurry and silage
effluents can be in the range of 10–80 g L−1 (Brookman, 1997) and
BOD reductions of around 70% can be reached after anaerobic diges-
tion (Smith et al., 2007), the values found in the present study can be
considered normal for digestates with the exception of CG samples
which were clearly higher.
3.2. Carbon mineralisation of digestates in soil
The addition of digestate to the soil caused a rapid development
of microbial activity, reflected by the high release of CO2 –C dur-
ing the first days of incubation (Fig. 1a). This was related to the
presence of an easily degradable organic fraction in the digestate
samples, already detected in the BOD test, with clear differences
among samples (the organic load and its microbial stability). Sub-
sequently, the CO2 –C production rates decreased rapidly during the
first two weeks, reaching nearly constant values at the end of the
incubation (<10 ␮g C g−1 soil and day), similar to those obtained in
the unamended soil as the easily mineralisable OM sources were
exhausted.
The amount of CO2 –C evolved from digestate-treated soil after
56 days of incubation (mineralised-C) increased significantly in the
order (mean value): 639 < 653 < 730 < 948 < 1027 < 1679 ␮g C g−1
soil for soil treated with CO, PS, PSB, CMS, CG4 and CG6, respec-
tively (Fig. 1a). The TOC mineralised from the digestate (Cm ) also
reflected the different biodegradability of the OM present in the
digestates (Fig. 1b). At the end of the incubation, CO showed the
lowest percentage of Cm (16% of TOC), followed by CMS and PS (30
and 34%, respectively), which were much lower than for the CG4
Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
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ARTICLE IN PRESS
(b)
110 CG4
Cumulative mineralised-C (% of TOC)
CG6
100
CMS
90 CO
PSB
80 PS
70
60
50
40
30
20
10
0
21 28 35 42 49 56
0 7 14 21 28 35 42 49 56
Incubation time (days)
and CG6 digestates (60 and 63% of TOC, respectively). Thus, CMS,
PS and, especially, CO showed a more recalcitrant nature than the
digestates from the glycerine mixtures, in agreement with the BOD
and DOC results. Also, more than 100% of the TOC added with PSB
had been mineralised after 56 days of incubation, indicating degra-
dation of native soil TOC during incubation (priming effect). Bernal
and Kirchmann (1992) found that the addition to soil of anaerobi-
cally treated pig manure led to 105% mineralised-C after 70 days,
related to the presence of a high amount of easily degradable OM.
The addition to soil of organic materials with a low C/N ratio, such
as PSB (Table 1), favours high C-mineralisation rates as noted by
Riffaldi et al. (1996).
Our results agree well with those obtained by Bernal and
Kirchmann (1992) and Kirchmann and Lundvall (1993) for soil
treated with fresh and aerobically or anaerobically treated ani-
mal slurry and manures, from similar incubation experiments.
The percentages of TOC evolved from these materials after 70
days of incubation ranged from 23 to 105%, depending on the
biodegradability of the OM and the presence of highly available
organic compounds to microorganisms under aerobic condi-
tions. Such compounds can accumulate in digested materials
when degradation of complex substrates into simple and water-
soluble compounds (hydrolysis) and their subsequent degradation
to produce methane (methanogenesis) are not completely bal-
anced during anaerobic digestion, leading to the production of
unstable materials (Drennan and DiStefano, 2010; Pesta, 2007;
Schievano et al., 2010). The presence of such intermediate products
could enhance soil microbial activity and oxygen demand when
digestates are added to soil, resulting in oxygen depletion and
N-immobilisation (Bernal and Kirchmann, 1992; Kirchmann and
Lundvall, 1993).
The dynamic of C-mineralisation from the CG4 and CG6 diges-
tates in the soil fitted to a combined first- and zero-order kinetic
model (Table 3), which suggests the presence of two different
pools of OM in these digestate samples of different degradability: a
labile pool, which was quickly decomposed in soil during an initial,
intense phase of microbial respiration (about 50% of the added TOC,
with high k values), and another, more resistant to microbial degra-
dation and hence mineralised at a low, constant rate with time. In
the CG samples, the addition of glycerine as co-digestion substrate
may have increased the labile pool of TOC, resulting in high insta-
bility with regard to microbial breakdown. The high percentage of
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Table 3
Parameters of the kinetic models used to describe C-mineralisation of the digestates (± standard error) and the statistical significance of the non-linear curve-fitting (RMS:
residual mean square, F-value of the ANOVA).

Combined first- and zero-order function: Cm = CR (1 − e−kR t ) + At

Digestate Cm (% of TOC) CR kR A RMS F

CG4 59.8 ± 2.4 52.0 ± 0.6 0.330 ± 0.011 0.143 ± 0.017 0.498 3015*
CG6 63.0 ± 0.8 46.5 ± 0.5 0.362 ± 0.013 0.305 ± 0.015 0.436 3577*

First order function: Cm = C0 (1 − e−kt )+B

Digestate Cm (% of TOC) C0 k B RMS F

CMS 30.1 ± 1.2 28.9 ± 0.6 0.078 ± 0.004 – 0.757 1447*

CO 16.1 ± 3.8 16.4 ± 0.3 0.272 ± 0.023 – 0.425 574*
PSB 108 ± 4.4 106 ± 10 0.025 ± 0.005 30.2 ± 1.9 4.625 568*
PS 34.2 ± 4.8 35.5 ± 1.5 0.289 ± 0.049 – 8.119 133*

CG4: cattle slurry + 4% glycerine, CG6: cattle slurry + 6% glycerine, CMS: cattle slurry + 4.3% cattle manure + 11.6% maize–oat silage, CO: cattle slurry + 5% orange peel waste,
PSB: pig slurry + 1.0% sludge from a slaughterhouse wastewater treatment plant + 6.5% biodiesel wastewaters, and PS: pig slurry + 0.6% pasteurised slaughterhouse waste. ‘–’:
parameter not included in the curve-fitting. Cm : mineralised-C (% of TOC) after 56 days of incubation, and t: incubation time (days). In the combined first- and zero-order
function: CR , rapid potentially mineralisable-C (% of TOC); kR , rapid rate constant (day−1 ); A (% of TOC day−1 ), slowly mineralisable-C rate (equivalent to “CS × kS ”; CS : slowly
mineralisable-C and kS : the slow rate constant). In the first order: C0 , potentially mineralisable-C (% of TOC) and k, rate constant (day−1 ). The constant term (B) indicates the
initial mineralisation flux detected in the PSB sample (% of TOC).
*
Significant at probability level P < 0.001.

TOC from CG digestates evolved as CO2 –C during the first stage
of incubation is typical of non-treated animal manure and slurries
(Dendooven et al., 1998; Kirchmann and Lundvall, 1993), identify-
ing them as unstable materials. The C-mineralisation of the CMS,
CO and PS digestates fitted better to a first-order kinetic model,
with a potentially mineralisable-C of 16.4–35.5% (Table 3), indicat-
ing the existence in these digestates of a predominant proportion
of OM that is hardly degradable under both anaerobic and aerobic
conditions. The PSB dynamic of C-mineralisation needed an inde-
pendent parameter (B) to show the initial flux of CO2 –C evolved
(30% of the TOC added), suggesting the elevated presence of an OM
fraction susceptible to rapid mineralisation (Riffaldi et al., 1996).
Although this digestate had the lowest OM concentration, it was
highly unstable according to the BOD5 d test (Table 2).
3.3. Nitrogen dynamics in digestate-treated soil
As shown in Fig. 2, all the digestates supplied NH4 –N to the
soil. During the first week of incubation, the inorganic-N concen-
tration decreased in most of the amended soils, due mainly to a
reduction in NH4 –N. However, during the initial stage of incuba-
tion, the NO3 –N concentration also decreased in soil treated with
CG samples (Fig. 2). The initial inorganic-N had decreased after 7
days and accounted for 8, 17, 22 and 60 ␮g N g−1 soil for soil treated
with CO, PS, PSB and CMS, respectively, while the decrease was
even faster after CG4 or CG6 addition: 74 and 115 ␮g N g−1 soil,
respectively (after 2 days, Fig. 2). Such initial decreases in inorganic-
N could be due to microbial immobilisation, since a concomitant
strong C-mineralisation was found in all treated soils (Fig. 1), which

350
Soil+CG4 Soil+CG6 Soil+CMS Soil+CO
300
250
µg N g -1

200
150
100
50
0
Soil+PSB Soil+PS Soil
Soil 14 28 42 56
300
250
µg N g -1

200 NO3-N

150 NH4-N

100 Inorganic-N (NH4-N + NO3-N)

50
0
0 14 28 42 56 0 14 28 42 56 0 14 28 42 56

Incubation time (days)

Fig. 2. Evolution of inorganic-N in a soil treated with digestates during incubation (mean value ±standard deviation; where absent, bars fall within symbols). CG4: cattle
slurry + 4% glycerine, CG6: cattle slurry + 6% glycerine, CMS: cattle slurry + 4.3% cattle manure + 11.6% maize–oat silage, CO: cattle slurry + 5% orange peel waste, PSB: pig
slurry + 1.0% sludge from a slaughterhouse wastewater treatment plant + 6.5% biodiesel wastewaters, and PS: pig slurry + 0.6% pasteurised slaughterhouse waste.

Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007
 ARTICLE IN PRESS
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6 J.A. Alburquerque et al. / Agriculture, Ecosystems and Environment xxx (2011) xxx–xxx

also indicates high activity of soil microorganisms. Kirchmann
and Lundvall (1993) also detected an initial period of inorganic-
N immobilisation, of up to 200 ␮g N g−1 soil, after amending soil
with animal slurries.
A significant, inverse correlation (r = −0.949 at P < 0.01) between
the net N-mineralisation and mineralised-C (microbial respiration)
was obtained in the present study (data at 56 days). The addition
to soil of high amounts of easily degradable OM in the digestates
should induce a quick development of the microbial population,
immobilising inorganic-N for tissue synthesis (Kirchmann and
Lundvall, 1993; Qiu et al., 2008). Although preferential microbial
immobilisation of NH4 over NO3 is generally accepted, microbial
assimilation of NO3 as a N source can occur (Drury et al., 1991;
Myrold and Posavatz, 2007). But, N-losses by denitrification cannot
be discounted in the soils treated with CG4 or CG6, which sup-
plied high amounts of unstable OM to the soil and produced intense
microbial respiration (CO2 –C production, Fig. 1a). This high respi-
ration during the first days of incubation could have reduced the
oxygen concentration in the soil system, leading to N-losses by den-
itrification (Clemens and Huschka, 2001; Dendooven et al., 1998).
After the immobilisation period, inorganic-N increased, indicating
re-mineralisation and nitrification.
The net N-mineralisation after 56 days of incubation was: +7,
−2, +18, −15, −39 and −77 ␮g N g−1 soil, for soil amended with CO,
PS, PSB, CMS, CG4 and CG6, respectively (Fig. 2). These values were
equivalent to net N-mineralisation from the digestate (Nm ) of −10,
−15, −8, −19, −68 and −96% for CO, PS, PSB, CMS, CG4 and CG6,
respectively, indicating the proportion of the TN from the digestate
samples which had been immobilised in the soil.
The PSB, CMS, CO and PS samples led to fast NO3 -N produc-
tion in the soil after an initial lag phase (related to adaptation
and/or immobilisation periods) and, concomitantly, to a decrease
in NH4 –N, due to nitrification (Fig. 2). The percentage of TN added
in digestates that had converted into nitrate after 56 days (nitrate
conversion: NC), was 44%, 50%, 59% and 84% for CO, CMS, PS-MW
and PSB, respectively. These results indicate the N-fertiliser value
of the digestates, as nitrate is the main form of N taken up by plants
from the soil.
3.4. Characteristics of the digestate defining the C and N
dynamics in soil
The digestate composition and stability parameters (BOD5 d
and C-mineralisation in soil) were highly interrelated (Table 4),
with highly significant correlations between DOC and BOD24 h and
BOD5 d and the 7-day C-mineralisation from digestate (C7 d ). The
latter is in agreement with the results presented by Trinsoutrot
et al. (2000), who related the amount of TOC mineralised from
crop residues after 7 days of incubation in soil mainly to the water-
soluble organic fraction added with the amendments. In addition,
DOC, BOD24 h , BOD5 d and C7 d correlated significantly with key
parameters which define C and N turnover in soil after digestate
addition, such as mineralised-C and both Nm and NC; however, for
the two latter parameters, the DOC/TN and C7 d /TN ratios showed
the most significant correlations (Table 4). These results suggest
that DOC, BOD5 d and C7 d , which represent the most labile organic
fraction of the digested materials, together with their ratios with
respect to the TN, can be considered reliable criteria to assess C
and N dynamics in soil after digestate addition. Although the com-
position of the digestates differed quite a lot in the present study,
significant regression equations were obtained, which allows cau-
tious prediction of the C-mineralisation and N-fertilising potential
of these materials in soil, especially from BOD data (in mg L−1 ),
avoiding time consuming procedures of mineralisation tests in soil:
Mineralised-C (␮g C g−1 soil) = 523.4 × log BOD24 h − 812.9; r2 = 0.903 (P < 0.01).
Nm (%) = −51.0 × logBOD24 h + 135.4; r2 = 0.970 (P < 0.001).
Nm (%) = −51.6 × logBOD5 d + 160.8; r2 = 0.915 (P < 0.01).
NC (%) = −69.8 × logBOD24 h + 262.0; r2 = 0.892 (P < 0.01).
NC (%) = −87.7 × log(DOC/TN) + 46.5; r2 = 0.915 (P < 0.01).
According to the BOD, DOC and DOC/TN data, digestates from
cattle slurry–glycerine mixtures (CG samples) constitute a group
clearly different from the rest of the digestates and represent highly
biodegradable materials, indicating that anaerobic co-digestion in
this case did not produce stable materials. The CG digestates were
characterised by the highest DOC concentrations (>10 g L−1 fresh
weight, accounting for >59% of TOC), DOC/TN > 5, BOD5 d > 37 g L−1
fresh weight and the lowest percentages of TN as NH4 –N (52 and
38% for CG4 and CG6, respectively). These digestates were charac-
terised by an intense initial period, with a high respiration activity,
after their addition to soil—during which N was mainly immo-
bilised. This limits their N-fertilising potential and hence their
possible use in agriculture, since further stabilisation is necessary
before their use.
In contrast, the CO and PS samples showed the lowest DOC con-
centrations (1.2 g L−1 each, accounting for only 12 and 21% of TOC
for CO and PS, respectively) and a high stability degree according
to the BOD5 d test (Table 1), which led to the lowest CO2 –C produc-
tion in the soil (Fig. 1a). These digestates showed DOC/TN ratios
<1, clearly lower than those of the CG samples, and high percent-

Table 4
Significant correlations found among parameters related to digestate composition, and C- and N-mineralisation dynamics (n = 6).

Parameters C7 d C7 d /TN Mineralised-C DOC DOC/TN BOD24 h BOD5 d Net N-min Nm NC

C7 d 1
C7 d /TN 0.932** 1
Mineralised-C 0.948** 0.889* 1
DOC 0.995*** 0.914* 0.943** 1
DOC/TN 0.957** 0.991*** 0.912* 0.951** 1
BOD24 h 0.969** 0.945** 0.950** 0.943** 0.944** 1
BOD5 d 0.975** 0.944** 0.880* 0.959** 0.953** 0.966** 1
Net N-min −0.937** −0.943** −0.949** −0.915* −0.940** −0.986*** −0.936* 1
Nm −0.946** −0.977** −0.918* −0.914* −0.961** −0.985*** −0.956** 0.969** 1
NC −0.884* −0.981** −0.878* −0.855* −0.956** −0.944** −0.908* 0.962** 0.975** 1

C7 d : 7-day C-mineralisation from digestate (mg L−1 fresh digestate weight), mineralised-C: the amount of CO2 –C evolved from digestate-treated soil after 56 days of incubation
(␮g C g−1 dry soil), DOC: dissolved organic carbon (mg L−1 fresh digestate weight), BOD24 h : 24-h biochemical oxygen demand (mg O2 L−1 fresh digestate weight), BOD5 d :
5-day biochemical oxygen demand (mg O2 L−1 fresh digestate weight), TN: total nitrogen (mg L−1 fresh digestate weight), net N-min: the net N-mineralisation in the amended
soils after 56 days of incubation (␮g N g−1 dry soil), Nm : nitrogen mineralisation after 56 days of incubation (% of TN from digestate) and NC: nitrate conversion as a percentage
of added TN that had been converted into nitrate from digestate after 56 days of incubation in soil (% of TN from digestate).
*
Significant at probability level P < 0.05.
**
Significant at probability level P < 0.01.
***
Significant at probability level P < 0.001.

Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007
 ARTICLE IN PRESS
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ages of TN as NH4 –N (>50%), which improve the soil N balance (44
and 59% of added TN for CO and PS, respectively, was converted
into nitrate with low N-immobilisation), representing an available
N source for plants. Based on the characteristics of such diges-
tates, DOC < 1.5 g L−1 (<25% of TOC), BOD5 d < 2.5 g L−1 , Cm < 35% and
DOC/TN < 1 can be used to define a high quality digestate appropri-
ate for use as a fertiliser.
The characteristics of the PSB and CMS digestates showed a
higher similarity to CO and PS (stable digested materials) than to the
CG samples (highly unstable). The PSB sample showed a relatively
low BOD5 d value, in accordance with its low TOC content (5.9 g L−1
fresh weight); however, it was highly labile (41% of TOC as DOC) and
had the highest percentage of TN as NH4 –N (87%). This, combined
with its low DOC/TN ratio (0.6), could explain the observed priming
effect on C-mineralisation when PSB was added to soil; however,
negative effects on the soil N dynamic were not seen. Finally, the
CMS sample showed an acceptable degree of stability considering
its much higher organic load relative to CO and PS (33.7 g TOC L−1
fresh weight, only 16% as DOC); however, it showed a relatively
high CO2 –C production at the end of the incubation, similar to that
resulting from CG4 addition to soil. The CMS sample had a DOC/TN
ratio of 1.4 with 61% of its TN as NH4 –N, leading to a percentage
conversion of TN into nitrate of about 50%. Therefore, digestate
properties such as DOC < 5.5 g L−1 (<45% of TOC), BOD5 d < 6.0 g L−1
and DOC/TN < 1.5 can be established as less restrictive quality cri-
teria compared with those defined previously for stable digested
materials. The suitable land application of these digestates may
require some precautions depending on the land use and manage-
ment. A curing or maturation period may be necessary in order to
increase digestate stability or to allow digestates to stabilise in soil
before sowing.
4. Conclusions
Our results demonstrate that digestate composition and stabil-
ity degree can vary greatly depending on the raw materials used for
co-digestion and process characteristics. Therefore, the anaerobic
co-digestion, including the selection of co-substrates, must guaran-
tee that a certain degree of OM stability of the digested material is
reached in order to avoid detrimental effects on the plant–soil sys-
tem. These are mainly conditioned by the biodegradability of the
digested material obtained, which determines whether the diges-
tate can be used directly (highly stable), after a curing/maturation
period (moderately stable) or after aerobic post-treatments to
achieve stabilisation (highly unstable).
In this context, the biochemical oxygen demand (BOD24 h or
BOD5 d ), dissolved organic-C (DOC) together with the DOC/TN can
be considered the most reliable parameters to describe digestate
biodegradability and hence provide a quick estimation of the main
effects on C and N turnover produced by digestate addition to soil.
Highly unstable and easily biodegradable materials, represented
in our study by digestate from cattle slurry–glycerine mixtures
(BOD5 d > 37 g O2 L−1 fresh weight, DOC > 10 g L−1 fresh weight and
DOC/TN > 5), gave rise to a strong CO2 –C production and a loss of
N-fertiliser value through N-immobilisation in soil. These materials
need more extensive biological treatment, such as aerobic stabilisa-
tion, in order to reduce potential detrimental impacts. In contrast,
ammonium in the more stable digested materials was oxidised by
nitrifiers in the very short term. These less biodegradable digested
materials (BOD5 d < 6.0 g O2 L−1 fresh weight, DOC < 5.5 g L−1 fresh
weight and DOC/TN < 1.5) showed a scaled degree of stability,
from highly stabilised materials, which can be used directly in
soils as fertilisers, to moderately stabilised materials, which may
need a curing/maturation period or stabilisation in the soil before
sowing.
Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007
7
Acknowledgements
This research was funded by the “Ministerio de Ciencia e Inno-
vación, Plan Nacional I+D+I 2008-2011” and FEDER Funds “Fondo
Europeo de Desarrollo Regional, una manera de hacer Europa”, in
the framework of the project “singular estratégico PROBIOGAS”:
sub-project 3 Agronomical evaluation of digestates; and subproject 8,
Co-digestion of citric and farm residues (Refs.: PSS-120000-2008-58;
PSS-120000-2008-62). The authors thank all the research groups
involved in the project PROBIOGAS (http://www.probiogas.es),
especially the GIRO and AINIA Technological Centres, the Univer-
sity of Oviedo, San Ramón Group and Treatments of Juneda Society
(Tracjusa) for providing the digested materials used in this work.
The authors wish to thank Dr. D.J. Walker for the English revision.
References
Abdullahi, Y.A., Akunna, J.C., White, N.A., Hallett, P.D., Wheatley, R., 2008. Investigat-
ing the effects of anaerobic and aerobic post-treatment on quality and stability of
organic fraction of municipal solid waste as soil amendment. Bioresour. Technol.
99, 8631–8636.
Adani, F., Confalonieri, R., Tambone, F., 2004. Dynamic respiration index as a descrip-
tor of the biological stability of organic wastes. J. Environ. Qual. 33, 1866–1876.
Álvarez, J.A., Otero, L., Lema, J.M., 2010. A methodology for optimising feed compo-
sition for anaerobic co-digestion of agro-industrial wastes. Bioresour. Technol.
101, 1153–1158.
Al Seadi, T., 2002. Good practice in quality management of AD residues from
biogas production. IEA Bioenergy, Task 24-Energy from Biological Con-
version of Organic Waste, January 2002 (Available at: http://web.sdu.dk/
bio/pdf/manage.pdf).
APHA, 2005. Standard Methods for the Examination of Water & Wastewater, 21st
ed. American Public Health Association, American Water Works Association and
Water Environment Federation, Washington, DC, USA.
Barrena, R., Vázquez, F., Sánchez, A., 2006. The use of respiration indices in the
composting process: a review. Waste Manage. Res. 24, 37–47.
Bernal, M.P., Kirchmann, H., 1992. Carbon and nitrogen mineralization and ammo-
nia volatilization from fresh, aerobically and anaerobically treated pig manure
during incubation with soil. Biol. Fert. Soils 13, 135–141.
Brookman, S.K.E., 1997. Estimation of biochemical oxygen demand in slurry and
effluents using ultra-violet spectrophotometry. Water Res. 31, 372–374.
Clemens, J., Huschka, A., 2001. The effect of biological oxygen demand of cattle
slurry and soil moisture on nitrous oxide emissions. Nutr. Cycl. Agroecosys. 59,
193–198.
Copperband, L.R., Stoner, A.G., Fryda, M.R., Ravet, J.L., 2003. Relating compost mea-
sures of stability and maturity to plant growth. Compost Sci. Util. 11, 113–124.
de la Fuente, C., Clemente, R., Martínez, J., Bernal, M.P., 2010. Optimisation of pig
slurry application to heavy metal polluted soils monitoring nitrification pro-
cesses. Chemosphere 81, 603–610.
Dendooven, L., Bonhomme, E., Merckx, R., Vlassak, K., 1998. N dynamics and sources
of N2 O production following pig slurry application to a loamy soil. Biol. Fert.
Soils 26, 224–228.
Drennan, M.F., DiStefano, T.D., 2010. Characterization of the curing process from
high-solids anaerobic digestion. Bioresour. Technol. 101, 537–544.
Drury, C.F., Voroney, R.P., Beauchamp, E.G., 1991. Availability of NH4 + -N to microor-
ganisms and the soil internal N cycle. Soil Biol. Biochem. 23, 165–169.
Fuchs, J.C., Berner, A., Mayer, J., Smidt, E., Schleiss, K., 2008. Influence of compost
and digestates on plant growth and health: potentials and limits. In: Fuch,
J.S., Kupper, T., Tamm, L., Schenk, K. (Eds.), Proceedings of the International
Congress CODIS 2008. 27–29 February 2008, Solothurn, Switzerland (Available
at: http://orgprints.org/13135/1/fuchs-etal-proceedings-codis-2008.pdf).
Holm-Nielsen, J.B., Al Seadi, T., Oleskowicz-Popiel, P., 2009. The future of anaerobic
digestion and biogas utilization. Bioresour. Technol. 100, 5478–5484.
Kirchmann, H., Lundvall, A., 1993. Relationship between N immobilization and
volatile fatty acids in soil after application of pig and cattle slurry. Biol. Fert.
Soils 15, 161–164.
Lasaridi, K.E., Stentiford, E.I., 1998. A simple respirometric technique for assessing
compost stability. Water Res. 32, 3717–3723.
MARM, 2010 Guía practica de la fertilización racional de los cultivos en España.
Ministerio de Medio Ambiente y Medio Rural y Marino.
Myrold, D.D., Posavatz, N.R., 2007. Potential importance of bacteria and fungi in
nitrate assimilation in soil. Soil Biol. Biochem. 39, 1737–1743.
Orzi, V., Cadena, E., D’Imporzano, G., Artola, A., Davoli, E., Crivelli, M., Adani, F.,
2010. Potential odour emission measurement in organic fraction of municipal
solid waste during anaerobic digestion: Relationship with process and biological
stability parameters. Bioresour. Technol. 101, 7330–7337.
Pesta, G., 2007. Anaerobic digestion of organic residues and wastes. In: Oreopoulou,
V., Russ, W. (Eds.), Utilization of By-products and Treatment of Waste in the Food
Industry. Springer, New York, pp. 53–72.
Ponsá, S., Gea, T., Sánchez, A., 2010. Different indices to express biodegradability in
organic solid wastes. J. Environ. Qual. 39, 706–712.
 ARTICLE IN PRESS
G Model
AGEE-3849; No. of Pages 8
8 J.A. Alburquerque et al. / Agriculture, Ecosystems and Environment xxx (2011) xxx–xxx
Qiu, S., McComb, A.J., Bell, R.W., 2008. Ratios of C,N and P in soil water direct microbial
immobilisation-mineralisation and N availability in nutrient amended sandy
soils in southwestern Australia. Agr. Ecosyst. Environ. 127, 93–99.
Riffaldi, R., Saviozzi, A., Levi-Minzi, R., 1996. Carbon mineralization kinetics as influ-
enced by soil properties. Biol. Fert. Soils 22, 293–298.
Salminen, E., Rintala, J., Härkönen, J., Kuitunen, M., Högmander, H., Oikari, A., 2001.
Anaerobically digested poultry slaughterhouse wastes as fertiliser in agriculture.
Bioresour. Technol. 78, 81–88.
Sánchez, M., Gomez, X., Barriocanal, G., Cuetos, M.J., Morán, A., 2008. Assessment
of the stability of livestock farm wastes treated by anaerobic digestion. Int.
Biodeter. Biodegr. 62, 421–426.
Schievano, A., D’Imporzano, G., Malagutti, L., Fragali, M., Ruboni, G., Adani, F., 2010.
Evaluating inhibition conditions in high-solids anaerobic digestion of organic
fraction of municipal solid waste. Bioresour. Technol. 101, 5728–5732.
Smith, K.A., Metcalfe, P., Grylls, J., Jeffrey, W., Sinclair, A., 2007. Nutrient value of
digestate from farm-based biogas plants in Scotland. Report for Scottish Exec-
utive Environment and Rural Affairs Department-ADA/009/06 (Available at:
http://www.scotland.gov.uk/Resource/Doc/1057/0053041.pdf).
Please cite this article in press as: Alburquerque, J.A., et al., Chemical properties of anaerobic digestates affecting C and N dynamics in amended
soils. Agric. Ecosyst. Environ. (2011), doi:10.1016/j.agee.2011.03.007
Sommer, S.G., Kjellerup, V., Kristjansen, O., 1992. Determination of total
ammonium nitrogen in pig and cattle slurry: sample preparation and anal-
ysis. Acta Agriculturae Scandinavica Section B. Soil and Plant Science 42,
146–151.
Tambone, F., Genevini, P., D’Imporzano, G., Adani, F., 2009. Assessing amendment
properties of digestate by studying the organic matter composition and the
degree of biological stability during the anaerobic digestion of the organic frac-
tion of MSW. Bioresour. Technol. 100, 3140–3142.
Trinsoutrot, I., Recous, S., Bentz, B., Linères, M., Chèneby, D., Nicolardot, B., 2000.
Biochemical quality of crop residues and carbon and nitrogen mineraliza-
tion kinetics under nonlimiting nitrogen conditions. Soil Sci. Soc. Am. J. 64,
918–926.
USEPA, 2007. Method 9210A. Potentiometric determination of nitrate in aque-
ous samples with an ion-selective electrode (Available at: http://www.
epa.gov/epawaste/hazard/testmethods/sw846/pdfs/9210a.pdf).
Williams, A.G., 1983. Organic acids, biochemical oxygen demand and chemical oxy-
gen demand in the soluble fraction of piggery slurry. J. Sci. Food Agric. 34,
212–220.