Bioresource Technology 227 (2017) 239–246

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Thermochemical pretreatment and anaerobic digestion of dairy cow

manure: Experimental and economic evaluation
Fabiana Passos b, Valentina Ortega a, Andrés Donoso-Bravo a,c,⇑
a
Escuela de Ingeniería Bioquímica, Pontificia Universidad Católica de Valparaíso, Chile
b
Environmental and Chemical Technology Group, Department of Chemistry, Universidade Federal de Ouro Preto, 35400-000 Ouro Preto, Minas Gerais, Brazil
c
INRIA-Chile, Avenida Apoquindo 2827, piso 12, Las Condes, Santiago, Chile

h i g h l i g h t s

Acid and alkali pretreatment were technically and economically evaluated.

1

A S/I ratio of 0.25 g g showed the best results in terms of degradation kinetic.

The methane potential increased 23.6% with a dose of 10% of NaOH at 100 °C for 5 min.

The methane potential increased 20.6% with a dose of 2% of HCl at 37 °C for 12 h.

Economically, anaerobic digestion without pretreatment outperforms the other options.

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

Article history: The aim of this study was to assess technically and economically the application thermochemical pre-
Received 8 November 2016 treatment in the anaerobic digestion of dairy cow manure. After selecting the optimum substrate to
Received in revised form 7 December 2016 inoculum (S/I) ratio in a preliminary BMP test, the following tests compared 20 different pretreatment
Accepted 9 December 2016
conditions varying temperature (100 and 37 °C), exposure time (5 and 30 min and 12 and 24 h) and
Available online 18 December 2016
chemical doses (0.5, 2, 6 and 10% of HCl or NaOH). The highest value of maximum production rate
was achieved at an S/I ratio of 0.25 g VSs g VS1
i . The major improvements of the methane potential were
Keywords:
23.6% with 10% of NaOH at 100 °C for 5 min and 20.6% with 2% of HCl at 37 °C. The technical-economic
Agricultural waste
Bioenergy
analysis showed that the implementation of neither thermal alkali nor thermal-acid pretreatment would
Biogas be feasible and the conventional one-step anaerobic digestion outperforms both alternatives.
Methane Ó 2016 Elsevier Ltd. All rights reserved.
Pre-treatment
Modeling

1. Introduction
Anaerobic digestion (AD) represents one of the most sustainable
ways of treating waste and wastewater due to the production of
biogas, which can be harnessed as electricity and/or heat replacing
the use of fossil fuels. Furthermore, the nutrient-rich digestate can
be used for soil amendment. Currently, AD is widely used in Europe
where the technology is considered consolidated. In Chile, around
50 MW of electricity are generated from biogas; however there are
plenty of other AD plants where the biogas is wasted and not well
energetically utilized.
⇑ Corresponding author at: Escuela de Ingeniería Bioquímica, Pontificia
Universidad Católica de Valparaíso (PUCV), Av. Brasil 2085, Valparaíso, Chile.
E-mail addresses: andres.donoso@pucv.cl, andres.donoso@inria.cl
(A. Donoso-Bravo).
Dairy cow manure (DCM) is the most used substrate for AD,
particularly in the south of Chile, where this type of livestock is
farmed. Therefore, it is crucial to count with a better understanding
of the anaerobic biodegradability of this substrate and the eco-
nomic and energetic viability of the technology application. In this
context, the biochemical methane potential (BMP) test is the most
used and accepted procedure to evaluate both the methane maxi-
mum production, as well as the hydrolysis rate when this step is
the limiting reaction in the AD process. The initial condition of a
BMP test, i.e. the ratio between substrate and inoculum (S/I ratio),
is the only variable that can be manipulated and it has been proven
that exerts a significant influence upon the results of BMP test.
Concerning the AD of cow manure, previous research obtained
methane yields between 200 and 340 mlCH4 g VS1 depending on
the applied S/I ratio (Kafle and Chen, 2016; Moset et al., 2015).
However, the majority of the studies have focused on the co-
digestion of cow manure with other substrates (Zheng et al., 2015).

http://dx.doi.org/10.1016/j.biortech.2016.12.034
0960-8524/Ó 2016 Elsevier Ltd. All rights reserved.
240 F. Passos et al. / Bioresource Technology 227 (2017) 239–246

Since the hydrolysis step is limiting the AD of particulate and Table 1

complex substrates, such as DCM, pretreatment methods may be Anaerobic digestion of dairy cow manure under different thermochemical pretreat-
ment conditions.
applied for solubilizing organic matter and, consequently, increas-
ing AD rate and extent. In fact, plenty of research have reported Trial Temperature (°C) Exposure time Chemical dose (mL/100 g VS)
improvements on the AD of several solid and semi-solid substrates Acid (HCl)/Alkali pretreatment
by employing pretreatment techniques (Carrere et al., 2015). C – – –
Nonetheless, for cow manure, few results have been carried out P1.0 100 5 min –
P1.1 100 5 min 0.5
so far, although from those all of them aimed at breaking down P1.2 100 5 min 2.0
the fiber present in the biomass. For instance, Jin et al. (2009) used P1.3 100 5 min 6.0
microwave and chemical pretreatments for evaluating the P1.4 100 5 min 10.0
biodegradability of dairy manure. In this case, acids and bases yield P2.0 100 30 min –
P2.1 100 30 min 0.5
the best results. Alkali along with mechanical pretreatment were
P2.2 100 30 min 2.0
assessed by Angelidaki and Ahring (2000). The results obtained P2.3 100 30 min 6.0
showed that an improvement on the methane potential was P2.4 100 30 min 10.0
achieved depending on the size of the pretreated fibers. Moreover, P3.0 37 12 h –
an important aspect besides the pretreatment effectiveness, P3.1 37 12 h 0.5
P3.2 37 12 h 2.0
regards the economic feasibility of its implementation. For chemi-
P3.3 37 12 h 6.0
cal pretreatment, the few studies that can be found are carried out P3.4 37 12 h 10.0
to evaluate waste activated sludge pretreatment (Cho et al., 2014; P4.0 37 24 h –
Ruffino et al., 2016). Results obtained indicated that the economic P4.1 37 24 h 0.5
P4.2 37 24 h 2.0
benefit is highly dependent on the chemical doses.
P4.3 37 24 h 6.0
The aim of this study was to compare acid and alkaline thermo- P4.4 37 24 h 10.0
chemical pretreatments prior to the anaerobic digestion of DCM in
BMP tests. For this, the first step consisted in optimising the S/I
ratio by investigating different amounts of DCM and digested sew-
Pretreatments were prepared in solutions and subsequently added
age sludge. After selecting the optimum S/I ratio, several tempera-
to BMP tests in a solid to liquid ratio of 0.05 g VS mL1 in order to
tures, exposure times and chemical doses were varied for studying
maintain the S/I ratio of 0.25.
the pretreatment effectiveness on the susbtrate anaerobic
For comparing the effect of only the temperature, a thermal pre-
biodegradability. Afterwards, the pretreated and raw DCM were
treatment without chemical addition was evaluated for the four
used as substrate for BMP test and the results were analyzed by
temperature-time pair conditions. Furthermore, a control trial with
using the Gompertz equation. Finally, a preliminary techno-
raw DCM without any pretreatment was also assessed. The pH of
economic assessment of full-scale application was carried out for
each test was measured right after the chemical addition at the
the best pretreatment conditions as well as the model parameters
beginning and at the end of the thermal pretreatment.
drawn from the BMP test.

2. Material and methods 2.3. Biochemical methane potential tests

2.1. Substrate, inoculum and S/I ratio assay
The DCM used as substrate in this study was obtained from two
small scale dairies located in the cities of Melipilla and Casablanca
(Chile), for S/I ratio and thermochemical pretreatment/BMP test,
respectively. The anaerobic inoculum was obtained from a contin-
uous mesophilic lab-scale stirred tank reactor, fed with primary
sludge and waste activated sludge at an organic load rate (OLR)
of 0.8 g VS L1 d1, and had a concentration of 5 g VS L1. The reac-
tor was made of plexiglass with a total volume of 5 L, placed in a
thermo-regulated chamber and mechanically stirred. The mixed
sewage sludge was taken from the wastewater treatment plant
of Santiago, Chile.
The S/I ratio influence was assessed at five different conditions,
namely: 0.25, 0.50, 0.75, 1.0 and 2.0 in terms of volatile solids
(g VSS g VS1
I ). This first step was used to obtain an optimum S/I
ratio, which was subsequently applied for thermochemical pre-
treatment in BMP tests.
2.2. Thermochemical pretreatments
BMP tests were carried out according to the guidelines pro-
posed by Angelidaki et al. (2009). To this aim, glass bottles of
120 mL with 100 mL of working volume were added with DCM
as organic substrate and digested sewage sludge as inoculum at
different proportions for the first part of this study and at an opti-
mum S/I ratio for the second part. Sodium bicarbonate was added
in a concentration of 1 g NaHCO3 g VS1 of inoculum. After the
addition of chemical (HCl or NaOH), pH was adjusted to neutral
values of 7.1 (±0.1). Blank trials with only inoculum were used to
quantify the amount of methane produced by endogenous respira-
tion. Each BMP was performed in triplicate. The temperature of the
assay was set at mesophilic temperature (37 °C) and was con-
trolled by a thermoregulated chamber.
The methane production was measured by liquid displacement
using NaOH solution (40 g L1), which absorbs CO2. Results were
expressed in methane yield, which was calculated by dividing
the accumulated volume of methane produced by the VS content
in each bottle (mlCH4 g VS1). The net value of methane yield
was obtained by subtracting the endogenous production of the
blank bottle and converted to normal conditions of temperature
(0 °C) and pressure (1 atm).

Thermochemical pretreatment was studied at twenty (20) dif-

ferent conditions for acid (HCl) and alkaline (NaOH) chemicals, as
detailed in Table 1. Four temperature and time pair conditions:
100 °C for 5 min, 100 °C for 30 min, 37 °C for 12 h and 37 °C for
24 h, were combined with four chemical doses: 0.5, 2.0, 6.0 and
10.0% in VS basis. For substrate heating, a Soxhlet equipment with
thermo-regulation at 100 °C and a water bath at 37 °C were used.
2.4. Parameter estimation
Experimental data from BMP tests were modeled using the
modified Gompertz equation, as described below (Eq. (1)), where
B is the methane yield at a time t (mlCH4 g VS1), P is the maxi-
mum methane production (mlCH4 g VS1), Rm is the maximum
 F. Passos et al. / Bioresource Technology 227 (2017) 239–246 241

yield rate (mlCH4 g VS1d1), k is the lag-time (d) and e corre- Table 2
sponds to exp(1) equal to 2.7183. Dairy cow manure characteristics. Mean values (standard deviation).

   Parameter S/I ratio assay Pretreatment assay

Rm  e
B ¼ P  exp exp ðk  tÞ þ 1 ð1Þ TS (g/L) 41.12 (1.29) 44.31 (1.31)
P
VS (g/L) 33.29 (1.17) 35.66 (0.91)
For parameters estimation, MatlabÒ and single least squares cri- VS/TS (%) 80.96 (0.30) 80.69 (0.34)
Ashes (% w/w) 19.04 (0.30) 19.50 (0.34)
teria between simulated and experimental data were used for the
Fiber (% w/w) 27.80 (1.87) 31.08 (0.51)
minimization procedure. The standard deviation was estimated Lipids (% w/w) 4.74 (0.11) 3.26 (0.04)
from the covariance matrix of the parameters obtained from the Proteins (% w/w) 8.41 (2.52) 26.76 (1.80)
inverse of the Fisher Information Matrix (FIM), which gives a lower Carbohydrates (% w/w)* 40.01 19.40
bound on the achievable parameter error covariance matrix. *
Note: Carbohydrates was estimated as the difference between ashes, fiber, lipid
and protein content from 100%, all in dry basis.
2.5. Analytical methods

Total (TS) and VS solids were measured by gravimetric meth- 300

ods. Ammonia (N-NH+4) was determined by selective electrode.

Content of ashes, fiber, lipid and total Kjeldahl nitrogen (TKN) were 250

Methane yield (mL CH4/g VS)

estimated in dry basis according to the methods 940,15; 920,169;
963,15 and; 2001,11; respectively, described in the AOAC (2012).
200
The protein content was calculated by assuming that protein con-
tains a 16% (w/w) of nitrogen. Therefore, it could be calculated
according to the equation below (Eq. (2)) (Donoso-Bravo et al., 150

2011).

Proteins ¼ TKN  NHþ4  6:25

100
ð2Þ 0.25 g VSs/g VSi
0.5 g VSs/g VSi
50 0.75 g VSs/g VSi
1.0 g VSs/g VSi
2.6. Economic analysis 2.0 g VSs/g VSi

0
A preliminary techno-economic analysis comparing three sce- 0 10 20 30 40
narios of full-scale implementation was incorporated. The three
Digestion time (days)
scenarios were: i) AD without a pretreatment step, ii) Thermal
alkali pretreatment followed by AD and, iii) Thermal acid pretreat- Fig. 1. Accumulated methane yield under different S/I ratios.
ment followed by AD. The best two pre-treatment combinations in
regards to temperature and time for both alkali and acid pre-
treatment obtained in the BMP test were chosen for the analysis. Table 3
In order to link the lab results with the techno-economic analysis pH values before and after thermal and thermochemical pretreatments.

the parameters obtained from the model fitting, i.e. Rm and P (see pH ini pH fin pH ini pH fin
2.4), were directly used for designing the anaerobic digesters and Acid (HCl) Alkaline (NaOH)
for estimating the methane yield of the process. Particularly, the P1.0 6.66 8.78 P1.0 7.17 8.62
hydraulic retention time (HRT) was assumed to be the ratio P1.1 6.58 8.49 P1.1 7.28 8.81
between P and Rm. P1.2 6.37 8.26 P1.2 8.25 9.03
P1.3 5.85 6.64 P1.3 9.68 9.33
The energy requirement for heating the DCM and the heat
P1.4 5.32 5.58 P1.4 10.41 9.68
losses were estimated by Eqs. (3) and (4), respectively. P2.0 6.67 8.75 P2.0 6.92 8.50
P2.1 6.61 8.48 P2.1 7.22 8.81
Q r ¼ qDCM  F p  C p  ðtop  t inlet Þ ð3Þ P2.2 6.37 8.22 P2.2 8.32 8.91
P2.3 5.82 6.48 P2.3 9.59 9.24
Q l ¼ U  A  DT  86:4 ð4Þ P2.4 5.29 5.53 P2.4 10.26 9.90
P3.0 6.63 7.28 P3.0 6.73 7.35
where Qr represents the heat required (kJ d ), qDCM represents the
1
P3.1 6.57 7.32 P3.1 6.93 7.45
DCM density (kg m3), Fp represents the DCM flow (m3 d1), Cp rep- P3.2 6.38 7.13 P3.2 7.81 7.98
P3.3 5.87 6.72 P3.3 9.45 8.76
resents the specific heat of DCM (kJ kg1°C1), top represents the
P3.4 5.38 5.64 P3.4 10.16 9.22
operating temperature of the digester and tinlet represents the inlet P4.0 6.54 7.57 P4.0 6.71 7.84
temperature of the DCM (°C), Ql represents the heat losses through P4.1 6.43 7.23 P4.1 6.88 7.73
the walls, roof and the floor (kJ d1), U represents the global heat P4.2 6.26 7.12 P4.2 7.65 7.90
transfer coefficient (W m2°C1), A represents the reactor area P4.3 5.79 6.96 P4.3 9.40 8.76
P4.4 5.31 5.91 P4.4 10.07 8.96
exposed to losses (m2) and DT represents the temperature differ-
ence between the reactor and the environment (°C).

3. Results and discussion
3.1. Dairy cow manure characterisation
The DCM was collected in two different periods and the main
characteristics are summarised in Table 2. As can be seen, both
characterization were very similar in terms of TS content (41–
44 g TS L1) and organic matter (81% VS TS1). In general, these
values indicate that DCM was suitable for anaerobic digestion,
which requires a concentration range of 2–10% TS. Lower concen-
trations (<2%) may compromise the microorganism metabolic
activity, inducing a decrease in biogas production, while higher
values (>10%) could cause an inadequate mass transfer (Raposo
et al., 2012).
Moreover, the content of ashes (19–19.5%), fiber (28–31%) and
lipids (3–5%) were also much alike. Fiber represented almost
242 F. Passos et al. / Bioresource Technology 227 (2017) 239–246
one-third of the total organic content which may corroborate the
potential improvement that can be expected by applying pre-
treatment, making this fraction, at least partially, biodegradable
in anaerobic conditions. Notwithstanding, the second biomass col-
lected had lower protein content, i.e. 8.41% compared to 26.76%
from the first case (Table 1). Such difference may be related to
the cow feeding in each period and may affect biomass anaerobic
biodegradability. In fact, theoretical values calculated from the
macromolecular composition suggests that the methane yield
could attain a maximum of 353 and 423 mlCH4 g VS1 in the first
and second period, respectively. This difference is due to the higher
energetic content of proteins (851 mlCH4 g VS1) in respect to car-
bohydrates (415 mlCH4 g VS1) (Angelidaki and Sanders, 2004).
3.2. Selection of optimum S/I ratio
The first assay was carried out to optimise the S/I ratio of DCM
and digested sewage sludge in BMP tests. To this aim, five different
S/I ratio conditions were evaluated as abovementioned. Results of
accumulated methane yield are shown in Fig. 1. As can be seen in
Table 4, the lower the amount of inoculum added in the test, the
higher the final methane yield. Indeed, the highest value was
attained for 2.0 S/I ratio, which was 220 mlCH4 g VS1, followed
by 1.0 (216 mlCH4 gVS1), 0.75 (192 mlCH4 g VS1), 0.50
(185 mlCH4 g VS1) and 0.25 g VSi g VS1 s (176 mlCH4 g VS1). This
result is expected since the substrate is more biodegradable than
the inoculum, which had been already previously digested.
Nonetheless, the inoculum consists on the microorganism com-
munity responsible for biodegrading the substrate and, therefore, it
aims at enhancing the process start-up and rate. Thus, higher rates
and lower lag times are preferable. In order to compare the differ-
ent cases, experimental data was modelled using the Gompertz
model (Table 4). According to the results obtained, the higher the
value of inoculum in the BMP test, the higher the value of Rm,
reporting the highest value for 0.25 g VSs g VS1 i (18.5 g mlCH4 -
g VS1 d1). For lag time (k), results were better for S/I ratios of
0.50 and 0.75, reporting values of 0.54 and 0.60 days, respectively.
Finally, the optimal S/I ratio selected was 0.25 g VSs g VS1
i . This
condition was adopted due to both parameters, i.e. the best
methane production rate (18.5 g mlCH4 g VS1 d1) and a low lag
time (0.95 d). Thus, this ratio was used for the subsequent BMP
tests with thermochemical pretreated substrate.
3.3. Effect of thermochemical pretreatment on DCM anaerobic
biodegradability
3.3.1. pH
Addition of chemicals during the pretreatment step causes
changes in the substrate pH. In order to evaluate the pretreatment
effect on DCM, pH values before and after all pretreatment
conditions assayed were measured and are summarised in Table 3.
The pH of the raw DCM (P1.0, P2.0, P3.0 and P4.0) was 6.88 ± 0.21 and
6.63 ± 0.06 for alkali and acid pretreatment, respectively. Both val-
ues were close to neutrality, however a slight reduction of the pH
value of the substrate was observed due to the 3-week timespan
between both assays and the probable production of acids during
storage. As can be seen, for thermal pretreatment without any
Table 4
Parameters values of modified Gompertz model fitted with the experimental data at different S/I ratios. Mean values (standard deviation).
S/I ratio 0.25 0.50
Parameter
P (mLCH4/g VS) 175.8 (3.6) 185.3 (3.7)
Rm (mLCH4/g VS d) 18.5 (1.8) 13.8 (0.97)
k (d) 0.95 (0.53) 0.54 (0.50)
r2 0.972 0.982
chemical addition there was an increment in the substrate pH for
all temperature and time combinations. This rise was clearly
greater for 100 °C than 37 °C, regardless the exposure time. This
can be explained by the fact that solubility of CO2 diminished as
temperature increases, which led to CO2 stripping and the equilib-
ria displacement of carbonic acid and consequent pH rise. The inor-
ganic carbon content of DCM was not measured in this study,
however it is usually high, with values around 5 g L1 (Yin et al.,
2014).
As expected, the addition of both alkali and acid led to an
increase and decrease in the initial pH, respectively. This variation
was greater as the doses increased (i.e. P1.1 < P1.2 < P1.3 < P1.4). In
the case of acid addition, the pH at the end of the pretreatment
conditions were higher compared to the initial pH; which were
lower compared to the test without acid addition. This result
demonstrated that at least part of the acid acted in the hydrolysis
of the fiber and that the acid/base equilibrium was partially dis-
placed towards the absorption of CO2. In the case of alkali addition,
the pH at the end of the pretreatment was higher than the initial
one for the two lowest doses; however, it was found to be lower
than the initial pH at the highest doses regardless the temperature
and time. This can be explained by the hydrolysis of protein (DCM
showed a high content of proteins). Indeed, it has been reported
that the addition of alkali increased the release of ammonia nitro-
gen after protein hydrolysis (Zhang et al., 2015). Therefore, at
higher doses of NaOH, the surplus of OH- that remained unreacted
can drive to the generation of ammonia due to the presence of this
base so that the pH drops. In general, the pH at the end of every
test, was higher compared to the pH at the end of the test without
alkali addition, which may be due to the remaining presence of
base in the solution and to the stripping of CO2.
3.3.2. Anaerobic digestion kinetics: methane yield increase
The effect of thermochemical pretreatment in the anaerobic
biodegradability of DCM was assessed in BMP tests and results
are shown in Figs. 2 and 3. For almost all cases, i.e. thermal pre-
treatment and thermochemical pre-treatment with acid (HCl)
and alkaline (NaOH) solutions, the final methane yield obtained
were not significantly different if compared with non-pretreated
DCM (control). This might be due to the high methane yield
obtained for the control DCM (292 and 334 mlCH4 g VS1), which
were 70 and 79% of the maximum theoretical methane yield calcu-
lated from the substrate macromolecular composition. Results
from this study were somewhat higher in respect to those previ-
ously reported in literature for the same substrate, i.e. 150–
200 mlCH4 g VS1 (Amon et al., 2007; Moller et al., 2004;
Zarkadas et al., 2015). Such differences may be related to many fac-
tors regarding environmental conditions, animal feeding, among
others.
For thermal pretreatments without chemical addition, the only
conditions that reached methane yield increments were those with
higher exposure times (i.e. 37 °C for 12 and 24 h), which were 3.6
and 20.5% higher than control, respectively. On the other hand,
thermal pretreatments at 100 °C for 5 and 30 min did not enhance
substrate final methane yield (Figs. 2 and 3). This indicated that
exposure time had more effect when compared to temperature
at this range. Up to date, no studies have been conducted dealing
0.75 1.0 2.0
192.0 (3.4) 215.9 (4.4) 219.9 (4.0)
11.6 (0.58) 10.8 (0.48) 11.4 (0.44)
0.60 (0.41) 1.33 (0.41) 2.77 (0.34)
0.990 0.991 0.994
 F. Passos et al. / Bioresource Technology 227 (2017) 239–246 243

500 500

400 400
P (mL CH4/g VS)

P (mL CH4/g VS)

300 300

200 200

100 100

0 0
C P1.0 P1.1 P1.2 P1.3 P1.4 P2.0 P2.1 P2.2 P2.3 P2.4 P3.0 P3.1 P3.2 P3.3 P3.4 P4.0 P4.1 P4.2 P4.3 P4.4 C P1.0 P1.1 P1.2 P1.3 P1.4 P2.0 P2.1 P2.2 P2.3 P2.4 P3.0 P3.1 P3.2 P3.3 P3.4 P4.0 P4.1 P4.2 P4.3 P4.4

(a) (d)
25 30

25
20

Rm (mL CH4/g VS·d))

Rm (mL CH4/g VS·d)

20
15

15

10
10

5
5

0 0
C P1.0 P1.1 P1.2 P1.3 P1.4 P2.0 P2.1 P2.2 P2.3 P2.4 P3.0 P3.1 P3.2 P3.3 P3.4 P4.0 P4.1 P4.2 P4.3 P4.4 C P1.0 P1.1 P1.2 P1.3 P1.4 P2.0 P2.1 P2.2 P2.3 P2.4 P3.0 P3.1 P3.2 P3.3 P3.4 P4.0 P4.1 P4.2 P4.3 P4.4

(b) (e)
12 5

10
4

8
Lag time (d)
Lag time (d)

2
4

1
2

0 0
C P1.0P1.1P1.2P1.3P1.4P2.0P2.1P2.2P2.3P2.4P3.0P3.1P3.2P3.3P3.4P4.0P4.1P4.2P4.3P4.4 C P1.0 P1.1 P1.2 P1.3 P1.4 P2.0 P2.1 P2.2 P2.3 P2.4 P3.0 P3.1 P3.2 P3.3 P3.4 P4.0 P4.1 P4.2 P4.3 P4.4

(c) (f)
Fig. 2. Values fitted with modified Gompertz model after cow dairy manure pretreatment with HCl (a–c) and with NaOH (d–f).

with thermal pretreatment of DCM. For other type of substrates, it (4.6%). Accordingly, results suggested that for thermal-alkaline pre-
has been seen that at 80–90 °C protein and carbohydrates are sol- treatment higher doses (10%) combined with higher temperatures
ubilised, increasing the anaerobic biodegradability of waste acti- (100 °C) were preferable for reaching methane yields increments.
vated sludge (Appels et al., 2010). Moreover, for microalgal For acid pre-treatment, the conditions that exhibited an
biomass at a low range temperature (<100 °C), exposure time seem improvement in the substrate biodegradability were less related
to play a more important role for increasing anaerobic biodegrad- to the HCl dose. The highest methane yield increase was obtained
ability (Passos et al., 2013). for 2.0% dose at 37 °C for 12 h (P3.2) (20.6%), which reached 95% of
Nonetheless, the highest methane yield increase was attained the theoretical methane yield calculated from the macromolecular
for thermochemical pretreatment with NaOH at 10.0% at 100 °C composition. Subsequently, the other conditions that had higher
for 5 min (P1.4), which was 23.6% higher compared to control methane yield in respect to control were 6.0% dose at 100 °C for
(361 mlCH4 g VS1). Following that condition, methane yield 5 min (P1.2) (20.2%), 0.5% dose at 37 °C for 12 h (P3.1) (15.4%) and
increments were obtained for 10.0% dose at 100 °C for 30 min 10.0% dose at 100 °C for 30 min (P2.4) (20.6%).
(P2.4) (15.9%), 6.0% dose at 100 °C for 30 min (P2.3) (13.4%), 2.0% dose However, as previously mentioned most thermochemical pre-
at 100 °C for 5 min (P1.2) (6.0%) and 0.5% dose at 37 °C for 12 h (P3.1) treatment conditions showed similar or lower values of methane
244 F. Passos et al. / Bioresource Technology 227 (2017) 239–246

500 500

Methane yield (mL CH4/g VS) 400 400

Methane yield (mL CH4/g VS)

300 300

200 C 200
P1.0 (100ºC, 5min) C
P2.0 (100ºC, 30 min)
P1.1 (100ºC, 5min, 0.5%HCl)
P2.1 (100ºC, 30 min, 0.5%HCl)
P1.2 (100ºC, 5min, 2.0%HCl)
100 100 P2.2 (100ºC, 30 min, 2.0%HCl)
P1.3 (100ºC, 5min, 6.0%HCl)
P2.3 (100ºC, 30 min, 6.0%HCl)
P1.4 (100ºC, 5min, 10.0%HCl)
P2.4 (100ºC, 30 min, 10.0%HCl)
0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60

Digestion time (days) Digestion time (days)

(a) (b)
500 500

400 400
Methane yield (mL CH4/g VS)

Methane yield (mL CH4/g VS)

300 300

200 C 200 C
P3.0 (37ºC, 12h)
P4.0 (37ºC, 24h)
P3.1 (37ºC, 12h, 0.5%HCl)
P4.1 (37ºC, 24h, 0.5%HCl)
P3.2 (37ºC, 12h, 2.0%HCl)
100 100 P4.2 (37ºC, 24h, 2.0%HCl)
P3.3 (37ºC, 12h, 6.0%HCl)
P4.3 (37ºC, 24h, 6.0%HCl)
P3.4 (37ºC, 12h, 10.0%HCl)
P4.4 (37ºC, 24h, 10.0%HCl)

0 0
0 10 20 30 40 50 60 0 10 20 30 40 50 60

Digestion time (days) Digestion time (days)

(c) (d)
500 500

400 400
Methane yield (mL CH4/g VS)

Methane yield (mL CH4/g VS)

300 300

200 200
C C
P1.0 (100ºC, 5min) P2.0 (100ºC, 30min)
P1.1 (100ºC, 5min, 0.5%NaOH) P2.1 (100ºC, 30min, 0.5%NaOH)
100 P1.2 (100ºC, 5min, 2.0%NaOH) 100 P2.2 (100ºC, 30min, 2.0%NaOH)
P1.3 (100ºC, 5min, 6.0%NaOH) P2.3 (100ºC, 30min, 6.0%NaOH)
P1.4 (100ºC, 5min, 10.0%NaOH) P2.4 (100ºC, 30min, 10.0%NaOH)
0 0
0 5 10 15 20 25 30 35 0 5 10 15 20 25 30 35

Digestion time (days) Digestion time (days)

(e) (f )
500 500

400 400
Methane yield (mL CH4/g VS)

Methane yield (mL CH4/g VS)

300 300

200 200
C C
P3.0 (37ºC, 12h) P4.0 (37ºC, 24h)
P3.1 (37ºC, 12h, 0.5% NaOH) P4.1 (37ºC, 24h, 0.5% NaOH)
100 P3.2 (37ºC, 12h, 2.0% NaOH) 100 P4.2 (37ºC, 24h, 2.0% NaOH)
P3.3 (37ºC, 12h, 6.0% NaOH) P4.3 (37ºC, 24h, 6.0% NaOH)
P3.4 (37ºC, 12h, 10.0% NaOH) P4.4 (37ºC, 24h, 10.0% NaOH)
0 0
0 5 10 15 20 25 30 35 0 5 10 15 20 25 30 35

Digestion time (days) Digestion time (days)

(g) (h)
Fig. 3. Accumulated methane yield of thermochemically preteatead DCM HCl (a–d) and NaOH (e–h).

yield when compared to non-pretreated substrate. Previous 0.45 and 2.25% doses of NaOH at 35 and 100 °C for 5–30 min
results, dealing with alkaline and acid pretreatment, showed con- showed lower values of methane yield in respect to the control for
troversy values. For instance, pretreatment of swine slurry with all pretreatment cases (Ortega-Martinez et al., 2016). In addition,
 F. Passos et al. / Bioresource Technology 227 (2017) 239–246 245

alkaline pretreatment of pig manure only improved the methane
yield in BMP tests for the condition at 190 °C and pH 10. For all
the other cases (i.e. 135 and 150 °C and pH 10 and 12), methane
yield was decreased after thermochemical pretreatment (Carrère
et al., 2009). According to the authors this result was linked to
the production of recalcitrant compounds and/or to the degrada-
tion of the liquid fraction during the pretreatment step. On the
other hand, alkaline pretreatment of pulp and paper sludge
enhanced methane yield obtained in BMP tests. In this study, the
higher the NaOH dose, the higher the methane yield, reaching an
optimal value for 1.2% of NaOH at 37 °C for 6 h (183.5% increase).
For microalgal biomass, acid and alkaline pretreatments were also
effective. Particularly, lower doses of chemical solutions (0.5% of
KOH or HCl) at 80 °C for 2 h increased methane yield by 82–86%
(Passos et al., 2016).
3.3.3. Anaerobic digestion kinetics: productivity and lag-phase
Pretreatment methods are applied prior to anaerobic digestion
not only to enhance biomass methane yield, but also to improve
the process kinetics, i.e. decrease lag time (k) and increase methane
production rate (Rm). Usually this means that biodegradation pro-
cess would take less time to start and would occur faster due to
solubilisation of organic matter that becomes more available to
anaerobic microorganisms.
The values of Rm (maximum methane yield rate) and lag time
(k) from the Gompertz equation are shown in Fig. 2 for thermoacid
(Fig. 2a, b and c) and thermoalkaline pretreatments
(Fig. 2d, e and f). As can be seen from the results obtained, process
rate (Rm) was faster only for five thermo-acid pretreatment condi-
tions: P1.1, P1.2, P2.4, P3.1 and P3.2 (from 3 to 15% increase). On the
other hand, thermo-alkaline pretreatment enhanced Rm for most
of the cases, reaching values from 2 to 36% higher compared to
control. In regard to the lag time, most pretreatment conditions
were effective. For thermochemical pretreatment with HCl, all con-
ditions decreased lag time, reaching values between 3.9 and 8.8 d,
while control trial was 10.8 d. For alkaline thermochemical
Table 5
Assumptions for the economic evaluation.
Value and unit
Size of the dairy 6500 head of cattle
Average ambient temperature 11 °C
DCM yield 105 L/cow/d
DCM density (qp) 958.6 kg/m3
DCM specific heat capacity (Cp) 4.2 kJ/kg °C
CHP electric efficiency (ge) 40%
CHP heat efficiency (gt) 30%
Electricity cost (industrial) 121.46 US$/MWh
Heat cost 36.1 US$/MWh
Selling price of electricity 63.6 US$/MWh
Acid agent price HCl 660 US$/ton
Alkaline agent price NaOH 330 US$/ton
pretreatment, almost all trials obtained lower lag time values,
especially for those at 100 °C. Among all pretreatment cases, low-
est lag time was reached for the condition P1.2, which was 1.9 d.
In any case, the importance of the lag-phase in BMP test may not
be that relevant due to the fact that it lacks of influence in contin-
uous operations, although it can be important during the start-up
of the reactor
In summary, alkaline pretreatment seem to be more efficient
compared to acid pretreatment. The best pretreatment condition
in terms of final methane yield in BMP test was 10% of NaOH,
100 °C and 5 min (P1.4), which was 23.6% higher compared to con-
trol (361 mlCH4 g VS1). However, in terms of kinetics, the selected
condition would be the one with 2% of NaOH, 100 °C and 5 min
(P1.2), with 25.9 mlCH4 g VS1 d1 and a lag time of 1.9 d. In this
case, methane yield reached a value of 310 mlCH4 g VS1. Although
in this case methane yield was lower, it could decrease process
costs since smaller reactor volumes could be used.
3.4. Economic assessment
Finally, in order to assess the viability of thermochemical pre-
treatment, an economic evaluation of process costs was estimated
for four of the studied conditions. Thus, the scenarios evaluated
were: i) EBAC anaerobic digestion of (non-pretreated) biomass; ii)
E1AC, 2% of HCl at 37 °C for 12 h; iii) E2AC, 0.5% of HCl at 37 °C for
12 h; iv) EBAlk anaerobic digestion of (non-pretreated) biomass;
v) E1Alk, 10% of NaOH at 100 °C for 5 min and; vi) E2Alk, 6% of NaOH
at 100 °C for 30 min. Even though the EB scenarios correspond to
the same conditions the experimental results for the anaerobic
degradation of the raw DCM were different since they were carried
out 3 weeks apart and, therefore, the two scenarios with their
respective parameters were assessed. The assumptions used for
the economic evaluation are summarised in Table 5. The average
prices for energy and chemicals were taken from the Chilean public
and private institutions.
The results obtained from all costs (revenues and expenses) are
shown in Table 6. According to the results, thermal-alkali pretreat-
ment showed a negative total cost since the surplus of energy
injected to the grid did not offset the expenses related to the extra
energy and the cost of the chemical agent. This result was due to
the high temperature required for the pretreatment, which cannot
be completely supplied by the thermal energy generated with the
biogas in the CHP unit. Despite the fact that the total income of the
of thermal-acid pretreatment was positive, it was still lower than
the income obtained by the conventional AD application.
The cost of the energy and the chemical agents may fluctuate
and it depends on the market conditions, as well as on the negoti-
ation terms between the companies. Therefore, a basic sensitivity
analysis based on other prices of the energy and chemical agents
was also assessed. The implementation of thermal-acid pretreat-
ment outperforms the conventional AD of the substrate with

Table 6
Results of the economic evaluation.

Costs (US$/year) Type of pretreatment

Thermal-Alkali Thermal-Acid
EBAlk E1Alk E2Alk EBAC E1AC E2AC
Income 821,414 172,333 197,851 1,033,506 1,044,948 968,282
Electricity surplus 628,802 572,870 588,275 756,662 799,551 754,387
Heat surplus 192,612 400,537 390,424 276,844 245,397 213,895
Expenses 0 811,036 636,724 0 126,048 31,512
Additional electricity 0 0 0 0 0 0
Additional heat 0 400,537 390,424 0 0 0
Chemical agents 0 410,499 246,300 0 126,048 31,512
Total income 821,414 638,703 438,873 1,033,506 918,899 936,770
246 F. Passos et al. / Bioresource Technology 227 (2017) 239–246
revenues of around 15.247 US$ y1 when the sell price injection to
the national grid was set as 162.7 US$ MW h1 (maximum value
reached in 2015 according to the Asociación Gremial de Generadoras
de Chile) and the cost of electricity for industries was 86.4 US
$ MW h1 (minimum market value reached during December
2015, according to Asociación Gremial de Generadoras de Chile). In
this context, the cost of the chemical agent must be, at least, half
of the average price presented in Table 5. Another aspect not con-
sidered in this study was the digestate handling and disposal cost.
It is assumed that this issue would not change compared to con-
ventional AD system and, therefore, the cost would remain the
same. Nonetheless, some pathogens might be removed by thermal
pretreatment, as well as the digestate may enhance its dewater-
ability (Donoso-Bravo et al., 2011). Ultimately, the pretreatment
may lead to some other economic benefits in this regard, although
it was not the focus of the present study. Further experiments
should analyze the effect of pretreatment in DCM digestate.
This economic analysis aimed to extend the assessment in com-
parison to the ones that can be found in literature. In general, the
basic design of the process units has not been taken into account
and the biogas yield obtained in the batch test are assumed to be
the same in continuous conditions. In the present study, a direct
link of the pretreatment conditions and BMP results was used
not only for the biogas production but also for the reactor and pre-
treatment units design itself. In the only study where the pretreat-
ment of cow manure has been assessed, Cano et al. (2014), found
that the implementation of thermal hydrolysis with steam explo-
sion (170 °C) yields an economic benefit between 13 and 29% com-
pared to the conventional AD. In this case, the high content of total
and organic solids of the substrate influenced the major positive
impact of the pretreatment. On the other hand, the cost paid for
the electricity produced from biogas in Europe is higher, due to
economic subsidies.
4. Conclusions
In some of the studied conditions thermal-alkali pretreatment
improved the methane potential compared to the test with raw
substrate. The methane potential was enhanced by 23.6% after pre-
treatment with 10% of NaOH at 100 °C for 5 min. The maximum
production rate was improved in all the studied conditions. In
the case of the thermal-acid pretreatment the best improvement
of the methane potential (20.6%) was observed with doses of 2%
of HCl at 37 °C. The technical-economic analysis showed that the
implementation of neither thermal alkali nor thermal-acid
pretreatment outperform the conventional one-step anaerobic
digestion of raw waste.
Acknowledgements
This study was funded by Fondecyt Initiation project n
°11130462. Fabiana Passos acknowledges her Post-Doctorate
scholarship founded by the National Council for Scientific and
Technological Development (CNPq) from the Brazilian Ministry of
Science, Technology and Innovation.
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