Journal of Environmental Chemical Engineering 4 (2016) 2033–2039

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Journal of Environmental Chemical Engineering

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

Improvement of anaerobic digestion of swine slurry by steam

explosion and chemical pretreatment application. Assessment based
on kinetic analysis
Eduardo Ortega-Martineza , Consuelo Zaldivara , Javiera Phillippia , Helene Carrerec ,
Andres Donoso-Bravoa,b,*
a
Escuela de Ingeniería Bioquímica, Pontificia Universidad Católica de Valparaíso (PUCV), Av. Brasil 2085, Valparaíso, Chile
b
INRIA-Chile, Avenida Apoquindo 2827, Piso 12, Las Condes, Santiago, Chile
c
INRA, UR0050, Laboratoire de Biotechnologie de l’Environnement, Avenue des Etangs, 11100 Narbonne, France

A R T I C L E I N F O A B S T R A C T

Article history: Swine slurry is generated in large quantities and anaerobic digestion may represent an appropriate
Received 7 January 2016 process to both treat and revalue this waste as an energy source. Pretreatment processes aim to increase
Received in revised form 21 February 2016 the anaerobic biodegradability of a specific waste. In this study, steam explosion and weak thermal-alkali
Accepted 21 March 2016
pretreatments are evaluated. The first series of Biochemical potential (BMP) test is set to establish the
Available online 22 March 2016
best substrate/inoculum ratio conditions, and then other tests are carried out to assess the impact of
these pretreatments on the methane potential and maximum production rate by using the modified
Keywords:
Gompertz equation. The S/I ratio exerts an influence on the BMP results and the optimum ratio was found
BMP
Biogas
to be between 0.1–0.25 gVSs gVSI
1. Steam explosion leads to an improvement of both the methane
Alkali Pre-treatment potential and the maximum production rate at the highest severity conditions. Weak alkali thermal
Substrate/inoculum ratio pretreatment improved the maximum production rate but affected negatively the methane potential of
Steam explosion pretreatment the substrate.
ã 2016 Elsevier Ltd. All rights reserved.

1. Introduction sector for energy conversion, such as biogas, is of increasing

Pig or Swine slurry (PS, hereafter) corresponds to the liquid
fraction that comes from the swine pen which comprises cleaning
water, urine and animal feces. Due to the large consumption of
pork meat, the generation of the waste poses a significant
challenge because of the quantity and the complexity of its
composition. A single animal may generate up to 20 l of slurry per
day which yields a stunning large amount of waste if, in the case of
Chile, we consider that there are around 5.5 million heads of pork.
Anaerobic digestion is a consolidated technology especially in
Europe, although increasingly in North and South America as well.
Recently the Chilean government released a 20/25 plan, which
obligates the electric companies to produce 20% of their electricity
from non-conventional renewable energy (ERNC) sources for 2025.
For this reason the utilization of waste from the agro-industrial
* Corresponding author at: Escuela de Ingeniería Bioquímica, Pontificia
Universidad Católica de Valparaíso (PUCV), Av. Brasil 2085, Valparaíso, Chile.
E-mail addresses: andres.donoso@ucv.cl, andres.donoso@inria.cl
(A. Donoso-Bravo).
importance. Problems related to the emissions from PS could be
avoided by using anaerobic digestion process, in addition, energy
could be produced. Waste to energy with high efficiency values
could be achieved thanks to the use of conventional combined
heat-power (CHP) units or more efficient technologies, such as, for
instance, the Solid oxide fuel cell (SOFC) energy generators [1]. The
anaerobic degradation of swine manure has been largely studied at
different scales and currently there are several full scale facilities
where the biogas is harnessed to produce electricity [2]. PS has
been commonly used in combination with another substrate in a
co-digestion process in order to balance the C/N ratio of the
substrate [3,4].
PS may contain high fractions of fibers and other compounds,
which are slowly degraded by anaerobic microorganisms [5].
Several pretreatment techniques have been implemented in order
to improve the methane conversion in anaerobic digestion by
making complex substrates more accessible to anaerobic biomass.
The type of pretreatment technique to choose will depend on the
type of feedstock that will be finally anaerobically digested. An up
to date list of suggested pretreatment procedures linked to the type
of feedstock is provided in Carrere et al. [6]. However, little

http://dx.doi.org/10.1016/j.jece.2016.03.035
2213-3437/ ã 2016 Elsevier Ltd. All rights reserved.
2034 E. Ortega-Martinez et al. / Journal of Environmental Chemical Engineering 4 (2016) 2033–2039

attention has been paid to the pretreatment of PS prior to
anaerobic digestion. Almost all the research has been focused on
the soft thermal pretreatment of this waste without steam
explosion [7] or with steam explosion [8]. Thermal/steam
explosion pretreatment has been implemented in full-scale
wastewater treatment plants to solubilize sewage sludge because
this technology has demonstrated its economical and energetic
benefit [9]. Chemical pretreatments, particularly thermal-alkali,
which breaks apart the particulate organic matter, especially
proteins ester solvation and saponification, has been applied to the
solid fraction of PS and high alkali doses were applied, above
0.1 gNaOH or Ca(OH)2 gTS
1 [10,11]. The latter entails the
incorporation of a dewatering process as well as the high cost of
alkali in full-scale installation and an upstream pH adjustment
before anaerobic digestion, in other words, an acid addition.
The Biochemical Methane Potential (BMP) test represents the
conventional procedure to evaluate anaerobic biodegradability of
any substrate fed into anaerobic digesters. The initial ratio between
substrate to inoculum, S/I, exerts a significant influence upon the
BMP results [12,13]. The optimum S/I of the anaerobic digestion of
PS, to our knowledge, has never been studied, and the influence of
this ratio is crucial in order to find the optimum conditions for a
BMP test. The aim of the study is to evaluate the impact of two
pretreatment technologies, steam explosion and weak thermal/
alkali, on both the methane production rate and the methane yield
in batch conditions from PS at the optimal initial substrate/
inoculum ratio.
2. Material & methods
2.1. Inoculum and substrate
Pig slurry samples were taken from the animal protein producer
Agrosuper Company located in Corneche Sector, around the city of
San Pedro, Chile. The anaerobic inoculum (I), (21.45  0.14 gTS kg
1
and 12.42  0.08 gVS kg
1) was obtained from a continuous lab-
scale stirred tank reactor maintained in mesophilic conditions and
fed with sewage sludge at an OLR of 0.8 KgVS m
3 d
1.
2.2. Pretreatments
2.2.1. Steam explosion (SE)
The experiment was carried out in an automatic pilot-scale
thermal system located in the Wastewater Treatment Plant
(WWTP, Santiago, Chile). This system consisted of a feeding tank,
a progressive cavity pump (Pmax = 12 bar), a steam boiler, a 20 l total
volume hydrolysis reactor (working volume = 10 l) connected to a
flash tank (V = 10 l) with also outlet pipes for steam and hydrolyzed
sludge. The pilot plant is equipped with automatic valves that
control the steam entrance from the boiler and the sludge exit from
the reactor to the flash. A data acquisition and control system is
used to measure pressure and temperature and to automatically
control the steam inlet and the hydrolyzed sludge exit to the flash.
The pump introduces 10 l of PS into the reactor, and then the steam
valve is opened until pressure and temperature reach the set point.
At the end of the reaction time, the decompression valve is
automatically opened and the hydrolyzed PS flows to the flash
tank. The pretreatment conditions applied are shown in Table 1.
The equation to calculate the severity factor (R0) of each evaluated
conditions is presented in Eq. (1):
 
T
100
LogR0 ¼ Log t  expð Þ ð1Þ
14:75
where t represents the time of thermal treatment part (min) and T
is the average temperature ( C). It has to be kept in mind that this
equation does not take into account the explosion effect but is
widely used for data analysis purposes.
2.2.2. Weak thermal-alkali pretreatment (wTA)
Weak-Alkali pretreatment was carried out with two different
NaOH solution doses. The pretreatment conditions (time and
temperature) were selected according to what has been
previously established for this type of pretreatment [14,15].
PS samples were pretreated at different conditions as it is
shown in Table 2. Reaction blanks (B) without NaOH solution
and blank test without PS addition were prepared. All the tests
were carried out in triplicate.
2.3. Biomethane potential test
The biochemical methane potential assay (BMP) was performed
according to [16] guidelines. The assay was carried out in a glass
bottle with 100 ml working volume. Given the small volume of the
BMP test and, consequently, the small amount of substrate and
inoculum to be added, both the substrate and inoculum additions
were carried out keeping the storage flask stirred continuously in
order to assure the homogeneity of the mixture as much as
possible. Sodium bicarbonate was added in a 1 g of NaHCO3 per
gram of volatile solid of substrate. The initial pH of the assay was
adjusted to 7.1  0.1. Blanks were carried out to subtract the biogas
produced from the inoculum. All the tests were carried out in
triplicate. The temperature of the assay was 37  C controlled by a
thermoregulated chamber. The methane production was measured
by liquid displacement using a NaOH solution which absorbs
carbon dioxide. The S/I optimum estimation was performed with
raw PS. A certain amount of substrate was added in order to have
the following S/I ratios: 0.1, 0.25, 0.5, 0.75 and 1 gVSS VSI
1. The
inoculum concentration was kept constant at 5 gVS/l. For
parameter determination the Modified Gompertz equation (GM)
(Eq. (2)), which has been used for methane production [17], was
applied
  
Rm  e
B ¼ P  exp
exp ðl
tÞ þ 1 ð2Þ
P

Table 2
Selected conditions for the weak alkali pretreatment.
Table 1
Condition NaOH (gNaOH gTS
1) Time (min) Temperature ( C)
Selected conditions for steam explosion pretreatment.
wTA1 0.0045 5 100
Notation Temperature ( C) Time (min) Severity Factor wTA2 0.0045 30 100
SE1 155 5 2.32 wTA3 0.0225 5 100
SE2 170 30 3.54 wTA4 0.0225 30 100
SE3 171 15 3.27 wTA5 0.0225 18 35
SE4 142 5 1.94 wTA6 0.0045 18 35
SE5 142 30 2.71 B1-3 – 5 100
SE6 134 15 2.18 B2-4 – 30 100
SE7 122 5 1.35 B5-6 – 18 35
 E. Ortega-Martinez et al. / Journal of Environmental Chemical Engineering 4 (2016) 2033–2039
where B is the biogas produced at time t,P is the maximum biogas
production (ml), Rm maximum biogas production rate (ml d
1), l
the lag time (d) and e corresponds to exp(1) equal to 2.7183. For
parameter estimation, Matlab 1 and a simple least squares
criterion, between the simulated and experimental, were used
for the minimization procedure. The standard deviation was
estimate from the covariance matrix of the parameters obtained
from the inverse of the Fisher Information Matrix (FIM), which
gives a lower bound on the achievable parameter error covariance
matrix.
2.4. Analytical methods
Total solids (TS) and volatile solids (VS) were measured by
gravimetric methods. Fat, ashes, fiber and Total Kjeldahl Nitrogen
(TKN) were estimated in dry basis through methods 963,15;
940,15; 920,169 and 2001,11 described in [18] AOAC, respectively.
Ammonia N-NH4+ and carbohydrates were estimated by selective
electrode and by mass balance, respectively. The protein content
(Pc) was calculated based on the assumption that the protein
contains 16% (w/w) nitrogen and taking into account TKN and
ammonia measurements, according to, protein = (TKN
NH4+)6.25
[19]. The accumulated methane volume was measured manually
by a NaOH solution displacement. The methane yield was
estimated by using the equation (Eq. (3)), proposed by Raposo
et al. [20].
Methaneyield ¼ 415  %Carbohydrates þ 496  %Proteins
þ 1014  %Lipids
3. Results and discussion
3.1. Substrate characteristics and substrate/inoculum ratio evaluation
Fig. 1 shows the PS characterization (composition of the dry
weight). The content of TS and VS was 43.4 and 16.6 g kg
1,
respectively. The high content of ashes in the PS stands out from
the characterization which reflects the amount of sand, and other
compounds that are dragged into the washing procedure that
finally tags along with the organic material. Within the organic
material, as expected, the proteins represent the largest fraction
followed by fat, fiber and carbohydrates in similar quantities. By
using this composition and the Eq. (3), the theoretical methane
potential may be estimated, being 470 ml CH4 gVS
1 without
considering the fiber degradation. After a pretreatment, where the
Fig. 1. Pig slurry characterization (dry weight).
2035
fiber is solubilized, the theoretical methane potential may reach
540 ml CH4 gVS
1.
Fig. 2 shows the results of the cumulative methane production
at different S/I ratios. The methane production profiles turned out
to be quite different depending on the S/I ratio used. Overall, it can
be noticed that the S/I ratio exerts an important influence on the
kinetic and extent of PS degradation. The greatest methane yield
and production rates were achieved at low S/I, specifically the best
results were obtained for the ratio of 0.1 and 0.25 (Table 3). The
error bars for the lowest S/I ratio tests are quite large, therefore a
significant difference between them cannot be established. These
results also match the ones obtained with swine slurry by
González-Fernández and García-Encina [21] who also found that
the lowest S/I ratio they used, yield the best results in term of
methane production. Nevertheless, it is clear that even for similar
substrates the results of the optimum S/I may vary depending on
the specific composition [22,23]. The lag-phase parameter is
unusually high for each conditions, which is mainly due to a
problem with the temperature of the room where the BM tests are
placed, which lasted for around 5 days.
The S/I ratio is quite influential on the results of BMP test;
however, it is not always evaluated for each substrate that is
analyzed. Most of the time, S/I = 0.5 (or inoculum to substrate ratio
(ISR) 2) is normally used for BMP tests. Indeed this value has been
widely used when sewage sludge is used as substrate and 0.5 ratio
has been identified as a sort of boundary of S/I ratio in order to
avoid product inhibition [24]. Several values of S/I ratio have been
evaluated for different substrates and, values lower than 0.5 have
ð3Þ been found to be the optimum, in terms of the attained methane
yield, with complex substrates such as algae residues [25] food
waste [23] or sunflower cake [13]. The majority of the research
focuses on the methane yield to determine the optimum S/I ratio
although other variables have also been taken into account such as
the VFA accumulation [26] or Escherichia coli disappearance [27].
The inoculum source and the S/I ratio have a significant effect on
the anaerobic biodegradation, due to different levels of microbial
activity and populations contained [20,28,29]. A S/I ratio of
0.25 was then chosen to carry out the BMP test of untreated and
pretreated PS.
3.2. Steam explosion
The results of the anaerobic degradation of raw and pretreated
PS at different conditions are depicted in Fig. 3. The parameter
values (P and Rm) of the Gompertz equation are shown in the
second column of Fig. 3. From the methane production profiles it
can be observed that two conditions, SE2 (170  C, 30 min) and SE3
(171  C, 15 min) exerted a positive influence on the BMP test results
in regards to the production rate and the methane potential. An
improvement of 44% and 16% of P was attained for SE2 and SE3,
respectively, as well as 25% of Rm for SE3 with respect to the test
with raw PS. The difference between SE2 and SE3 demonstrates
that the cooking time also exerts a positive influence in increasing
the methane potential. At 4 bar (130–140  C, i.e. SE5, SE6 and SE7),
none of the tests showed an improvement in comparison to the
raw PS test. A pretreatment for at least 30 min at 170  C is required
to improve methane yield and at least for 15 min at 170  C to
enhance the digestion rate. Despite the other conditions show a
trend to decrease the methane potential it has to be pointed out
that the standard deviation of the parameters is obtained by using
the mean value of the BMP test and its disagreement with the
model at the optimum point, it is a method that does not take into
account the experimental error depicted in the methane produc-
tion profiles. Therefore a more in-depth discussion about this was
not included. The lag-phase parameters are not shown due to the
fact that the majority of the values were close to zero, in addition,
2036 E. Ortega-Martinez et al. / Journal of Environmental Chemical Engineering 4 (2016) 2033–2039

Fig. 2. Methane production profiles at different S/I ratios.

the ones that were different to zero did not reflect properly the 3.3. Chemical pretreatment
experimental data.
The only study found in literature in which steam explosion is The results for the BMP test of the thermal-alkali pretreated PS
applied to swine waste is presented by Ferreira et al. [30]. In that are shown in Fig. 4. These results are presented in order to
study the solid fraction of swine manure is pretreated at different highlight the effect of the reaction blanks (only thermal treatment)
combinations of temperature and time. The best results in regards and assessing the whole methane production kinetic of the
to both Rm and P are obtained at 7 bar (around 170  C) and 30 min, anaerobic degradation. Both aspects have not been usually taken
although the values of the parameters in comparison to the ones into account in the existing related literature. In regards to the
obtained in the present study differ. The Rm obtained by Ferreira methane production profiles, it is clearly noticed that the final
et al. [8] was double that which was obtained in this study, but in production of methane was always less than the one reached with
turn, P is 39% less than the value obtained with PS. This important raw PS. In the second column of Fig. 4, a comparison of the
difference may be explained by the nature of the substrate and its estimated parameters for each condition is presented. As
solid content. Ferreira et al., [6] used concentrated pig slurry with a mentioned, the final methane production, P, was lowered after
solid content of around 3-fold the one we used. These results show the pretreatment which is in agreement with the results obtained
that the steam explosion will have a variable impact depending, on for the steam explosion at low severity factors. Rincon et al., [31]
the substrate concentration in regards to the macromolecular observed a diminishing of the methane potential at 120  C and
compositions. For the case of sewage sludge, it has been then, an increase at 180  C in a BMP test using olive pomace as
demonstrated that in order to make a process feasible, the waste substrate. Likewise, Carrère et al., [32] only attained a significant
needs to be concentrated [9]; however, that cannot be extrapolated improvement of the methane potential at 190  C treating pig
to any industry and these results show that for raw PS, it may be manure. Refractory compounds have been reported to be produced
more appropriate to undergo thermal and steam explosion in alkali pretreatment that may impair the anaerobic biodegrad-
pretreatment as such, than only the solid fraction, although a ability [33]. Particularly sodium salts of free amino acids from
lot of the heat will be used to boil water instead of focusing on the proteins and glycerol and soap/salts of fatty acids from lipids,
dry matter. Temperature may not be a limitation when combined which are formed during the alkali hydrolysis may impair the final
CHP units are used to harness the biogas. Furthermore, pumping anaerobic biodegradability of the substrate [34,35]. Likewise the
and clogging, common problems with concentrated waste are also lag-phase in general was increased after the pretreatment from
lowered thanks to steam explosion which leads to the reduction of 8 to around 12 days. Similar results in regards to the worsening of
waste viscosity. the methane production was also obtained by Carrere et al. [32]
who evaluated the biodegradability of both the liquid and
particulate fractions of the raw and thermal-alkali pretreated
swine manure when they used high soda doses (0.4 g NaOH gTS
1).
In contrast, they obtained an increase of manure methane potential
Table 3 using a lower soda dose (0.15 g NaOH gTS
1) and temperature
Parameter values from Gompertz modified equation fitted with BMP test results at
ranging from 150 and 190  C, whereas the liquid fraction
different S/I ratios.
biodegradability was decreased. Ferreira et al. [8], who used
S/I ratio Parameters dewatered pig manure for the thermal-alkali pretreatment
P (ml CH4 gVS
1) Rm (ml CH4 gVS
1 d
1) l (d) r2 evaluation, obtained variable results depending of the temperature
1 105  1 10.5  0.3 8.5  0.1 0.99
of the alkali pretreatment. These two studies used a much greater
0.75 126  70 23.0  5.3 12.4  0.6 0.98 alkali dose between 10 and 20 fold.
0.5 160  8 10.8  0.9 8.2  0.6 0.99 On the other hand, a high enhancement of the maximum
0.25 289  4 27.7  0.9 7.4  0.2 0.99 production rate (Rm) was attained at WTA3 (0.0225 g NaOH gTS
1,
0.1 357  19 24.5  2.1 8.9  0.5 0.99
5 min and 100  C) and WTA2 (0.0045 gNaOH gTS
1, 30 min and
 E. Ortega-Martinez et al. / Journal of Environmental Chemical Engineering 4 (2016) 2033–2039
Fig. 3. Experimental methane production profiles (symbols) and simulated values (solid line) at different steam explosion conditions. Figs. (a) and (b) Methane production
profiles of raw PS and pretreated PS. Second column fitted parameters from the modified Gompertz equation.
100  C), although a real improvement was due to the alkali addition
which was only observed at WTA2, which indicates the importance
of the contact time between the substrate and the alkali. A high Rm
entails, in the end, that a lower retention time will be needed to
achieve a specific methane yield in a continuous full scale plant.
Strong alkali pretreatment of the solid fraction of swine manure
(pH = 14, 32  C for 24 h) was carried out by Pérez-Elvira and Fdz-
Polanco [9] who observed an increase in both in the production
rate and the final methane production.
In regards to the lag-phase parameter, there was an increase in
comparison the lag-phase of PS, which must be related to the slight
increase of pH that occurs after the pretreatment. The lag-phase
parameter does not possess the same importance than the other
two parameters due to the fact that strongly depends on the initial
conditions of the test, which are much less influential in a
continuous operation at full-scale.
3.4. Overall pretreatment comparison
In regards to the final methane yield (P), steam explosion
allowed to reach higher values than chemical pretreatment, even
higher than the value obtained for raw PS during the chemical
pretreatment. It is worth to mention, the unexpected high value of
this parameter for raw PS during this experiment, which may have
been influenced by changes occurred during storage. With respect
to the maximum production rate (Rm), in turn, higher values were
attained after chemical pretreatment than steam explosion, taking
2037
only into account the best found pretreatment conditions for both
tests. In general, these results are indicating that steam explosion
helps to increase the biodegradability by converting some of the
non-degradable organic matter into accessible for the anaerobic
biomass (fibers, for instance), whereas weak-alkali pretreatment
converts slowly degradable organic matter to readily degradable
compounds. Regardless, a wider characterization of the pretreated
substrates would give more insight on what really occurs after this
pretreatment, however, BMP tests offer a good glance of the what
this process leads to.
Regarding a possible full-scale implementation of these
processes, in the case of steam explosion, the improvement of
the methane yield and production rate entail a better performance
of a digester in a shorter time, in other words, an important
increment of the process productivity. At a full-scale plant, a
smaller reactor that allows to reach a high methane production
would mean a decreasing in the investment and operational cost
compared to a conventional anaerobic digestion configuration.
This may offset, in some extent, the cost of the pretreatment unit.
In the case of the chemical pretreatment, the improvement of the
production rate will reduce the residence time, or reactor volume
in a full-scale continuous reactor, however the productivity of the
digester may not vary since the methane production is lowered.
The potential benefit of this alternative is less clear than the steam
explosion, although, in any case, a detailed economic and technical
assessment is always necessary to decide whether a pretreatment
procedure is necessary and worthwhile.
2038 E. Ortega-Martinez et al. / Journal of Environmental Chemical Engineering 4 (2016) 2033–2039
Fig. 4. Experimental methane production profiles (symbols) and simulated values (solid line) at different weak thermal-alkali pretreatment conditions. Fig. (a): raw PS and
blanks. Fig. (b): raw PS and pretreated samples. Second column, fitted parameters from the modified Gompertz equation.
4. Conclusion
The impact of steam explosion and weak thermal-alkali
pretreatment upon the kinetic and the extent of the anaerobic
biodegradation of pig slurry was assessed. The S/I ratio exerts an
influence on the BMP results and the optimum ratios were found to
be between 0.1–0.25 gVSs gVSI
1. Steam explosion led to an
improvement of both the methane potential and the maximum
production rate at the highest severity conditions investigated
(170  C, 30 min). On the other hand, weak alkali thermal
pretreatment improved the maximum production rate but affected
negatively the methane potential of the substrate. For a potential
real application, steam explosion showed more promising results
that would allow to at the same time reduce the residence time of
the anaerobic system and increase the methane productivity.
Further research should focus in going deeper into the changes of
the substrate properties during pretreatment as well as the
development of a modeling procedure that links the results
obtained in batch with the operation of a full-scale (semi)
continuous anaerobic reactor.
Acknowledgements
This study was funded by Fondecyt Initiation project no
11130462 and supported by CIRIC-INRIA-Chile (EP BIONATURE)
through Innova Chile Project.
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