Algal Research 15 (2016) 5964

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Algal Research
jou r nal hom epa ge: ww w. elsevi er. com / lo cat e / algal

Use of Spirulina biomass produced from treatment of

aquaculture wastewater as agricultural fertilizers
Shy Chyi Wuang , Mar Cho Khin, Pei Qiang Danny Chua, Yanpei Darren Luo
School of Applied Science, Temasek Polytechnic, 21 Tampines Ave 1, 529757, Singapore

a r t i c l ei n f o
Article history:
Received 4 August 2015
Received in revised form 27
October 2015 Accepted 8 February
2016
Available online xxxx
Keywords:
Water remediation
Spirulina platensis
Agricultural
fertilizers Fish
water

a b s t r a c
t
Microalgal research has been an area of great interest as microalgae have higher productivities
than land plants and can be used for the production of valuable commodities such as biofuel,
animal feeds and agricultural fertil- izers, among others. To enhance the economic feasibility of
algal-based commodities, the growth of microalgae can be coupled to wastewater remediation.
The technical feasibility of cultivating Spirulina platensis with sh water for production of algae
fertilizers was investigated. The remediation potential of S. platensis was found to be good for
ammonia and nitrate removal, but inadequate for nitrite removal. Its specic growth rate of
1
0.026 h and the nutrient reduction times compare well with various literature reports. This
work provides
insight into the potential of algal biomass as agricultural fertilizers, when coupled with
aquaculture wastewater remediation. The ability of Spirulina-based fertilizers to enhance plant
growth was demonstrated in leafy vegeta- bles such as Arugula (Eruca sativa), Bayam Red
(Ameranthus gangeticus) and Pak Choy (Brassica rapa ssp. chinensis). The germination of
Chinese Cabbage (B. rapa ssp. chinensis) and Kai Lan (Brassica oleracea alboglabra) also
improved signicantly in terms of seedlings' dry weight.
2016 Elsevier B.V. All rights reserved.

1. Introduction
The global seafood consumption is increasing and the
new World Bank Report 83177-GLB [1] has projected that
aquaculture will produce half of the world's supply of sh in
2030. Intensive aquaculture utilizes large quantities of clean
freshwater, and generates nutrient-rich waste- water streams
which can cause eutrophication of coastal waters and
negatively impact downstream biological communities.
Sustainable water treatment technologies are needed to
reduce nutrient and chem- ical discharge into receiving
waters. Currently, chemical fertilizers are extensively used
for agricultural crops as they are inexpensive, have low
labor application, and provide immediate availability of the
nutri- ents. However, the excessive use of chemical
fertilizers contribute to greenhouse gas emission and can
lead to on-site soil degradation [2], nutrient pollution and
eutrophication [3]. Organic fertilizers, such as farmyard
manure and crop residues are possible alternatives to reduce
dependency on synthetic chemical fertilizers [4], as well as to
counter- act soil degradation [5]. However, these are
inadequate in meeting the nutrient demands for high
yielding crops as essential macro- and micronutrients may
not be present in desired amounts.
Microalgae research has gained interest in recent decades
and is cen- tered mainly in the exploitation of microalgae for
extraction of products of value, and the utilization of
microalgae in wastewater treatment sys- tems. A key area of
interest in microalgae biotechnology is the economic
* Corresponding author.
E-mail address: scwuang@tp.edu.sg (S.C. Wuang).

benets associated with the mass-culture exploitation of

microalgae. The use of microalgae to remove certain
excess nutrients from aquacul- ture efuents algal
bioremediation has been widely discussed [69]. A
signicant reduction of nutrient loads in aquaculture
efuents
through algae bioremediation requires the
selection of appropriate spe- cies. This selection will also
impact the applicability of algal biomass for various uses.
Microalgal biomass contains high concentrations of
protein, lipids, and vitamins. Therefore, besides being a
valuable alternative to the conventional purication
treatments, the use of microalgae offers several advantages
[10]. Firstly, the algal biomass can be recycled or used as fertilizers
and
livestock
feeds.
Furthermore,
under
photosynthetic condi- tions, algal growth allows oxygen to
be released, thus enhancing the auto-depuration potential of
water. In agricultural countries, algal bio- mass could be
directly employed in soil inoculation to increase crop productivities. They offer an alternative to the vicious cycle of
chemical fertilizers, soil exhaustion and dependency on
imports. Algae fertilizers can also boost the protability of
aquaculture by generating additional income from fertilizers'
sale. The use of algal fertilizers is not well- established,
though there are numerous reports documenting its feasibility in rice plants [1113]. In India, Tripathi et al. [12] found
signicant enhancement of growth in rice plants growing on
amended soils enhanced
with
blue-green
algae
biofertilizer. Similarly, Watanabe et al. [14] used blue-green
algae as a fertilizer in approximately 40 rice varieties and
noted remarkable increases in rice yields in Japan. In Iran,
Saadatnia et al. [13] have found that the germination of
rice seeds treated with cyanobacteria was faster than that of
control. These

http://dx.doi.org/10.1016/j.algal.2016.02.009
2211-9264/ 2016 Elsevier B.V. All rights reserved.

S.C. Wuang et al. / Algal Research 15

(2016) 5964

reports evidenced the potential use of algae fertilizers when

applied to other agricultural crops.
Spirulina platensis, a lamentous cyanobacterium, has
been widely used as a food supplement due to its high protein
content and nutrition- al value. Spirulina thrives in alkaline
environments and this preference
prevents external
contamination [15], suggesting its suitability for environmental applications. Spirulina has also been suggested
as a good alternative to chemical fertilizers as well as a good
protein supplement in livestock feeds [16]. The objectives of
this work are to assess the suit- ability of S. platensis as a
bioremediation agent for aquaculture waste- water and to
examine the applicability of its biomass as agricultural
fertilizers to leafy vegetables. The water quality of the
treated water
was studied and the excess algae were harvested and
studied for their
usefulness as crop fertilizers with different types of leafy
vegetables such as Arugula, Bayam Red, Pak Choy, Chinese
Cabbage, Kai Lan and White Crown. The nutrient composition
of the algae fertilizer was also
compared to a commercial chemical
fertilizer.
2. Material and methods
2.1.

Materials

2.3. Harvesting and analytical methods

Stipulated amounts of algae suspension (50 mL) were
withdrawn from the cultivation asks and subjected to
centrifugation at 2650 g for 10 min. The supernatants were
withdrawn and analyzed for ammo- nia, nitrite and nitrate.
Ammonia, nitrite and nitrate concentrations
were
determined using standard methods [17]. Absorbance
readings were obtained using the HACH DR2800
spectrophotometer. The per- centage reduction in
ammonia, nitrite and nitrate was calculated by the
following formula:

% reduction

concentration at t 0concentration at time t

100:
concentration at t
2
0

The analysis of the constituents of S. platensis was

performed by
Pacic Lab Services, Singapore. The N, P, K percentages in
the chemical fertilizer (Triple Pro 15-15-15) are calculated
based on its inorganic constituents, and metal contents
were determined via inductively
coupled plasma
spectrometry.

S. platensis (LB 1928) was obtained from the University

of Texas (UTEX) collection and routinely cultured in the
Guillard F/2 commercial media, Micro Algae Grow, according to
2.4.
Potted
plants
the manufacturer's recommend- ed dilution (1:1000) in tap
experiment
water with 30 g/L marine salt. The Nessler reagent set,
nitrite and nitrate testing reagents (Nitriver 3, Nitraver 5),
Potted plants experiments were conducted with Arugula
and chemical oxygen demand (COD) vials were purchased
(Eruca sativa), Bayam Red (Ameranthus gangeticus), and
from Fluke South East Asia Pte Ltd. Microscopy images of the
Pak Choy (Brassica rapa ssp. chinensis) plants. For each
algae cells were ac- quired using an Olympus IX51
plant, four trials were conducted Control, Spirulina (5
microscopy (low magnication) and a Nikon-E upright
g/pot), chemical fertilizer (Triple Pro 15-15-15,
microscope (high magnication).
1.3 g/pot/week), and Spirulina (5 g/pot) plus chemical
2.2.

Cultivation of S. platensis

Fish water was obtained during water changes for catsh

(Pangasius
hypophthalmus)
reared
in
Temasek
Aquaculture Facility, Temasek Polytechnic (TP), Singapore.
The rearing of catsh was approved by TP Institutional
Animal Care and Use Committee (IACUC control number:
2012-50A). 30 g/L NaCl was subsequently added to the
sh water. Batch cultivation (2 L) of the microalgae was
performed using the ob- tained sh water in glass bottles
and air-aerated with a ow rate of 1015 mL/min. An
4
average of 2 10 algae cells per milliliter of sh water
was used as the starting concentration. The growth and
nutrient proles of the water were monitored for 3 days.
Illumination was pro- vided by uorescent lamps with a
luminance of 5001000 lx, typical of indoor lighting and
similar to the natural light intensity on an over- cast day.
Temperature has a diurnal range of 2830 C. The asks
were placed on a tray with the uorescent light placed 8
cm away from the side of the asks. The various trays
were covered with opaque boxes, with articial light as the
only light source. pH was not controlled but closely
monitored during the experiments. Throughout the study,
the only form of agitation was provided by air aeration via
single tubing in each cultivation bottle.
For algal density quantication, 3 mL of algae suspension
was col- lected and the cell optical density was measured at
660 nm using a spec- trophotometer (HACH DR2800). The
relative cell density was calculated by the following formula:
absorbanceAt at time t

fertilizer (0.3 g/pot/week). Each pot consists of 500 g of

soil. The duration of ex- periments ranged between 21 and
40 days. All trials were performed in triplicates. Weekly
measurement of plant growth was recorded in terms of
leaf number per plant and plant height. At the end of the
trials, the plants were harvested and their leaf number,
plant height, chloro- phyll content, root length, fresh
weight and dry weight were deter- mined. Chlorophyll
content was measured using the Hansatech meter (Model
CL-01) which has a range of 02000 CCI (Chlorophyll
Content Index) units.
2.5.
Seed
germination study
Seed germination studies were performed for three
types of vegetable Chinese Cabbage (B. rapa ssp.
chinensis), Kai Lan (Brassica oleracea alboglabra) and
White Crown (B. rapa ssp. chinensis, F1 hybrid). For each
type of vegetables, Spirulina inoculation on seed
germination trials was conducted using 100 seeds per
treatment for 6 treatments; T1 to T5, containing various
concentrations of Spirulina (2 g/L, 4 g/L, 6 g/L, 8 g/L and
10 g/L respectively) in tap water and control T0 (tap water
only). The seeds were soaked in the respective solution
overnight before germination on tissue towel. Triplicate runs
of 100 seeds per run were performed for each treatment
group and the average results were described. Germination
rate, shoot length, root length, vigor index and dry weight
of 100 seedlings were measured. Germination rate is the
av- erage number of seeds that germinate over a 7-day
period. Seed vigor index is obtained by multiplying the
germination rate with the shoot length [18].

S.C. Wuang et al. / Algal Research 15

(2016) 5964
2.6.
Statistical

Relative cell
density

A0 at t 0

:
1

Specic growth rate (, in h ) was calculated based on

the equation
l

Ay

A
x
ty t
x

3. Results
discussion

analysis
All experiments were performed in triplicate sets and the
data pre- sented are means standard deviation. The
differences in the results
obtained with two treatment groups were analyzed
statistically using two-sample t-test. The differences
observed between the treatment groups were considered
signicant for P b 0.05.

and

3.1.
Cultivation of S.
platensis
S. platensis is microscopic blue-green algae and appears as
spiral l- aments under a Nikon Ni-E upright microscope (see
Fig. 1). It is charac- terized by cylindrical, multicellular
trichomes in an open helix. As the lament length varies
between different cells, counting the laments is not an
accurate way to assess algal growth. The green pigment within
the Spirulina cells absorbs strongly at 660 nm, and this
absorbance is used as a gauge of cell growth.
Fig. 2 shows the relative algal density and pH proles of
the sh water used for Spirulina cultivation. The alga grows
1
well in sh water with a specic growth rate of 0.026 h
1
(0.623 day ) and a doubling time of 28 h. These growth
parameters compare favorably with those
reported
elsewhere [1920], indicating the suitability of cultivating
S. platensis in sh water. Throughout the experiments, the pH
remained approximately consistent between 8.238.46.
As cells assimilate nutrients during growth, the
phytoremediation potential of algae is closely linked to its
growth. Greater algal growth will promote better nutrient
removal from the cultivation media. In sh water, the main
nutrients to be removed to maintain good sh health are
ammonia and nitrite. The acceptable level for total ammonia
is less than 4 mg/L while that for free ammonia should be
less than
1.4 mg/L [21]. The desired level of nitrite is 01 mg/L and the
acceptable range is less than 4 mg/L. The acceptable concentration
of nitrates is much higher at 50 mg/L in freshwater and 100
mg/L in seawater [22]. The starting sh water concentrations
of ammonia, nitrite and ni- trate average about 2.56 mg/L,
0.245 mg/L and 15.3 mg/L respectively. As there were
signicant variations between different batches of sh
water, the percentage reductions of the nutrient (rather than
the abso- lute values) were used for comparison. Fig. 3 shows
the relative ammo- nia, nitrite and nitrate proles of sh
water cultivated with S. platensis. This alga assimilates
ammonia and nitrate quickly, with these concen- trations
rapidly decreasing with cultivation time. This is in line with a
previous report [23]. However, nitrite was not assimilated and
its level increased with growth, possibly due to its byproduction during nitrate metabolism. This implies the
necessity of a further nitrite reduction step to achieve the
recycling of sh water, when S. platensis is used as the
phytoremediation agent.
In the conventional biological treatment of aquaculture
wastewater, ammonia is rst converted to nitrite and then to
nitrate by different ni- trifying bacteria which typically has
doubling times of 24 h [24]. Nitrate is a stable end product
which does not harm the sh in the concentra- tions typically
present. In comparison, with the algal-based treatment in
this work, ammonia was also efciently reduced to non-toxic
levels. However, the alga was unable to remove nitrite. It
should be noted that Spirulina cultivated in marine sh
wastewater exhibit similar per- formance (Wuang et al.,
unpublished work). A different algae species

Fig. 2. Relative algal density (line graph) and pH (point graph) proles of sh
water used in cultivation of S. platensis.

or a combination of various species will be required to achieve

adequate ammonia and nitrite reduction simultaneously.
3.2.

Efcacy of S. platensis as fertilizers

Table 1 illustrates the efcacies of S. platensis in

enhancing the growth of Bayam Red, Arugula and Pak Choy.
For Bayam Red, the plants cultivated in soil enriched with
Spirulina showed increased plant height by 58.3% as well as
greater fresh and dry weights, by 110.1% and 155.8%
respectively, when compared with the control group. There
were no sig- nicant differences in chlorophyll content, root
length and leaf number. These results are in agreement to
previous reports [12, 13] which main- ly compared the
efcacy of blue-green algae to controls with no supple- ments
added. In this study, the efcacy of S. platensis was also
compared to that of a commercial fertilizer, Triple Pro 15-1515. Bayam Red culti- vated with S. platensis showed no
signicant differences in terms of leaf number, plant height,
root length and chlorophyll content, when com- pared to
those cultivated using Triple Pro 15-15-15. However, the
fresh and dry weights were signicantly smaller than those
achieved with Triple Pro 15-15-15. The combination of
Spirulina and Triple Pro 15-15-15 resulted in greater fresh
and dry weights when compared to the Spirulina group.
When compared to the control, Arugula plants cultivated
in soil enriched with Spirulina showed increased plant
height (by 55.3%), in- creased chlorophyll content (by
30.2%), greater fresh and dry weights (by 18.7% and
21.1% respectively). When compared to plants cultivated
using Triple Pro 15-15-15, the plants grown in Spirulinaenriched soil showed a 71.8% increase in plant height, and
comparable performance
in all the other growth
parameters. The addition of S. platensis to Triple

S.C. Wuang et al. / Algal Research 15

(2016) 5964

Fig. 1. Microscopy image of S. platensis at (a) 10 magnication and (b) 100 magnication.

Table 2
Constituents of S. platensis and chemical fertilizer.
Constituents

S. platensis

Carbohydrate (%)
Protein (%)
Fat (%)
Nitrogen (%)
Phosphorus (%)
Potassium (%)
Calcium (%)
Iron (ppm)
Manganese (ppm)
Zinc (ppm)
Selenium (ppb)

Fig. 3. Relative concentrations of ammonia, nitrite and nitrate in sh

water used in cultivation of S. platensis.

Pro 15-15-15 showed no enhancement in most growth

parameters, when compared to the trials with either
Spirulina or Triple Pro alone. The signicant enhancements
were in root length and fresh weight. For Pak Choy, there
were no signicant differences in between plants cultivated
in Spirulina-enriched soil and control for all the growth parameters except root length which is higher for the plants
cultivated in Spirulina-enriched soil. The plants cultivated in
soil enriched with Tri- ple Pro 1515-15 performed better in
terms of greater leaf number and 29.1% increase in dry
weight, when compared to those in Spirulina- enriched
soil. The addition of S. platensis to Triple Pro 1515-15
showed no enhancement in growth parameters. Prior work
in the comparison study of algae-based and chemical-based
fertilizers is very limited. The results of our work suggest
that the efcacy of S. platensis in enhancing plant growth is
species-dependent and in some cases (e.g. for Arugula) it
can be a better growth supplement when compared to
chemical fertil- izers. Its efcacy can be explained by the
presence of micro- and macro- nutrients in the microalgae.

1
6
4
84
.7
.
0.8
1
.0
.
1057
4
1
155.4
1
7

Chemical
fertilizer (Triple
Pro 1515-15)
12.4
6.6
12.5
0.1
455
26.1
-

Table 2 shows the constituents of the microalgae and

chemical fertil- izers. Comparatively, the algal biomass
contain less nitrogen, phospho- rus and potassium (NPK)
which are the primary nutrients found in fertilizers. Primary
nutrients are usually applied at higher rates as they are
utilized in the largest amounts by crops, followed by secondary
nu- trients and micronutrients. Other than NPK, all the
analyzed nutrients (calcium, iron, manganese, zinc and
selenium) were found in greater quantities in the algal
biomass. Calcium is a constituent of cell walls and is
involved in production of new growing points and root tips. It
provides elasticity and expansion of cell walls, which keeps
growing points from becoming rigid and brittle. The algal
biomass can supply a 3-fold higher calcium concentration and
a 2-fold higher Iron concentra- tion to the crops, as compared
to the chemical fertilizer. Iron is a struc- tural component of
several essential enzymes and its shortage is a common
problem in many plant species. Manganese content of the
algal biomass is also signicantly higher than the chemical
fertilizer. This element acts as an enzyme activator for
nitrogen assimilation and is essential for the synthesis of
chlorophyll. Zinc, which is not detected in the chemical
fertilizer, is involved in the synthesis of growth substances
and enzyme systems. It is necessary for the production of
chlorophyll and carbohydrates and essential for promoting
certain metabolic reactions. Selenium, another element not
detected in the chemical fertilizer, was found in trace
amount (17.8 ppb) in the algae. Selenium is important, not
to the crops, but more to alleviate selenium deciency in the
animals and humans further up the food chain [25].

Table 1
Efcacy of Spirulina in enhancing Bayam Red, Arugula and Pak Choy growth.
Plant parameter

Control

Spirulina

Chemical fertilizer

Bayam Red (21 days at

harvest)
Number of leaf
Plant height (cm)
Root length (cm)

Spirulina + chemical
fertilizer

9.33 2.08
16.03 3.05
22.57 0.50

11.00 1.00
25.37 5.90**
26.00 5.57

12.33 1.53
29.50 4.71**
22.87 3.67

13.33 0.58
30.90 4.11**
19.77 4.25

Chlorophyll
content Fresh
weight (g)
Dry weight
Arugula (30 days at
harvest)
Number of
leaf Plant
height (cm)
Root length
Chlorophyll
content Fresh
weight
(g) (g)
Dry weight
Pak Choy (40 days at
harvest)
Number
of leaf
Plant height
(cm)

5.72 2.48
12.52 0.82
4.28 0.40

S.C. Wuang et al. / Algal Research 15

(2016)
5964
7.65
0.85
8.98 2.22
, ,#
,
26.31 4.65** *
38.11 5.16** *
, ,#
,
10.95 1.07** *
21.16 3.11** *

7.67 0.67
31.33 5.93
6.00 1.73
13.67 0.28
10.67 0.67
7.48 0.38

9.00 0.58
, ,#
48.67 3.67** *
6.17 0.60*
17.80 1.87
,
12.67 0.33** *
9.06 0.36**

9.67 0.67
28.33 3.33*
#
7.33 0.33
18.60 0.32
13.67 1.45
9.52 1.05

10.33 0.88
15.83 1.59
2.33 0.33
13.17 1.37
12.67 1.20
3.71 0.60

13.00 0.00*
17.5 2.50
7.00 1.00**
14.25 2.85
14.00 3.00
4.79 0.26*

14.00 0.00** *
19.17 0.73
6.17 1.59**
12.77 0.79
17.33 1.76
,
5.91 0.22** *

Root length (cm)

Chlorophyll content
Fresh
weight (g)
Dry weight
#
* and denote statistical signicance between test groups.
** denotes statistical signicance with control group.

8.80 2.16
,#
36.33 1.74**
,#
22.74 1.05**

9.67 0.33
#
32.00 8.00
,#
9.67 0.33*
18.70 0.87
,
14.67 0.33** *
10.00 0.05**
,

13.50 0.50
21.75 0.75
7.00 1.00**
11.10 0.80
17.50 0.50
5.30 0.23**

Despite a lower NPK content, the algae fertilizers

enhanced plant growth to similar extent as the chemical
fertilizers. This is likely due to the higher amounts of other
secondary and micro-nutrients which help to moderate the
amounts of primary nutrients required.
3.3.
Efcacy of S. platensis inoculation on
seed germination
Many studies [14, 2627] have reported the use of dried
cyanobacteria to inoculate soils as a means of aiding
fertility in rice plants. On average, these results have
shown increases in rice grain yield of 1520% in eld
experiments. While these prior studies have concentrated on
rice plants, the effects of Spirulina inoculation on seed
germination of leafy vegetables were studied in this work. A
germina- tion test determines the maximum seed viability.
The germination rate of a seed lot is a key indicator to its
performance in the eld. The pre-soaking of rice in bluegreen algae culture has decreased losses from sulfatereducing processes and this has been attributed to the enhancement of germination and faster seedling growth [2829].
Further- more, vigor tests are used to understand the ability
of seeds to emerge from soils under stress conditions or to
maintain viability during
storage. Table 3 lists the
performance
of
S.
platensis
inoculation
on
seed
germination of Chinese Cabbage, Kai Lan and White Crown.
For Chinese Cabbage, no improvement to germination
rate was ob- served at all the inoculated Spirulina
concentrations. The only signicant enhancement was
observed with an inoculation of 8 g/L Spirulina, where the
dry weight of the germinated seedlings increased by more
than 3-fold that of the control. In the case of Kai Lan,
signicant in- creases in dry seedling weight were
observed with 8 g/L and 10 g/L Spirulina inoculations.
Similarly, there were no signicant improve- ments for the
other plant growth parameters. Spirulina inoculations at all
concentrations did not show signicant effects on the
germination of White Crown. These suggest that Spirulina
may not be a universal plant growth and germination
enhancer, as its effects varies consider- ably between
different plant species.
Previous studies on bio-fertilizers have reported
enhanced germina- tion rates for sunower (Helianthus
annuus L.) [30], Dill (Anethum graveolens) [31], Cumin
(Cuminum cyminum), Marigold (Calendula ofcinalis) and
Roma tomato (Solanum lycopersicum var. Roma) [32]. In
the application of algae fertilizers, Sanaz and Ragaa [33]
have report- ed the enhancement of growth to varying
extents of maize (Zea mays L.) plants with three different
types of red marine algae. The application of

Chlorella vulgaris was also found to be benecial to the growth

of lettuce (Lactuca sativa) [34]. Also, it has been shown that
Spirulina, when used as a biofertiliser, increases the growth
and yield of green gram [35]. From these studies, there
seem to be a general consensus on the useful- ness of algae
fertilizers, though specic benets would require comprehensive investigation. Our study has demonstrated that the
application of S. platensis can enhance the growth of Bayam
Red, Arugula and Pak Choy to similar extents when compared
to chemical fertilizers. In addi- tion, the germination of
Chinese Cabbage and Kai Lan improved signi- cantly in
terms of seedlings' dry weight.

4. Conclusi
on
The usefulness of S. platensis in aquaculture wastewater
treatment was studied and the subsequent application of algal
biomass in fertilizer studies was demonstrated. The
cultivation of S. platensis was done indoors, under an
illuminance of not more than 1000 lx. At these condi- tions,
the algae were able to remove the ammonia and nitrate
concen- trations in sh water, indicating its ability to treat
the water despite its inadequacy in removing nitrite.
Potentially, the efcacy of water treatment can be much
higher under sunlight where illuminance is typically about
100,000 lx. The supplementation of S. platensis for leafy
vegetables found enhanced plant growth in all tested
vegetables,
when compared to the controls. When
compared to the performance of chemical fertilizer, the
Spirulina-based fertilizer performed compara- bly in most
plant growth parameters, and favorably for one tested
species Arugula. Seed germination (when measured by
seedling's
dry weight) also improved for all tested
vegetables except White Crown. This work has evidenced
the usefulness of S. platensis in sh water treatment and its
applicability as agricultural fertilizers.

Authors' contributions and competing

interests
DPQ Chua, DYP Luo and MC Khin conducted the
experiments and participated in the collection and assembly
of data. SC Wuang and MC Khin were involved in the study
design and the analysis and interpreta- tion of data. SC
Wuang also conceived the study, wrote and revised the
manuscript. All authors read and approved the nal
manuscript. The authors declare that they have no
competing interests.

Table 3
Efcacy of Spirulina inoculation on seed germination of Chinese Cabbage, Kai Lan and White Crown.
Treatment
group
Chinese
Cabbage
T0 (control)
T1 (2 g/L)
T2 (4 g/L)
T3 (6 g/L)
T4 (8 g/L)
T5 (10 g/L)

Germination rate
(%)

Shoot Length
(cm)

Root Length
(cm)

Dry weight of 100 seedlings

(g)

Vigor
Index

93.3 5.8
88.3 12.6
98.3 2.9
100.0 0.0
100.0 0.0
100.0 0.0

1.07
1.02
1.22
0.92
1.08
0.95

0.19
0.08
0.05
0.14
0.19
0.13

1.82
2.01
2.28
2.71
2.05
2.31

0.24
0.15
0.31
0.35
0.40
0.17

3.45
3.30
3.85
2.75
11.15
3.09

99.8
12.5
90.1
6.4
119.9
8.0
92.0
13.9
108.0
18.6
95.0
12.5

1.88
2.21
2.02
2.02
1.83
1.96

0.15
0.17
0.17
0.23
0.12
0.14

1.51
1.56
2.08
1.65
1.70
1.83

0.15
0.11
0.40
0.06
0.12
0.13

2.60 0.13
3.93 1.48
5.00 1.20
6.53 2.68
10.22 2.78**
7.15 2.41**

Kai Lan
T0
T1
T2
T3
T4
T5

70.0
56.7
63.3
75.0
66.7
81.7

White Crown
T0

98.3 2.9

8.7
5.8
10.4
5.0
16.1
17.6

2.18 0.05

1.71 0.21

0.59
0.3
0.44
0.26
1.38**
0.59

4.80 0.06

131.6
25.6
125.3
17.3
127.9
22.4
151.5
6.9
122.1
20.8
160.1
26.0

214.3
7.6

T1
T2
T3
T4
T5

100.0 0.0
100.0 0.0
98.3 2.9
98.3 2.9
96.7 2.9

** denotes statistical signicance with control group.

1.98
2.07
2.05
1.97
1.91

0.16
0.11
0.11
0.17
0.22

1.98
2.07
2.05
1.97
1.91

0.11
0.31
0.03
0.16
0.02

4.67
4.32
4.64
4.90
4.22

0.93
0.35
0.40
0.33
0.43

198.0
15.3
207.0
10.9
201.5
15.3
193.7
11.3
184.7
20.4

Acknowledgments
The work was supported nancially by the Social and
Innovation Research Fund (M54), awarded by the TOTE
Board, Singapore (Project
M54). High magnication
microscopy images for the study were ac- quired in the
Singapore Bioimaging Consortium (SBIC)-Nikon Imaging
Centre at Biopolis (Singapore) using a Nikon Ni-E upright
microscope.
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