Energy balance and environmental impact analysis of marine

microalgal biomass production for biodiesel generation in

a photobioreactor pilot plant
E. Sevigne Itoiz
a,d,
*, C. Fuentes-Gru newald
b,c
, C.M. Gasol
a,d
, E. Garces
b
, E. Alacid
b
,
S. Rossi
c
, J. Rieradevall
d
a
Ine`dit, Carretera de Cabrils, Km. 2, IRTA, 08348 Cabrils, Spain
b
Department of Marine Biology and Oceanography, Marine Science Institute, CSIC, Passeig Martim de la Barceloneta,
37-49 E-08003 Barcelona, Spain
c
Institute of Environmental Science and Technology (ICTA), Universitat Auto`noma de Barcelona (UAB), Building C Campus UAB,
08193 Cerdanyola del Valles (Barcelona), Spain
d
SOSTENIPRA, Department of Chemistry Engineering, Universitat Auto`noma de Barcelona (UAB), Building Q UAB,
08193 Cerdanyola del Valle`s (Barcelona), Spain
a r t i c l e i n f o
Article history:
Received 21 May 2011
Received in revised form
11 January 2012
Accepted 12 January 2012
Available online 3 February 2012
Keywords:
Alexandrium minutum
Karlodinium venecum
Heterosigma akashiwo
Pilot plant photobioreactor
Life cycle assessment
Energy balance
a b s t r a c t
A life cycle assessment (LCA) and an energy balance analysis of marine microalgal biomass
production were conducted to determine the environmental impacts and the critical points
of production for large scale planning. The articial lighting and temperature conditions of
an indoor bubble column photobioreactor (bcPBR) were compared to the natural conditions
of an equivalent outdoor system. Marine microalgae, belonging to the dinoagellate and
raphidophyte groups, were cultured and the results were compared with published LCA
data obtained from green microalgae (commonly freshwater algae). Among the species
tested, Alexandrium minutum was chosen as the target marine microalgae for biomass
production under outdoor conditions, although there were no substantial differences
between any of the marine microalgae studied. Under indoor culture conditions, the total
energy input for A. minutum was 923 MJ kg
1
vs. 139 MJ kg
1
for outdoor conditions.
Therefore, a greater than 85% reduction in energy requirements was achieved using
natural environmental conditions, demonstrating the feasibility of outdoor culture as an
alternative method of bioenergy production from marine microalgae. The growth stage
was identied as the principal source of energy consumption for all microalgae tested, due
to the electricity requirements of the equipment, followed by the construction material of
the bcPBR. The global warming category (GWP) was 6 times lower in outdoor than in indoor
conditions. Although the energy balance was negative under both conditions, this study
concludes with suggestions for improvements in the outdoor system that would allow up-
scaling of this biomass production technology for outdoor conditions in the Mediterranean.
2012 Elsevier Ltd. All rights reserved.
* Corresponding author. Ine` dit, Carretera de Cabrils, Km. 2, IRTA, 08348 Cabrils, Spain. Tel.: 34 93 581 37 60; fax: 34 93 581 33 31.
E-mail address: eva.sevigne@uab.cat (E. Sevigne Itoiz).
Available online at www.sciencedirect.com
ht t p: / / www. el sevi er. com/ l ocat e/ bi ombi oe
b i oma s s a nd b i oe ne r g y 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5
0961-9534/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biombioe.2012.01.026
1. Introduction
The next decade will be crucial in solving many of the envi-
ronmental issues of our planet, especially those regarding the
increase in greenhouse gases (GHG), water shortages, and the
depletion of fossil fuels. Issues related to CO
2
emissions and
fossil fuel depletion are linked, due to the large amounts of
CO
2
released into the atmosphere from the industrial,
transportation, and energy sectors [1]. To avoid further
increases in GHG emissions and to increase the energy
reserves of different countries, governments, policy stake-
holders and research groups are investing in and developing
projects related to the production of biofuels from terrestrial
biomass feedstock, known as the rst generation biodiesel,
including corn, rapeseed, sunowers, and sugarcane plants.
There are advances in the production of second generation
biodiesel, using residues from trees or lignocellulosic mate-
rial as feedstock for bio-ethanol production. However, the use
of these feedstocks for biodiesel production is controversial
because the processing and commercialization of terrestrial
plants are associated with several environmental and social
problems, including a loss of biodiversity, increased fresh-
water consumption, higher prices of edible plants, and the
resulting social inequalities [2e4]. Alternatively, one of the
most promising feedstocks for the third generation of bio-
diesel production involve microalgae, due to their photo-
synthetic conversion efciency, fast growth, sustainable
biomass production, and high content of triacylglycerols
(TAG), which is the oil that is commonly used as a raw
material for biodiesel production [5,6]. To date, freshwater
microalgae have been the main microalgal species
researched for biomass and biodiesel production purposes.
Of particular interest are the green algae, or Chlorophycean,
including Chlorella vulgaris, Chlorella protothecoides, Chlamydo-
monas reinhardtii, and Neochloris oleoabundans, due to their
high growth rates and their well-studied life cycle [7,8].
However, a drawback to their use is the permanent need for
large quantities of freshwater in the continuous production
of sufcient microalgal biomass, independent of the culture
system. Use of sea/wastewater as the culture medium would
signicantly reduce the water footprint [9]. This implies the
need to isolate seawater strains from the same place where
they will later be grown. The efcient use of these strains
requires that they have high TAG concentrations in addition
to other energetically or commercially favorable cellular
metabolites. Several advantages of the use of seawater as the
medium for microalgae are that it leaves freshwater supplies
free for other human and ecosystem uses, avoids ecological
problems associated with the introduction of exotic micro-
algal species, maintains the system without any alteration to
the local ecology, and avoids the loss of biodiversity [10,11].
The use of seawater microalgae strains allows the installa-
tion and operation of industrial scale plants in coastal
countries, use non-arable land, and avoids or at least reduces
freshwater consumption.
Based on these considerations, our group has explored the
growth rates, lipid proles, and TAG concentrations of various
marine microalgal species and involved culturing the strains
of interest in enclosed systems and improving these cultures
for energetic purposes [12,13]. Most of the microalgae evalu-
ated by our group in previous studies belong to the dinoa-
gellates and raphidophytes classes [12]. Dinoagellates are
well known because of their extensive bloom-forming prolif-
erations in natural marine environments throughout the
world [14,15]; in terms of the production of biomass for bio-
energy, this harmful trait becomes an opportunity and an
advantage. Previous studies [16,17] determined that dinoa-
gellates and raphidophytes readily adapt to growth in
enclosed systems and that their natural capacity of prolifer-
ation can be exploited to establish long-term biomass culture
facilities in various coastal countries [17,18]. The strains used
in this study are present globally and can be considered
strategic species because they can be isolated readily from
local seawater spots around the world [14]. Alexandrium min-
utum is a tecate dinoagellate with a high cell biovolume
(>2800 mm
3
) with a high biomass and lipid productivity. The
dinoagellate Karlodinium venecum and the raphidophyte
Heterosigma akashiwo are atecate cells and are advantageous in
terms of lipid extraction by the ease of breaking the cells and
avoidance of a higher energy input for the extraction of the
lipids [13].
The biotechnology used for biomass production from
microalgae principally involves two types of culture congu-
ration: open and enclosed systems. Open systems, including
raceways or open ponds, have a low initial cost of construc-
tion and maintenance, with a relatively low volumetric
productivity, and parameters including temperature, evapo-
ration, and contamination cannot be totally controlled [5].
Enclosed systems, including horizontal photobioreactors,
bubble columns, or at panels, produce a higher volumetric
biomass (13-fold greater than raceways or ponds), allow the
growth of a single microalgal cell type (monoculture), and
have fewer contamination problems than open systems.
However, the initial cost of construction is higher for enclosed
systems than for open systems [5]. The energy cost of micro-
algal biomass productionin enclosed systems suffers fromthe
current need for materials and procedures that require high
amounts of energy, including the different plastics used in the
construction of the photobioreactor in bubble column photo-
bioreactors and the concrete needed for open pond systems.
Electricity consumption during the microalgal growth stage
(water, air pumping, CO
2
injection, etc.) or in the ltration
systems used to extract the biomass from the seawater in the
dewatering stage is also high. Bothopenand enclosed systems
are used to grow microalgae under autotrophic conditions,
with sunlight as the energy source, nutrients obtained from
a liquid medium, and inorganic carbon, as CO
2
, provided in
pure form or as injected air with atmospheric CO
2
concen-
trations. With these inputs, chemical energy is formed via
photosynthesis [19]. Presently, most of the studies that use
microalgae for biofuel purposes have been implemented in
the lab or pilot scale, pending industrial scaling to demon-
strate the production feasibility [7,8].
In this study, an enclosed system was chosen to achieve
high marine microalgae biomass production because it allows
the control of abiotic parameters and its biomass production
per volumetric area is higher than in open systems. Addi-
tional considerations in establishing open system facilities
b i o ma s s a nd b i o e ne r gy 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 325
are the high price of land in the Mediterranean area and the
stable weather conditions in this area. The local strains of
dinoagellates and raphidophytes produce extensive natural
proliferations in the Mediterranean basin [20], so these
conditions were reproduced in controlled systems [12,13],
together with the same abiotic parameters and seawater
encountered by natural populations, following the suggestion
of built around algae facilities for long-term microalgal
biomass production [21].
Life cycle assessment (LCA) is a tool that allows the
potential impacts along the life cycle of a product, process, or
activity to be evaluated. LCA studies in microalgal biomass
production for biodiesel purposes are principally based on
models or laboratory data; however, most of the data are
assumptions or refer to a hypothetical system based on
extrapolations from lab-scale studies [9,22,23]. In this study,
data for the LCA were obtained from a previous study [18], in
which microalgal cultures were run in a bubble column pho-
tobioreactor (bcPBR) pilot plant under controlled conditions
(indoors) and in a natural environment (outdoors). Energy
balance is the key consideration in the design and develop-
ment of a new methodology/feedstock aimed at energy
production. Accordingly, measuring and evaluating the
energy consumption of a newly proposed system simplies
improvements and facilitates increases in its efciency.
The aims of the present study can be dened as follows:
1) To determine the energy balance of dry marine microalgal
production (A. minutum, K. venecum and H. akashiwo) in
a bcPBR pilot plant under indoor and outdoor conditions.
2) To evaluate and determine the principal environmental
and energy impacts in the production of marine microalgal
biomass under articial (indoor) and natural (outdoor)
conditions of temperatureandlightinginabcPBRpilot plant.
3) To assess the relative energy and environmental contri-
butions of LCA stages, to detect the weak also in addition to
the critical points of an outdoor system, with the goal of
obtaining a viable and scalable design for an industrial
scale biodiesel facility.
4) To discuss the feasibility of microalgal biomass production
facilities for biodiesel generation in the Mediterranean
basin using outdoor conditions without the need of energy
inputs using articial light and temperature control.
2. Materials and methods
2.1. Description of the microalgal cultivation in the pilot
plant
The study was conducted at the Institut de Cie` ncies del Mar
(ICM-CSIC), Barcelona, Spain, under ambient Mediterranean
climate conditions (41

23
0
16.5
00
N; 02

10
0
11.71
00
E). Three
species of microalgae, two belonging to Dinophyceae (AMP4 A.
minutum and ICMB252 K. venecum) and one to Raphidophy-
ceae (ICMB830 H. akashiwo) were grown in bubble columns
under indoor and outdoor environmental conditions.
The experimental design consisted of a bcPBR, which has
a supporting structure of wood and polymethylmethacrylate
tubes, as depicted in Fig. 1. The polymethylmethacrylate tubes
(height 2.0 m and diameter 0.15 m) each had a volume of
33 dm
3
. Three tubes were used for each microalgal species,
both for indoor and outdoor conditions; therefore, the indoor
system had a total workload of 0.297 m
3
as did the outdoor
system. The bcPBR was 2.65 m in length and 0.75 m in width.
The separation between the tubes was 0.11 m, with a total
surface utilized of 1.98 m
2
and a volume-surface ratio of
0.15 m
3
m
2
. For both growth conditions, the microalgae were
cultured in triplicate.
Under indoor conditions, the microalgal strains were
grown in a temperature-controlled room at 20

C 1

C. All
cultures were grown in ltered (0.21 mm) seawater (salinity of
37 kg m
3
and neutral pH) obtained from the ICM culture
facilities and supplemented with a full L1-enriched medium
without added silicates [24]. Pre-ltered air (Iwaki lter, 0.2 mm
pore size) with a CO
2
concentration of 420 mL L
1
16 mL L
1
(measured by a Qubitsystem S151 CO
2
Analyzer) was injected
from the bottom of the tubes at a ow of 50c m
3
s
1
, which
allowed gentle agitation inside the bubble column.
For outdoors conditions, a bcPBR with the same layout,
seawater salinity, pH, injected air, and growth medium as
used for the indoor conditions was placed on the terrace of the
ICM-CSIC. The experiment started in mid November 2009 and
was terminated at the end of May 2010 (autumn, winter, and
spring in the northern hemisphere). Cultures were run in
a semi-continuous mode because 50% of the biomass was
harvested depending on the duplication time of each species
(Fig. 2). Throughout the experiment, light and temperature
Fig. 1 e Photograph of the bubble column photobioreactor (bcPBR) under outdoor (left) and indoor (right) conditions.
b i oma s s a nd b i oe ne r g y 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 326
were recorded under the outdoor conditions from the Cata-
lonia meteorological station net [25].
To obtain dry biomass, the samples were centrifuged at
471 rad s
1
for 420 s in a Sigma 3e16 K centrifuge to separate
the seawater fromthe microalgae. The supernatant water was
discarded and a wet biomass pellet was recovered.
2.2. Life cycle assessment (LCA) of the microalgal
biomass production in a bcPBR pilot plant
The energy and environmental assessment of the proposed
experimental design was carried out using the LCA method-
ology. The LCA evaluates the potential impacts along the life
cycle of a product, process, or activity, from raw material
extraction to production, use, and disposal [26]. The ISO 14040
provides guidance on the four steps of the LCA: goal and
scope, inventory analysis, life cycle impact assessment, and
life cycle interpretation.
2.2.1. Functional unit and boundary system
The functional unit of this study is the production under
indoor and outdoor conditions of 1 kg of dry microalgal
biomass from each of the species studied. The biomass ob-
tained would be used for biodiesel production. Fig. 3 depicts
the studied system and its limits. The system includes all the
steps necessary to obtain dry biomass from microalgae:
culture medium production, bcPBR structure production,
energy consumption during the lling and dewatering stages,
Fig. 2 e Growth curve of the different microalgae tested under outdoor conditions. indicates the harvest time of the
culture.
Fig. 3 e Life cycle system of microalgal biomass production for biodiesel production.
b i o ma s s a nd b i o e ne r gy 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 327
growth of the microalgae (indoors and outdoors), and bcPBR
maintenance (cleaning). Lipid extraction and trans-
esterication are not considered in the limits of biomass
production of this LCA.
2.2.2. Life cycle inventory
Table 1 shows the life cycle inventory and the data, which
were collected and classied throughout the experiment
(November 2009eMay 2010). All data are expressed per func-
tional unit, i.e., the production of 1 kg of dry microalgal
biomass, except for the equipment, is expressed in terms of
power. Table 2 details the dry biomass obtained per liter [18].
Inows to the system included equipment power (kW),
operating rates (s kg
1
), photobioreactor material (acrylic
kg kg
1
), culture medium doses (kg kg
1
), and seawater
consumption (m
3
kg
1
). Outows from the system were dry
biomass (kg) and the waste seawater with L1 culture medium
obtained following centrifugation (kg m
3
). In the dewatering
process, 98.5% of the water is lost as a result of the centrifu-
gation dewatering [12]. The production inventory of the
culture medium was taken from the literature and the
ecoinvent database [27,28]. Data for the electricity was ob-
tained from the ecoinvent database as well [29].
The water and air needed for the experiment were supplied
by general pumps located in the ICM which in turn supply
water and air to various experiments of the research center.
The total energy consumption from the water pump was
calculated from the hours of working required for the exper-
iment and pump power. The same procedure was followed for
the energy consumption of the dewatering, although specic
equipment was used for the experiment. Air was pumped into
a tank with a owof 202 dm
3
s
1
and then was provided to the
experiment with a ow of 50 cm
3
s
1
. The total pump energy
consumption was calculated considering time for tank lling
and air pump power.
The total volume of the chamber used is greater than the
volume required for this experiment; therefore, the total
energy consumption of the chamber (28.8 m
3
) was adapted to
the volume of the growing tubes (0.3 m
3
), taking into account
the space needed between the tubes (the volume fraction is
14%). The same procedure used for the chamber was adopted
to determine the energy consumption due to the uorescent
lights. To calculate the bioenergy production from the
biomass obtained the lipid extraction and the oil trans-
esterication should be considered. A production rate of 25%
lipids was measured for each microalgal species in a previous
study [13,19] and a transformation of 90% was considered.
2.2.2.1. Assumptions for life cycle inventory. In the life cycle
inventory the following assumptions were made:
For the bioenergy production calculation, the experimental
low caloric value of 39 MJ kg
1
was used [30].
The useful life of the bcPBR was estimated to be 10 years,
and its total weight 80 kg.
2.2.3. Life cycle impact assessment (LCIA)
The SimaPro 7.1.8 software was used for the environmental
evaluation together with the method detailed in CML base-
line 2001. The impact categories include are: abiotic depletion
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s
.
b i oma s s a nd b i oe ne r g y 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 328
(AD) in kg Sb eq.; acidication (A) in kg SO
2
eq.; eutrophication
(E) in kg PO
4
eq.; global warming potential (GWP) in kg CO
2
eq.;
ozone layer depletion (ODP) in mg CFC-11 eq.; human toxicity
(HT) in kg 1,4-DB eq.; freshwater aquatic ecotoxicity (FWAE) in
kg 1,4-DB eq.; marine aquatic ecotoxicity (MAE) in kg 1,4-DB
eq.; terrestrial ecotoxicity (TE) in kg 1,4-DB eq.; and photo-
chemical oxidation (PO) in kg C
2
H
4
eq.
2.2.4. Energy assessment
Simapro 7.1.8 software and the Cumulative Energy Demand v
1.4 method were used in the energy assessments at all stages
of the LCA. This method was used to estimate the direct
energy consumption, including the use of seawater and the
freshwater needed for the maintenance, production of culture
medium and the production of bcPBR. In addition, the net
energy balance was determined, calculated as the difference
between energy output and energy input.
2.3. Sensitivity analysis
A sensitivity analysis was conducted using the variables of
energy consumption and lipid content of dry biomass to
observe when positive balances would be achieved. The
analysis used results obtained for outdoor production from A.
minutum because this dinoagellate species presented the
best energy results. Five scenarios where dened as A, B, C, D
and E. The base case for all results reported in this LCA is
calculated for the algae composition of 25% lipids so the
percentage of lipid content was increased at intervals of 10%
from the base case represented by scenario A. Energy
consumption was reduced at intervals of 50% from the base
results obtained in the study. Both variables were modied in
each scenario, so in scenario B the energy consumption was
reduced by 50%over scenario A and lipid content increased by
10%; in scenario C energy consumption was reduced by 50%
over scenario B and lipid content was increased again by 10%;
and so on for scenarios D and E.
3. Results
The following sections describe the energy balances obtained
for indoor andoutdoor productionsystems andthe energy and
environmental assessment of the different stages considered
in the LCA. Finally, the data from the sensitivity analyses
determined from the best results (A. minutum) is presented.
3.1. Energy results
Table 3 lists the total energy consumption by each species of
marine microalgae for both production systems and the
output of bioenergy production from microalgae based on the
inventory and the assumptions described in Section 2.2.2. The
energy balances obtained are also presented. The results are
expressed in MJ per kg of dry microalgae species biomass.
3.1.1. Energy results of production systems
First, it is observed from Table 3 that negative balances were
obtained for both productions systems. In addition, the energy
balance results demonstrated large differences between the
indoor and outdoor systems in contrast to the biomass results
displayed in Table 2, in which the two systems did not differ
substantially. The outdoor systemconsumed signicantly less
energy than the indoor system with differences between 721
and 783 MJ kg
1
. Specically, A. minutum grown in the outdoor
system had the best energy balance (139 MJ kg
1
) while
indoor production of this same microalgae had the worst
balance (923 MJ kg
1
).
Table 2 e Dry biomass per liter for each microalgal specie
and growth system.
Heterosigma
akashiwo (g L
1
)
Alexandrium
minutum (g L
1
)
Karlodinium
venecum (g L
1
)
Indoor Outdoor Indoor Outdoor Indoor Outdoor
1.25 0.97 1.18 1.03 1.2 0.98
Table 3 e Energy consumption, output and balance per kg of dry biomass for each life cycle stage and for each microalgal
species and growth system.
Heterosigma akashiwo Alexandrium minutum Karlodinium venecum
Indoor Outdoor Indoor Outdoor Indoor Outdoor
Input (MJ kg
1
) bcPBR lling and culture 30.60 39.60 32.15 36.50 32.15 37.98
Filling (water pump) 0.13 0.17 0.13 0.16 0.13 0.17
Filling (seawater) 0.24 0.31 0.26 0.29 0.25 0.31
Culture 0.26 0.30 0.34 0.32 0.27 0.34
Growing of microalgae
Chamber 598.37 0.00 633.87 0.00 623.30 0.00
Air pump 73.47 94.98 77.83 89.17 76.54 93.72
Fluorescents 158.09 0.00 167.47 0.00 164.68 0.00
Dewatering
Centrifuge 6.21 8.00 6.57 7.53 6.46 7.92
Maintenance
Washing pump 2.80 3.61 2.97 3.40 2.92 3.57
Water 0.31 0.40 0.32 0.37 0.32 0.39
Total 872 148 923 139 908 146
Output (MJ kg
1
) 8.78 8.78 8.78 8.78 8.78 8.78
Balance (MJ kg
1
) 863 139 914 130 899 137
b i o ma s s a nd b i o e ne r gy 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 329
3.1.2. Energy results of microalgae
Minor differences were found for the energy results of the
different microalgal strains grown in the same production
system. In the case of outdoor production, energy consump-
tion differences were less than 7.5%and for indoor production
the energy demands differed by less than 6.0%. This means
that for each type of microalgae and for both systems,
biomass production was robust, and in future experiments
and applications any microalgal species could be used.
3.1.3. Energy results of life cycle stages
The analysis of life cycle stages of both types of production
and species indicated that the largest contributors to the
energy demand were the microalgal growth and the
construction of the bcPBR stages.
In the indoor system, the growing life stage required high
energy demands for light and temperature maintenance,
which need to be articially provided and controlled to
maintain constant environmental conditions for growth
(values highlighted in gray in Table 3) and using more than
85% of the electricity consumption of the entire system. The
elimination of these operations reduces the overall electricity
consumption by 90%, as observed in the outdoor system, in
which temperature and light were provided naturally, with no
need for additional electricity input. However, the outdoor
system air pumping involves considerable electricity
consumption in the growth stage, approximately 60% of the
entire system, constituting an energy demand of approxi-
mately 90 MJ. Notably that the equipment used for lighting,
temperature and air pumping at the growthstage was adapted
and not specially designed for the experiment, the ecodesign
of the equipment could signicantly reduce the electricity
consumption and therefore improve the energy balance. In
addition, the production of the bcPBR involves a signicant
energy demand in both systems because the chosen material
has a high energy requirement in its production. The poly-
methylmethacrylate tubes were chosen because they allow
a good light penetration for photosynthesis activity and
prevent the aging of the material by the action of UV rays. The
replacement of this material by other with same characteris-
tics or the bcPBR ecodesign could contribute to reduce the
energy inputs and improve the energy balances.
Other stages including dewatering, water consumption or
L1 culture production to promote microalgal growth involve
lower energy consumption in both systems; however, they
should be considered in further research.
3.2. Environmental results
The environmental impacts of bioenergy production per
functional unit were determined for ten impact categories.
The total environmental impact by production system and by
type of marine microalgae, particularly compared with the
global warming category, is presented followed by an evalu-
ation of the relative contributions of the life cycle stage.
3.2.1. Total environmental impacts
For all impact categories and microalgal species, outdoor
systems had lower environmental impacts (see Table 4).
Specically, A. minutum outdoor production had the lowest
environmental impact in all categories (marked in black in
Table 4). By contrast, A. minutum indoor production had the
highest impact (indicated in gray in Table 4) for all categories.
The outdoor system had signicantly fewer environmental
impacts than the indoor systems with differences between
85% and 88%, indicating that in environmental terms the
outdoor system had superior results and it is therefore pre-
sented as the preferable choice. Similar to energy results,
there were few differences between the types of microalgae,
for outdoor and indoor systems the environmental impacts
differ less than 6% between them in all impact categories.
Compared with the global warming (GWP) category, the
indoor systemproduction yielded an average of 146.3 kg 4 kg
of CO
2
eq. per functional unit (kg of dry biomass). The outdoor
production in the same category resulted in an average of
23.24 kg 0.7 kg of CO
2
eq. Thus, the GWP was 6 times lower
under outdoor than indoor conditions.
3.2.2. Environmental impacts of life cycle stage
To analyze in greater detail the environmental impacts by
impact category, it is necessary to assess the impacts by life
cycle stages. Fig. 4 shows the relative contributions of the
life cycle stages of A. minutum indoor production which
has the worst environmental impact results. The higher
Table 4 e Environmental impacts for microalgal species and impact category. Abiotic depletion (AD); acidication (A),
eutrophication (E), global warming potential (GWP); ozone layer depletion (ODP); human toxicity (HT); freshwater aquatic
ecotoxicity (FWAE); marine aquatic ecotoxicity (MAE); terrestrial ecotoxicity (TE) and photochemical oxidation (PO).
Impact category (eq. Units) Heterosigma akashiwo Alexandrium minutum Karlodinium venecum
Indoors Outdoors Indoors Outdoors Indoors Outdoors
A.D (kg SB eq.) 1.06E00 1.75E-01 1.12E00 1.69E-01 1.10E00 1.73E-01
A.C (kg SO
2
eq.) 1.36E-00 2.01E-01 1.44E00 1.94E-01 1.42E00 1.99E-01
E (kg PO
4
eq.) 7.02E-02 1.14E-02 7.45E-02 1.09E-02 7.32E-02 1.13E-02
GWP (kg CO
2
eq.) 1.44E02 2.38E01 1.53E02 2.29E01 1.51E02 2.35E01
ODP (kg CFC-11 eq.) 7.59E-06 9.82E-07 8.66E-06 1.63E-06 7.99E-06 9.72E-07
HT (kg 1,4-DB eq.) 4.29E01 5.82E00 4.56E01 5.64E00 4.47E01 5.77E00
FWAE (kg 1,4-DB eq.) 9.57E00 1.35E00 1.02E01 1.30E00 9.97E00 1.33E00
MAE (kg 1,4-DB eq.) 2.42E04 3.19E03 2.57E04 3.11E03 2.52E04 3.16E03
TE (kg 1,4-DB eq.) 2.41E-00 3.10E-01 2.56E00 3.04E-01 2.51E00 3.07E-01
PO (kg C
2
H
4
eq.) 5.05E-02 7.74E-03 5.37E-02 7.47E-03 5.27E-02 7.65E-03
b i oma s s a nd b i oe ne r g y 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 330
environmental impacts under indoor conditions for A. minu-
tum were due to the microalgal growth stage, which accoun-
ted for more than 95% of all of the environmental impacts and
is a totally function of electricity consumption, i.e., tempera-
ture, light conditions requirements and air pumping. The
impacts are mainly due to the electricity production which
depends on the Spanish energy mix considered which had
a contribution of 57% fossil fuel energy and 20% renewable
energy. The relative contribution of lling and centrifugation
were less than 2% and were dependent on the electricity
consumption and water and nutrient consumption for the
lling stage; thus, more than 96% of all of the environmental
impacts are due to electricity consumption and therefore due
to the Spanish mix. A change in the contributions of fossil
energies would contribute to decrease the environmental
impacts. The remaining environmental impacts from the
indoor production were a consequence of the bcPBR produc-
tion. A material change could involve a reduction of the
environmental impacts.
As was the case for the indoor production of A. minutum,
the outdoor production of H. akashiwo had the worst envi-
ronmental results; therefore, its breakdown of life cycle stages
was chosen to analyze the environmental impacts of the
outdoor system and to dene the principal environmental
impact. The results and its relative percentages for each life
cycle stages are depicted in Fig. 5. The electric consumption is
considerably lower in this system; therefore, the impacts due
to other stages implied a higher relative contribution for
certain categories. This demonstrates that these stages are
also a source of impacts and should be considered.
The electricity consumption yielded results of 71% (AD)
and 95% (ODP-TE) in all environmental impacts where the
growth stage accounted for 65% (AD) and 87% (ODP-TE) and
the centrifuge represented approximately 7% of impacts in all
categories. As for the indoor system, these impacts are due to
the energy mix considered. The production of the bcPBR
constitutes the second stage with higher impacts, and as in
the indoor production, the consumption of fossil fuels implies
that in AD, AC, E, GWP and PO, the contribution was between
14% and 24% indicating again that the reactor material
substitution could involve great environmental
improvements.
The lowest environmental impacts in all of the categories
were during the stage of lling which depends on electricity
for pumping, water and nutrients consumption. Fig. 6 pres-
ents their relative contributions showing that the L1 culture
Fig. 4 e Relative contributions of different life stages of A. minutum under indoor conditions.
Fig. 5 e Relative contribution of different life cycle stages of H. akashiwo under outdoor conditions.
b i o ma s s a nd b i o e ne r gy 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 331
consumption had the highest contribution in the categories of
E and GWP due to the nutrient consumption of nitrogen or
phosphorous.
3.3. Sensitivity analysis
Sensitivity analysis of the outdoor production of A. minutum
was performed by changing the energy consumption and lipid
content of the dry biomass. Table 5 displays the results ob-
tained for the scenarios dened. Positive balances were ob-
tained for scenarios D and E, which implies an energy
reduction of 88%fromthe base results presented in scenario A
and a content lipid of 55%. These results demonstrate that
great efforts should be made to achieve positive balances of
this production system. However, as noted in Section 3.1,
there is a great potential for energy reduction if ecodesign and
specically adapted equipment is used for the microalgae
production and/or if the bcPBR or the material itself is
replaced. The environmental impacts of scenario D would be
reduced by 63e84%; so the emissions of CO
2
eq. would be
8.2 kg per functional unit.
4. Discussion
The production of microalgae in an outdoor rather than an
indoor system results in a slight decrease in biomass
production; nevertheless, it involves a signicant decrease in
the total energy consumption, thus outdoor systems are pre-
sented as a preferable option. This study was conducted on
experimental data froma pilot plant and a key aspect was that
the equipment used was not specically designed for the
experiment. However, this is the rst step to properly scale an
experiment and the joint analysis of production, energy and
environmental impacts allows us to establish what the
weakest points are on which further research or greater effort
must be applied. The results of the pilot plant production
indicate that outdoor production is possible and that the
differences are notably small with controlled productions.
However, future studies should take into account that
biomass productivities in outdoor photobioreactors naturally
illuminated would depend on the prevailing weather condi-
tions in a particular locality [31]. Under Mediterranean climate
conditions, our outdoor production system yielded similar or
superior results as obtained for green algae in others studies
based on the same geographical area [32,33], and the differ-
ences between the marine microalgal species studied in this
study were so small that the production of any of them would
be possible.
In recent years, many LCA and energy balance studies on
the microalgae production for energetic purposes have been
conducted [34e43]; however, there is an enormous variety of
microalgae species that can be used to produce biodiesel and
many different methods of microalgal cultivation. In addition,
the life cycle stages included in each study may vary, thus,
while certain studies have analyzed the entire cycle [34,41]
others have only considered the culture process [38]. The
results of several of these studies are presented in Table 6.
However, due to methodological and life cycle differences,
general comparisons and extrapolations are difcult.
The energy assessment indicates negative balances for
both indoor and outdoor production systems; however, for the
latter, positive balances can be gained by reducing energy
consumption. In addition, for all the studies complied in Table
5 [37e40], negative balances are obtained except for [38] when
raceway pond and at-plate PBR are considered. These types
of reactors consume considerably less energy than tubular
PBRs [38,44,45] or open ponds [40], thus an alternative strategy
Fig. 6 e Relative contribution of electricity, water and L1 culture consumption of H. akashiwo under the outdoor conditions
during the lling stage.
Table 5 e Sensitivity analysis after modifying energy
consumption and lipid content for scenarios A, B, C, D
and E.
MJ kg
1
input MJ kg
1
output MJ kg
1
balance
Scenario A 139 9 130
Scenario B 69 12 57
Scenario C 35 16 19
Scenario D 17 19 2
Scenario E 9 23 14
b i oma s s a nd b i oe ne r g y 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 332
to decrease energy consumption would be to use an outdoor
system based on a raceway pond inside a greenhouse. None-
theless, in places in which evaporation is high, raceway ponds
require more frequent water pumping than tubular bioreac-
tors [41], which would increase energy consumption, and this
needs to be taken into consideration. In addition, raceway or
open ponds should be implemented in those countries with
extensive non-arable or inexpensive land (e.g., North African
countries). In contrast, in those countries in which high land
prices limit the system (EU Mediterranean countries), bcPBRs
or other enclosed systems is a reasonable choice. In addition,
the production of bcPBR has been observed to be the second
highest source of energy consumption due to material elec-
tion. As indicated by [40], one of the disadvantages of such
reactors is that their construction requires sophisticated
materials. Thus, innovations and ecodesign in the layout and
construction materials would signicantly reduce the energy
consumption associated with its production and decrease the
overall energy requirements. These innovations include the
combination of advanced designs of synthetic bags oating
partially submerged in an articial pond (a combination of
open and enclosed systems), or a single reactor module con-
sisting of one large translucent plastic bag containing multiple
vertical panels [21].
Downstream processing, i.e., dewatering and lipid extrac-
tion, have been observed as important stages and should be
considered in energy balances [46,47]. In a previous study [39],
dewatering constitutes the largest energy input, consuming
54 MJ per kg of dry biomass due to natural gas consumption.
However, a different study [40] carried out a comparative LCA
on dry and wet dewatering, and the dry process consumed
4.7 MJ per kg of dry biomass due to a centrifuge (similar to our
study) in which energy consumption resulting from dew-
atering is 6 and 8 MJ kg
1
for outdoor and indoor systems,
respectively. The lipid extraction is not discussed; however,
certain authors found the highest energy consumption as
a result of this stage [42,43]. Further studies must be con-
ducted to establish the best options for the dewatering alter-
natives and lipid extraction processes.
The use of a culture mediumto promote microalgal growth
is the life cycle stage with the lowest energy consumption,
which contrasts with results found in a previous study [37]
and with terrestrial crops for biofuel purposes, in which
energy consumption related to crop fertilization and to
production could be the highest in the entire cycle. Fertilizer
manufacture itself amounts to 46%in the establishment of the
crop and 32% in the rst cycle [48] for an LCA conducted of
a Populus spp. crop.
Relative to environmental impacts, the use of microalgae
production has been promoted in part as a means to reduce
CO
2
emissions and improve sustainability [49,50]. Certain
previously reported LCA studies have also conducted envi-
ronmental analyses [39,41]. The environmental results of our
study demonstrated that main environmental impacts are
due to electricity consumption and for the global warming
category (GWP) the emission of 0.16 kg CO
2
eq. per MJ were
found. Lower results of 0.07 kg and 0.06 kg per MJ were re-
ported by other studies [39,41]. However, results from the
sensitivity analysis demonstrate that positive balances could
be achieved by reducing the GWP to 0.06 kg MJ
1
.
Finally, there is a need to standardize data quality for the
inventory used, especially for the purpose of comparing
studies. Our study used experimental data, whereas in most
cases, the data were obtained from a bibliographic inventory
or were extrapolated from industrial processes used for other
modes of generic biofuel production. In this sense, the energy
balances obtained may not be consistent.
5. Conclusions
In Mediterranean outdoor conditions, marine microalgae
productionfor biodiesel is a good optionand a feasible route to
obtain bioenergy. We recommend that production and
research under indoor conditions be rejected based on the
energy results obtained. However, for outdoor systems, efforts
should be made to decrease energy consumption. As revealed
herein, the highest energy consumption occurs during the
growing stage due to the mechanical requirements of the
pumps and the need for air injection. Thus, for industrial scale
improvements, more efcient equipment is needed. In the
same manner, more energy-conserving bcPBR material or its
ecodesign could signicantly reduce energy consumption.
Any of the three microalgae analyzed can be cultivated and
exploited on a large scale as there were no substantial
differences in biomass production between them. In addition,
Table 6 e Schemes of various LCA studies of bioenergy from microalgae.
Author Microalgae Reactor E. consumption (MJ kg
1
) Balance
Reactor Growing Dewatering
Razon et al. (2011) [37] Haematococcus pluvialis (freshwater) PBR raceway pond e 83.1 17 134
Nannochloropsis sp. (seawater) Raceway pond e 151 e 465
Nannochloropsis sp. (seawater) Raceway pond 4.5a 3.8b e 23.3(ab)/27.7b
Jorquera et al. (2010) [38] Nannochloropsis sp. (seawater) Flat-plate PBR 7.3a 7.0b e 17.3(ab)/24.6b
Nannochloropsis sp. (seawater) Tubular PBR e 159.0b e 127b
Sander et al. (2010) [39] e PBR and raceway pond e 0.1 53.9 49
Xu et al. (2011) [40] Chlorella vulgaris (freshwater) Open pond dry route 0.8 3.3 4.7 5.2
Open pond wet route 1.0 2.2 0.40 5.8
This work Alenxandrium minutum (seawater) bcPBR 36.5 89.17 7.53 130
a
Energy required for reactors production.
b
Only included the energy consumption required for air pumping.
b i o ma s s a nd b i o e ne r gy 3 9 ( 2 0 1 2 ) 3 2 4 e3 3 5 333
the use of any of these marine microalgae leaves freshwater
for other human uses and thus helps to overcome the critical
issue of freshwater consumption in the production of micro-
algae. This would improve the feasibility of bioenergy in terms
of its large scale production and the scarcity of freshwater in
the Mediterranean area.
Other experiments should be conducted to assess
productivities in Mediterranean climates for spring-summer
periods to evaluate whether higher productivities are ach-
ieved and less energy is needed. Besides biodiesel production,
additional research is needed to identify the coproducts for
bioenergy and other purposes.
Acknowledgments
The authors would like to thank to Comisio n Nacional de
Investigacio n Ciencia y Tecnologa (CONICYT) from Chile for
supporting the scholarship Beca de Gestio n Propia, which
nances the PhD studies of C. Fuentes-Gru newald; and to
Spanish Ministry of Science and Innovation for supporting the
work of E. Garce s and S. Rossi by the Ramon and Cajal award.
The authors would like also to thank S. Fraga for providing the
clonal culture AMP4, Laura del R o and Xavi Leal for their help
with the experiments, and the Zona Acuarios Experimentales
(ZAE) of the ICM-CSIC for the use of their facilities. The
authors would like also to thank to project Ecotech Sudoe
SOE2/P2/E377 for its nancial support.
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