Você está na página 1de 8

Applied Energy 108 (2013) 168175

Contents lists available at SciVerse ScienceDirect

Applied Energy
journal homepage: www.elsevier.com/locate/apenergy

Biogas production from microalgae grown in wastewater:


Effect of microwave pretreatment
Fabiana Passos, Maria Sol, Joan Garca, Ivet Ferrer
GEMMA Group of Environmental Engineering and Microbiology, Department of Hydraulic, Maritime and Environmental Engineering, Universitat Politcnica de
Catalunya.BarcelonaTech, c/Jordi Girona 1-3, Building D1, E-08034 Barcelona, Spain

h i g h l i g h t s
" Microwave irradiation enhanced the disintegration and digestibility of microalgae.
" Algal biomass solubilisation increased by 800% with microwave pretreatment.
" The main parameter inuencing biomass solubilisation was the applied specic energy.
" Increased biogas production rate (2775%) and yield (1278%) with pretreated biomass.
" Linear correlation between microalgae solubilisation and biogas yield.

a r t i c l e

i n f o

Article history:
Received 2 October 2012
Received in revised form 14 February 2013
Accepted 17 February 2013

Keywords:
Anaerobic digestion
Biofuel
High rate algal pond
Methane
Renewable energy

a b s t r a c t
The aim of this study was to evaluate the effect of microwave pretreatment on the solubilisation and
anaerobic digestion of microalgaebacterial biomass cultivated in high rate algal ponds for wastewater
treatment. The microwave pretreatment comprised three specic energies (21,800, 43,600 and
65,400 kJ/kg TS), combining three output power values with different exposure times. Response surface
analysis showed that the main parameter inuencing biomass solubilisation was the applied specic
energy. Indeed, a similar solubilisation increase was obtained for the same specic energy, regardless
of the output power and exposure time (280350% for 21,800 kJ/kg TS, 580610% for 43,600 kJ/kg TS
and 730800% for 65,400 kJ/kg TS). In biochemical methane potential tests, the initial biogas production
rate (2775% increase) and nal biogas yield (1278% increase) were higher with pretreated biomass. A
linear correlation was found between biomass solubilisation and biogas yield. It can be concluded that
microwave irradiation enhanced the disintegration and digestibility of microalgae.
2013 Elsevier Ltd. All rights reserved.

1. Introduction
During the last decade, there has been a growing interest in
investigating the energy potential of biofuels obtained from microalgae cultures [1]. The high lipid content of microalgae makes them
an alternative to terrestrial energy crops for biodiesel production.
However, microalgae cultures and energy production are at an initial research phase. According to the literature, the cultivation of
microalgae to produce biofuels has a number of requirements that
limit its current implementation at industrial scale [2]. To make it
economically feasible, massive biomass production and energy
generation technologies must be addressed.
The cultivation of certain specic strains of microalgae is not
viable in economic and environmental terms, since freshwater
and fertilizers are needed. In contrast, if microalgal biomass is
Corresponding author. Tel.: +34 934016463; fax: +34 934017357.
E-mail address: ivet.ferrer@upc.edu (I. Ferrer).
0306-2619/$ - see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.apenergy.2013.02.042

grown as a by-product of high rate algal ponds (HRAPs) operated


for wastewater treatment, the economic and ecological footprint
are more realistic if used at large-scale [3,4]. High rate ponds are
shallow, open raceway ponds, with continued mixing provided
by paddle-wheels. This system works by a symbiosis between heterotrophic bacteria, which oxidize organic matter contained in
wastewater, and the phytoplankton, which by photosynthesis consumes the CO2 derived from organic matter mineralization. Microalgaebacterial biomass grown in this environment assimilates
nutrients and subsequently, its separation from the nal efuent
eliminates nutrients from the wastewater [5].
Anaerobic digestion of microalgae was rst studied in the 1950s
[6,7]. These authors used microalgal biomass from HRAP, pointing
out biomass separation from the liquor as a major limitation of the
process. Up to date, the literature on microalgae digestion is very
limited. The review by Gonzlez-Fernndez et al. [1] reports a specic methane production of 0.10.5 L CH4/g VS, with 6080% CH4
in biogas, depending on process temperature (1552 C) and

169

F. Passos et al. / Applied Energy 108 (2013) 168175

hydraulic retention time (HRT) (364 days). This value is commensurate with biogas production from other substrates. For instance,
the specic methane production of waste activated sludge, another
by-product of wastewater treatment, ranges between 0.15 and
0.3 L CH4/g VS [8]; and that of lignocellulosic agricultural crops,
such as maize, wheat, rice and sugarcane wastes, between 0.28
and 0.34 L CH4/g VS [9].
The complex cell wall structure of microalgae, composed by cellulose, hemicellulose and pectin, makes bacteria attack difcult
[1]; suggesting that biomass pretreatment is necessary for the feasibility of microalgae anaerobic digestion [10]. The pretreatment of
substrates to increase the anaerobic biodegradability has been the
subject of intense research in recent years. Physical, chemical and
biological processes have proven successful at improving the disintegration and anaerobic biodegradability of lignocellulosic biomass
[11]. Furthermore, waste activated sludge pretreatment using
mechanical, thermal and biological processes increased the specic
methane production, leading to positive energy balances, and is
currently applied in full scale facilities [12]. The few studies that
have so far been conducted with microalgae show an increased
methane yield after thermal and ultrasound pretreatments [13
16].
The electromagnetic radiation of microwaves has also been
investigated as a pretreatment process [1719]. Microwaves are
short waves of electromagnetic energy varying in a frequency from
300 MHz to 300 GHz, which can increase the kinetic energy of the
water leading to a boiling state [17]. The quantum energy applied
by microwave irradiation is not capable of breaking down chemical
bonds, however hydrogen bonds are or can be broken [20]. Induction heating and dielectric polarization result in changes in the secondary and tertiary structure of proteins and cause cell hydrolysis.
The polarization of macromolecules occurs by a consistent rotation
through an alternating electric eld. This process is inuenced by
microwave frequency, radiation time, biomass concentration and
penetration depth [18].
The literature on microwave pretreatment of waste activated
sludge under different conditions [18,2125], shows how the process enhanced sludge solubilisation and/or cumulative gas production in anaerobic batch tests (Table 1). Microwave irradiation can
produce both thermal and athermal effects. Some authors have
compared microwave and thermal pretreatments, observing higher volatile solids and hemicellulose solubilisation [19] and biogas
production [26] with the former. So far, the effect of microwave
irradiation on microalgae remains unexplored.
Thus, the objective of this study was to evaluate the effect of
microwave pretreatment on the disruption and anaerobic biodegradability of microalgal biomass from HRAP for wastewater
treatment.

2. Materials and methods


2.1. Microalgal biomass production system
The experimental set-up was located at the laboratory of the
GEMMA research group (Universitat Politcnica de Catalunya. BarcelonaTech, Spain). The system had been in operation since March
2010. The microalgae production system was composed of a
hydrolytic up-ow sludge blanket reactor (HUSB), a high rate algal
pond and a settler. Urban wastewater was pumped from a municipal sewer and stored in a tank (1.2 m3), which was continuously
stirred to avoid solids sedimentation. From the storage tank, pretreated wastewater was conveyed to the primary treatment: a
cylindrical PVC HUSB, with an internal diameter of 0.3 m, a total
height of 1.9 m and an effective volume of 0.105 m3; operated at
an HRT of 5 h. The sludge blanket inside the HUSB reactor was kept
at a total volatile solids (VS) concentration below 10 g/L by periodical purge. A detailed description of the operation and performance
of the HUSB reactor can be found in Pedescoll et al. [27].
The primary efuent of the HUSB reactor was stored in a 50 L
regulation tank and pumped to the HRAP by means of peristaltic
pumps. The experimental HRAP was a PVC raceway pond with a
paddle wheel for stirring the mixed liquor. The HRAP had a nominal volume of 0.5 m3, a surface area of 1.5 m2 and a water depth of
0.3 m. The HRAP treated 62.5 L/day corresponding to an HRT of
8 days. Average surface loading rates were 24 g COD/m2 day and
4 g NH4N/m2 day. The daily biomass production potential was
calculated from Eq. (1), where TSS is the concentration of total
suspended solids in the mixed liquor of the HRAP, A is the HRAP
surface area and Q is the ow rate.

Biomass production TSS g=L=A m2   QL=d

Microalgal biomass was harvested from the HRAP by means of a


conventional settling tank, with a nominal volume of 0.01 m3
(0.16 days HRT). Purged biomass was then settled in laboratory
Imhoff cones stored at 4 C for 24 h. The HRAP performance was
monitored by taking weekly samples at 2 PM from March 2011
to March 2012. Average characteristics of the HRAP inuent, efuent and harvested biomass are summarized in Tables 2 and 3.
2.2. Microwave pretreatment
To study the effect of microwave pretreatment on microalgal
biomass a household type microwave (Samsung M1914,
2450 MHz frequency) was used. The output power range of the
equipment was 1001000 W (Microwave test procedure IEC705). Three target specic energies were applied: 21,800; 43,600
and 65,400 kJ/kg TS. Considering the following output power

Table 1
Effect of microwave pretreatment on activated sludge solubilisation and biochemical methane potential tests.
Pretreatment conditions
14.3 min; 400 W; 102 C; 2.3% TS
5 min; 800 W; 13,000 kJ/kg SS
5 min; 96 C; 5.5% TS
Progressive heating 1.21.4 C/min;
175 C
0.83 kJ/ml; 1000 W; 78% TS
1168 W; 90 C; 4% TS

Batch test
conditions
55 C; 32 d
33 C; 23 d
33 C; 18 d
35 C; 2025 d
35 C; 22 d

Results
Increase
Increase
rate*
Increase
Increase

References
of 17.9% in the CODs/COD ratio*
of 311% in the VSs/VS ratio and no difference in biogas production and production

[18]
[21]

of 143% in the CODs/COD ratio and 211% in the cumulative biogas production*
of 74.3% in COD solubilisation and 34% in biogas production*

[22]
[23]

Decreased solubilisation (CODs/COD) and increase of 15.4% in methane production*


Increase of 2.5% in the CODs/COD ratio, 37% in the digestion rate and no impact on methane
production*

[24]
[25]

Note: TS: total solids, SS: suspended solids, VS: volatile solids, VSs: soluble volatile solids, COD: chemical oxygen demand, CODs: soluble chemical oxygen demand.
Compared to control.

170

F. Passos et al. / Applied Energy 108 (2013) 168175

Table 2
High rate algal pond performance from March 2011 to March 2012. Mean values (standard deviation) from samples taken at 2 PM.
Parameter

Inuent (HUSB primary efuent)

Efuent (HRAP ltrated efuent)

% Removal

pH
Temperature (C)
DO (mg/L)
Turbidity (NTU)
TSS (mg/L)
COD (mg/L)
TKN (mg/L)
NH4N(mg/L)

6.8 (0.8)
22 (4)
0.5 (0.7)
60 (34)

250 (57)
50 (12)
35 (11)

8 (1.1)
20 (7)
8 (4)
12 (9)
400 (130)*
80 (49)
20 (6)
2 (1.2)

87

68
60
95

Note: DO: dissolved oxygen, TSS: total suspended solids, COD: chemical oxygen demand, TKN: total Kjeldahl nitrogen, NH4N: ammonia nitrogen.
Not ltrated sample.

Y b0 b1 X 1 b2 X 2 b11 X 21 b22 X 22 b12 X 1 X 2

Table 3
Harvested microalgal biomass characteristics.
Parameter

Mean value (standard deviation)

pH
TS (% (w/w))
VS (% (w/w))
VS/TS (%)
VSs (% (w/w))
VSs/VS (%)
COD (g/L)
CODs (g/L)
CODs/COD (%)
TKN (mg/kg)
NH4N (mg/L)
Lipids (% (w/gTS))
Proteins (% (w/g TS))
Carbohydrates (% (w/g TS))

7.5 (0.8)
1.65 (0.2)
0.98 (0.2)
60 (0.9)
0.012 (0.002)
0.89 (0.11)
16.7 (0.42)
0.12 (0.02)
0.72 (0.44)
800 (180)
12 (7)
17.4 (1.50)
49.3 (1.23)
19.5 (1.87)

The polynomial function was estimated by the least squares


method and checked with MatLab software. Experimental data
were collected in triplicate.
2.4. Biochemical methane potential tests

Note: TS: total solids, VS: volatile solids, VSs: soluble volatile solids, COD: chemical
oxygen demand, CODs: soluble chemical oxygen demand, TKN: total Kjeldahl
nitrogen, NH4N: ammonia nitrogen.

values: 300, 600 and 900 W; the time needed to achieve


each target specic energy was calculated from the following
equation:

Specific energy kJ=kg TS power W


 time s=sample weight kg TS
2
For each pretreatment condition, a volume of 150 mL thickened
microalgal biomass was pretreated in a beaker and cooled to room
temperature.
2.3. Microlgal biomass solubilisation
Biomass solubilisation was evaluated by the soluble to total volatile solids ratio (VSs/VS) and by the increase in this ratio with respect to the control (untreated biomass), calculated according to
Eq. (3), were sub-indexes refer to samples before (o) and after pretreatment (p).

VSs =VS increase% VSs =VSp  VSs =VSo =VSs =VSo


 100

The response surface methodology was used to analyse the relation between pretreatment conditions and biomass solubilisation
by means of a second order polynomial expression, as proposed
by other authors [18,21]. Experimental results were evaluated by
tting the second order polynomial expression (Eq. (4)), where Yi
corresponds to experimental results (VSS/VS ratio); and X1 and X2
to the microwave output power (300, 600 and 900 W) and exposure time (19 min), respectively.

The anaerobic biodegradability of pretreated microalgal biomass was compared to the control (untreated biomass) by means
of mesophilic (35 C) batch tests. Pretreatment conditions are summarized in Table 4. The inoculum was mesophilic sludge from a
full-scale anaerobic digester located in a municipal wastewater
treatment plant near Barcelona (Spain). Each treatment was performed in duplicate.
Serum bottles had a total volume of 160 mL and a useful volume
of 100 mL. The concentration of substrate was 16.7 g COD/L, corresponding to 29.94 g microalgal biomass/bottle. As recommended
by Cho et al. [28], the substrate/inoculum ratio was 0.5 g COD/g
VS, corresponding to 43.48 g sludge/bottle. The bottles where
ushed with Helium gas (He), sealed with butyl rubber stoppers
and incubated at 35 C until biogas production ceased.
Biogas production was periodically determined by measuring
the pressure increase with an electronic manometer (Greisinger
GMH 3151). After each measurement gas was released until atmospheric pressure. Samples from the headspace volume were taken
every 23 days, to determine biogas composition (CH4/CO2) by gas
chromatography.
Accumulated volumetric methane production (mL) was calculated from the pressure increase and methane content in biogas,
expressed under standard conditions. The net values of methane
production and yield were obtained by subtracting the endogenous
production of the blank bottle.
2.5. Analytical methods
All analyses were triplicated and results are given as mean values. The following parameters were determined from the inuent
and efuent of the HRAP: total suspended solids (TSS), total chemical oxygen demand (COD), ammonia nitrogen (NH4N) and total
Kjeldahl nitrogen (TKN), according to Standard Methods [29].
Water temperature and dissolved oxygen (DO) were measured
in situ with an YSI 58 oxymeter. Turbidity was determined with
a Hanna Microprocessor Turbidity Meter HI93703 and pH was
determined with a Crison Portable 506 pH-meter.
Microalgal biomass was characterised by the concentration of
total solids (TS), VS, COD, CODs, NH4N, TKN, determined according
to Standard Methods [29]. The lipid content was determined by the
Soxhlet extraction method [29]. A TKN to protein conversion factor
of 5.95 was used [30]. Carbohydrates were determined by phenol
sulfuric acid method after acid hydrolysis and measured by spectrophotometry (Spectronic Genesys 8). Microalgae identication

171

F. Passos et al. / Applied Energy 108 (2013) 168175


Table 4
Microwave pretreatment conditions and microalgal biomass solubilisation.
Trial

Target specic energy (kJ/kg TS)

Output power (W)

Exposure time (min)

Temperature (C)

VSs/VS ratio

VSs/VS increase (%)

Control
T1
T2
T3
T4
T5
T6
T7
T8
T9

21,800
21,800
21,800
43,600
43,600
43,600
65,400
65,400
65,400

300
600
900
300
600
900
300
600
900

3.0
1.5
1.0
5.0
3.0
2.0
9.0
4.5
3.0

50
58
56
81
86
92
95
98
98

0.0089
0.034
0.040
0.040
0.061
0.062
0.063
0.075
0.074
0.081

280
345
346
580
597
606
742
728
799

was carried out by microscopical examination (Nikon Optiphotpol, Japan).


Batch test samples were analyzed for TS, VS, VSs, COD and CODs,
according to Standard Methods [29]. The methane content in biogas
and volatile fatty acids (VFA) were measured with a gas chromatograph (GC), following the procedure described by Ferrer et al. [31].
3. Results and discussion
3.1. Microalgal biomass production
The HRAP performance was monitored from March 2011 to
March 2012 (Table 2). The HRAP inuent corresponds to the primary efuent of the HUSB reactor, which had a turbidity (60
NTU) and COD (250 mgO2/L) within the typical range of a conventional primary efuent [27]. The pH in the HRAP (8.0) was always
higher than in the inuent (6.8), due to CO2 consumption by microalgal photosynthetic activity. The HRAP performed correctly during the whole experimental period with an HRT of 8 days: COD
removal (68%), TKN removal (60%) and NH4N removal (95%) efciencies (Table 2) were in accordance with previous studies [5,32].
Indeed, Garca et al. [32] found a TKN removal efciency of 73% and
57%, and a NH4N removal efciency of 97% and 91%, with an HRT
in the HRAP of 7 and 4 days, respectively.
The HRAP biomass production potential was 16 g TSS/m2/d,
which ts in the range from 12 to 40 g TSS/m2/d reported by Park
et al. [4]. However, harvestable biomass was approximately 9 g
TSS/m2/d, since 45% of the produced biomass escaped from the settler. Up to date, microalgal biomass harvesting continues to be a
bottleneck of this technology.
The main properties of the produced biomass are summarised
in Table 3. It was characterized by an organic content around
60% VS/TS, most of it in particulate form as indicated by the low
VSs/VS (0.89%) and CODs/COD (0.72%) ratios. This means that almost all organic matter was not soluble and, therefore, retained inside microalgal cell walls. For this reason, the use of pretreatment
techniques to disintegrate microalgal cell walls seems appropriate
to improve the anaerobic digestion of this substrate.
The biomass grown in the system was a consortia of microalgae
and bacteria, especially from the group Chlorophyta. The main
microalgae species identied in the mixed liquor were Scenedesmus and Chlorella. The average macromolecular composition of
microalgal biomass was 49% proteins, 17% lipids and 20% carbohydrates. These results are commensurate with those found in pure
cultures of Scenedesmus obliquus: 5056% proteins, 1214% lipids
and 1017% carbohydrates; and Chlorella vulgaris: 5158% proteins, 1422% lipids and 1217% carbohydrates [33]. In microalgae
pure cultures, higher lipids accumulation is obtained by applying
techniques such as nutrient starvation or light deciency. On the
other hand, mixed cultures grown in HRAP for wastewater treatment tend to have lower lipid content, mainly because operating
conditions are less controlled. For instance, Gonzlez-Fernndez

et al. [34] found a 3% lipid content for Scenedesmus sp. grown in


swine slurry.
3.2. Solubilisation of microalgal biomass
Microwave irradiation enhanced microalgae solubilisation under all pretreatment conditions assayed. The VSs content increased
from 0.05% to 0.11%; corresponding to a VSs/VS increase between
280% and 800% depending on the microwave output power, exposure time and temperature reached (Table 4).
To our knowledge, this is the rst study dealing with the disintegration of microalgal biomass by microwave pretreatment. For
the sake of comparison, literature results on the effect of microwave irradiation on waste activated sludge are summarised in
Table 1. Almost all authors report an increase on sludge solubilisation; the solubilisation degree increases together with the applied
specic energy, up to a limit value which is related to the sludge
boiling point [21]. However, there is some controversy regarding
the effect of the output power and exposure time. For the
same specic energy (13,000 kJ/kg SS), Climent et al. [21] found
higher VSs/VS increase with an output power of 800 W for 5 min
(311%) compared to an output power of 400 W for 10 min
(289%). On the other hand, Park et al. [18] found higher
CODs/COD increase (17.5%) with a low output power and a long
exposure time (400 W for 14.3 min) for the same target specic
energy.
In our study, both output power and exposure time inuenced
biomass solubilisation (Table 4). For the same exposure time (T1,
T5 and T9) biomass solubilisation increased with the microwave
output power. Similarly, for the same output power (i.e. T1, T4
and T7; or T2, T5 and T8; or T3, T6 and T9), biomass solubilisation
increased with the exposure time. The combination of both variables represents the applied specic energy. Indeed, the solubilisation increase was similar for the same specic energy regardless of
the output power and exposure time: between 280% and 350% for
21,800 kJ/kg TS, between 580% and 610% for 43,600 kJ/kg TS and
between 730% and 800% for 65,400 kJ/kg TS (Table 4). Like this,
the highest specic energy (65,400 kJ/kg TS) resulted in 21% higher
solubilisation compared to the second specic energy (43,600 kJ/kg
TS) and 58% higher solubilisation compared to the lowest specic
energy (21,800 kJ/kg TS). As shown in Fig. 1, the solubilisation degree increased linearly with the applied specic energy, in accordance with previous results with sewage sludge [35]. Such an
increase in readily digestible organic compounds should accelerate
and/or increase the anaerobic biodegradability of microalgal biomass. For this reason, the effect of microwave irradiation on the
methane production was evaluated in biochemical methane potential (BMP) tests.
In order to represent the inuence of pretreatment variables on
the solubilisation degree, the following quadratic equation was
determined by response surface analysis (Eq. (5)), where Yi corresponds to experimental results (VSs/VS ratio); and X are indepen-

Biomass solubilisation (VSs/VS ratio)

172

F. Passos et al. / Applied Energy 108 (2013) 168175

0,10

0,08

0,06

R2= 0,96

0,04

0,02

0,00
0

10000

20000

30000

40000

50000

60000

70000

Specific energy (kJ/kg TS)


Fig. 1. Microalgal biomass solubilisation (VSs/VS) versus applied specic energy in
the microwave pretreatment according to Eq. (7).

dent variables (output power and exposure time). The statistic


parameters of the coefcients adjusted for the quadratic polynomial are summarised in Table 5.

VSs =VS ratio 2:579  108 X 2power  1:422  107 X 2time

Experimental results together with the model output are shown


in Fig. 1. These results were found for specic energies from 21,800
to 65,400 kJ/kg TS. Notice that biomass solubilisation may reach an
asymptote at higher specic energy values (i.e. 100,000 kJ/kg TSS)
[21].
Previous studies point out a potential microwave thermal effect
[19,26]. According to Hu et al. [19], the pretreatment of cattail (Typha latifolia) by microwave irradiation (500 W for 14 min) increased VS and hemicellulose solubilisation by 24.5% and 23.5%,
respectively; while conventional heating at the same temperature
(100 C) increased VS and hemicellulose solubilisation by 15.4%
and 7.4%, respectively. This highlights the microwave athermal effect. In our study, the temperature reached increased with the applied specic energy, which also led to the highest biomass
solubilisation (Table 4). In order to consider the thermal effect,
microalgae solubilisation with conventional heating at 5595 C
was evaluated. After 1 h of pretreatment the solubilisation degree
(0.027 VSs/VS (data not shown)) was lower than in all microwave
pretreatment conditions, with exposure times below 9 min
(0.0340.081 VSs/VS) (Table 4). This suggests that microwave
athermal effect enhanced biomass solubilisation with a shorter
exposure time (1.09.0 min) if compared with conventional heating (1 h).
3.3. Biogas production in biochemical methane potential tests

2:043  107 X power  X time 6:726


 105 X power 1:519  104 X time  1:627
 102

Previous studies have adjusted a quadratic equation for describing the inuence of pretreatment conditions on biomass solubilisation, for instance the sludge thermal pretreatment [21] and
microwave pretreatment [18]. However, in our quadratic model
Pr values were all higher than 0.05. Since the interaction between
output power and exposure time coefcient (Xpower  Xtime) was the
main factor inuencing biomass solubilisation, a simplied model
was proposed instead (Eq. (6)). In this simple model, both coefcient terms had Pr values lower than 0.05, indicating a signicant
tting (Table 5).

VSs =VS ratio 3:589  107 X power X time 2:022  102

As summarized in Table 5, R2 values were similar for the quadratic and simplied models (0.98 and 0.96, respectively), indicating that the interaction between output power and exposure time
(Xpower  Xtime) (i.e. simplied model) explained most of the variability. Moreover, the simplied model p-value (4.12  106) was
much lower than the quadratic model p-value (9.29  103).
According to this, the applied specic energy (Eq. (2)) was the main
factor inuencing biomass solubilisation. Therefore, the simplied
model could be expressed in terms of specic energy (Eq. (7)) by
introducing the total solids into Eq. (6), where Xspecic energy is the
applied specic energy and XTS the total solids content in biomass.

VSs =VS ratio 3:589  107 X TS X specific energy 2:022  102

Cumulative biogas production with pretreated and untreated


microalgal biomass is shown in Fig. 2. All trials were completed
within 46 days of incubation. The methane content was around
68% in all cases (Table 6).
The initial biogas production rate was signicantly higher (27
75% increase) for all pretreatment conditions compared to the control (Table 6). The highest biogas production rate corresponded to
biomass pretreated at 65,400 kJ/kg TS (the highest applied specic
energy), followed by biomass pretreated at lower specic energies
and untreated biomass (control). For instance, trial T9 had a production rate of 88 mL biogas/g VSday, while the control produced
57% of this value (50 mL biogas/g VSday). This suggests that
microwave pretreatment may improve the methane production
in anaerobic reactors operated at short HRT, reducing the required
reactor volume and investment cost, although this hypothesis
should be addressed in future research studies. The sonication of
Scenedesmus biomass also increased the initial methane production
rate, and has been pointed out as a way of reducing the HRT [14].
The nal biogas yield was signicantly higher (1278% increase) for microalgae pretreated with all microwave conditions
compared with the control (Fig. 2). The highest biogas yield was
achieved by trial T9, 307 mL biogas/g VS (78% increase), followed
by T8, 296 mL biogas/g VS (72% increase) and T7, 276 mL biogas/
g VS (60% increase); corresponding to the highest applied specic
energy (65,400 kJ/kg TS) with output powers of 900 W, 600 W
and 300 W, respectively (Table 5). The same pattern occurred for
lower specic energies (43,600 kJ/kg TS and 21,800 kJ/kg TS), with
somewhat lower biogas production increase (below 42% for all pretreatment conditions). For the same specic energy, the nal bio-

Table 5
Comparison between quadratic and simplied model coefcients.
Parameters

Estimate value
Std. error
t value
Pr

Quadratic model

Simplied model

Intercept

X 2power

X 2time

Xpower

Xtime

Xpower  Xtime

Intercept

Xpower  Xtime

1.627e02
3.100e02
0.525
0.636

2.579e08
4.200e08
0.614
0.583

1.422e07
1.316e07
1.081
0.359

6.726e05
7.002e05
0.961
0.408

1.519e04
1.337e04
1.136
0.338

2.043e07
1.406e07
1.453
0.242

2.022e02
3.272e03
6.18
0.000454

3.589e07
2.805e08
12.80
4.12e06

Quadratic model: R2 = 0.98, p-value = 0.009289, F = 29.7; simplied model: R2 = 0.96, p-value = 4.122e06, F = 163.8.

173

F. Passos et al. / Applied Energy 108 (2013) 168175

desmus biomass thermal pretreatment at 90 C and 33 days of


incubation. More recently, different microalgae mixed cultures
were pretreated using ultrasounds (10,00057,000 kJ/kg TS), low
temperature (55 C) and thermal hydrolysis (110170 C) [16].
While the biological pretreatment at 55 C had no impact on the
methane yield, the thermal hydrolysis at 110 C increased the
methane yield by 62% compared to untreated biomass. These results can be compared with our study, where temperatures in
the range of 9598 C increased biomass solubilisation by 730
800% VSs/VS and the methane yield by 6078% in respect to the
control. Considering the positive effect of microwave irradiation
in enhancing biomass solubilisation and anaerobic digestion, future research studies should evaluate the methane production in
continuous reactors, and compare the energy balance of the process with and without pretreatment step.
Fig. 2. Cumulative biogas production after microalgal biomass microwave pretreatment, under the conditions shown in Table 4.

gas yield was signicantly higher with an output power of 600


900 W compared to 300 W (Table 6), putting forward the effect
of the output power on the methane production potential.
The biogas production showed a linear correlation with the solubilisation degree (Fig. 3): the higher the biomass solubilisation,
the higher the biogas production. A linear correlation between biogas production and solubilisation after thermal pretreatment was
previously reported by other authors. In the case of sunower oil
cake, the methane production potential and organic content
solubilisation after pretreatment showed a linear correlation
(R2 = 0.87) for temperatures between 130 and 170 C and acid
addition between 0.74% and 5% [36]. A comparison between ozonation, sonication and thermal sludge pretreatment showed faster
and/or higher biogas production depending on the amount of particles solubilised [37]. Our results with microalgae microwave pretreatment suggest that the higher the applied specic energy, the
higher the organic matter solubilisation, biogas production rate
and biogas yield. Bearing in mind the low soluble organic content
of microalgal biomass (<10%), the anaerobic biodegradability increase may be explained by the disruption of microalgae cell wall
structure, which lead to degradation of organic material that was
not yet solubilised after the pretreatment process.
The anaerobic digestibility of untreated microalgal biomass was
relatively low (172 mL biogas/g VS). The digestibility was higher
for S. obliquus biomass (0.21 L CH4/g VS) and Phaeodactylum
tricornutum (0.35 L CH4/g VS) [38], grown as pure cultures with
nutrients supply. Chen and Oswald [13] investigated the thermochemical pretreatment in the sole study dealing with the pretreatment and anaerobic biodegradability of microalgae and bacteria
biomass grown in wastewater developed so far. According to these
authors, biomass pretreatment at 100 C for 8 h increased the
methane production by 33%. Furthermore, Gonzlez-Fernndez
et al. [15] reported a biogas production increase by 48% after Scene-

3.4. Energy considerations


Energy considerations are a key issue for evaluating the viability
of pretreatment techniques at large-scale [10,19]. In principle the
energy requirements should (at least) be covered by the extra
methane produced as a result of implementing the pretreatment
step. Based on our experimental results, the energy balance of microalgae microwave pretreatment was estimated. The energy input
(Ei) corresponded to the specic energy applied to organic biomass
(Eq. (8)).

EikJ=kg VS power W  times=volatile solids kg VS 8


The energy output (Eo) was calculated from the difference between the methane yield of trials with pretreated (T1  T9) and untreated biomass (control), according to the following equation (Eq.
(9)), where DP CH4 is the methane yield increase after biomass pretreatment (mL/g VS) and n is the lower heating value of methane
(35,800 kJ/m3CH4) [39].

EokJ=kg VS DPCH4 mL=g VS  nkJ=m3 =1000

In this manner, the energy balance of the process was estimated


by the energy input to the energy output (Ei/Eo) ratio. In Table 7,
energy ratios for all pretreatment conditions are summarized. As
can be seen, the Ei/Eo ratio ranged from 32 to 70, attesting that
the energy input was always higher than the energy output of
the pretreatment. This is in accordance with a previous study on
lignocellulosic biomass microwave pretreatment, where Ei/Eo ratios between 5 and 130 were found [19]. In our research, however,
such high energy consumption was rstly attributed to the low VS
concentration of harvested biomass. Tang et al. [40] mentioned the
water content of sewage sludge as the most important factor inuencing the energy balance of microwave pretreatment. Moreover,
hydrothermal pretreatment of cotton stalks was only feasible with
10% TS content [41]. Microalgae harvesting is a challenge for massive microalgae cultivation and different techniques are currently

Table 6
Microwave pretreatment conditions and biogas production in biochemical methane potential tests. Mean values (standard deviation).
Trial

Applied specic energy (kJ/


kg TS)

Output power
(W)

Temperature
(C)

Initial biogas production rate (mL/


day/g VS)

Biogas yield (mL/g


VS)

Methane content in biogas


(%CH4)

Control
T1
T2
T3
T4
T5
T6
T7
T8
T9

21,800
21,800
21,800
43,600
43,600
43,600
65,400
65,400
65,400

300
600
900
300
600
900
300
600
900

50
58
56
81
86
92
95
98
98

50.08
63.69
68.50
73.25
66.06
68.16
67.44
78.65
84.11
87.76

172.48
194.06
207.47
219.54
222.63
238.83
244.50
275.76
295.97
307.11

68.2
68.4
68.4
68.5
68.3
68.3
68.4
68.3
68.2
68.4

(1.16)
(1.70)
(0.34)
(0.83)
(0.39)
(3.36)
(0.78)
(4.67)
(0.24)
(1.70)

(2.95)
(5.84)
(4.67)
(3.36)
(3.06)
(2.04)
(1.31)
(0.58)
(1.17)
(2.92)

(0.11)
(0.31)
(0.21)
(0.03)
(0.07)
(0.03)
(0.01)
(0.08)
(0.06)
(0.23)

174

F. Passos et al. / Applied Energy 108 (2013) 168175

320

Biogas yield (mL/g VS)

300
280

R= 0,85

260
240
220
200
180
200

300

400

500

600

700

800

900

biomass cultivated in HRAP. The main parameter inuencing biomass solubilisation was the applied specic energy. A similar solubilisation increase was obtained for the same specic energy,
regardless of the output power and exposure time (280350% for
21,800 kJ/kg TS, 580610% for 43,600 kJ/kg TS and 730800% for
65,400 kJ/kg TS). This pretreatment increased the biogas production rate (2775%) and nal biogas yield (1278%) in BMP tests.
Therefore, microwave irradiation enhanced microalgae solubilisation, accelerating the anaerobic digestion rate and increasing the
methane production potential. However, the pretreatment of concentrated biomass seemed imperative to obtain a positive energy
balance. Besides, less energy consuming pretreatment techniques
ought to be investigated.
Acknowledgements

Solubilisation (% VSs/VS increase)


Fig. 3. Microalgal biomass solubilisation versus biogas yield under microwave
pretreatment.

Table 7
Energy ratio of microalgal biomass under different microwave pretreatment
conditions.
Trial

VS content (%
(w/w))

Energy input
(kJ/kg VS)

Energy output
(kJ/kg VS)

Energy ratio
(Ei/Eo)

T1
T2
T3
T4
T5
T6
T7
T8
T9

0.98
0.98
0.98
0.98
0.98
0.98
0.98
0.98
0.98

36,700
36,700
36,700
73,400
73,400
73,400
110,200
110,200
110,200

525.3
851.8
1145.6
1220.9
1615.2
1753.3
2514.2
3006.2
3277.4

69.9
43.1
32.0
60.1
45.4
41.9
43.8
36.7
33.6

being investigated. This is a matter of concern, since pretreatment


technologies such as microwave irradiation may require concentrated biomass to improve the energy balance of biogas production. Thus, effective low-cost thickening techniques are required
for competitive biofuel production from microalgae.
To our knowledge, this is the rst study including the energy balance of microalgae pretreatment. Nevertheless, literature results
were used to compare the energy balance of microwave and thermal
pretreatment, at temperatures within the range of the microwave
pretreatment (5595 C). The energy balance was calculated as proposed by Ferrer et al. [42]. According to this, energy ratios for Scenedesmus biomass pretreated at 70 and 80 C were 97.3 and 18.7 Ei/Eo,
respectively (data by Gonzlez-Fernndez et al. [14]); and the energy ratio for a mixed microalgae culture pretreated at 55 C was
60.2 (data by Alzate et al. [16]). This suggests that microalgae
pretreatment succeeds in enhancing the methane yield, even though
the energy consumed seems higher than the extra energy produced
by the pretreatment. In light of the results, future studies should
focus on less energy consuming pretreatment techniques.
It is worth mentioning that the methane yield of microalgae
may have been underestimated, because BMP tests were not carried out using inoculum acclimated to microalgae digestion. While
the results of BMP tests are useful to compare pretreatment conditions, continuous reactors with a microbial community acclimated
to microalgal biomass should better be used to verify the energy
balance of the process.
4. Conclusions
This study assessed the effect of microwave pretreatment on
the solubilisation and anaerobic digestion of microalgaebacterial

This research was funded by the Spanish Ministry of Science


and Innovation (Project BIOALGAS CTM2010-17846). Fabiana Passos is grateful to the Coordination for the Improvement of Higher
Level Personal (CAPES) funded by the Brazilian Ministry of Education for her PhD scholarship. The contribution of Javier Carretero
and Sabina Cordn from the Universitat Politcnica de Catalunya.BarcelonaTech is appreciated.
References
[1] Gonzlez-Fernndez C, Sialve B, Bernet N, Steyer JP. Impact of microalgae
characteristics on their conversion to biofuel. Part II: Focus on biomethane
production. Biofuels, Bioprod Bioren 2011;6(2):20518.
[2] Rawat I, Kumar RR, Mutanda T, Bux F. Biodiesel from microalgae: a critical
evaluation from laboratory to large scale production. Appl Energy 2013;103:
44467.
[3] Sturm BSM, Lamer SL. An energy evaluation of coupling nutrient removal from
wastewater with algal biomass production. Appl Energy 2011;88:3499506.
[4] Park JBK, Craggs RJ, Shilton AN. Wastewater treatment high rate algal ponds for
biofuel production. Bioresour Technol 2011;102:3542.
[5] Garca J, Green BF, Lundquist T, Mujeriego R, Hernndez-Marin M, Oswald WJ.
Long term diurnal variations in contaminant removal in high rate ponds
treating urban wastewater. Bioresour Technol 2006;97:170915.
[6] Golueke CG, Oswald WJ, Gotaas H. Anaerobic digestion of algae. Appl Microbiol
1957;7:21927.
[7] Oswald WJ, Golueke CG. Biological transformation of solar energy. Adv Appl
Microbiol 1960;2:22362.
[8] Ferrer I, Vzquez F, Font X. Comparison of the mesophilic and thermophilic
anaerobic sludge digestion from an energy perspective. J Residuals Sci Technol
2011;8:817.
[9] Chandra R, Takenchi H, Haregawa T. Methane production from lignocellulosic
agricultural crop wastes: a review in context to second generation of biofuel
production. Renew Sust Energy Rev 2012;16(3):146276.
[10] Gonzlez-Fernndez C, Molinuevo-Salces B, Garca-Gonzlez MC. Evaluation of
anaerobic codigestion of microalgal biomass and swine manure via response
surface methodology. Appl Energy 2011;88:344853.
[11] Liu C-Z, Wang F, Stiles AR, Guo C. Ionic liquids for biofuel production:
opportunities and challenges. Appl Energy 2012;92:40614.
[12] Crrere H, Dumas C, Battimelli A, Batstone DJ, Delgens JP, Steyer JP, et al.
Pretreatment methods to improve sludge anaerobic degradability: a review. J
Hazard Mater 2010;183:115.
[13] Chen PH, Oswald W. Thermochemical treatment for algal fermentation.
Environ Int 1998;24:88997.
[14] Gonzlez-Fernndez C, Sialve B, Bernet N, Steyer JP. Comparison of ultrasound
and thermal pretreatment of Scenedesmus biomass on methane production.
Bioresour Technol 2012;110:6106.
[15] Gonzlez-Fernndez C, Sialve B, Bernet N, Steyer JP. Thermal pretreatment to
improve methane production of Scenedesmus biomass. Biomass Energy
2012;40:10511.
[16] Alzate ME, Muoz R, Rogalla F, Fdz-Polanco F, Prez-Elvira SI. Biochemical
methane potential of microalgae: inuence of substrate to inoculum ratio,
biomass concentration and pretreatment. Bioresour Technol 2012;123:
48894.
[17] Marin J, Kennedy KJ, Eskicioglu C. Effect of microwave irradiation on anaerobic
degradability of model kitchen waste. Water Manage 2010;30:17729.
[18] Park W-J, Ahn J-H, Hwang S, Lee C-K. Effect of output power, target
temperature, and solid concentration on the solubilisation of waste activated
sludge using microwave irradiation. Bioresour Technol 2010;101:S136.
[19] Hu Z-H, Yue Z-B, Yu H-Q, Liu S-Y, Harada H, Li Y-Y. Mechanisms of microwave
irradiation pretreatment for enhancing anaerobic digestion of cattail by rumen
microorganisms. Appl Energy 2012;93:22936.

F. Passos et al. / Applied Energy 108 (2013) 168175


[20] Kaatze U. Fundamentals of microwaves. Radiation Phys Chem
1995;45:53948.
[21] Climent M, Ferrer I, Baeza MM, Artola A, Vzquez F, Font X. Effects of thermal
and mechanical pretreatments of secondary sludge on biogas production
under thermophilic conditions. Chem Eng J 2007;133:33542.
[22] Eskicioglu C, Kennedy KJ, Droste RL. Characterization of soluble organic matter
of waste activated sludge before and after thermal pretreatment. Water Res
2006;40:372536.
[23] Eskicioglu C, Kennedy KJ, Droste RL. Enhanced disinfection and methane
production from sewage sludge by microwave irradiation. Desalination
2009;248:27985.
[24] Slyom K, Mato RB, Prez-Elvira SI, Cocero MJ. The inuence of the energy
absorbed from microwave pretreatment on biogas production from secondary
wastewater sludge. Bioresour Technol 2011;102:1084954.
[25] Zheng J, Kennedy KJ, Eskicioglu C. Effect of low temperature microwave
pretreatment on characteristics and mesophilic digestion of primary sludge.
Environ Technol 2009;30:31927.
[26] Eskicioglu C, Terzian N, Kennedy KJ, Droste RL, Hamoda M. A thermal
microwave effects for enhancing digestibility of waste activated sludge.
Water Res 2007;41:245766.
[27] Pedescoll A, Corzo A, Alvarez E, Puigagut J, Garca J. Contaminant removal
efciency depending on primary treatment and operational strategy in
horizontal subsurface ow treatment wetlands. Ecol Eng 2011;37:37280.
[28] Cho YT, Young JC, Jordan JA, Moon HM. Factors affecting measurement of
specic methanogenic activity. Water Sci Technol 2005;52:43540.
[29] APHA-AWWA-WPCF. Standard methods for the examination of water and
wastewater. 20th ed. Washington; 1999.
[30] Lpez CVG, Cron Garca MC, Acin Fernandz FG, Segvia Bustos C, Chisti Y,
Fernndez Sevilla JM. Protein measurements of microalgal and cyanobacterial
biomass. Bioresour Technol 2010;101:758791.
[31] Ferrer I, Pons S, Vsquez F, Font X. Increasing biogas production by thermal
(70 C) sludge pretreatment prior to thermophilic anaerobic digestion.
Biochem Eng J 2008;42:18692.

175

[32] Garca J, Mujeriego R, Hernandez-Marine M. High rate algal pond operating


strategies for urban wastewater nitrogen removal. J Appl Phycol
2000;12:3319.
[33] Becker EW. Microalgae in human and animal nutrition. In: Richmond A, editor.
Handbook of microalgal culture. Oxford: Blackwell Publishing; 2004.
[34] Gonzlez-Fernndez C, Molinuevo-Salces B, Garca-Gonzlez MC. Open and
enclosed photobioreactor comparison in terms of organic matter utilization,
biomass chemical prole and photossynthetic efciency. Ecol Eng
2010;36:1497501.
[35] Carrre H, Bougrier C, Castets D, Delgens JP. Impact of initial biodegradability
on sludge anaerobic digestion enhancement by thermal pretreatment. J
Environ Sci Heal A 2008;43:15515.
[36] Monlau F, Latrille E, Costa AC, Steyer JP, Carrre H. Enhancement of methane
production from sunower oil cakes by dilute acid pretreatment. Appl Energy
2013;102:110513.
[37] Bougrier C, Albasi C, Delgens JP, Carrre H. Effect of ultrasonic, thermal and
ozone pretreatment on waste activated sludge solubilisation and anaerobic
digestion. Chem Eng Process 2006;45:7118.
[38] Zamalloa C, Boon N, Verstraete W. Anaerobic digestion of Scedesmus obliquus
and Phaeodactylum tricornutum under mesophilic and thermophilic conditions.
Appl Energy 2012;92:7338.
[39] Metcalf, Eddy. Wastewater engineering. Treatment and reuse. 4 ed. New
York: McGraw-Hill; 2003.
[40] Tang B, Yu LF, Huang SS, Luo JZ, Zhuo Y. Energy efciency of pretreating excess
sewage
sludge
with
microwave
irradiation.
Bioresour
Technol
2010;101:50927.
[41] Adl M, Sheng K, Gharibi A. Technical assessment of bioenergy recovery from
cotton stalks through anaerobic digestion process and the effects of
inexpensive pretreatments. Appl Energy 2012;93:25160.
[42] Ferrer I, Serrano E, Pons S, Vzquez F, Font X. Enhancement of thermophilic
anaerobic sludge digestion by 70 C pretreatment: energy considerations. J
Residuals Sci Technol 2009;6(1):118.

Você também pode gostar