Waste Management 77 (2018) 140–146

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Waste Management
journal homepage: www.elsevier.com/locate/wasman

Biological stability of multi-component agri-food digestates and

post-digestates
Irena Wojnowska-Baryła, Katarzyna Bernat ⇑, Sabina Sartowska
Department of Environmental Biotechnology, University of Warmia and Mazury in Olsztyn, Sloneczna 45G, 10-709 Olsztyn, Poland

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

Article history: The use of digestate in agriculture has been an efficient way to mitigate greenhouse gas emissions
Received 22 November 2017 through the recycling of organic materials. However, harmful effects can arise if the organic matter is
Revised 9 May 2018 unstable. The goal of this study was to determine the biological stability (4-day oxygen demand for
Accepted 10 May 2018
degradation of readily biodegradable organic matter (AT4), 21-day anaerobic biogas potential (GP21),
Available online 14 May 2018
and organic matter (VS) content) of six digestates after mesophilic digestion, and that of the correspond-
ing post-digestates after psychrophilic post-digestion. Moreover, the kinetics of the changes in biological
Keywords:
stability during post-digestion were determined. Mesophilic digestion of six multi-component agri-food
Biological stability
Post-digestates
feedstocks consisting of maize silage, bovine manure, mallow silage, pig slurry, glycerin, and spent wash
Biogas productivity from distillation was carried out at an organic loading rate of 2–3 kg VS/(m3
d), and at a hydraulic reten-
Aerobic respiration activity tion time of 45–60 days. Digestates were left in stirred reactors, imitating storage digesters, and kept for
Anaerobic post-treatment the next 120 d under anaerobic psychrophilic conditions (20 ± 1 °C) for further stabilization.
The additional biogas yields during post-digestion (50.9–114.9 dm3/kg TS) accounted for 8.5–27.4% of
the biogas productivity of the feedstocks and 40–80% of that of the digestates. The efficiency of the loss of
organic matter content was 22.5–40.2%. The decrease in the values of AT4, GP21 and VS content made the
post-digestates more biologically stable than the digestates (digestates: AT4 = 13.7–67.0 mg O2/g TS,
GP21 = 71.5–130.1 dm3/kg TS; post-digestates: AT4 = 6.6–37.4 mg O2/g TS, GP21 = 15.7–79.2 dm3/kg
TS). For digestates and post-digestates, AT4 values strongly correlated with GP21 values.
Ó 2018 Elsevier Ltd. All rights reserved.

1. Introduction
The large quantities of biodegradable sources produced by the
intensive livestock production systems, agro-food industry and
from dedicated crops can be anaerobically digested for biogas pro-
duction as an energy recovery process and for digestate production
to recover nutrients and carbon for fertilization. The variability in
the biochemical properties of anaerobic digestates is considerable,
reflecting the diversity of biomass input (Teglia et al., 2011). Anaer-
obic digestion (AD) can be used to transform organic wastes, such
as cattle manure (Gomez et al., 2007), livestock manure and agri-
cultural residues (Amon et al., 2007; Tambone et al., 2010), and
organic solid wastes and sewage sludge (Gomez et al., 2007), into
digestate, which contains organic matter and nutrients in forms
available to plants (Tambone et al., 2010). The properties of the
resulting digestate are affected by the feedstock composition and
the technological parameters of the process (Alburquerque et al.,
2012a; Holm-Nielsen et al., 2009). Unfortunately, optimizing AD
⇑ Corresponding author.
E-mail address: bernat@uwm.edu.pl (K. Bernat).
for methane production mostly entails the use of a shorter hydrau-
lic retention times or higher organic loadings than those necessary
for stabilization of the organic matter. Thus, the unstabilized diges-
tate that is produced often needs to be stored in aboveground,
uncovered tanks. This method of digestate storage can create a
number of problems, including odor emission, as well as produc-
tion of toxic compounds and pathogen re-growth, which can cause
the digestate to have unfavorable effects on the soil, thus limiting
its potential fertilizing value (Abdullahi et al., 2008). Using unsta-
ble digestates can immobilize nitrogen or exhaust oxygen due to
an excessive increase in soil microbial activity (Alburquerque
et al. 2012a, Holm-Nielsen et al. 2009). Rather than just a simple
disposal method, the agricultural use of digestates is also a recov-
ery process. Thus, technological processes should not only produce
the maximum possible yield of biogas, but also safe, biologically
stable digestate that is suitable for application to agricultural soil.
There is no formal definition of biological stability. It can be
defined as the extent to which biodegradable organic matter has
decomposed or the resistance of the organic matter to further
biodegradation. Brewer and Sullivan (2003) determined the stabil-
ity and maturity indices for yard trimmings compost. Those

https://doi.org/10.1016/j.wasman.2018.05.016
0956-053X/Ó 2018 Elsevier Ltd. All rights reserved.
 I. Wojnowska-Baryła et al. / Waste Management 77 (2018) 140–146
authors stated that maturity is a general term describing the suit-
ability of the compost for a particular end use, while stability refers
exclusively to the resistance of the organic matter content in com-
post to further degradation. With the use of respirometric tests,
they identified a period of high respiration rates during active com-
posting and a period of relatively low respiration rates during the
latter part of the process. This could relate to the low stability
and high biodegradability of the substrate before and during active
composting, and the high stability of final product, compost. Gen-
erally, during biological processes (composting/aerobic stabiliza-
tion, anaerobic digestion) the biodegradability of organic
substrates should be reduced, and thus their stability should be
increased.
The stability of organic substrates can be evaluated using non-
biological methods, including determination of TS (total solids),
organic matter (as VS) and total organic carbon (TOC) content,
and by biological methods, including aerobic and anaerobic tests
(Astals et al., 2012; Barrena et al., 2014). Nonetheless, there is little
agreement about which indicators and threshold limits should be
used to assess the biological stability of digestate. For example,
to evaluate the biological stability of lignocellulosic substrates
(agriculture and forest by-products), Liu et al. (2015) suggested
using the aerobic biodegradability (BDaero, % of CODtot), expressed
as the BOD/CODtot ratio, and the anaerobic biodegradability
(BDana, % of CODtot), expressed as the ratio of methane yield to
CODtot. For the same purpose, Schievano et al. (2008) determined,
among others, the following stability indicators of the feedstocks
and digestates from full scale biogas plants: the anaerobic biogasi-
fication potential (ABP), the VS content (% of TS), the TOC/TKN
ratio, and the oxygen demand in a 20-h respirometric test (OD20).
There is a large, significant correlation between the results of
the aerobic tests and those of the anaerobic tests (Ponsá et al.,
2008), and these results can be used to determine the best meth-
ods for treatment and post-treatment of organic waste composting
(Barrena et al., 2009; Liu et al., 2015). However, little research
(Maynaud et al., 2017) is available on the relationships between
the digestate that serves as the input for post-digestion and the
biological stability of the output, i.e. the post-digestate. Not many
studies have focused on the stability of digestates after anaerobic
digestion and the possibility of further stabilization of these diges-
tates via psychrophilic post-digestion. The additional biogas yield
during post-digestion is especially interesting, not necessarily from
an economic point of view, but rather because of the possibility of
reducing uncontrolled methane emissions.
Thus, the purpose of the present study was to provide informa-
tion on the change in the biological stability of six multi-
component agri-food digestates during 120 days of psychrophilic
anaerobic post-digestion, with a particular focus on additional bio-
gas production. Furthermore, during post-digestion, the kinetics of
the changes in indicators of the biological stability of the digestates
were determined.
2. Materials and methods
2.1. Technological conditions of anaerobic psychrophilic post-digestion
After mesophilic anaerobic digestion, six multi-component agri-
food digestates (the composition of these digestates is explained
below) were left in stirred reactors, simulating storage digesters,
and kept for the next 120 d under anaerobic psychrophilic condi-
tions (20 ± 1 °C) for further stabilization.
The characteristics of the process of mesophilic anaerobic diges-
tion (35 ± 2 °C) of the six multi-component agri-food feedstocks
were as follows: hydraulic retention times (HRT) of 45 or 60 d
and organic loading rates of (ORL) of 2 or 3 kg VS/(m3
d). The vol-
141
ume of the reactors was 10 L, and their working volume was 6 L.
The reactors were equipped with water-jacket heaters to maintain
the required temperature. Mesophilic anaerobic digestion pro-
ceeded for 75–200 days to allow the microorganisms to acclima-
tize to the feedstocks and to achieve stable conditions. During
this period, stable biogas production and organic matter degrada-
tion took place (data not shown). For mesophilic anaerobic diges-
tion, the following feedstocks were used:
a two-component feedstock comprised of maize silage (M) and
bovine manure (B) (MB) (50% of VS derived from M, and 50% of
VS derived from B),
three-component feedstocks comprised of maize silage (M) or
mallow silage (P), with pig slurry (S) and glycerine (G) (MSG
and PSG) (87.5% of VS derived from M or P, 6.25% of VS derived
from S, and 6.25% of VS derived from G),
a four-component feedstock comprised of maize silage (M), pig
slurry (S), glycerine (G), and spent wash after distillation (D)
(MSGD) (87.5% of VS derived from M, 4.2% of VS derived from
S, 4.2% of VS derived from G and 4.1% of VS derived from D).
All of these substrates came from working farms and plants
located in north–east Poland.
The digestates that were produced by mesophilic anaerobic
digestion and used in anaerobic psychrophilic post-digestion are
referred to with abbreviations that indicate the composition of the
feedstock and the OLR and HRT of mesophilic anaerobic digestion:
MB/2/45, MB/3/60, MSG/2/45, MSG /3/45, PSG/2/45, MSGD/3/45.
For example, MB/2/45 indicates the two-component feedstock with
maize silage (M) and bovine manure (B) at an ORL of 2 kg VS/(m3
d)
and an HRT of 45 d, and MSG/3/45 indicates the three-component
feedstock with maize silage (M), pig slurry (S), and glycerine (G) at
an ORL of 3 kg VS/(m3
d) and an HRT of 45 d, etc.
2.2. Chemical analyses
Total solids (TS) and volatile solids (VS) in digestates, in samples
that were taken every 3–7 days during post-digestion, and in post-
digestates were determined with standard procedures according to
APHA (Greenberg et al., 1992). Then, the same samples were cen-
trifuged for 25 min at 9000 rpm. In the liquid phase, the content
of organic compounds as COD, and as dissolved organic carbon
(DOC) (after filtration with a 0.45 mm filter), were determined.
2.3. Biological stability measurements
Two respirometric tests were used to determine the biological
stability of the digestates, of the samples taken during 120 days
of post-digestion, and of the post-digestates. Firstly, the aerobic
respiration test (AT4) was carried out during 4 days under aerobic
conditions by measuring the oxygen consumed for degradation of
readily biodegradable organic matter using a manometric Oxi-Top
Control set (German Standard AbfAblV, Anhang 4 (Anon, 2001)).
Secondly, the GP21 anaerobic test (gas potential) was carried out
during 21 days under anaerobic conditions by measuring biogas
production per kg of total solids in the tested sample, with the
use of an inoculum consisting of fermented sludge from the anaer-
obic digester of a wastewater treatment plant (WWTP) (German
Standard AbfAblV, Anhang 4 (Anon, 2001)). AT4 and GP21 values
were determined in triplicate for each sample.
2.4. Calculations
The efficiency of the loss of organic matter content (EVS) during
120 day post-digestion was calculated according to Paredes et al.
(2000):
142 I. Wojnowska-Baryła et al. / Waste Management 77 (2018) 140–146
 
A1
ð100  A2 Þ
EVS ¼ 100  100
A2 ð100  A1 Þ
where A1 – ash content in digestate (%), A2 – ash content in post-
digestate (%).
2.5. Kinetic and statistical analyses
The kinetics of changes in the values of AT4, GP21 and VS con-
tent during 120 days of post-treatment of the digestates proceeded
according to zero-order kinetics, involving linear changes in AT4,
GP21 and VS content with time. In zero-order reactions, the pro-
cess rate (rAT4 – the rate of AT4 decrease, rGP21 – the rate of GP21
decrease, rVS – the rate of VS content decrease) is equal to the rate
constant (the rate constant for the decreases of AT4 (kAT4), GP21
(kGP21) and VS content (kVS)). In these reactions, the values of
AT4, GP21 and VS content after the time (t) of post-digestion can
be estimated with the following formulas:
AT4 ¼ kAT4
t þ AT40 ; GP21 ¼ kGP21
t þ GP210 ;
VScont ¼ kVS
t þ VS0
where the abbreviations with the subscript 0 (e.g. AT40) refer to the
values at the beginning of post-digestion.
Linear regression was used to determine the relationships
between (1) the values of AT4 and organic matter content (VS),
(2) the values of GP21 and organic matter content (VS), and (3)
the values of AT4 and GP21 of the multi-component digestates
and post-digestates. The fit of the models was assessed with the
coefficient of determination (R2).
In addition, the correlations between the rate constants for the
decreases of AT4 (kAT4), GP21 (kGP21) and VS content (kVS) in the
samples taken every 3–7 days during the 120 days of post-
digestion were determined using Pearson’s r. For all calculations,
STATISTICA 10.0 (StatSoft, Tulsa, USA) was used. A result was con-
sidered significant at p  0.05.
3. Results and discussion
3.1. Biological stability of digestates
Recently, both the amount of studies published and the number
of full-scale plants operating in Europe testify to the increased
interest in anaerobic digestion. Besides producing biogas rich in
methane, anaerobic digestion with various organic substrates,
including organic waste, also produces digestate that can be used
as a fertilizer. Many studies have investigated anaerobic treatment
of agro-industrial wastes such as animal manures, sewage sludge
and biowaste (Mata-Alvarez et al., 2014). The effectiveness of VS
removal was commonly in the range of 50–70% (Trzcinski and
Stuckey, 2011; Kim et al., 2011). However, during anaerobic diges-
tion, conditions for effective production of biogas with a high con-
tent of methane are given precedence, and thus there is a risk of
producing unstable digestate. For example, a shortening of the
time that organic substrates reside in the digester, which improves
biogas productivity, may produce digestate in which labile organic
matter has not been completely exhausted (Astals et al., 2012;
Alburquerque et al. 2012b).
The usefulness of digestates as fertilizer may depend on their
chemical properties, biological stability, and hygienization. In the
present study, six multi-component agri-food digestates that had
been produced via mesophilic anaerobic digestion of two-, three-
and four-component feedstocks (HRT of 45–60 d, and ORL of 2–3
kg VS/m3
d) were used. The biological stability of these digestates
was evaluated on the basis of the VS content (% of TS, g VS/dm3),
the 4-day oxygen demand for degradation of readily biodegradable
organic matter (AT4), and a 21-day anaerobic test for determina-
tion of the biogas potential (GP21) (Table 1).
Linear regression showed strong relationships between the con-
tent of organic matter in digestates and the values of AT4 and GP21
of the digestates (Fig. 1a and b). There was good agreement
between the linear regression model and the observed values of
VS content and AT4 (R2 = 0.96), and those of VS content and
GP21 (R2 = 0.86). There was also a strong linear relationship (R2
= 0.86) between the AT4 and GP21 values of the digestates (Fig. 1c).
The digestate with the highest content of organic matter,
PSG/2/45, had the highest AT4 and GP21 values. With regard to
the two-component digestates (MB), the AT4 and GP21 values
were higher when mesophilic anaerobic digestion proceeded at
an OLR of 3 kg VS/(m3
d) and an HRT of 60 d than when it pro-
ceeded at an OLR of 2 kg VS/(m3
d) and an HRT of 45 d. These val-
ues were also higher when there were three or four components in
the feedstocks than when there were only two components. As
shown by the GP21 values, the MB/2/45 and MSG/2/45 digestates
had the lowest biogas productivity, whereas PSG/2/45 and
MSGD/3/45 had the highest. The digestates of MSG/3/45 and
PSG/2/45 were the least biologically stable.
Similarly, when Schievano et al. (2008) analyzed feedstocks and
the resulting digestates at a biogas plant, their regression analysis
showed significant positive relationships between biogas produc-
tivity (ABP) and VS content (% of TS), TOC/TKN and OD20 (in mg
O2/g TS
h) (P < 0.001).
To summarize, on the basis of the AT4 and GP21 values, and the
organic matter content, the biological stability of the digestates
decreased in the following order: MB/2/45 > MSG/2/45 >
MB/3/60 > MSG/3/45 > MSGD/3/45 > PSG/2/45.
Despite the differences in the composition of the feedstocks,
and in the OLR and HRT of mesophilic anaerobic digestion (Table 1),
all of the investigated digestates had VS/TS ratios of over 0.70,
except for MSG/2/45 (0.64). A higher content of organic matter
(as shown by a higher VS/TS ratio) reflects greater biodegradability
and biogas potential, and lower stability. However, the biogas
potential of a substrate depends not only on VS content, but also
on the degradability of the organic matter (Mottet et al., 2009;
Schievano et al., 2008). Lignocellulosic materials usually have low
biodegradability due to the complex structures of lignin and other
cell-wall polysaccharides, and these materials produce less biogas
than other kinds of substances (e.g. carbohydrates, lipids, proteins)
(Li et al., 2013a, 2013b). In the three- and four-component feed-
stocks, the content of lignocellulosic substrates was much higher
than in the two-component feedstocks, and thus it is not surprising
that less stable digestates were produced and their biogas produc-
tivity was higher (Table 1). The biogas production of the digestates
investigated in the present study ranged from 71.5 to 167.5 dm3
biogas/kg TS (100.6–226.9 dm3/kg VS), which was 18–21.4% of
the biogas production of the feedstocks (data not shown).
It is common that the organic matter in digestates is less
degradable than that in the feedstocks, and thus the biogas produc-
tivity of digestates is much lower. Tambone et al. (2010) used an
ABP test to determine the biological stability of composts, digested
sludge, and feedstocks and their corresponding digestates. The ABP
production of the feedstocks, which are the most biodegradable of
these substrates, was highest (396–732 dm3/kg VS). In contrast,
the biologically treated composts (322–517 dm3/kg VS), digested
sludge (359–438 dm3/kg VS), and digestates (184–399 dm3/kg
VS) produced less biogas. The digestates had ABP values lower than
those of compost and digested sludge, and similar to the values
also reported for digestates by Schievano et al. (2008), which aver-
aged 261.7 dm3/kg. Tambone et al. (2010) stated that digestates
had high biological stability, despite the fact that their biogas pro-
duction values were higher than those obtained in the present
study (184–399 dm3/kg VS vs 100.6–226.9 dm3/kg VS).
 I. Wojnowska-Baryła et al. / Waste Management 77 (2018) 140–146 143

Table 1
Indicators of biological stability of the multi-component digestates.

Indicator Unit MB/2/45 MB/3/60 MSG/2/45 MSG/3/45 PSG/2/45 MSGD/3/45

VS % TS 71.1 ± 1.1 74.0 ± 1.2 64.1 ± 0.8 71.0 ± 0.9 73.8 ± 1.0 73.1 ± 0.9
g VS/dm3 17.2 ± 0.5 39.4 ± 0.3 26.0 ± 0.5 41.0 ± 0.7 57.6 ± 1.2 54.6 ± 1.1
AT4 mg O2/g TS 13.7 ± 1.2 35.3 ± 1.3 24.9 ± 1.4 48.0 ± 1.4 67.0 ± 1.6 56.5 ± 1.4
GP21 dm3/kg TS 71.5 ± 4.5 124.3 ± 5.2 99.1 ± 6.1 111.0 ± 6.7 167.5 ± 9.1 130.1 ± 8.2
Organics* mg COD/dm3 2100.0 ± 125.0 3950.0 ± 140.0 8320.0 ± 320.5 13120.0 ± 240.0 24200.0 ± 250.5 30580.0 ± 360.0
mg DOC/dm3 840.0 ± 90.0 1780.0 ± 86.0 3200.0 ± 120.0 5510.0 ± 140.0 9920.0 ± 150.0 13150.0 ± 240.0
*
Organic compounds (COD, DOC) in the liquid fraction of digestates, DOC for the compounds below 0.45 mm.

100 200 PSG/2/45

a) b)
y = 1.91x + 42.29

GP21 [dm /kg TS ]

AT4 [mg O 2/g TS ]

80 160 R2 = 0.86
MB/3/60
y = 0.83x - 5.44 MSGD/3/45
60 R2 = 0.96 120 MSG/2/45

3
PSG/2/45
MSG/3/45
40 MSG/3/45 80
MSGD/3/45
MSG/2/45 MB/2/45
20 MB/2/45 40
MB/3/60
0 0
0 10 20 30 40 50 60 70 0 10 20 30 40 50 60 70
VS [g/dm3] VS [g/dm3]
100
c)
AT4 [mg O 2/g TS ]

80 y = 0.38x - 17.34
R2 = 0.84
60
MSGD/3/45 PSG/2/45
40
MB/3/60
20 MSG/3/45
MB/2/45 MSG/2/45
0
0 40 80 120 160 200
GP21 [dm3/kg TS]

Fig. 1. The relationships between (a) AT4 values and organic matter content (VS) in digestates, (b) GP21 values and organic dry matter content (VS) in digestates, and (c) AT4
and GP21 values of digestates; linear regression equations and the coefficients of determination (R2) are given.

3.2. 120 d post-digestion and biological stability of post-digestates for MSG/3/45 were similar to those for PSG/2/45, but higher than
those for MSG/2/45, indicating that MSG/2/45 is less stable than
If they are intended for agricultural use, digestates are required the other two post-digestates. The rates of decrease of AT4 and
to be biologically stable with organic matter that is not a source of GP21 for MSGD/3/45 were 0.176 mg O2/(g TS
d) and 0.389 dm3/
emissions of methane, ammonium and carbon dioxide. The biogas (kg TS
d), respectively (Fig. 2).
productivity of the digestates during post-treatment depends Decreases in the values of AT4 and GP21 resulted from the fact
chiefly on their organic matter content and quality, as well as on that organic matter content (as g VS/dm3) decreased during 120
the internal tank temperature. In general, biogas productivity in days of post-treatment. Similarly, as with the decrease in AT4 val-
storage digesters in the winter months is lower than in the sum- ues, the rates of the decrease of VS content in the two-component
mer months (Hansen et al., 2006). To reduce the influence of the digestates during 120 d of post-treatment were the lowest of all
temperature in the present study, post-digestion was carried out the digestates (0.033 g VS/(dm3
d) and 0.062 g VS/(dm3
d) respec-
in psychrophilic conditions at 20 ± 1 °C in a covered reactor. In tively for MB/3/60 and MB/2/45). The rates of the decrease of VS
the literature, information on the kinetics of post-treatment is content for the other digestates ranged from 0.124 g VS/(dm3
d)
scarce. Thus, in the present study, the kinetics of changes in the to 0.301 g VS/(dm3
d) (Fig. 3).
values of AT4, GP21 and VS content during 120 days of post- Fig. 4 presents the rates of the decrease of AT4 (mg O2/(g TS
d)),
treatment were determined. The rate constants and the rates of GP21 (dm3/(kg TS
d)) and VS content (g VS/(dm3
d)) during 120
the decreases of AT4 (kAT4, rAT4) and GP21 (kGP21, rGP21) values, days of post-digestion and the composition of multi-component
and of VS content (kVS, rVS) were found (Figs. 2 and 3). digestates. There were large, significant positive correlations
With regard to the two-component digestates, although between rAT4, rGP21, and rVS (rAT4 and rVS r = 0.97, p = 0.06; rAT4
MB/3/60 had higher initial values of AT4 and GP21, the rates of and rGP21 r = 0.87, p = 0.06; rGP21 and rVS r = 0.94, p = 0.06).
decrease of AT4 and GP21 for this digestate were lower (0.037 In the literature, post-digestates are rarely characterized in
mg O2/(g TS
d), 0.367 dm3/(kg TS
d)) than the corresponding values terms of their biological stability, thus, we also focused on this sub-
for MB/2/45 (0.050 mg O2/(g TS
d), 0.425 dm3/(kg TS
d)). It should ject in the present study. The efficiency of the reduction of organic
be emphasized that the rates of AT4 decrease for the two- matter content (EVS) was determined after 120 days of post-
component digestates during 120 d of post-treatment were about treatment. EVS during post-treatment of MB/2/45 was the highest
2 to 10 times lower than the corresponding rates for the other (40.2%), and as a result, this post-digestate was the most stable
multi-component digestates. As far as the three-component diges- of all the post-digestates (Table 2). This post-digestate had lower
tates were concerned, PSG/2/45 had the highest initial values of AT4 and GP21 values than all the three and four component
AT4 and GP21, and the fastest rates of decrease of these values. post-digestates. Organic matter content in all post-digestates ran-
The final values (after 120 d of post-digestion) of AT4 and GP21 ged from 9.9 g VS/L (MB/2/45) to 36.3 g VS/L (MSGD/3/45).
144 I. Wojnowska-Baryła et al. / Waste Management 77 (2018) 140–146

75 200
AT4 [mg O 2/g TS ] a1) a2)

GP21 [dm /kg TS]

60 160
AT4 = -0.037t + 34.10 GP21 = -0.367t + 113.42
45 120 R2 = 0.87

3
R2 = 0.77
30 80
AT4 = -0.050t + 12.83 GP21 = -0.425t + 61.77
15 R2 = 0.95 40 R2 = 0.92

0 0
0 20 40 60 80 100 120 0 20 40 60 80 100 120
time [d] time [d]
75 b1 ) AT4 = -0.369t + 65.06 200 b2 )
AT4 [mg O2/g TS]

GP21 = -0.754t + 154.47

GP21 [dm /kg TS]

60 R2 = 0.98 160
AT4 = -0.216t + 41.63 R2 = 0.95 GP21 = -0.461t + 110.80
45 R2 = 0.88 120 R2 = 0.95

3
30 80
15 AT4 = -0.113x + 26.33 GP21 = -0.446t + 91.66
40
R2 = 0.88 R2 = 0.96
0 0
0 20 40 60 80 100 120 0 20 40 60 80 100 120
time [d] time [d]
200 c2)
75
c1)

GP21 [dm /kg TS]

AT4 [mg O 2/g TS]

AT4 = -0.176t + 55.47 160 GP21 = -0.389t + 134.33

60
R2 = 0.92 R2 = 0.93
45 120

3
30 80

15 40

0 0
0 20 40 60 80 100 120 0 20 40 60 80 100 120
time [d] time [d]

MB/2/45 MB/3/60 PSG/2/45 MSG/3/45 MSG/2/45 MSGD/3/45 0. order kinetic model

Fig. 2. Linear trends in changes in the values of AT4 and GP21 of the digestates during 120 days of post-digestion (a1-a2) two-component digestates, (b1-b2) three-component
digestates, and (c1-c2) four-component digestate; zero order kinetic models and the coefficients of determination (R2) are given.

b) VScont = -0.301t + 58.38

60 a) 60
R2 = 0.96 VScont = -0.141t + 45.51
VScont = -0.033t + 36.20 R2 = 0.84
45 45
VS [g/dm ]
VS [g/dm ]

3
3

R2 = 0.82
30 30 VScont = -0.124t + 24.84
VScont = -0.062t + 17.33
R2 = 0.89
R2 = 0.91
15 15

0 0
0 20 40 60 80 100 120 0 20 40 60 80 100 120
time [d] time [d]

60 c) VScont = -0.126t + 51.50

MB/2/45
R2 = 0.88
MB/3/60
45
VS [g/dm ]
3

PSG/2/45
30 MSG/3/45
MSG/2/45
15
MSGD/3/45
0. order kinetic model
0
0 20 40 60 80 100 120
time [d]

Fig. 3. Linear trends in changes in VS content of the digestates during 120 days of post-digestion (a) two-component digestates, (b) three-component digestates, and (c) four-
component digestate; zero order kinetic models and the coefficients of determination (R2) are given.

MB/2/45 and MSG/2/45 had the lowest VS/TS ratios (both were food industry. They found the annual cumulative emissions from
below 0.6). For the rest of the post-digestates, the VS/TS ratios ran- stored digestate amounted to 12% of the CH4 produced within
ged from 0.63 to 0.68, with the highest value for PSG/2/45. the digester. The biogas yields obtained in the present study during
Covering storage tanks not only increases the biological stabil- post-digestion of multi-component digestates ranged from 50.9
ity of the digestate, but also reduces gaseous emissions to the dm3/kg TS (MSGD/3/45) to 114.9 dm3/kg TS (PSG/2/45). These
atmosphere and enables the capture of residual methane yield values amounted to 8.5–27.4% of the biogas productivity of the
from the digestate (Kaparaju and Rintala, 2006). Weiland (2003) feedstocks, and could be additional methane yields. Moreover,
found that the additional methane yields during post-digestion additional biogas yields during post-digestion accounted for 40–
are as much as 15%. Baldé at al. (2016) measured methane emis- 80% of the biogas productivity of the digestates, which means the
sions over two years from stored digestate from a mesophilic post-digestates will have much higher biological stability and emit
biodigester processing dairy manure and co-substrates from the much less biogas. Weiland (2003) stated that if only 5% of the
 I. Wojnowska-Baryła et al. / Waste Management 77 (2018) 140–146 145

0.8 The present results indicate strong linear relationships between

rate of the process (r)

the content of organic matter in post-digestates and their AT4 and

0.6
0.4
0.2
0.0
MB/3/60 MB/2/45
rAT4
Fig. 4. The rates of the decrease of AT4 (rAT4, mg O2/(g TS
d)) and GP21 (rGP21, dm3/
(kg TS
d)) values, and of VS content (rVS, g VS/(dm3
d)) during 120 days of post-
digestion of multi-component digestates.
produced methane is emitted from the storage tank, the positive
effect on the climate of the energetic use of biogas is completely
canceled out because the global warming effect of methane is at
least of 23 times higher than that of CO2. Therefore, it is suggested
that a closed secondary digestion tank should be used after anaer-
obic digestion. This practice is not common to all anaerobic diges-
tion plants because it is not economically valuable, and according
to the plant operators, the biogas potential of the digestate is
exhausted.
GP21 values (Fig. 5 a,b). There was good agreement between the
linear regression model and the observed values of VS content
and AT4 (R2 = 0.95), and between the model and the values of VS
content and GP21 (R2 = 0.96). This was also true when modeling
the relationship between AT4 and GP21 (R2 = 0.86).
Taking into account the indicators of the biological stability of
MSG/2/45 MSGD/3/45 MSG/3/45 PSG/2/45 the post-digestates and the relationships presented in Fig. 5a–c,
the stability of the post-digestates can be ranked in the following
rVS rGP21 order: MB/2/45 > MSG/2/45 > PSG/2/45 > MSG/3/45 > MB/3/45 >
MSGD/3/45. This means that after 120 days of post-digestion,
MB/2/45 was the most biologically stable and MSGD/3/45 was
the least biologically stable of the six tested post-digestates. It
should be emphasized that post-digestion improved the relative
stability of PSG/2/45 digestate, which was the least stable of the
digestates, but after post-digestion, was the third most stable
post-digestate.
There is little information available on the biological stability of
post-digestates. Tambone et al. (2009) tested biological stability
(ABP and OD20) of feedstock for a biogas plant, the corresponding
digestate and the outgoing material from the post-digester (after
50 days of post-digestion). They also found that post-digestion
increased the stability of digestate: ABP values decreased from
235 to 77 dm3/kg TS, and OD20 values decreased from 101 to 30

Table 2
Indicators of the biological stability of the multi-component post-digestates.

Indicators Units MB/2/45 MB/3/60 MSG/2/45 MSG/3/45 PSG/2/45 MSGD/3/45

EVS % 40.2 ± 1.2 32.3 ± 1.1 22.9 ± 0.9 22.5 ± 0.6 26.4 ± 0.5 35.8 ± 0.4
VS % TS 58.3 ± 1.1 63.0 ± 1.1 57.1 ± 1.2 66.2 ± 1.1 68.1 ± 1.4 65.2 ± 1.2
g VS/dm3 9.9 ± 0.5 30.9 ± 0.8 11.7 ± 0.8 26.2 ± 1.0 22.3 ± 0.9 36.3 ± 1.2
VS removed during 120 d g/dm3 7.3 ± 0.5 8.5 ± 0.6 14.3 ± 0.7 14.8 ± 0.7 35.3 ± 0.8 18.3 ± 0.5
% 45.3 ± 1.2 22.2 ± 1.0 55.0 ± 1.4 48.1 ± 1.5 61.6 ± 1.4 34.1 ± 1.1
AT4 mg O2/g TS 6.6 ± 1.2 28.8 ± 1.4 10.2 ± 1.4 19.8 ± 1.2 16.8 ± 1.2 37.4 ± 1.4
BY** dm3/kg TS 55.8 ± 5.2 57.9 ± 4.2 65.8 ± 4.8 52.2 ± 5.4 114.9 ± 7.5 50.9 ± 4.6
DBY** dm3/(kg TS
d) 0.47 ± 0.1 0.48 ± 0.1 0.55 ± 0.1 0.44 ± 0.1 0.96 ± 0.2 0.42 ± 0.1
GP21 dm3/kg TS 15.7 ± 4.5 66.4 ± 4.8 33.3 ± 3.5 58.8 ± 5.4 52.6 ± 4.6 79.2 ± 7.2
Organics* mg COD/dm3 1407.0 ± 80.0 2646.0 ± 75.0 5574.0 ± 65.0 8790.0 ± 90.0 16214.0 ± 120.0 19805.0 ± 220.0
mg TOC/dm3 520.0 ± 34.0 1103.0 ± 50.5 1984.0 ± 65.0 3416.0 ± 50.5 6150.0 ± 50.5 8153.0 ± 90.0
*
Organic compounds (COD, DOC) in the liquid fraction of post-digestates after 120 d of post-treatment, DOC for the compounds below 0.45 mm.
**
BY (biogas yield) = GP21digestate  GP21post-digestate, DBY (daily biogas yield)=BY/120 days.

60
a) 100
b)
GP21 [dm /kg TS ]
AT4 [mg O 2/g TS ]

y = 2.15x + 1.74
y = 1.07x - 4.58
80 R2 = 0.96 MSGD/3/45
45 R2 = 0.95 MB/3/60
MSGD/3/45
60
3

PSG/2/45 MSG/3/45
30 MB/3/60
40
PSG/2/45 MSG/3/45 MSG/2/45
15 MSG/2/45
20
MB/2/45
MB/2/45
0 0
0 10 20 30 40 50 0 10 20 30 40 50
VS [g/dm3] VS [g/dm3]
60
c)
AT4 [mg O 2/g TS ]

45 y = 0.48x - 4.38
R2 = 0.90 MSGD/3/45
30 MB/3/60
MSG/3/45
15 PSG/2/45
MSG/2/45
MB/2/45
0
0 20 40 60 80 100
GP21 [dm3/kg TS]

Fig. 5. The relationships between (a) AT4 values and organic matter content (VS) in post-digestates, (b) GP21 values and organic dry matter content (VS) in post-digestates,
and (c) AT4 and GP21 values of post-digestates; linear regression equations and the coefficients of determination (R2) are given.
146 I. Wojnowska-Baryła et al. / Waste Management 77 (2018) 140–146
mg O2/(g TS
20 h). They also showed that the values of biogas pro-
duction determined with the ABP test correlated with the values of
the OD20 test (r = 0.95, P < 0.05). The ABP values that those authors
reported after 50 days of post-digestion were similar to those of
the least biologically stable post-digestates in the present study
(MB/3/60, MSGD/3/45), although post-digestion lasted for 120
days; in contrast, the other post-digestates in the present study
were more stable.
Maynaud et al. (2017) showed the relationship between the
anaerobic (BDana) and aerobic (BDaero) biodegradability, which they
estimated according to Liu et al. (2015), and between CH4 yield and
respirometric test results (BOD, AT4 and DRI24) of post-treated
digestates (composted or dried solid fractions, and solid fractions
of digestate obtained after liquid-solid separation). The authors
observed large correlations between BDana and BDaero and between
CH4 yield and BOD (R2 = 0.8), whereas correlations between CH4
yield and AT4 and between CH4 yield and DRI24 were somewhat
smaller (R2 of 0.7 and 0.6, respectively). They concluded that the
composted solid fractions contained organic matter that was diffi-
cult to biodegrade because it contained complex molecules, which
implies that organic matter was more stable than before post-
treatment. The availability of such information on the stability of
digestates and post-digestates should contribute to the improve-
ment of agricultural re-use of these substances as fertilizers.
4. Conclusions
Anaerobic post-treatment of digestates seems to be valuable
and cost effective because it further degrades VS, making the
post-digestates more biologically stable. Psychrophilic post-
digestion seems to be especially advantageous when considering
the requirements for agricultural use of agri-food post-digestates
as fertilizers, which should be as biologically stable as possible,
and contain organic matter that is not a source of uncontrolled
methane emissions. It is suggested that a closed post-digestion
tank should be used after anaerobic digestion systems. Although
the amount of additional biogas produced during post-digestion
may not be important from an economic point of view, it reduces
uncontrolled methane emissions, which is important from an envi-
ronmental point of view because methane is a potent greenhouse
gas. Because the results of the AT4 test were highly correlated with
those of the GP21 test, both tests can be used to successfully deter-
mine the biological stability of digestates and post-digestates.
Acknowledgement
The work was supported by a Ministry of Science and Higher
Education in Poland (Statutory Research, 18.610.006–300).
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