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Greenhouse Gas Emissions and Growth of Wheat Cultivated in Soil Amended

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Pedosphere 27(2): 318–327, 2017
doi:10.1016/S1002-0160(17)60319-9
ISSN 1002-0160/CN 32-1315/P
⃝c 2017 Soil Science Society of China
Published by Elsevier B.V. and Science Press

Greenhouse Gas Emissions and Growth of Wheat Cultivated

in Soil Amended with Digestate from Biogas Production

Liliana PAMPILLÓN-GONZÁLEZ1 , Marco LUNA-GUIDO2 , Victor Manuel RUÍZ-VALDIVIEZO2 ,

Olivia FRANCO-HERNÁNDEZ3 , Fabián FERNÁNDEZ-LUQUEÑO4 , Octavio PAREDES-LÓPEZ5 ,
Gerardo HERNÁNDEZ1 and Luc DENDOOVEN2,∗
1 PhD Program on Science, Technology and Society, Cinvestav, IPN 2508, Mexico DF 07360 (Mexico)
2 Laboratory of Soil Ecology, Laboratorio de Matemáticas Aplicadas y Cómputo de Alto Rendimiento, Cinvestav-Edomex (ABACUS),
Cinvestav, IPN 2508, Mexico DF 07360 (Mexico)
3 Department of Chemistry, Unidad Profesional Interdisciplinaria de Biotecnologı́a-Instituto Politécnico Nacional (UPIBI-IPN), Me-

xico DF 07340 (Mexico)

4 Sustainability of Natural Resources and Energy Program, Cinvestav-Saltillo 1062, Coahuila 25900 (Mexico)
5 Department of Biotechnology and Biochemistry, Cinvestav, Irapuato 36821 (Mexico)

(Received June 16, 2016; revised January 18, 2017)

ABSTRACT
Digestate, the product obtained after anaerobic digestion of organic waste for biogas production, is rich in plant nutrients and
might be used to fertilize crops. Wheat (Triticum spp. L.) was fertilized with digestate, urea, or left unfertilized and cultivated in the
greenhouse for 120 d. Emissions of greenhouse gasses (carbon dioxide (CO2 ), methane (CH4 ), and nitrous oxide (N2 O)) were monitored
and plant growth characteristics were determined at harvest. The digestate was characterized for heavy metals, pathogens, and C and
N mineralization potential in an aerobic incubation experiment. No Salmonella spp., Shigella spp., or viable eggs of helminths were
detected in the digested pig slurry, but the number of faecal coliforms was as high as 3.6 × 104 colony-forming units (CFU) g−1
dry digestate. The concentrations of heavy metals did not surpass the upper limits established by US Environmental Protection
Agency (EPA). After 28 d, 17% of the organic C (436 g kg−1 dry digestate) and 8% of the organic N (6.92 g kg−1 dry digestate)
were mineralized. Emissions of CO2 and CH4 were not significantly affected by fertilization in the wheat-cultivated soil, but digestate
significantly increased the cumulative N2 O emission by 5 times compared to the urea-amended soil and 63 times compared to the
uncultivated unfertilized soil. It could be concluded that digestate was nutrient rich and low in heavy metals and pathogens, and did
not affect emissions of CH4 and CO2 when applied to a soil cultivated with wheat, but increased emission of N2 O.
Key Words: biodigester, C and N mineralization potential, faecal coliform, heavy metal, pathogen, pig slurry

Citation: Pampillón-González L, Luna-Guido M, Ruı́z-Valdiviezo V M, Franco-Hernández O, Fernández-Luqueño F, Paredes-López

O, Hernández G, Dendooven L. 2017. Greenhouse gas emissions and growth of wheat cultivated in soil amended with digestate from
biogas production. Pedosphere. 27(2): 318–327.

INTRODUCTION EPA, 2006; IPCC, 2007). If CH4 emissions increase

Livestock production is one of the major causes
of the world’s most pressing environmental problems,
including global warming, land degradation, air and
water pollution, and loss of biodiversity (FAO, 2006).
Greenhouse gas (GHG) emission from livestock is one
of the most significant contributors to global warming
(FAO, 2006; US EPA, 2006; IPCC, 2007). The main
gases emitted during agricultural and livestock produc-
tion are carbon dioxide (CO2 ), methane (CH4 ), and
nitrous oxide (N2 O), with global warming potentials
(GWP) of 1, 25, and 298, respectively (IPCC, 2007).
Nitrous oxide and CH4 account for nearly 70% of the
total GHG emissions from agricultural practices (US
in direct proportion to increases in livestock numbers,
then global livestock-related CH4 production is expec-
ted to increase by 60% in 2030 (FAO, 2003), while
agricultural N2 O emissions are projected to increase
by 35%–60% due to increased nitrogen (N) fertilizer
use and animal production (FAO, 2003).
All around the world, anaerobic digestion of farm
waste to produce bioenergy has become an established,
robust technique, and it is predicted that in the future
up to 25% of all bioenergy will be biogas (Holm-Nielsen
and Oleskowicz-Popiel, 2007; Hu et al., 2012). China
and India are the countries with most number of di-
gesters installed so far (Ding et al., 2012). The anaero-
bic digestion generates a biogas rich in CH4 , which is

∗ Corresponding author. E-mail: dendooven@me.com.

GHG EMISSIONS FROM SOIL AMENDED WITH DIGESTATE
suitable for energy production due to its high calori-
fic value. This biogas can be used as fuel in a hob or
stove, an internal combustion engine, lighting, heaters
for homes, or grain dryers (Murphy et al., 2004). A-
part from producing CH4 and reducing farm organic
wastes, an organic product is left after anaerobic di-
gestion that is nutrient rich and generally known as
digestate (Möller and Müller, 2012; WRAP, 2013).
Large amounts of fertilizers are used to increase
crop yields. Organic residue left after biogas produc-
tion can easily be used as organic fertilizer (Odlare et
al., 2011). Application of digestate to crops would re-
duce the use of inorganic fertilizers and GHG emissions
during its production. Application of the digestate also
sequesters carbon (C) in soil (Møller et al., 2009). Ad-
ditionally, anaerobic digestion of organic wastes re-
duces the number of pathogens (Franco-Hernández et
al., 2003). Consequently, the digestate can be applied
safely to soil (Holm-Nielsen et al., 2009; Massé et al.,
2011; Abbasi et al., 2012).
In this study, residual organic waste from biogas
production at a pig farm in Huejotzingo, Puebla, Me-
xico was applied to wheat (Triticum spp. L., var. Tem-
poralera M87) cultivated in the greenhouse. The diges-
tate was characterized for heavy metals and pathogens,
and the N and C mineralization potential was deter-
mined in an aerobic incubation. Emissions of GHG
were monitored during crop growth while plant deve-
lopment and yields were determined at harvest. This
study aimed to investigate how the application of
residue from a biodigester affected wheat growth and
emissions of GHG. It was hypothesized that biogas
residue would stimulate wheat growth while reducing
emissions of GHG compared to the urea-amended soil.
Finding a sustainable, economical, and safe strategy
for the reuse and disposition of these special products
is a matter of environmental importance.
Fig. 1 Flow diagram of biodigester used to anaerobically digest swine faecal wastes.
319
MATERIALS AND METHODS
Residue of biogas production
The organic residues left over from biogas produc-
tion were obtained from Granja Porcı́cola Topoyanes
(Puebla, Mexico) with a total pig population of 7 000
animals. The bioreactor (lagoon type) was located be-
side the stables at the farm. The digestate comprised
solid material and water. The anaerobic digestion pro-
cess had a hydraulic retention time that varied between
45 and 55 d. The operation temperature for the bio-
gas reactor was mesophilic-thermophilic (35–55 ◦ C). A
sample of the digestate was taken via an evacuation
tube from the biodigester in three consecutive weeks
and characterized (Fig. 1). As such, three samples of
digestate were characterized and used in this study.
Details of the digestate characterization can be found
in Franco-Hernández et al. (2003).
Soil sampling
Soil was sampled near the farm where the digestate
was produced in Huejotzingo, Puebla, Mexico (2 230 m
above the sea level, 19◦ 07′ N, 98◦ 22′ W), 85 km south-
east of Mexico City. The climate is temperate with dry
winters (Peel et al., 2007), with a mean annual tem-
perature of 10 ◦ C and precipitation of 600 mm. The
field where the soil was collected was mostly cultivated
with maize (Zea mays L.).
The 0–15 cm top-layer of three plots (approximate-
ly 0.5 ha) was sampled with a 7-cm auger (Eijkelkamp,
The Netherlands). In each plot, 90 soil cores were taken
(approximately 100 kg soil). The soil taken at each plot
was pooled so that three soil samples were obtained,
air-dried, passed through a 5-mm sieve, and characte-
rized. The loamy sand soil had a pH of 6.7, electrolytic
conductivity (EC) of 1.34 dS m−1 , total C content of
3.51 g C kg−1 soil, total N content of 0.33 g N kg−1
320
soil, and water-holding capacity (WHC) of 311 g kg−1
dry soil.
Incubation experiment
The potential amount of CO2 emitted and mine-
ral N that would be released when digestate was ap-
plied to soil (C and N mineralization potential) was
determined in an aerobic incubation for 28 d. A 28-d
incubation period was selected as most of the easily
decomposable organic material is mineralized within
this period and C mineralization rates can be deter-
mined. Briefly, sub-samples of soil of each plot were
amended with or without 1 g digestate and incubated
separately and aerobically at 22 ± 2 ◦ C for 28 d in
1-L glass jars that contained a vial with distilled water
to avoid desiccation of the soil and a vial with 20 mL
0.5 mol L−1 NaOH to trap CO2 evolved. Each treat-
ment was performed in triplicate. After 1, 3, 7, 14, and
28 d, the glass jars were opened, the vial with NaOH
was removed and analysed for CO2 trapped and the
−
soils were extracted for mineral N (NH+ 4 , NO2 , and
−
NO3 ) measurement with 200 mL K2 SO4 . The remai-
ning glass jars were opened, aired for 10 min to avoid
anaerobic conditions, air-tight closed, and incubated
further.
Greenhouse experiment
A greenhouse experiment was conducted at Cinves-
tav (Mexico City, Mexico). The experimental layout
was a completely randomized 1 × 4 factorial design
with three replications from each sampled plot. Four
different treatments were applied to the soil: uncul-
tivated unfertilized soil (control), soil cultivated with
wheat but not fertilized (wheat), soil cultivated with
wheat and fertilized with urea at 150 kg N ha−1 (wheat
+ urea), and soil cultivated with wheat and fertilized
with anaerobically digested pig slurry at a concentra-
tion of 150 kg N ha−1 (wheat + digestate), taking in-
to account the mineral N in the waste and conside-
ring that 40% of the organic N was mineralized within
the experimental period (120 d from mid-June 2013 to
September 2013) (Parnaudeau et al., 2004). Details of
the cultivation of wheat in the greenhouse that started
on June 13, 2013 can be found in Aguilar-Chávez et al.
(2012).
At grain maturity, i.e., 120 d, the entire soil column
was removed from the PVC tube taken care not to
damage the root structure. The roots were separa-
ted from the shoots and the soil was washed from
the roots. The fresh weight and length of shoots and
roots were measured. Afterwards, shoots and roots
were dried at 45 ◦ C and weighed. The weight of grains
L. PAMPILLÓN-GONZÁLEZ et al.
dried at room temperature and number of grains per
plant were determined.
Measurements of the CO2 , CH4 , and N2 O emitted
The closed chamber technique was used to deter-
mine the GHG emissions from the soil surface to the
atmosphere for the first 53 d of the greenhouse ex-
periment (Dane and Hopmans, 2002; Cardenas et al.,
2010). Details of the closed chamber technique and
measurements of the GHG can be found in Aguilar-
Chávez et al. (2012). Details of the settings of the gas
chromatograph and the standards used to calculate the
flux of the GHG on each sampling day can be found in
Ruı́z-Valdiviezo et al. (2010).
Global warming potential
Cumulative GHG emissions were assessed as CO2 -
equivalent emissions using GWP: 298 for N2 O and 25
for CH4 (IPCC, 2007). The cumulative N2 O and CH4
emissions over the cultivation period of 53 d of the
greenhouse experiment were calculated by linear inter-
polation of data points between each successive sam-
pling event and numerical integration of underlying
area using the trapezoid rule (Ussiri et al., 2009).
Statistical analysis
Significant differences in plant characteristics and
GHG emissions as a result of the different treatments
were determined by analysis of variance (ANOVA) and
based on the least significant difference using the gene-
ral linear model procedure (PROC GLM) (SAS Insti-
tute, 1989). Plant growth characteristics were sepa-
rately explored with a principal component analysis
(PCA) using PROC FACTOR (SAS Institute, 1989).
RESULTS
Digestate characteristics
The water content of the digestate was 902 g kg−1
with pH 8.0 and EC 15.5 dS m−1 . The total C content
was 436 g kg−1 and the total N content 6.92 g kg−1 ,
−1
while the concentration of NH+ 4 was 1 065 mg N kg ,
− −1 − −1
NO3 18 mg N kg , and NO2 3.4 mg N kg , all ex-
pressed on dry matter basis.
The digestate contained total coliforms of 240 ×
104 colony-forming units (CFU) g−1 dry digestate and
faecal coliforms of 3.6 × 104 CFU g−1 dry digestate
(Table I). No Salmonella spp., Shigella spp., or viable
eggs of helminths were detected in the digested pig slu-
rry. The concentrations of the metals determined in the
digestate were < 50 mg kg−1 except for calcium (Ca),
magnesium (Mg), copper (Cu), iron (Fe), manganese
GHG EMISSIONS FROM SOIL AMENDED WITH DIGESTATE 321

(Mn), and Zn (Table II). NO− −

3 . After 28 d, the NO3 concentration reached 109
−1
mg N kg . As such, 8% of the organic N applied with
TABLE I
Selected microbiological parameters of the anaerobically diges- TABLE II
ted pig slurry (digestate) and the corresponding upper limits in Concentrations (dry-weight basis) of different metals in the
digestate in class A and class B as stipulated by US EPA (1994) anaerobically digested pig slurry (digestate) and the correspon-
Parameter Digestate Upper limit ding upper limits for normal class in US EPA (1994) and maxi-
mal contents in Swedish Government (2008) in digestate (n = 3)
Class A Class B
Metal Digestate Upper limit Maximal content
CFUa) g−1
mg kg−1
Fungi 1.6 × 104b) NMc) NM
Total coliforms 2.40 × 106 NM NM Be 4.6 ± 0.1a) NGb) NG
Faecal coliforms 3.6 × 104 < 1 × 103 < 2 × 106 Li 5.2 ± 0.2 NG NG
Pb 7.2 ± 0.3 840 100
Shigella spp. NDd) NM NM
Sb 0.9 ± 0.3 NG NG
Salmonella spp. ND <3 < 300
Se 8.5 ± 0.4 100 NG
Viable eggs of helminths ND < 1 × 104 < 3.5 × 104
Ca 37 300 ± 3 669 NG NG
a) Colony-forming units. Mg 24 266 ± 2 048 NG NG
b) Data are means (n = 3). Mo 12.4 ± 0.2 75 NG
c) Not mentioned. Sr 44.1 ± 0.7 NG NG
d) Not detectable. Ti 22.3 ± 2.1 NG NG
V 17.9 ± 0.3 NG NG
Digestate effect on soil C and N mineralization poten- Cd 3.8 ± 0.1 85 1
tial Co 0.8 ± 0.2 NG NG
Cr 10.1 ± 0.3 3 000 100
The application of digestate to soil increased the Cu 1 796 ± 118 4 300 600
Fe 5 860 ± 424 NG NG
production of CO2 and the fastest C mineralization
Mn 1 953 ± 115 NG NG
took place in the first 14 d. After 28 d, 17% of or- Ni 16.9 ± 0.2 420 50
ganic material C applied with the added digestate was As 3.1 ± 0.2 75 NG
mineralized (Fig. 2). Application of digestate increased Zn 3 873 ± 72 7 500 800
the NH+4 concentration in soil compared to the una- a) Means ± standard deviations (n = 3).
mended soil, but it was quickly oxidized via NO− 2 to
b) Not given.

− −
Fig. 2 Emissions of CO2 and concentrations of NH+ 4 , NO2 , and NO3 in soil amended with (+D) or without (–D) digestate incubated
aerobically at 22 ± 2 ◦ C for 28 d. Values are means with standard deviations shown by vertical bars (n = 3).
322 L. PAMPILLÓN-GONZÁLEZ et al.

the digestage (6.92 g kg−1 ) was mineralized. The
concentration of NO− 2 in the digestate-amended soil
reached a maximum of 2.95 mg N kg−1 on day 7, but
decreased again thereafter.
Greenhouse gas emissions
The CO2 emission rates showed a peak on day 3,
but remained < 70 µg C m−2 h−1 thereafter (Fig. 3).
Treatment had no significant effect on the CO2 emis-
sion rate (Table III).
The emission rates of CH4 ranged between –0.1 and
0.1 µg C m−2 h−1 (Fig. 3), with no significant difference
between treatments (Table III). The cumulative CH4
emission over a 53-d period was significantly higher in
the wheat + digestate soil compared to the wheat +
urea soil (P < 0.05).
The emission rates of N2 O were generally < 0.1
µg N m−2 h−1 except in the wheat + digestate soil
(Fig. 3). Between days 6 and 20, the N2 O emission rates
from the wheat + digestate soil were higher than from

Fig. 3 Emissions of CO2 , CH4 , and N2 O from uncultivated unfertilized soil (control) and soils cultivated with wheat unfertilized
(wheat) or fertilized with urea (wheat + urea) or digestate (wheat + digestate) at 150 kg N ha−1 over the first 53 d of a greenhouse ex-
periment. Values are means with standard deviations shown by vertical bars (n = 3).

TABLE III
Average emission rates and cumulative emissions of CO2 , CH4 , and N2 O from uncultivated unfertilized soil (control) and soils cultivated
with wheat unfertilized (wheat) or fertilized with urea (wheat + urea) or digestate (wheat + digestate) at 150 kg N ha−1 over the
first 53 d of a greenhouse experiment

Treatment Emission rate Cumulative emissiona)

CO2 CH4 N2 O N2 O CH4 GHGb)

µg C m−2 h−1 µg N m−2 h−1 µg CO2 m−2
Control 20.7c) Ad) −0.008A −0.001B 34B −5.1AB 14B
Wheat 33.5A 0.044A 0.003B 29B −4.7AB 81B
Wheat + urea 30.0A −0.018A 0.025B 422B −14.5B 408B
Wheat + digestate 44.4A 0.008A 0.123A 2 149A 3.2A 2 152A
P value 0.118 0.539 < 0.001 < 0.000 0.128 < 0.001
a) Calculated as CO2 -equivalent emission using global warming potentials of 298 for N2 O and 25 for CH4 (IPCC, 2007) over 53 d.
b) Greenhouse gases.
c) Data are means (n = 53).
d) Values with the same letter in a column are not significantly different (P < 0.05).
GHG EMISSIONS FROM SOIL AMENDED WITH DIGESTATE 323

the other treatments and ranged from 0.22 to 0.58 µg
N m−2 h−1 . The N2 O emission rate and cumulative
N2 O emission over a 53-d period were both significant-
ly higher from the wheat + digestate soil than from
the other treatments (P < 0.05) (Table III).
Wheat growth
Root length as well as ear fresh and dry weight
increased significantly in wheat + digestate soil com-
pared to wheat + urea soil (P < 0.05) (Table IV). The
PCA results indicated that application of digestate
improved wheat development compared to the other
treatments (Fig. 4).
DISCUSSION
Digestate characteristics
The digestate used in this study had a high EC.
The high salt content in the digestate derives from the
food supplements given to the animals (Velthof et al.,
2005) and the soluble salts remain mainly in the li-
quid fraction of the manure (Burton, 2007). Applying
the digestate to soil will increase its salt content and
a high salt content is known to inhibit plant growth
(Alburquerque et al., 2012a). It will thus be important
to monitor the salt content in soil when this kind of di-
gestate is applied regularly so as to avoid plant growth
inhibition.
No Salmonella spp., Shigella spp., or eggs of he-
lminths were detected in the digestate so they did not
survive the anaerobic digestion, while the amount of
faecal coliforms was 3.6 × 104 CFU g−1 dry diges-
tate. Goberna et al. (2011) found no Salmonella sp.
in digested slurry. Scaglia et al. (2014) reported lower
numbers of faecal coliforms (< 10 × 103 most probable
number (MPN) g−1 ), while Bustamante et al. (2012)

TABLE IV
Growth characteristics of wheat cultivated in soils unfertilized (wheat) or fertilized with urea (wheat + urea) or digestate (wheat +
digestate) at 150 kg N ha−1 in the 120-d greenhouse experiment

Treatment Length Fresh weight Dry weight Number of

grains
Shoot Root Shoot Root Ear Shoot Root Ear
cm g
Wheat 77.5a) Ab) 10.8B 1.5B 0.2B 1.8B 0.8A 0.14B 1.6B 31A
Wheat + urea 74.1B 15.0A 1.7AB 0.6A 2.1AB 0.9A 0.25A 1.9AB 32A
Wheat + digestate 77.4A 15.9A 1.8A 0.5A 2.2A 0.9A 0.24A 2.0A 33A
LSDc) 2.9 2.5 0.25 0.2 0.3 0.1 0.09 0.3 4
F value 5.12 12 3.69 9.01 6.63 0.87 5.40 6.31 1.17
P value 0.007 < 0.001 0.027 < 0.001 0.002 0.422 0.006 0.002 0.315
a) Data are means (n = 3).
b) Values with the same letter in a column are not significantly different (P < 0.05).
c) Least significant difference (P < 0.05).

Fig. 4 Principal component (PC) analysis of growth characteristics of wheat cultivated in soils unfertilized or fertilized with urea or di-
gestate at 150 kg N ha−1 in the 120-d greenhouse experiment.
324
reported that the number of faecal coliforms (Es-
cherichia coli) decreased from 7.9 × 104 MPN g−1
in cattle slurry to 4.0 × 102 MPN g−1 in the diges-
tate. These results confirm that the thermophilic con-
ditions during the anaerobic digestion eliminated most
pathogens, but did not reduce faecal coliforms suffi-
ciently.
The digestate can be classified as a class B biosolid
according to US EPA part 503 (US EPA, 1994) due
to the high number of faecal coliforms. The biosolid
can be applied safely to land, but can not be used to
fertilize crops. Consequently, other measures have to
be taken, e.g., composting, application of lime or ver-
micomposting, drying, thickening, ammonia recovery,
or pelletizing (WRAP, 2013) to further reduce the nu-
mber of coliforms so that it could become a class A
biosolid and can be applied to arable soil (Möller and
Müller, 2012; WRAP, 2013). Considering the high nu-
trient value of the digestate, an additional treatment,
such as application of lime, might be economically vi-
able.
The metal concentrations in the digestate did not
exceed the upper limits established by US EPA (1994)
and can be considered as normal quality (Table II). Ne-
vertheless, the concentrations of Mg, Fe, Cu, Ca, and
Zn were 3 times higher than values reported for other
biosolids (Alburquerque et al., 2012a). The concentra-
tions of Cu and Zn in the digestate (1 796 and 3 873
mg kg−1 , respectively) were 3 to 4 times the maximal
content as stipulated by the Swedish Government for
organic wastes, but no such limits exist in Mexico. It
has been reported that the high concentrations of Cu,
Fe, and Zn in the digestate are presumably the result of
additives that are used in livestock feed to prevent pig
and cattle diseases and also to stimulate their growth
(Alburquerque et al., 2012a).
C and N mineralization potential
In this study, 17% of the organic C in the diges-
tate was mineralized within 28 d. Lower mineralization
rates of 6.4% have been reported for biogas residue of
anaerobically digested maize plants in soil after 21 d
(Chen et al., 2012), while 30% of anaerobically digested
cattle slurry mineralized in soil after 56 d (de la Fuente
et al., 2013). Nearly 8% of organic N in the digestate
was mineralized within 28 d. Goberna et al. (2011) re-
ported that 58% of organic N in the biogas digestate
was mineralized within 30 d, while de la Fuente et al.
(2013) found that 55% of organic N in digested cat-
tle slurry was mineralized after 56 d. Difference in the
percentage of organic C and N mineralized when the
digestate is applied to soil will depend on the orga-
L. PAMPILLÓN-GONZÁLEZ et al.
nic material used in the digestate and/or the residence
time in the digester (Alburquerque et al., 2012b). For
instance, the longer the anaerobic digestion takes, the
lower the subsequent C and N mineralization when the
digestate is applied to soil.
In general, digestates from feedstock with a high
degradability and a low C/N ratio are characterized by
a high NH+ 4 content (Möller and Stinner, 2009; Fou-
da, 2011; Alburquerque et al., 2012b). The NH+ 4 in
the digestate is the result of the hydrolysis of urea and
mineralization of N-rich organic components (mostly
amino acids and proteins) during the anaerobic diges-
tion process (Jørgensen and Jensen, 2009). The mine-
ral N in the digestate used in this study contained 98%
−1
NH+ 4 or 1.065 g N kg . Clemens et al. (2006) reported
similar values for NH+ 4 (1.41 g kg−1 ) in cattle slurry di-
gested for 29 d. However, much higher concentrations
of NH+ 4 have been reported ranging from 3.3 to 5.3 g
−1
NH+ 4 kg for digestate of slaughter house waste, or-
ganic household waste, and distiller’s waste (Abubaker
et al., 2012).
During storage of the digestate, mineral N can
get lost. Ammonium can get lost through NH3 vo-
latilisation as a result of the alkaline pH of the di-
gestate. Ammonium can be oxidized to NO− 3 (Albur-
querque et al., 2012b), and if anaerobic conditions pre-
vail in parts of the digestate, NO− 3 is reduced to N2 O
and N2 (denitrification). Consequently, storage of the
digestate should be limited and anaerobic conditions
during storage should be avoided so as not to lose a
valuable plant nutrient.
Emissions of CH4 , CO2 , and N2 O from wheat-culti-
vated soil
The emission of CO2 increased after application
of the digestate to the wheat-cultivated soil. Although
the organic residues had been fermented for 45 d, they
still contained enough degradable organic material to
increase CO2 emission at the onset of the experiment
(López-Valdez et al., 2011; Sänger et al., 2011). The
easily decomposable organic material in the digestate
served as C substrate for soil microorganisms increa-
sing the CO2 production (Fangueiro et al., 2010; Od-
lare et al., 2012).
Odlare et al. (2012) reported N2 O emission at a
rate of 0.5 µg N kg−1 h−1 when digestate was ap-
plied to a clay soil, a value within the range found
in this experiment (0.22 to 0.58 µg N m−2 h−1 ). Pro-
duction of N2 O in soil is a by-product of the oxidation
− − −
of NH+ 4 to NO2 and NO2 to NO3 under aerobic con-
ditions (nitrification) (Burton, 2007) and the result of
anaerobic reduction of NO− 3 (Sänger et al., 2011). The
GHG EMISSIONS FROM SOIL AMENDED WITH DIGESTATE
emission of N2 O increased in the urea-amended soil
compared to the unfertilized soil between days 7 and
20. It can be assumed that the increased emission of
N2 O was mostly due to oxidation of NH+ 4 as irrigation
of the cultivated wheat plants was controlled so as to
avoid anaerobic conditions. In the digestate-amended
soil, the emitted N2 O was even larger than in the urea-
amended soil. A similar amount of NH+ 4 was applied to
soil with urea and the digestate (considering the NH+ 4
content of the digestate and N mineralization rate of
the organic material in the digestate), so nitrification
could not explain the increased emission of N2 O in the
digestate-amended soil compared to the urea-amended
soil. It can be assumed that the mineralization of the
organic material applied with the digestate to the soil
created more anaerobic micro-sites than in the urea-
amended soil. These anaerobic microsites induce deni-
trification and an increased emission of N2 O (Chad-
wick et al., 2000).
Although agricultural soils are normally a sink for
CH4 , they can become a source when easily mine-
ralizable organic material is applied (Smith et al.,
2011). For instance, soil amended with swine effluent
became a source of CH4 compared to the unfertilized
soil (Sistani et al., 2010). The mineralization of the
organic material applied creates anaerobic microsites
favouring production of CH4 that diffuses out of the
soil without being oxidized (Fangueiro et al., 2010;
Sänger et al., 2011). In this study, CH4 emissions were
low and similar to values reported by Odlare et al.
(2012) when filtered and unfiltered biogas residues were
applied to a clayey and an organic soil. Application of
digestate did not stimulate CH4 emissions significantly
compared to the other treatments.
The cumulative emissions of GHG over a 53-d pe-
riod were significantly higher when digestate was ap-
plied to soil (5 times) compared to the urea-amended
soil. Emission of N2 O was the major contributor to
GHG emissions in the greenhouse experiment. Similar
results were reported by Odlare et al. (2012).
Plant growth characteristics
Application of digestate increased yields of oat
and barley compared to compost amendment (Odlare,
2005). The PCA results indicated that application of
digestate stimulated plant development more than ap-
plication of urea. Application of digestate might affect
plant growth in several ways (Möller and Müller, 2012).
The digestate improves soil fertility and soil structure
(Thangaranjan et al., 2013) and is an effective nutrient
source (Liu et al., 2009; Odlare et al., 2012), providing
crops with a significant amount of mineral N and other
325
necessary micro-elements (Alburquerque et al., 2012a).
CONCLUSIONS
The hydrolysis of urea and mineralization of the
organic N in pig slurry waste released large amounts
of NH+ 4 . This high mineral and organic N content
makes the digestate an ideal organic N fertilizer. The
anaerobic storage of the pig slurry killed most of the
pathogens, so that no Salmonella spp. or eggs of he-
lminths were detected. The number of faecal coliforms,
however, remained high. Therefore, the digestate was
classified as a type B biosolid that can be applied safely
to land, but repeated applications should be avoided
due to the high salt content and cannot be applied
to arable land. Application of the digestate promo-
ted wheat growth and did not affect emissions of CH4 ,
but increased strongly the emission of N2 O compared
to urea. The cumulative emission of GHG from the
digestate-amended soil was 5 times larger than that
from the urea-amended soil.
ACKNOWLEDGEMENTS
This research was funded by Cinvestav (Mexico).
The National Council for Science and Technology, Me-
xico awarded a PhD. grant (No. 300785) to Drs. L.
Pampillón-González and V. M. Ruı́z-Valdiviezo. We
thank ‘The Shared Risk Trust (FIRCO)’ for giving
permission to access the farm Granja Porcı́cola Topo-
yanes (Puebla, Mexico) for providing the anaerobically
digested sludge and the Research Centre for Agricul-
tural and Forestry of the State of Mexico, Campo Ex-
perimental Valle de Mexico for wheat seeds. Dr. L. M.
Rivera assisted with the microbiological characteriza-
tion of the digestate and we thank the analytical sec-
tion at Cinvestav-Saltillo, Coahuila, Mexico, for metal
analyses.
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