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Assessment of the fertiliser potential of digestates from farm


and agroindustrial residues

José Antonio Alburquerque a,*, Carlos de la Fuente a, Alicia Ferrer-Costa b, Lucı́a Carrasco a,
Juan Cegarra a, Manuel Abad b, Marı́a Pilar Bernal a
a
Department of Soil and Water Conservation and Organic Waste Management, Centro de Edafologı́a y Biologı́a Aplicada del Segura,
CSIC, P.O. Box 164, 30100 Murcia, Spain
b
Instituto Agroforestal Mediterráneo, Universidad Politécnica de Valencia, P.O. Box 22012, 46071 Valencia, Spain

article info abstract

Article history: The sustainability of biogas production systems depends greatly on the appropriate
Received 22 February 2011 disposal of the digestates produced. The main agrochemical characteristics of 12 digestates
Received in revised form from the anaerobic co-digestion of farm and agroindustrial residues were determined and
11 February 2012 compared with quality standards to assess their potential use as fertilisers. The digestates
Accepted 22 February 2012 have a high fertilising potential, associated mainly with their contents of NH4-N; however,
Available online 14 March 2012 their recycling in agriculture might be restricted by their Cu and Zn contents, salinity,
biodegradability, phytotoxicity and hygiene characteristics, which must be addressed to
Keywords: obtain the maximum benefits. Such characteristics determine the need for applying pre- or
Anaerobic co-digestion post-treatments to increase digestate quality until acceptable levels. Therefore, digestate
Digestate composition quality must be taken into account when managing the co-digestion process, including
Agroindustrial residues substrate selection, in order to use digestates as fertilisers without the additional cost of
Biodegradability post-digestion conditioning treatments.
Phytotoxicity ª 2012 Elsevier Ltd. All rights reserved.

1. Introduction implementation of the anaerobic co-digestion of animal


manures and slurries with other residues, as put forward in
The non-renewable nature of fossil energy sources and their the Spanish Slurry Biodigestion Plan [3].
contribution to global warming have caused a great, world- Anaerobic digestion produces biogas and a very-wet
wide interest in the development and implementation of residue called digestate which is a mixture of partially-
renewable energy programmes. In this context, technologies degraded organic matter (OM), microbial biomass and inor-
such as anaerobic digestion - for which the share of the ganic compounds. The direct application of digestates to soil
renewable energy market is increasing - play a major role, is currently considered an inexpensive means for their
leading to benefits for residue management, energy supply disposal and for recovery of their mineral and organic
and the environment [1,2]. constituents for agricultural systems. During anaerobic
The vast amounts of biodegradable residues and by- digestion, labile organic constituents are mostly degraded,
products produced by the livestock and agroindustrial leading to an increase in the stability of the remaining OM
sectors have a large potential for biogas production through contained in the digestate [4e6]. However, the prevalence of
co-digestion. This constitutes a great incentive for the efficiency criteria for energy production (biogas) at an

* Corresponding author. Tel.: þ34 968 396313; fax: þ34 968 396213.
E-mail addresses: jalburquerquemendez@yahoo.es, jalburquerque@cebas.csic.es (J.A. Alburquerque).
0961-9534/$ e see front matter ª 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biombioe.2012.02.018
182 b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9

industrial scale can lead to a limited residence time of the As shown in Table 1, these samples included two diges-
material in the digester (after which energy efficiency starts to tates derived from industrial processes (PS-AB3 and CS-AW3),
decline); this can produce a digestate that is not completely while the rest came from laboratory-scale experiments run in
exhausted in terms of easily-degradable organic compounds. order to optimise biogas production by using co-substrates
This may generate problems during storage (odour emission, such as glycerine or orange peel residues (for CS) and
production of toxic compounds, pathogen re-growth and slaughterhouse or energy crop residues (for PS). The diges-
phytotoxicity) and cause unfavourable impacts on the soil- tates were sampled directly after anaerobic digestion
plant system, thus limiting the potential fertilising value of processes (without post-treatments), stored at a temper-
digestate when unstable materials are added to soil [7,8]. ature < 4  C and processed quickly to prevent chemical or
These can cause an extensive range of deleterious effects on biological alterations.
crops, such as prevention or delay of seed germination, plant
death or marked reductions in growth, which must be
2.2. Analytical methods for digestate characterisation
addressed. Bioassays incorporating plant material are simple,
reproducible, rapid and economic tests that identify phyto-
The following parameters were determined in the fresh
toxic materials.
digestate samples: electrical conductivity (EC) and pH; dry
On the other hand, the agricultural use of digestates should
matter content (DM) after drying the digestate sample at
be considered as a recovery process instead of a simple
105  C for 24 h; the volatile solids, which reflect the OM
disposal method. But, the market demand for digestates as
content, by loss on ignition at 500  C for 24 h. The total organic
soil conditioners or fertilisers depends on the compliance with
carbon (TOC) and total nitrogen (TN) were measured in freeze-
quality standards, as regulated by European and national
dried samples, by automatic microanalysis (EuroVector
guidelines [9,10] as well as by quality protocols for digestate
elemental analyser). The dissolved organic carbon (DOC) was
application in Germany [11] and the United Kingdom [12]. In
measured after filtration of the fresh digestate (through
Spain, no specific, applicable standards for digestates, which
a synthetic filter with a pore diameter of 0.45 mm), using an
would favour their use in agriculture, have been developed -
automatic analyser for liquid samples (TOC-V CSN Analyzer,
which represents a major barrier for the development of
Shimadzu). Ammonium was determined by steam-distillation
anaerobic digestion.
from alkalised fresh samples with MgO, and chloride (Cl) by
In this study, the main agrochemical characteristics of
potentiometry with silver nitrate. After HNO3/HClO4 (2:1 v/v)
a number of digestates, including phytotoxicity, have been
digestion, the following elements were determined by induc-
determined, by analysing twelve samples from several,
tively coupled plasma-optical emission spectrometry (ICP-
representative co-digestion processes carried out in Spain,
OES, Thermo Elemental Co. Iris Intrepid II XDL): P, K, S, Na, Ca,
and then compared to the quality criteria established in order
Mg, Fe, Cu, Mn, Zn, B, Pb, Cd, Cr and Ni. The 5 d biochemical
to assess their potential use as fertilisers in agricultural
oxygen demand (BOD5) was determined, with respirometric
systems.
Oxitop IS 6 equipment (WTW, Germany), based on pressure
measurement, which is automatically computed as an oxygen
value with units of mg L1. In the Oxitop equipment, the
2. Materials and methods
cumulative oxygen consumption was recorded each day
during a period of 5 d. Salmonella spp. and Escherichia coli were
2.1. Digestate origin and sampling
determined according to the modified method of the USEPA

After the anaerobic co-digestion of twelve mixtures incorpo-


rating pig (PS) or cattle (CS) slurries as major components, the
corresponding, representative digestate samples were
Table 1 e The main technical aspects of the anaerobic co-
collected for analysis. According to the origin of the digestate, digestion processes which produced the 12 digestates
the samples were classified into the following four groups studied.
(on a fresh mass basis): Digestate Operation Scale Temperature ( C) HRT (d)

- From PS plus energy crop residues (PS-EC): þ9.6% rape PS-EC1 CONT LS 35 30
PS-EC2 CONT LS 35 30
residue (PS-EC1), þ4.5% sunflower residue (PS-EC2) and
PS-EC3 CONT LS 35 30
þ5.4% corn residue (PS-EC3).
PS-AB1 CONT LS 35 20
- From PS plus animal by-products (PS-AB): þ0.6% pasteurised PS-AB2 CONT LS 35 30
slaughterhouse residues (PS-AB1), þ3.8% pasteurised PS-AB3 CONT IS 37 21
slaughterhouse residues (PS-AB2) and þ1.0% sludge from CS-G1 DISCONT LS 35 40
a slaughterhouse wastewater treatment plant þ6.5% bio- CS-G2 DISCONT LS 35 40
diesel wastewaters (PS-AB3). CS-G3 CONT LS 55 22
CS-AW1 DISCONT LS 38 28
- From CS plus glycerine (CS-G): þ4% glycerine (CS-G1) and
CS-AW2 DISCONT LS 38 40
þ6% glycerine (CS-G2 and CS-G3). CS-AW3 CONT IS 38.5 25
- From CS plus agroindustrial residues (CS-AW): þ5% orange
peel residue (CS-AW1), þ10% orange peel residue (CS-AW2) CONT: continuous operation, DISCONT: discontinuous operation,
LS: laboratory-scale (2 L to 6 L digester), IS: industrial-scale (3000 m3
and þ4.3% cattle manure þ11.6% maize-oat silage
digester), and HRT: hydraulic residence time.
(CS-AW3).
b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9 183

[13] and the most probable number (MPN) method [14], K2O 24:21:27, at 3 or 1.5 mL per L distilled water for cress and
respectively. lettuce, respectively) was added (in 3 successive applications)
to the digestate solutions to maintain seedling growth. At the
2.3. Potential phytotoxicity end of the experiment (4 weeks from sowing), 5 seedlings per
tray were harvested and their total dry mass (shoot þ roots)
To obtain homogeneous samples, the 12 digestate samples determined after drying at 105  C.
were sieved (0.25 mm mesh-size) and then centrifuged. The
supernatants thus obtained were shaken and tested in
2.4. Statistical analyses
bioassays involving seed germination and seedling growth,
with both cress (Lepidium sativum ‘Alenois’) and lettuce (Lac-
Basic statistical analyses of data, correlation coefficients and
tuca sativa ‘Bionda degli Ortelani’).
regression models were produced using the SPSS 18.0 program
for Windows. The normal distribution of the data was checked
2.3.1. Seed germination bioassays
by the Shapiro-Wilk’s test, and failed data were adjusted to
In these bioassays, the Petri dishes used (8.5 cm diameter)
a normal distribution through logarithmic transformation.
each contained 2 filter papers (base and cover), which were
moistened with 1 mL of the digestate. Digestate samples were
tested at concentrations of 100% (pure), 20%, 10%, 1% and 0.1%
(volume fraction), which were prepared with distilled water. 3. Results and discussion
Ten seeds per dish were sown and each experimental treat-
ment was replicated 5 times. The dishes were transferred to 3.1. Physico-chemical characterisation
a germination chamber under controlled conditions of
temperature (17  C and 23  C for lettuce and cress, respec- All digestate samples had low dry matter contents (Table 2),
tively) and darkness for 3 d (cress) or 5 d (lettuce). After this these being classified as liquid products (DM mass
period, the number of germinated seeds was counted, the fraction  15% [11]). They were characterised by slightly-
radicle lengths of these seeds measured and the germination alkaline pH values (> 7.5), except in the case of CS-G1 and
index (GI) calculated as a percentage of the control (distilled CS-G3. The trend is for the pH to increase as anaerobic
water), according to Zucconi et al. [15]. digestion progresses, due to volatile fatty acid degradation
and ammonia production [16]. In addition, the pH is also
2.3.2. Plant growth bioassays conditioned by the addition of strong bases or carbonates to
Plastic cell trays (10 cells/tray and 33 mL/cell), with a drainage control pH and the buffer capacity of the system during
hole in the bottom of each cell, were filled with horticultural anaerobic digestion [2]. Generally, the pH of digestates derived
perlite (1 mme2 mm in diameter) and used for these bioas- from animal slurries is in the alkaline range, around 8 [5,17].
says. Trays (in triplicate for each treatment tested) were The highest EC values were found in the PS digestates and
placed in a holder vessel (18 cm  6.5 cm  5 cm) - containing the CS-AW3 sample (> 20 dS m1, Table 2), which also had the
400 mL of distilled water - for 24 h, until the perlite was highest concentrations of Cl, while CS-G1, CS-G2 and PS-AB2
saturated by capillarity. Then, 2 seeds of cress or lettuce per exhibited the highest Na concentration (Table 3). Therefore,
cell were sown and maintained under the same wetting special care must be taken since excessive doses or continued
conditions for 9 d or 13 d, respectively, until seed germination applications of digestates rich in Cl and Na to soils could lead
and seedling emergence occurred (and only 1 seedling/cell to an increase in soil salinity and inhibit plant growth.
was left). After this period, the water in the holder vessel was Regarding the plant nutrient content and hence the fertil-
replaced completely by 200 mL (equivalent to the perlite iser value, the most-significant property of the digestates was
container volume) of the digestate concentrations to be that a large proportion of the N occurred as inorganic forms
examined: 20%, 10%, 1% and 0.1%, keeping trays without (Table 2), representing NH4-N more than 70% of the TN (on
digestate addition as a control. Pure digestate was not tested a mass basis) in the PS mixtures and 39%e61% in mixtures
for plant growth bioassays since GI was 0% for all the digestate with CS. This form of N can be easily lost by ammonia vola-
samples. In these bioassays, and due to the limited availability tilisation during storage and land spreading due to the alka-
of adequate volumes of the samples obtained from the raw line pH of the digestates [18]. In addition, NH4-N is nitrified
digestates, only 7 co-digestion mixtures were studied: PS-EC1, rapidly in soil under favourable conditions, this form being
PS-EC2, PS-EC3, PS-AB2, PS-AB3, CS-G3 and CS-AW3. Imme- highly available to crops but also subjected to leaching
diately following the sowing, the trays and their holder vessels through the soil profile, which may result in groundwater
were transferred to a growth chamber with controlled condi- pollution. Therefore, storage and land spreading operations
tions: 17  C (lettuce) and 23  C (cress) during the first with digestates must be carefully controlled to avoid negative
week, followed by 19  C (lettuce) and 21  C (cress) environmental impacts.
during the remaining 3 weeks. During the bioassay, the daily The TOC/TN ratio was highest for CS-AW3, CS-G1, CS-G2
photoperiod was 16 h at a photosynthetic irradiance of and CS-G3 (> 8), clearly higher than the mean value of 4
250 mmol m2 s1 provided by fluorescent tubes (Sylvania Cool reported by Siebert et al. [11] for digestates from different
White VHO). Solution losses from the holder vessels (by plant sources. Unbalanced C/N ratios in digestates can limit their
uptake þ evaporation) were restored periodically using use in agriculture, as an excess of degradable organic
distilled water. From the second week, a liquid, mineral substrate for microorganisms with respect to N leads to
compound fertiliser (Hesi Coco, The Netherlands; N-P2O5- immobilisation of this nutrient in the microbial biomass and
184 b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9

Table 2 e The main characteristics and composition of the 12 digestate samples (mean value, expressed on a fresh mass
basis). EC: electrical conductivity, DM: dry matter, TOC: total organic carbon, DOC: dissolved organic carbon, BOD5: 5
d biochemical oxygen demand, TN: total nitrogen and CV: coefficient of variation.
Digestate pH EC DM TOC DOC BOD5 TN NH4-N P K TOC/TN
(dS m1) (g L1) (g L1) (g L1) (g L1) (g L1) (g L1) (g L1) (g L1)

Pig slurry
PS-EC1 7.82 26.0 43.9 14.7 4.3 6.5 3.6 2.9 1.1 3.1 4.1
PS-EC2 7.92 24.1 38.3 12.2 3.7 4.0 3.5 2.6 1.1 3.1 3.5
PS-EC3 7.90 23.3 28.3 8.3 3.7 4.7 3.4 2.7 1.2 2.7 2.4
PS-AB1 7.95 21.1 21.0 5.8 1.2 2.3 2.9 2.2 0.5 2.2 2.0
PS-AB2 7.86 30.8 29.5 8.4 3.5 6.2 4.9 3.4 0.8 3.1 1.7
PS-AB3 8.20 30.3 19.5 5.9 2.4 2.2 4.0 3.5 0.2 2.0 1.5
Median 7.91 25.0 28.9 8.4 3.6 4.4 3.6 2.8 0.9 2.9 2.2
(CV) (1.7) (15.2) (31.8) (38.6) (35.6) (43.7) (18.2) (16.9) (50.9) (18.2) (40.9)
Cattle slurry
CS-G1 5.64 14.5 38.3 17.8 10.6 37.5 1.9 1.0 0.5 1.8 9.5
CS-G2 7.35 11.7 72.9 42.8 27.6 52.5 2.3 0.9 0.4 1.6 18.5
CS-G3 6.35 5.2 17.6 8.3 8.2 10.6 0.6 0.4 0.1 0.8 13.6
CS-AW1 7.86 8.7 24.4 9.4 1.2 1.3 1.4 0.8 0.2 1.1 6.6
CS-AW2 7.90 10.0 17.6 5.8 1.0 1.2 1.5 0.9 0.2 1.2 3.8
CS-AW3 7.50 25.7 90.1 33.7 5.4 5.9 4.0 2.4 0.8 3.1 8.5
Median 7.42 10.9 31.4 13.6 6.8 8.3 1.7 0.9 0.3 1.4 9.0
(CV) (12.8) (56.2) (71.0) (77.5) (109.6) (118.9) (58.0) (67.1) (71.9) (49.8) (52.2)
ANOVAa * * NS NS * ** NS ** * NS ***

NS: not significant. *, ** and ***: significant at probability level P < 0.05, P < 0.01 and P < 0.001, respectively.
a considering the following grouped mixtures: pig slurry þ energy crop residues (PS-EC1, -2 and -3), pig slurry þ animal by-product (PS-AB1, -2
and -3), cattle slurry þ glyceryne (CS-G1, -2 and -3) and cattle slurry þ agroindustrial residues (CS-AW1, -2 and -3).

to crop deficiency, while excess N causes losses through CS-AW3 showed the highest concentrations of P, K, Ca and
ammonia volatilisation, nitrate leaching, etc. [19]. Mg, whereas the S concentration was generally higher in PS
The nutrient concentrations of the digested materials were samples than in those from CS mixtures (Table 3). From the
within the range found by other authors [5,17]. As a whole, the standpoint of quality criteria for digested materials [11,12], no
major elements were N and K, followed by P and Ca (Tables 2 limits have been established for their nutrient contents,
and 3). PS-EC1, PS-EC2 and PS-EC3 together with PS-AB2 and although a number of parameters should be declared,

Table 3 e The concentrations (mg LL1) of elements in the 12 digestate samples (mean value, expressed on a fresh mass
basis). CV: coefficient of variation.
Digestate S Ca Mg Na Cl Fe Mn Zn Cu B

Pig slurry
PS-EC1 401 1993 633 666 1574 155 22.9 49.2 8.4 3.2
PS-EC2 367 1970 721 699 1495 143 23.4 45.9 7.0 3.2
PS-EC3 417 1863 698 697 1613 224 31.0 62.5 7.8 2.7
PS-AB1 219 799 324 696 1598 51 11.4 84.4 14.3 2.2
PS-AB2 302 828 365 995 2120 63 15.4 140.2 15.1 3.1
PS-AB3 680 218 67 726 1993 22 2.9 34.7 4.0 2.3
Median 384 1345 499 698 1606 103 19.1 55.8 8.1 2.9
(CV) (39.3) (59.4) (55.4) (16.5) (14.9) (69.9) (56.1) (55.6) (46.2) (16.5)
Cattle slurry
CS-G1 180 1550 267 1164 685 117 13.7 18.1 10.8 1.8
CS-G2 265 1753 333 1842 665 165 17.1 28.3 13.0 4.8
CS-G3 48 192 79 66 448 95 3.2 10.6 1.4 1.3
CS-AW1 113 1008 257 276 366 30 6.0 7.7 2.8 1.7
CS-AW2 131 1035 314 303 452 39 6.9 8.0 3.1 3.5
CS-AW3 457 4026 698 746 1418 301 27.5 27.7 10.8 3.4
Median 155 1293 290 525 558 106 10.3 14.4 6.9 2.6
(CV) (73.2) (82.1) (62.8) (91.6) (57.6) (80.4) (72.8) (56.8) (72.8) (49.4)
ANOVAa NS NS NS NS ** NS NS * NS NS

NS: not significant. * and **: significant at probability level P < 0.05 and P < 0.01, respectively.
a considering the following grouped mixtures: pig slurry þ energy crop residues (PS-EC1, -2 and -3), pig slurry þ animal by-product (PS-AB1, -2
and -3), cattle slurry þ glyceryne (CS-G1, -2 and -3) and cattle slurry þ agroindustrial residues (CS-AW1, -2 and -3).
b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9 185

including: bulk density, DM, OM, pH, salt content, TN, P2O5, 2 and 3) of most of the parameters analysed reveal, in general,
K2O, CaO, MgO, S, NH4-N, NO3-N, micronutrients, Cl and Na. a high degree of variability. Thus, digestates should be ana-
In the current Spanish legislation for fertilisers [10,20], lysed fully before use. However, highly-significant correla-
minimum limits for N, P and K contents are specified for tions between the DM and parameters such as TOC, Ca and Fe
marketable liquid organo-mineral fertilisers. However, were found, while TN, NH4-N, K, S, Zn and Cl were correlated
according to the legal requirements, digestates cannot be significantly with EC (Table 4), indicating their preferential
considered balanced fertiliser products and they must be association with the solid or liquid digestate fraction,
complemented with other inorganic fertilisers. Another respectively.
significant limitation is the digestate TOC content, generally The use of regression equations based on highly-
lower than the required concentration (> 4% on a mass basis). significant correlations between EC and ionic species (NHþ 4,
Therefore, the inclusion of a new fertiliser category in the Kþ, etc.) resulted in a reliable tool for providing a quick esti-
current Spanish legislation, specifically considering digestate mation of nutrient contents in animal slurries [22,23]. As
composition, may be advisable. a result, the EC and DM of digested materials can be used
The most-abundant micronutrient in the digestates was Fe successfully to estimate other valuable and useful parameters
(Table 3), CS-AW3 having the highest value due to the use of Fe which are much-more difficult and time-consuming to
salts during its anaerobic digestion. High concentrations of Cu analyse (Table 5) and which, together with BOD or DOC data,
and Zn were found in the digestates due to the use of pig and characterise the quality of digested materials, as discussed in
cattle slurry as the major co-digestion substrates, since these the following section.
two elements are frequently used as additives - to prevent pig
and cattle diseases and to stimulate livestock growth. The Cu 3.2. Digestate biodegradability
and Zn concentrations were especially high in digestates
arising from PS mixtures (on a dry mass basis): (76e682) The digestate samples showed a great variability in their
mg kg1, with a median value of 173 mg kg1 for Cu, and degree of stability, according to the BOD5 values (Table 2).
(222e4757) mg kg1 with a median value of 446 mg kg1 for Zn. These results indicate the great influence on digestate
Some values were higher than the established limits [11,12], so stability not only of the raw materials used as co-digestion
special care must be taken with respect to Cu and Zn substrates but also of the co-digestion process, as shown by
concentrations, especially for digestate produced from PS. the clear differences in the organic load and its biodegrad-
With respect to heavy metals, the concentrations on a dry ability among digestate samples from cattle slurry þ glycerine
mass basis in some digestate samples for Ni (n ¼ 7, 6 mg kg1 mixtures (CS-G). The CS-G digestates showed BOD5 values
to 36 mg kg1), Pb (n ¼ 7, 11 mg kg1 to 46 mg kg1), Cr (n ¼ 2, that were clearly higher than those of the other digestates
3 mg kg1 and 55 mg kg1) and Cd (n ¼ 6, 0.1 mg kg1 to tested (Table 2), indicating that they had the lowest degree of
1.0 mg kg1) were lower than both the limits established by the microbial stability, associated with the presence of easily-
cited Spanish legislation and the quality protocols for the degradable compounds. The industrial digestate sample
production and use of digestates [11,12]. The concentrations of (CS-AW3) had the highest DM content of all the digestates
heavy metals found in this study were similar to those produced from CS mixtures, but only 16% of the TOC occurred
reported previously for digested materials from substrates of as DOC, which led to a higher degree of stability (less biode-
different origin [6,11,21]. gradable material) than for the digestates from CS-G mixtures.
Our results reveal the influence of the raw materials on the This is related to the use of cattle manure and, especially,
digestate characteristics. The coefficients of variation (Tables silage (solid lignocellulosic materials) as co-substrates for

Table 4 e Correlation matrix between selected parameters related to the digestate composition (n [ 12).
Parameters DM EC TOC TN NH4-N P K Ca Mg Fe Mn Zn Cl

DM 1
EC NS 1
TOC 0.946*** NS 1
TN NS 0.973*** NS 1
NH4-N NS 0.984*** NS 0.951*** 1
P NS 0.607* NS 0.644* 0.599* 1
K NS 0.875*** NS 0.900*** 0.837** 0.878*** 1
Ca 0.856*** NS 0.725** NS NS 0.612* 0.577* 1
Mg 0.584* NS NS NS NS 0.923*** 0.780** 0.800** 1
Fe 0.789** NS 0.705* NS NS 0.655* NS 0.886*** 0.752** 1
Mn 0.684* NS NS NS NS 0.932*** 0.787** 0.806** 0.941*** 0.859*** 1
Zn NS 0.814** NS 0.836** 0.813** 0.649* 0.793** NS NS NS NS 1
Cl NS 0.968*** NS 0.936*** 0.979*** NS 0.814** NS NS NS NS 0.888*** 1

NS: not significant. *, ** and ***: significant at probability level P < 0.05, 0.01 and 0.001, respectively.
EC: electrical conductivity (dS m1). DM: dry matter, TOC: total organic carbon, TN: total nitrogen. NH4-N, P, K, Ca, and Mg in g L1 on a fresh
mass basis. Fe, Mn, Zn and Cl in mg L1 on a fresh mass basis. The DM, TOC and Zn data were adjusted to a normal distribution (Shapiro-Wilk’s
test) through logarithmic transformation.
186 b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9

anaerobic digestion, which enriched the digestate in OM of


Table 5 e The linear regression equations (y [ Ax D B)
low biodegradability.
calculated for selected parameters determined in the
The CS-AW1 and CS-AW2 samples had the lowest BOD5
digestates (n [ 12).
values and percentages of DOC with respect to TOC (<16%),
y x A B r
indicating a high stability degree. By contrast, digested
materials where PS was the main substrate showed an inter- lgTOC lgDM 1.176*** 0.708** 0.946***
mediate behaviour, with BOD5 values between 2 g L1 and Ca 3.807*** 4.286** 0.856***
7 g L1 and DOC values between 1 g L1 and 4 g L1, repre- Fe 289.6** 318.4* 0.789**

senting from 20% to 40% of the TOC in the digestates.


TN EC 0.141*** 0.117NS 0.973***
Therefore, differences in stability degree between diges- NH4-N 0.124*** 0.431* 0.984***
tates were related directly to the concentration of DOC, readily K 0.084*** 0.543NS 0.875***
available for microorganisms. The BOD5 was correlated lgZn 0.037** 0.764** 0.814**
significantly with both TOC (r ¼ 0.722; P < 0.01) and, especially, Cl 0.070*** 0.144NS 0.968***
DOC (r ¼ 0.960; P < 0.001), which demonstrates the strong NS: not significant, *, ** and ***: significant at P < 0.05, 0.01 and 0.001,
influence of DOC on digestate biodegradability characteristics. respectively.
A significant regression equation was obtained for BOD5 and TOC: total organic carbon, TN: total nitrogen, DM: dry matter, NH4-
DOC (g L1 fresh digestate mass): lgBOD5 ¼ 1.172lgDOC þ 0.057, N, K, Ca, and Cl in g L1 (fresh mass basis). Fe and Zn in mg L1
which allows cautious prediction of BOD5 from DOC data. (fresh mass basis). EC: electrical conductivity in dS m1.
Considering the BOD data after 24 h, which corresponds to
the period of maximum biological activity, and expressing
them as the average oxygen uptake rate (on an organic caused a high CO2-C production and led to N-immobilisation
matter basis), only the CS-AW3 sample satisfied the respira- in soil, thus greatly limiting their N-fertilising potential in
tion index (<1 mg g1 h1) proposed for stabilised biowastes the short-term.
[9]. On the other hand, using the criterion established by
Ponsá et al. [24], the digestates can be classified as follows 3.3. Evaluation of the potential phytotoxicity of digestates
(on a dry mass basis): 1) low biodegradability (<2 mg g1 h1): and their hygiene
PS-AB3, CS-AW1, CS-AW2 and CS-AW3 (1.4, 0.9, 1.0
and 0.8 mg g1 h1, respectively); 2) moderate biodegradability The effects of the 12 digestates and 5 digestate concentrations
(2 mg g1 h1e5 mg g1 h1): PS-AB1, PS-AB2, PS-EC1, PS-EC2 studied (100% -pure-, 20%, 10%, 1% and 0.1%) on the GI,
and PS-EC3 (2.1, 3.2, 2.4, 2.2 and 2.6 mg g1 h1, respectively); expressed as a percentage of the control (distilled water), of
and 3) high biodegradability (> 5 mg g1 h1): digestates from cress and lettuce seeds (germination bioassays) are shown in
CS-G mixtures (8.2, 20.0 and 11.8 mg g1 h1 for CS-G1, CS-G2 Fig. 1(a) and (b), respectively. Notable and highly-significant
and CS-G3, respectively). The procedure for certifying the effects of both digestates and concentrations were found.
quality of digestates in the United Kingdom [12] establishes Among the 12 digestates examined, the average GI ranged
a dissolved chemical oxygen demand (COD) lower than from 34% (CS-G3) to 75% of the control (CS-AW1) for cress and
0.43 g g1 OM as a stability requirement. In our study, the CS-G from 34% (CS-G3) to 82% (CS-AW2) for lettuce and, with regard
digestates did not meet this requirement, assuming that DOC to the concentration tested, from 0% (100% digestate concen-
contributes greatly to COD, thus underlining the high biode- tration, both cress and lettuce) to 102% (1% digestate
gradability of the CS-G digestates. In such cases, a longer concentration, cress) or 110% (1% digestate concentration,
residence time during anaerobic co-digestion, further pro- lettuce). At a digestate concentration of 1%, various digestates,
cessing or post-treatment stabilisation is recommendable particularly PS-AB1 and CS-AW3, exceeded the threshold
before digestate application to soil, in order to obtain the value for GI of 125% (of the control), in both the cress and
maximum agricultural and environmental benefits. lettuce bioassays; thus, these two digestate solutions could be
Among the quality criteria defined for digestates, biode- considered to have plant nutrient or plant growth stimulant
gradability or the degree of stability is the parameter for attributes, as suggested by Emino and Warman [26] and
which there is least agreement. As a result, different Moldes et al. [27].
threshold values have been established as quality criteria, The GI of both cress and lettuce at the digestate concen-
based on respiration indices [9], volatile fatty acid contents trations of 20% and 10% were inversely correlated with the EC
[11] or chemical oxygen demand [12]. In our previous work values (P < 0.01) of the digestates (an indicator of their salt
[25], digestate composition parameters such as DOC, BOD content) and also with the concentrations of TN (P < 0.01),
and DOC/TN were related to the C and N dynamics in a soil NH4-N (P < 0.05), K (P < 0.01), S (P < 0.05), Zn (P < 0.01) and Cl
amended with digestates. In this case, a digestate suitable (P < 0.01). McLachlan et al. [28] obtained similar results when
for use as a fertiliser was defined based on DOC < 1.5 g L1, they studied the potential phytotoxicity of digestates
BOD5 < 2.5 g L1 and DOC/TN < 1, while digestates showing produced from municipal solid wastes, high soluble salt
characteristics such as DOC < 5.5 g L1, BOD5 < 6.0 g L1 and concentrations being the major reason for phytotoxicity. Tam
DOC/TN < 1.5 were established as less-appropriate for fer- and Tiquia [29] related the phytotoxic effects of spent pig litter
tilisation purposes, with a curing or maturation period being to its salt, NH4-N, Cu and Zn contents.
required in order to increase digestate stability. Highly- For concentrations of 1% and, especially, 0.1%, TN, NH4-N,
biodegradable digested materials, such as the CS-G K, S, Zn and Cl were not correlated significantly with the GI for
samples, were not suitable for agricultural use as they cress, but the latter was correlated positively with EC, TN,
b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9 187

Fig. 1 e The effects of digestates and concentrations on the germination index (GI, expressed as percentage of the control) of
cress (a) and lettuce (b) seeds. Each value is the mean of five replications ± the standard error of the mean. GI was 0% for all
the pure (100%) digestate samples studied.

Fig. 2 e The effects of digestates and concentrations on the biomass accumulation (expressed as total dry mass, shoot D roots,
in mg) of cress (a) and lettuce (b) seedlings. Each value is the mean of three replications ± the standard error of the mean. Control
(without digestate addition): CO.
188 b i o m a s s a n d b i o e n e r g y 4 0 ( 2 0 1 2 ) 1 8 1 e1 8 9

NH4-N and S for lettuce at 1% concentration (P < 0.05). Indeed,


parameters related to digestate stability (DOC, DOC/TN and 4. Conclusions
BOD) correlated negatively with GI, especially for cress at both
concentrations of 1% and 0.1% (P < 0.05). Several authors [7,30] The digestates had a high potential fertiliser value due to their
related decreases in the phytotoxic effects of digestates to contents of N, P, K and micronutrients. However, their high
reductions in the amount of easily-biodegradable organic variability and unbalanced contents necessitate their analysis
compounds. before they can be integrated into fertilisation programmes.
The influence of digestates PS-EC1, PS-EC2, PS-EC3, PS-AB2, Together with the sanitary quality of the digestates,
PS-AB3, CS-G3 and CS-AW3, at the concentrations of 20%, 10%, a minimum degree of stability of their OM is required to obtain
1% and 0.1%, and of the control (without digestate addition), on the maximum benefits of digestate recycling in agriculture,
biomass accumulation in cress and lettuce plants - expressed which must be harmonised at an international level. There-
as total dry mass (shoot þ roots) per seedling - is presented in fore, digestate characterisation is an unavoidable task, to
Fig. 2(a) and (b), respectively. Significant differences in seedling determine the application rates, phytotoxicity risk and need
dry mass among both digestates and concentrations were for a safety period, before sowing, or a stabilisation treatment,
found, the best results (on average) being obtained with PS- before soil application.
AB3, PS-EC1, PS-EC2, PS-EC3 and CS-AW3 in cress and with
PS-EC1, PS-EC2 and PS-EC3 in lettuce; the concentrations of 1%
(cress and lettuce) or 0.1% (cress) yielded the greatest seedling
Acknowledgements
biomass. In addition, most of the best treatments indicated
above were as efficient as the untreated controls, or even more
This research was funded by the “Ministerio de Ciencia e
so, with regard to seedling biomass accumulation. This indi-
Innovación, Plan Nacional IþDþI 2008-2011” and EU through
cates - as do the seed germination results - plant nutrient,
FEDER Funds “Fondo Europeo de Desarrollo Regional, una
growth stimulant or even phytohormone-like effects of these
manera de hacer Europa”, in the framework of the project
digestate solutions.
“singular estratégico PROBIOGAS” (Refs.: PSS-120000-2008-58;
Therefore, agricultural uses of stable digestates can be
PSS-120000-2008-62). The authors thank all the research
conditioned by their phytotoxic effects during early growth
groups involved in the project PROBIOGAS (http://www.
(germination), due mainly to salinity. Thus, digestate appli-
probiogas.es), especially the GIRO and AINIA Technological
cation rates must consider the concentrations of Na and Cl,
Centres, the University of Oviedo, San Ramón Group and
and also heavy metals (especially Cu and Zn), in order to
Treatments of Juneda Society (Tracjusa), for providing the
avoid any risk of metal accumulation in soil as well as sali-
digested materials used in this work. The authors also thank
nisation or phytotoxic effects, already detected following
Dr. D.J. Walker for the English revision.
excessive application of animal manure and slurries [31,32].
In order to avoid phytotoxicity, digestate application to soil
should be done well in advance of sowing, avoiding direct
references
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maximum benefit from the nutrients provided can be
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