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Algal Research
a r t i c l e i n f o a b s t r a c t
Article history: Microalgal research has been an area of great interest as microalgae have higher productivities than land plants
Received 4 August 2015 and can be used for the production of valuable commodities such as biofuel, animal feeds and agricultural fertil-
Received in revised form 27 October 2015 izers, among others. To enhance the economic feasibility of algal-based commodities, the growth of microalgae
Accepted 8 February 2016
can be coupled to wastewater remediation. The technical feasibility of cultivating Spirulina platensis with fish
Available online xxxx
water for production of algae fertilizers was investigated. The remediation potential of S. platensis was found to
Keywords:
be good for ammonia and nitrate removal, but inadequate for nitrite removal. Its specific growth rate of
Water remediation 0.026 h−1 and the nutrient reduction times compare well with various literature reports. This work provides
Spirulina platensis insight into the potential of algal biomass as agricultural fertilizers, when coupled with aquaculture wastewater
Agricultural fertilizers remediation. The ability of Spirulina-based fertilizers to enhance plant growth was demonstrated in leafy vegeta-
Fish water 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 significantly in terms of seedlings' dry weight.
© 2016 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.algal.2016.02.009
2211-9264/© 2016 Elsevier B.V. All rights reserved.
60 S.C. Wuang et al. / Algal Research 15 (2016) 59–64
reports evidenced the potential use of algae fertilizers when applied to 2.3. Harvesting and analytical methods
other agricultural crops.
Spirulina platensis, a filamentous cyanobacterium, has been widely Stipulated amounts of algae suspension (50 mL) were withdrawn
used as a food supplement due to its high protein content and nutrition- from the cultivation flasks and subjected to centrifugation at 2650 g
al value. Spirulina thrives in alkaline environments and this preference for 10 min. The supernatants were withdrawn and analyzed for ammo-
prevents external contamination [15], suggesting its suitability for envi- nia, nitrite and nitrate. Ammonia, nitrite and nitrate concentrations
ronmental applications. Spirulina has also been suggested as a good were determined using standard methods [17]. Absorbance readings
alternative to chemical fertilizers as well as a good protein supplement were obtained using the HACH DR2800 spectrophotometer. The per-
in livestock feeds [16]. The objectives of this work are to assess the suit- centage reduction in ammonia, nitrite and nitrate was calculated by
ability of S. platensis as a bioremediation agent for aquaculture waste- the following formula:
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 concentration at t ¼ 0−concentration at time t
% reduction ¼ 100:
usefulness as crop fertilizers with different types of leafy vegetables concentration at t ¼ 0
such as Arugula, Bayam Red, Pak Choy, Chinese Cabbage, Kai Lan and ð2Þ
White Crown. The nutrient composition of the algae fertilizer was also
compared to a commercial chemical fertilizer.
The analysis of the constituents of S. platensis was performed by
Pacific Lab Services, Singapore. The N, P, K percentages in the chemical
2. Material and methods fertilizer (Triple Pro 15-15-15) are calculated based on its inorganic
constituents, and metal contents were determined via inductively
2.1. Materials coupled plasma spectrometry.
Fig. 1. Microscopy image of S. platensis at (a) 10× magnification and (b) 100× magnification.
62 S.C. Wuang et al. / Algal Research 15 (2016) 59–64
Table 2
Constituents of S. platensis and chemical fertilizer.
Table 1
Efficacy of Spirulina in enhancing Bayam Red, Arugula and Pak Choy growth.
Despite a lower N–P–K content, the algae fertilizers enhanced plant Chlorella vulgaris was also found to be beneficial to the growth of lettuce
growth to similar extent as the chemical fertilizers. This is likely due (Lactuca sativa) [34]. Also, it has been shown that Spirulina, when used
to the higher amounts of other secondary and micro-nutrients which as a biofertiliser, increases the growth and yield of green gram [35].
help to moderate the amounts of primary nutrients required. From these studies, there seem to be a general consensus on the useful-
ness of algae fertilizers, though specific benefits would require compre-
3.3. Efficacy of S. platensis inoculation on seed germination hensive investigation. Our study has demonstrated that the application
of S. platensis can enhance the growth of Bayam Red, Arugula and Pak
Many studies [14, 26–27] have reported the use of dried Choy to similar extents when compared to chemical fertilizers. In addi-
cyanobacteria to inoculate soils as a means of aiding fertility in rice tion, the germination of Chinese Cabbage and Kai Lan improved signifi-
plants. On average, these results have shown increases in rice grain cantly in terms of seedlings' dry weight.
yield of 15–20% in field 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- 4. Conclusion
tion test determines the maximum seed viability. The germination
rate of a seed lot is a key indicator to its performance in the field. The The usefulness of S. platensis in aquaculture wastewater treatment
pre-soaking of rice in blue-green algae culture has decreased losses was studied and the subsequent application of algal biomass in fertilizer
from sulfate-reducing processes and this has been attributed to the en- studies was demonstrated. The cultivation of S. platensis was done
hancement of germination and faster seedling growth [28–29]. Further- indoors, under an illuminance of not more than 1000 lx. At these condi-
more, vigor tests are used to understand the ability of seeds to emerge tions, the algae were able to remove the ammonia and nitrate concen-
from soils under stress conditions or to maintain viability during trations in fish water, indicating its ability to treat the water despite
storage. Table 3 lists the performance of S. platensis inoculation on its inadequacy in removing nitrite. Potentially, the efficacy of water
seed germination of Chinese Cabbage, Kai Lan and White Crown. treatment can be much higher under sunlight where illuminance is
For Chinese Cabbage, no improvement to germination rate was ob- typically about 100,000 lx. The supplementation of S. platensis for leafy
served at all the inoculated Spirulina concentrations. The only significant vegetables found enhanced plant growth in all tested vegetables,
enhancement was observed with an inoculation of 8 g/L Spirulina, when compared to the controls. When compared to the performance
where the dry weight of the germinated seedlings increased by more of chemical fertilizer, the Spirulina-based fertilizer performed compara-
than 3-fold that of the control. In the case of Kai Lan, significant in- bly in most plant growth parameters, and favorably for one tested
creases in dry seedling weight were observed with 8 g/L and 10 g/L species — Arugula. Seed germination (when measured by seedling's
Spirulina inoculations. Similarly, there were no significant improve- dry weight) also improved for all tested vegetables except White
ments for the other plant growth parameters. Spirulina inoculations at Crown. This work has evidenced the usefulness of S. platensis in fish
all concentrations did not show significant effects on the germination water treatment and its applicability as agricultural fertilizers.
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. Authors' contributions and competing interests
Previous studies on bio-fertilizers have reported enhanced germina-
tion rates for sunflower (Helianthus annuus L.) [30], Dill (Anethum DPQ Chua, DYP Luo and MC Khin conducted the experiments and
graveolens) [31], Cumin (Cuminum cyminum), Marigold (Calendula participated in the collection and assembly of data. SC Wuang and MC
officinalis) and Roma tomato (Solanum lycopersicum var. Roma) [32]. Khin were involved in the study design and the analysis and interpreta-
In the application of algae fertilizers, Safinaz and Ragaa [33] have report- tion of data. SC Wuang also conceived the study, wrote and revised the
ed the enhancement of growth to varying extents of maize (Zea mays L.) manuscript. All authors read and approved the final manuscript. The
plants with three different types of red marine algae. The application of authors declare that they have no competing interests.
Table 3
Efficacy of Spirulina inoculation on seed germination of Chinese Cabbage, Kai Lan and White Crown.
Treatment group Germination rate (%) Shoot Length (cm) Root Length (cm) Dry weight of 100 seedlings (g) Vigor Index
Chinese Cabbage
T0 (control) 93.3 ± 5.8 1.07 ± 0.19 1.82 ± 0.24 3.45 ± 0.59 99.8 ± 12.5
T1 (2 g/L) 88.3 ± 12.6 1.02 ± 0.08 2.01 ± 0.15 3.30 ± 0.3 90.1 ± 6.4
T2 (4 g/L) 98.3 ± 2.9 1.22 ± 0.05 2.28 ± 0.31 3.85 ± 0.44 119.9 ± 8.0
T3 (6 g/L) 100.0 ± 0.0 0.92 ± 0.14 2.71 ± 0.35 2.75 ± 0.26 92.0 ± 13.9
T4 (8 g/L) 100.0 ± 0.0 1.08 ± 0.19 2.05 ± 0.40 11.15 ± 1.38** 108.0 ± 18.6
T5 (10 g/L) 100.0 ± 0.0 0.95 ± 0.13 2.31 ± 0.17 3.09 ± 0.59 95.0 ± 12.5
Kai Lan
T0 70.0 ± 8.7 1.88 ± 0.15 1.51 ± 0.15 2.60 ± 0.13 131.6 ± 25.6
T1 56.7 ± 5.8 2.21 ± 0.17 1.56 ± 0.11 3.93 ± 1.48 125.3 ± 17.3
T2 63.3 ± 10.4 2.02 ± 0.17 2.08 ± 0.40 5.00 ± 1.20 127.9 ± 22.4
T3 75.0 ± 5.0 2.02 ± 0.23 1.65 ± 0.06 6.53 ± 2.68 151.5 ± 6.9
T4 66.7 ± 16.1 1.83 ± 0.12 1.70 ± 0.12 10.22 ± 2.78** 122.1 ± 20.8
T5 81.7 ± 17.6 1.96 ± 0.14 1.83 ± 0.13 7.15 ± 2.41** 160.1 ± 26.0
White Crown
T0 98.3 ± 2.9 2.18 ± 0.05 1.71 ± 0.21 4.80 ± 0.06 214.3 ± 7.6
T1 100.0 ± 0.0 1.98 ± 0.16 1.98 ± 0.11 4.67 ± 0.93 198.0 ± 15.3
T2 100.0 ± 0.0 2.07 ± 0.11 2.07 ± 0.31 4.32 ± 0.35 207.0 ± 10.9
T3 98.3 ± 2.9 2.05 ± 0.11 2.05 ± 0.03 4.64 ± 0.40 201.5 ± 15.3
T4 98.3 ± 2.9 1.97 ± 0.17 1.97 ± 0.16 4.90 ± 0.33 193.7 ± 11.3
T5 96.7 ± 2.9 1.91 ± 0.22 1.91 ± 0.02 4.22 ± 0.43 184.7 ± 20.4
Acknowledgments [16] M.A.B. Habib, M. Parvin, T.C. Huntington, M.R. Hasan, A review on culture, produc-
tion and use of Spirulina as food for humans and feeds for domestic animals and
fish, FAO Fish. Aquac. Circ. (2008) (No. 1034).
The work was supported financially by the Social and Innovation [17] Standard Methods for the Examination of Water and Wastewater 4500-NH3 B & C,
Research Fund (M54), awarded by the TOTE Board, Singapore (Project 20th ed. American Public Health Association (APHA), 2000.
[18] P.C. Gupta, Germination testing, in: P.K. Agrawal (Ed.), Handbook of Seed Testing,
M54). High magnification microscopy images for the study were ac- Seednet, India 2000, pp. 242–243.
quired in the Singapore Bioimaging Consortium (SBIC)-Nikon Imaging [19] T. Göksan, A. Zekeriyaoğlu, İ. Ak, The growth of Spirulina platensis in different culture
Centre at Biopolis (Singapore) using a Nikon Ni-E upright microscope. systems under greenhouse condition, Turk. J. Biol. 31 (2007) 47–52.
[20] F.F. Madkour, A.E.-W. Kamil, H.S. Nasr, Production and nutritive value of Spirulina
platensis in reduced cost media, Egypt. J. Aquat. Res. 38 (2012) 51–57.
References [21] N.M. Stone, H.K. Thomforde, Understanding Your Fish Pond Water Analysis Report,
Cooperative Extension Program, University of Arkansas at Pine Bluff Aquaculture/
[1] World Bank, Fish to 2030: prospects for fisheries and aquaculture, Agriculture and Fisheries, 2004.
Environmental Services Discussion Paper No. 3, World Bank Group, Washington [22] Ornamental Aquatic Trade Association (OATA), Water Quality Criteria—Ornamental
DC, 2013. Fish, 2008 (Wiltshire, BA13 3JN, UK).
[2] J.H. Guo, X.J. Liu, Y. Zhang, J.L. Shen, W.X. Han, W.F. Zhang, P. Christie, K.W.T. [23] G. Xu, X. Fan, A.J. Miller, Plant nitrogen assimilation and use efficiency, Annu. Rev.
Goulding, P.M. Vitousek, F.S. Zhang, Significant acidification in major Chinese crop- Plant Biol. 63 (2012) 153–182.
lands, Science 327 (2010) 1008–1010. [24] W.J. Golz, Biological treatment in recirculating aquaculture systems, Recirculating
[3] R. Howarth, F. Chan, D.J. Conley, J. Garnier, S.C. Doney, R. Marino, G. Billen, Coupled Aquaculture in the Classroom: a Training Workshop for Agricultural Science
biogeochemical cycles: eutrophication and hypoxia in temperate estuaries and Teachers, a Proceedings of a Workshop Sponsored by the Louisiana Sea Grant Col-
coastal marine ecosystems, Front. Ecol. Environ. 9 (2011) 18–26. lege Program, Louisiana State University, and the Louisiana Department of Educa-
[4] B. Yang, Z. Xiong, J. Wang, X. Xu, Q. Huang, Q. Shen, Mitigating net global warming tion, 6–7 December 1995, Louisiana State University, Baton Rouge, Louisiana, 1995.
potential and greenhouse gas intensities by substituting chemical nitrogen fertil- [25] R.V.S. Lavu, V. De Schepper, K. Steppe, P.N.V. Majeti, F. Tack, G.D. Laing, Use of sele-
izers with organic fertilization strategies in rice–wheat annual rotation systems in nium fertilizers for production of Se-enriched Kenaf (Hibiscus cannabinus): effect on
China: a 3-year field experiment, Ecol. Eng. 81 (2015) 289–297. Se concentration and plant productivity, J. Plant Nutr. Soil Sci. 176 (2013) 634–639.
[5] J.J.O. Odhiambo, V.N. Magandini, An assessment of the use of mineral and organic [26] G.S. Venkataraman, S. Neelakantan, Effect of the cellular constituents of the nitrogen
fertilizers by small holder farmers in Vhembe district, Limpopo province, South fixing bluegreen algae Cylindrospermum muscicola on the root growth of rice seed-
Africa, Afr. J. Agric. Res. 3 (2008) 357–362. lings, J. Gen. Appl. Microbiol. 13 (1967) 53–61.
[6] J.J. Bolton, D.V. Robertson-Andersson, D. Shuuluka, L. Kandjengo, Growing Ulva [27] U. Mishra, S. Pabbi, Cyanobacteria: a potential biofertilizer for rice, Resonance
(Chlorophyta) in integrated systems as a commercial crop for abalone feed in (2004) 6–10.
South Africa: a SWOT analysis, J. Appl. Phycol. 21 (2009) 575–583. [28] B. Nanda, S.K. Tripathy, S. Padhi, Effect of algalization on seed germination of vege-
[7] M.H. Abreu, R. Pereira, C. Yarish, A.H. Buschmann, I. Sousa-Pinto, IMTA with table crops, World J. Microbiol. Biotechnol. 7 (1991) 622–623.
Gracilaria vermiculophylla: productivity and nutrient removal performance of the [29] Z. Shariatmadari, H. Riahi, S. Shokravi, Study of soil blue-green algae and their effect
seaweed in a land-based pilot scale system, Aquaculture 312 (2011) 77–87. on seed germination and plant growth of vegetable crops, Bot. J. Iran 12 (2011)
[8] P.H. de Paula Silva, S. McBride, R. de Nys, N.A. Paul, Integrating filamentous ‘green 101–110.
tide’ algae into tropical pond-based aquaculture, Aquaculture 284 (2008) 74–80. [30] M. Jalaluddin, M. Hamid, Effect of adding inorganic, organic and microbial fertilizers
[9] R.J. Lawton, L. Mata, R. de Nys, N.A. Paul, Algal bioremediation of waste waters from on seed germination and seedling growth of sunflower, Pak. J. Bot. 43 (2011)
land-based aquaculture using ulva: selecting target species and strains, PLoS One 8 2807–2809.
(2013), e77344. [31] Y. Miri, S.B. Kochebagh, B. Mirshekari, Effect of inoculation with bio-fertilizers on
[10] G. Markou, D. Georgakakis, Cultivation of filamentous cyanobacteria (blue-green germination and early growth, Dill (Anethum graveolens), Fennel (Foeniculum
algae) in agro-industrial wastes and wastewaters: a review, Appl. Energy 88 vulgare), Cumin (Cuminum cyminum) and Marigold (Calendula officinalis), Int. J
(2011) 3389–3401. Agron. Plant Prod. 4 (2013) 104–108.
[11] R. Wang, B. Peng, K. Huang, The Research Progress of CO2 Sequestration by Algal [32] J. Garcia-Gonzalez, M. Sommerfeld, Biofertilizer and biostimulant properties of the
Bio-Fertilizer in China, J. CO2 Util, 2015, http://dx.doi.org/10.1016/j.jcou.2015.01. microalga Acutodesmus dimorphus, J. Appl. Phycol. (2015), http://dx.doi.org/10.
007. 1007/s10811-015-0625-2.
[12] R.D. Tripathi, S. Dwivedi, M.K. Shukla, S. Mishra, S. Srivastava, R. Singh, U.N. Rai, D.K. [33] A.F. Safinaz, A.H. Ragaa, Effect of some red marine algae as biofertilizers on growth
Gupta, Role of blue green algae biofertilizer in ameliorating the nitrogen demand of maize (Zea mayz L.) plant, Int. Food Res. J. 20 (2013) 1629–1632.
and fly-ash stress to the growth and yield of rice (Oryza sativa L.) plants, [34] F.A. Faheed, A.A.-E. Fattah, Effect of Chlorella vulgaris as bio-fertilizer on growth pa-
Chemosphere 70 (2008) 1919–1929. rameters and metabolic aspects of lettuce plant, J. Agric. Soc. Sci. 4 (2008) 165–169.
[13] H. Saadatnia, H. Riahi, Cyanobacteria from paddy fields in Iran as a biofertilizer in [35] K.L.N. Aung, Effect of Spirulina biofertilizer suspension on growth yield of Vigna
rice plants, Plant Soil Environ. 55 (2009) 207–212. radiata (L.) Wilczek, Univ. Res. J. 4 (2011) 351–363.
[14] A. Watanabe, R. Ito, C. Konishi, Effect of nitrogen-fixing blue-green algae on the
growth of rice plants, Nature 168 (1951) 748–749.
[15] E.J. Olguín, S. Galicia, R. Camacho, G. Mercado, T. Peŕez, Production of Spirulina sp. in
sea water supplemented with anaerobic effluents in outdoor raceways under tem-
perate climatic conditions, Appl. Microbiol. Biotechnol. 48 (1997) 242–247.