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J Appl Phycol

DOI 10.1007/s10811-015-0775-2

The use of microalgae as a high-value organic slow-release


fertilizer results in tomatoes with increased carotenoid
and sugar levels
Joeri Coppens 1 & Oliver Grunert 1 & Sofie Van Den Hende 2 & Ilse Vanhoutte 3 &
Nico Boon 1 & Geert Haesaert 4 & Leen De Gelder 3

Received: 17 August 2015 / Revised and accepted: 7 December 2015


# Springer Science+Business Media Dordrecht 2015

Abstract The heightened awareness concerning environ- an increase in sugar and carotenoid content, although a
mental preservation, resource scarcity, food safety, and nu- lower tomato yield was obtained. An economic evaluation
trition has engendered the need for a more sustainable and indicates the economic feasibility of the microalgae-based
resource-efficient agricultural production system. In this fertilizers. Further research is required to optimize the
context, microalgae offer the potential to recover nutrients microalgae-based fertilizer composition.
from waste streams and subsequently use the microalgal
biomass as a sustainable slow-release fertilizer. The aim Keywords Nutrient recovery . Wastewater treatment .
of this study was to assess microalgal bacterial flocs Biofertilizer . Horticulture . Nannochloropsis . MaB-flocs
treating aquaculture wastewater and marine microalgae as
organic slow-release fertilizers for tomato cultivation.
Comparable plant growth was observed using microalgal Introduction
and commercial organic fertilizer treatments. Furthermore,
the microalgal fertilizers improved the fruit quality through In light of the growing global population, rising resource scar-
city, and environmental preservation, the transition toward a
Joeri Coppens and Oliver Grunert contributed equally to this work. more sustainable food production system is becoming increas-
ingly important (Sutton et al. 2013). The implementation of
Electronic supplementary material The online version of this article
innovative nutrient recycling technologies, green fertilizers,
(doi:10.1007/s10811-015-0775-2) contains supplementary material,
which is available to authorized users. and advanced cultivation practices is needed to further in-
crease agricultural outputs, improve nutrient use efficiencies,
* Leen De Gelder and reduce nutrient losses. Modern greenhouse horticulture is
leen.degelder@ugent.be characterized by high crop yields and a stable year-round sup-
ply of high-quality fruits and vegetables. This high crop out-
1
Laboratory of Microbial Ecology and Technology (LabMET),
put is often accompanied by the intensive and unsustainable
Department of Biochemical and Microbial Technology, Faculty of use of biocides, inorganic fertilizers, and soilless cultivation
Bioscience Engineering, Ghent University, Coupure Links 653, techniques (Vox et al. 2010). The heightened social awareness
9000 Gent, Belgium concerning environmental preservation, food safety, and nu-
2
Laboratory of Industrial Water and Ecotechnology (LIWET), trition has engendered an increasing consumer interest in sus-
Department of Industrial Biological Sciences, Faculty of Bioscience tainable and organic products (Kondo et al. 2010). Conven-
Engineering, Ghent University, Graaf Karel de Goedelaan 5,
8500 Kortrijk, Belgium
tional greenhouse practices are therefore being reassessed to
3
include the use of high-quality organic fertilizers and growing
Laboratory for Environmental Technology, Department of Applied
Biosciences, Faculty of Bioscience Engineering, Ghent University,
media that enable the production of sustainable, safe, and nu-
Valentin Vaerwyckweg 1, 9000 Gent, Belgium tritional fruits and vegetables.
4
Laboratory for Crop Production, Department of Applied Biosciences,
The cultivation of microalgae has been studied extensively
Faculty of Bioscience Engineering, Ghent University, Valentin within the field of biotechnology. Microalgae are a rich source
Vaerwyckweg 1, 9000 Gent, Belgium of carbohydrates, lipids, proteins, pigments, and vitamins and
J Appl Phycol

are therefore a valuable feedstock for cosmetics, biofuels, and Materials and methods
food and feed supplements (Spolaore et al. 2006). Moreover,
they allow for the efficient recovery of the nitrogen (N) and Microalgal biomass
phosphorus (P) present in wastewaters by concentrating these
nutrients in algal biomass (Cai et al. 2013). The valorization of MaB-flocs were harvested during August 2013 from an out-
wastewater-grown microalgae for food or feed purposes is, door raceway pond (28 m2). The raceway pond was operated
however, in general legally restricted (Van Den Hende et al. as a sequencing batch reactor and treated wastewater from a
2014b). Although much less studied, the microalgal biomass recirculating freshwater aquaculture system for the cultivation
can instead be employed as an organic slow-release fertilizer of pikeperch (Sander lucioperca L.; Inagro, Roeselare, Bel-
(Mulbry et al. 2005, 2007). As an organic fertilizer, gium) (Van Den Hende et al. 2014a). After gravitational set-
microalgae have the potential to prevent nutrient losses tling, MaB-flocs were dewatered in filter bags (150–250 μm
through a gradual release of N, P, and potassium (K), which pore size; Lampe, Belgium) by a hydropress (4 bar;
is attuned to the plant requirements (Mulbry et al. 2007). Be- Enotecnica Pillan, Italy). MaB-flocs were dominated by fila-
sides macronutrients, phototrophic microorganisms also con- mentous microalgae (Ulothrix sp. and Klebsormidium sp.)
tain trace elements and plant growth-promoting substances (Van Den Hende et al. 2014a).
such as phytohormones, vitamins, carotenoids, amino acids, Nannochloropsis oculata was obtained from Proviron
and antifungal substances (Spolaore et al. 2006). The benefi- (Hemiksem, Belgium) and was produced on a 700-m2 algae
cial characteristics of phototrophic biomass as a biofertilizer, pilot plant, located on a landfill site (Hooge Maey, Antwerp,
i.e., bioactive growth-promoting amendments, have already Belgium).The microalgae were cultivated in outdoor closed
been demonstrated for cyanobacteria, anoxygenic flat panel photobioreactors (25 m2) in modified Guillard’s f/
phototrophic bacteria, and macroalgae (Kobayashi and 2 marine medium, using flue gas CO2 and residual heat from
Kobayashi 1995; Tripathi et al. 2008; Kumari et al. 2011). the landfill biogas cogeneration installation (Proviron, person-
The application of these biofertilizers stimulated plant growth al communication). After cultivation, the algae were concen-
and increased crop yields. In addition, the cultivation of per- trated using microfiltration followed by centrifugation. The
simmon and tomato fruits resulted in improved fruit quality, dewatered MaB-flocs and Nannochloropsis biomass were
through higher sugar and carotenoid concentrations in the pasteurized and dried at 70 °C for 14 h and subsequently
fruits (Kobayashi and Kobayashi 1995; Kumari et al. 2011). pulverized. The elemental biomass compositions are de-
This indicates that nutrients recovered through microalgae scribed in Table 1.
cultivation can be recycled as microalgae-based fertilizers to
improve the quality and market value of high-value fruits such Organic growing medium
as tomatoes.
The aim of this study was to assess the potential of The organic growing medium (Peltracom, Gent, Belgium)
microalgae as an organic slow-release fertilizer for tomato used during the mineralization and tomato cultivation experi-
cultivation. Two types of microalgal biomass were evalu- ments was a nutrient-poor mixture of sod peat (40 % v/v), Irish
ated: microalgal bacterial flocs (MaB-flocs) from a race- peat (40 % v/v), and coconut fiber (20 % v/v). Calcium and
way pond treating aquaculture wastewater (Van Den Hende magnesium carbonate (Dolokal Extra PG, The Netherlands)
et al. 2014a) and a marine culture of Nannochloropsis sp. were added at 2.5 kg m−3 in order to reach an optimal pH
cultivated on a landfill site. In a first stage, the microalgal (H2O) of 5.5. Chemical characteristics of the growing medium
biomass composition and N and P mineralization rates (pH, electrical conductivity (EC), ammonium, nitrate, phos-
were determined. Subsequently, a greenhouse tomato cul- phorus, potassium, calcium, magnesium, sulfate, sodium,
tivation experiment was performed, wherein the fertilizer chloride, iron, and manganese) were measured according to
potential of both types of microalgal biomass was com- Gabriels et al. (1998). Physical characteristics of the growing
pared with conventional inorganic and organic horticulture media were measured according to EN 13041 (DIN 2012).
fertilizer systems. The growth rate of the tomato plants and The physical and chemical characteristics of the soil can be
the tomato yield were assessed for each fertilizer treatment, found in Supplementary Information (Table S1).
as well as the composition of the leaves and the water and
sugar and carotenoid concentrations in the tomato fruits. Microalgal N and P mineralization rate analysis
Afterward, the economic feasibility of microalgal fertil-
izers was determined through a comparison with conven- The N and P mineralization rates of the dried MaB-flocs and
tional inorganic and organic greenhouse cultivation sys- Nannochloropsis biomass were determined according to
tems. To the authors’ knowledge, this study is the first to Chaves et al. (2004). The organic growing medium was mixed
assess the application of microalgal biomass as fertilizers with the microalgal biomass to obtain a final concentration of
for greenhouse tomato production. 555 mg biomass-TN L−1 soil. The microalgae-soil mixtures
J Appl Phycol

Table 1 Elemental composition of the dried MaB-floc and extractant/gram soil) were performed to determine the concen-
Nannochloropsis biomass
tration of plant-extractable P (Mehlich 1984).
MaB-flocs Nannochloropsis

Ash (%) 67.12 23.22 Tomato cultivation setup


N (%) 2.44 8.07
P (%) 0.59 1.29 Solanum lycopersicum cv. BMaxifort^ was sown on 29 Octo-
K (%) 0.18 1.36 ber 2013, and Solanum lycopersicon cv. BMerlice^ was
N:P:K 14:3:1 6:1:1 grafted on the stems on 15 November 2013. The plants were
Ca (%) 20.40 0.20 grown on slabs filled with non-sterilized organic growing me-
Mg (%) 0.21 0.40 dium (Peltracom, Belgium). The cultivation experiment
S (%) 0.24 0.05 consisted of four fertilizer treatments: (1) a liquid inorganic
Na (%) 0.03 1.34 fertilizer, (2) solid organic fertilizer, (3) MaB-floc fertilizer,
Fe (mg kg−1) 328 143 and (4) Nannochloropsis fertilizer (Fig. 1).
Cu (mg kg−1) 7.86 11.8 The fertilizer demand of the plants was calculated accord-
Zn (mg kg−1) 143 66.7 ing to commercial greenhouse tomato practices (Sonneveld
B (mg kg−1) 5.65 1.16 and Voogt 2009). For the inorganic fertilizer control treatment,
Mn (mg kg−1) 42.5 113 a standard fertilizer solution suitable for the growth of tomato
plants (Sonneveld and Voogt 2009) was dosed with a standard
drip irrigation system. The organic fertilizer control treatment
were transferred to PVC tubes (diameter = 4.63 cm; filling consisted of a blend of two commercial solid fertilizers SF1
height = 12 cm; bulk density = 140 g L−1), and distilled water (66 % of N demand) and SF2 (33 % of N demand) (Frayssinet,
was added to obtain a field capacity of 80 %. The tubes were France), which were blended in the growing medium prior to
incubated at 20 °C, and samples were taken after 0, 7, 14, 21, the start of the cultivation experiment. SF1 was characterized
35, 67, and 95 days. At each sampling time, tubes were col- by a nutrient content of 4 % N, 2.2 % P, and 5 % K and a
lected in quadruplicate. Soil extractions using distilled water mineralization coefficient of 90 % mineralization after
(10:1 water/gram soil) were performed for the determination 3 months. SF2 had a nutrient content of 8 % N, 2.2 % P, and
of pH, EC, total ammonium nitrogen (TAN), nitrite, nitrate, 5 % K and a mineralization coefficient of 80 % after 3 months.
phosphate, and sulfate. Mehlich-3 extractions (10:1 The nutrient demand of the plants for a cultivation period of

Fig. 1 Overview of the tomato cultivation setup. The setup consists of a liquid inorganic fertilizer control treatment, a solid organic fertilizer control
treatment, a MaB-floc fertilizer treatment, and Nannochloropsis fertilizer treatment
J Appl Phycol

3 months was added to the growing medium prior to the start Tomato fruits were harvested, and the fresh weight, dry matter
of the experiment (Table 2). For both microalgal treatments, content, sugars, and carotenoid concentrations were
the microalgal fertilizer was blended with the organic growing determined.
medium, considering an N mineralization rate of 33 % after
3 months. The organic and microalgal fertilizer treatments
Analytical techniques
were amended with kali vinasses (38 % K2O; Rendapart, Bel-
gium) and Patentkali (30 % K2O, 10 % MgO, and 42 % SO3;
Elemental soil and biomass compositions were determined
Pillaert Meststoffen, Belgium), respectively, to obtain a com-
using ICP-OES (Vista-MPX, Varian, Australia) as described
parable macronutrient ratio in the growing media (Table 2).
by Greenberg et al. (1992). Nitrate, nitrite, and phosphate were
For each fertilizer treatment, three slabs (1.0 × 0.2 × 0.085 m)
analyzed after sample filtration using anion chromatography
with organic growing medium were used. Three plants were
(Metrohm 761 Compact IC, Switzerland). Ammonium
grown per slab, with an interspacing of 50 cm between plants
(Nessler method), total Kjeldahl nitrogen (TKN), and total
to obtain nine plant replicates. The fertilized slabs were incu-
phosphorus (TP) (molybdate/vanadium method) were deter-
bated for 1 week prior to the plant cultivation experiment to
mined according to standard methods (Greenberg et al.
initiate microbial activity and mineralization in the growing
1992). The chlorophyll and carotenoid content of the tomato
medium. The 134-day-old tomato plants were planted on top
fruits were determined according to Lichtenthaler (1987).
of the different growing media on the 12 March 2014. A drip
Sugars were extracted according to De Swaef et al. (2012).
irrigation system was used for a controlled dosage of rainwa-
Glucose and fructose were determined using HPAEC-PAD
ter. The amount of irrigation water was the same for each
(ICS 3000, Dionex, USA) with a 3 × 150 mm CarboPac
treatment and was set to achieve a minimum drain in order
PA20 analytical column.
to prevent nutrient losses through leaching; although, in com-
mercial growing systems, generally a drain of 30 % is applied.
The greenhouse was equipped with an automated ventilation Statistical analysis
and heating system and accommodated with movable screens
for shading (ILVO, Melle, Belgium). Temperature was main- A non-parametric non-paired Kruskal-Wallis test was per-
tained at 17 °C during the night and above 22 °C during the formed and combined with an all-pairwise multiple compari-
day. Additional artificial lighting was provided from 8 am to son Dunn’s test to compare tomato yields and fruit quality
9 pm at 50 μmol photons m−2 s−1 PAR. parameters (Prism 5.0, Graphpad Software, USA). Homoge-
Tomato plant growth was monitored by determining the neity of variances was verified with Levene’s test. For the
plant height and diameter three times per week, just below physicochemical characteristics of the leaves and the growing
the first leaf on the bottom of the plants. Side shoots were medium, a one-way ANOVA repeated measures analysis of
removed according to commercial practice while tomato variance was conducted to evaluate the null hypothesis that
trusses were left unpruned. Pollination of the flowers was there is no significant difference between the treatments.
done by hand. Tomato leaves were harvested on regular time Sphericity and normality of dependent variables were
intervals and analyzed for their length, weight, elemental com- assessed using a Mauchly test and Shapiro-Wilk test, respec-
position, dry matter, and ash content. Soil samples were taken tively. In the case that sphericity was not obtained, corrections
on regular time intervals, and a water extraction was per- were performed according to Greenhouse-Geisser. P values
formed to determine the pH, EC, and elemental composition. were adjusted with the Bonferroni confidence interval.

Table 2 Composition of the


different fertilized growing media Inorganic fertilizer Organic fertilizer MaB-flocs Nannochloropsis
at the start of the tomato
cultivation experiment (n = 3) pH 5.79 ± 0.04 5.77 ± 0.01 7.23 ± 0.05 5.13 ± 0.12
EC (μS cm−1) 58 ± 4 462 ± 112 205 ± 44 875 ± 356
NH4+-N (mg L−1) 1.14 ± 0.13 55.63 ± 24.21 47 ± 4 6.22 ± 3.52
P (mg L−1) 11.05 ± 0.75 38.03 ± 7.70 268 ± 66 239 ± 68
K+ (mg L−1) 56 ± 3 1006 ± 288 189 ± 40 784 ± 282
Ca2+ (mg L−1) 589 ± 53 578 ± 55 15,634 ± 2587 636 ± 136
Mg2+ (mg L−1) 132 ± 9 193 ± 28 199 ± 22 277 ± 55
SO42− (mg L−1) 129 ± 12 888 ± 168 126 ± 86 1239 ± 635
Na+ (mg L−1) 40 ± 2 167 ± 40 48 ± 5 360 ± 118
Cl− (mg L−1) 85 ± 7 201 ± 42 66 ± 10 648 ± 223
J Appl Phycol

Pairwise differences between treatments were considered sig-


nificant at p < 0.05.

Economic analysis

An economic evaluation was performed to assess the econom-


ic feasibility of microalgae-based fertilizers. Different fertiliz-
er scenarios were investigated, in which microalgae were
combined with conventional inorganic and organic fertilizer
systems. The costs for the outdoor cultivation of microalgae in
a raceway pond were calculated according to Norsker et al.
(2011). Production data of greenhouse tomato cultivation was
obtained from a government survey on greenhouse tomato
farmers in Flanders, Belgium (Jourquin et al. 2013) and from
the agricultural consultancy agency DLV Plant (the Nether-
lands; personal communication). Supplementary Information
(S.I.) contains detailed referencing on the different assump-
tions, data sources, and calculations (Tables S4, S5, and S6).

Results and discussion

Mineralization of the microalgal biomass

A batch incubation study was performed to determine the N


and P mineralization rate for the two types of microalgal bio-
Fig. 2 N and P mineralization profile of dried MaB-floc and
mass. The MaB-flocs and Nannochloropsis had an initial N
Nannochloropsis biomass (n = 4). Error bars indicate standard deviation
availability of 7 and 5 %, respectively (Fig. 2a). After 21 days,
11 and 16 % of the biomass N was mineralized, respectively.
A final plant-available N fraction of 25 and 31 % was obtained Furthermore, plant growth and root activity support the re-
after 95 days for the MaB-flocs and Nannochloropsis bio- lease of nutrients from fertilizers through the excretion of mobi-
mass, respectively. An initial plant-extractable P availability lizing substances such as organic acids (Ahmed et al. 2015). The
of 53 % was obtained for both biomass types. Also, an initial prediction of the plant available nutrients is therefore more ac-
orthophosphate availability of 11 % P for MaB-flocs and 7 % curately described by plant uptake than by chemical extraction.
P for Nannochloropsis was measured (Fig. 2b). These values In greenhouse production systems, where irrigation and temper-
did not increase significantly during the incubation period. ature can be controlled, fertilizer management that considers
Both algal biomass types showed a similar mineralization both the nutrient source and soil environment can further im-
profile. However, the N mineralization rates are lower than prove the effectiveness of the chosen organic fertilizer materials.
previously described, as algal biomass grown on manure efflu-
ents showed an N availability of 25–33 % after 21 days and a Effect of microalgal biomass on tomato plant performance
maximal N availability of 41 % after 63 days (Mulbry et al.
2005, 2007). The mineralization rates are nonetheless within Tomato plant growth
the range of commercially available slow-release fertilizers
which are applied in organic horticulture (Stadler et al. 2006). The high concentration of calcite (CaCO3) precipitates present
The release of N and P from organic nutrient sources is strongly in the MaB-flocs (Van Den Hende et al. 2014a) resulted in an
controlled by the soil environment. Discrepancies in minerali- increase in the calcium concentration and pH of the growing
zation rates may therefore be explained by differences in soil medium compared to the control treatments. The marine
richness, moisture content, temperature, and pH during the in- Nannochloropsis biomass resulted in elevated sodium, sulfate,
cubations (Agehara and Warncke 2005). The lower microbial and chloride concentrations and an increased EC of the growing
activity in our study is reflected in the low nitrification activity medium (Tables 2 and 3). The ammonium concentration of the
obtained during the mineralization test, as less than 2 % of the Nannochloropsis and the MaB-flocs treatment were compara-
available ammonium nitrogen was oxidized to nitrite or nitrate. ble to the organic control treatment, but higher than the
J Appl Phycol

Table 3 Characteristics of the


organic growing medium for the Inorganic fertilizer Organic fertilizer MaB-flocs Nannochloropsis
different fertilizer treatments
during the experiment pH 5.73 ± 0.07c 5.58 ± 0.07c 7.25 ± 0.07a 6.11 ± 0.07b
−1 b
EC (μS cm ) 86.3 ± 50.8 504.1 ± 50.8a 243.3 ± 50.8b 657.9 ± 50.8a
NO2−-N (mg L−1) 0.16 ± 0.08b 0.22 ± 0.08ab 0.56 ± 0.08a 0.18 ± 0.08b
NO3−-N (mg L−1) 7.2 ± 0.5a 1.9 ± 0.5c 5.3 ± 0.5ab 3.7 ± 0.5bc
NH4+-N (mg L−1) 2.8 ± 1.6b 34.4 ± 1.6a 36.3 ± 1.6a 37.0 ± 1.6a
P (mg L−1) 10.5 ± 10.8b 43.6 ± 10.8b 243.7 ± 10.8a 167.0 ± 10.8a
K+ (mg L−1) 25.2 ± 37.6c 302.8 ± 37.6ab 140.5 ± 37.6bc 465.7 ± 37.6a
Ca2+ (mg L−1) 678 ± 1113b 665 ± 1928.2b 12,986 ± 1113a 603.7 ± 1113b
Mg2+ (mg L−1) 184.5 ± 28.8b 232.3 ± 20.4ab 197.5 ± 16.6b 319.8 ± 20.4a
SO42− (mg L−1) 202.7 ± 114.2b 1131.1 ± 114.2a 180.5 ± 114.2b 983.4 ± 114.2a
Na+ (mg L−1) 45.5 ± 18.5b 122.5 ± 13.1b 73.0 ± 10.7b 256.5 ± 13.1a
Cl− (mg L−1) 32.4 ± 16.4b 57.1 ± 16.4b 41.4 ± 16.4b 258.2 ± 16.4a

Mean values and standard error of means over the different treatments are displayed of five sampling points (day
1, 22, 36, 62, and 90). Bonferroni-corrected p values are calculated for the effect of fertilization strategy and time
on the chemical characteristics of organic growing media in tomato plants (n = 3) per treatment and per time
point. Factors which do not share superscripts are significantly different from each other (p = 0.05)

inorganic control. For both microalgal fertilizers, a higher phos- stem diameter and the number of trusses among the different
phorus concentration was obtained in the growing medium. treatments (Table S2).
There was no significant difference between the growth Leaf analysis showed a significantly higher nitrogen
rate of plants grown on the algal fertilizers and the organic content for the algal fertilizer treatments, which demon-
fertilizer. Compared to the organic and algal fertilizers, the strates the good nitrogen fertilizer properties of the algal
conventional inorganic growing system revealed a faster ini- fertilizers (Table 4). Additionally, a lower ash content of
tial plant growth (Fig. 3), which is attributed to the readily the tomato plant leaves was observed for both algal fertil-
available inorganic nutrients and the presence of nitrate as izer treatments throughout the experiment. The MaB-floc
the predominant nitrogen source (Sonneveld and Voogt fertilizer resulted in a significantly lower magnesium and
2009). The faster plant growth is confirmed by the higher potassium content of the leaves (Table 4). This deficiency
mean plant length (162.5 ± 3.8 cm), i.e., the average plant was visually noticeable as interveinal chlorosis and can be
length over the duration of the cultivation experiment, in com- attributed to the relatively high calcium concentration in
parison to the organic treatment (145.6 ± 3.8 cm), MaB-flocs the algal biomass, which can inhibit the uptake of magne-
treatment (143.2 ± 17 cm) and Nannochloropsis treatment sium and potassium (Jakobsen 1993). A minor magnesium
(139.2 ± 3.8 cm) (Table S2). Nevertheless, there was no sig- deficiency is nevertheless common in greenhouse horticul-
nificant difference in the final plant height for the inorganic ture and rarely results in yield reduction. Although the high
fertilizer, MaB-flocs, and Nannochloropsis treatment (Fig. 3). pH of the MaB-floc fertilizer could lower the plant avail-
Growth suppression due to ammonia toxicity therefore did not ability of micronutrients, no symptoms of micronutrient
occur. Also, no significant difference was observed in mean deficiencies were observed.

Fig. 3 Growth curve of the


tomato plants (n = 9) for the
different fertilizer treatments.
Error bars indicate standard
deviation
J Appl Phycol

Table 4 Leaf characteristics for


the different fertilizer treatments Inorganic fertilizer Organic fertilizer MaB-flocs Nannochloropsis
during the experiment
Leaf length (cm) 40.7 ± 1.1 40.6 ± 1.1 37.8 ± 1.1 42.2 ± 1.1
Leaf fresh weight (g) 20.7 ± 1.2 19.8 ± 1.2 19.3 ± 1.2 23.5 ± 1.2
Leaf dry weight (%) 18.2 ± 0.6a 19.0 ± 0.6a 20.8 ± 0.6a 16.3 ± 0.6b
Ash (% DM) 17.78 ± 0.38b 16.11 ± 0.38b 13.56 ± 0.38a 12.64 ± 0.38a
N (% DM) 2.83 ± 0.11c 3.84 ± 0.11b 4.7 ± 0.11ab 5.05 ± 0.11a
P (% DM) 0.42 ± 0.26b 0.72 ± 0.26a 0.79 ± 0.26a 0.71 ± 0.26a
K (% DM) 46.00 ± 3.96 30.95 ± 3.96 21.46 ± 3.96 37.50 ± 3.96
Ca (% DM) 27.21 ± 1.09a 21.56 ± 1.09bc 25.96 ± 1.09ab 12.54 ± 1.09a
Mg (% DM) 6.2 ± 0.2b 7.2 ± 0.2a 5.2 ± 0.2b 7.1 ± 0.2a
Zn (% DM) 0.040 ± 0.004 0.039 ± 0.004 0.039 ± 0.004 0.034 ± 0.004
Mn (% DM) 0.15 ± 0.16a 0.12 ± 0.16ab 0.11 ± 0.16bc 0.05 ± 0.16c
Fe (% DM) 0.064 ± 0.004 0.053 ± 0.004 0.050 ± 0.004 0.055 ± 0.004
Cu (% DM) 0.014 ± 0.005 0.005 ± 0.005 0.003 ± 0.005 0.002 ± 0.005
B (% DM) 0.02 ± 0.02b 0.32 ± 0.02a 0.29 ± 0.02a 0.01 ± 0.02b

Mean values and standard error of means over the different treatments are displayed of four sampling points (day
22, 36, 60 and 90). Bonferroni-corrected p values are calculated for the effect of fertilization strategy and time on
leaf length, fresh and dry weight, and chemical characteristics of leaves in tomato plants (n = 3) per treatment and
per time point. Factors which do not share superscripts are significantly different from each other (p = 0.05)

The Nannochloropsis treatment resulted in lower calcium the developing fruits. Also, differences in nitrogen availability
concentrations in the tomato leaves. Calcium deficiency is often can impose stress, as plants grown on ammonium tend to
the result of water stress and can therefore be attributed to the grow smaller fruits and are more affected by blossom-end
elevated salt concentrations of the marine microalgal biomass, rot than are plants grown on nitrate (Borgognone et al.
since absolute calcium levels did not differ from the controls 2013). The adverse effects of an increased supply of ammo-
(Table 3). Also, the high magnesium content can induce calci- nium as a nitrogen source are also associated with a lower
um deficiency, as the same plant absorption sites are shared for calcium and magnesium uptake, which was observed in the
the uptake of calcium, magnesium, and potassium elemental leaf composition for the Nannochloropsis and
(Papadopoulos 1991). No significant difference in iron (Fe), MaB-floc fertilizers (Table 4).
copper (Cu), and zinc (Zn) content was observed among treat-
ments. These results show that with regard to plant growth, both
the MaB-flocs and marine Nannochloropsis biomass can re-
place conventional inorganic and organic fertilizers.

Tomato fruit yield

There was no significant difference in the number of fruits be-


tween treatments. A lower tomato yield was nevertheless obtain-
ed for the algal treatments compared to the inorganic and organic
systems (Fig. 4). Also, a higher incidence of blossom-end rot
was observed for both the organic and algal fertilizers. The fresh
weight of the red tomatoes was significantly higher for the con-
ventional inorganic fertilizer treatment, but no difference was
observed among the organic and algal treatments (Fig. 5a).
The lower marketable yield and fruit size obtained for the
organic and algal fertilizer treatments can be explained by
different phenomena. Lower tomato yields have been com-
monly observed under salt stress conditions (Magán et al.
2008). The lower yields in this study can therefore be attrib- Fig. 4 Tomato yield per plant (n = 9) for each fertilizer treatment
uted to the osmotic pressure caused by an increased salinity of subdivided in the weight of green, red, and blossom-end rot-affected
the organic and algal fertilizers, which limits the water flux to tomatoes per plant. Error bars indicate standard deviation
J Appl Phycol

Tomato quality inclusion of MaB-flocs as an aquaculture fodder supplement


resulted in an increased red pigmentation of white Pacific
Tomato fruits grown on Nannochloropsis and MaB-flocs had a shrimps (Van Den Hende et al. 2014b).
34 and 20 % higher dry weight content, respectively, compared
to the inorganic fertilizer treatment; whereas no difference be- Potential of microalgal fertilizers in greenhouse
tween the inorganic and organic fertilizer treatment was ob- horticulture
served (Fig. 5b). Furthermore, both algal fertilizers resulted in
significantly higher sugar concentrations in the fruits (Fig. 5c). Globally, tomato is one of the most produced vegetables, ranking
The Nannochloropsis-grown tomato fruits had a glucose con- second after potato (Kumari et al. 2011). This illustrates the
centration that was 18 % higher than the organic and 33 % economic and nutritional importance of this crop. The increased
higher than the inorganic fertilizer treatment. The MaB-floc sugar and carotenoid concentrations obtained with the algal fer-
fertilizer resulted in a 23 % higher glucose concentration com- tilizers indicate the potential of microalgae-based fertilizers to
pared to the inorganic fertilizer. For the Nannochloropsis- increase the quality and economic value of tomato fruits. Carot-
grown tomatoes, a 21 % higher fructose concentration was enoids play an important role in many plants during photosyn-
observed compared to those grown on inorganic fertilizers. thesis, the protection against photooxidative stress, and attraction
The higher sugar concentrations observed confirm the impor- of insects. The amendment of microalgal biomass can therefore
tance of reduced nitrogen forms such as ammonium or organic also have beneficial effects for other high-value plants. Compa-
nitrogen to improve the tomato fruit taste (Heeb et al. 2005). rable to tomatoes, microalgal fertilizers can improve the value of
The most remarkable improvement in fruit quality was ob- peppers (Capsicum annuum), while they can also be implement-
served for the carotenoid content (Fig. 5d). Tomatoes grown ed in flower cultivation, as carotenoids induce the typical yellow
on MaB-flocs contained 70 % more carotenoids than in the and orange color in, for instance, roses (Lachman et al. 2001).
inorganic fertilizer treatment and 44 % more than in the or- Nevertheless, the difference in fruit yield compared to the
ganic fertilizer treatment. The effect of the Nannochloropsis conventional horticulture fertilizers indicates that a more op-
fertilizer was less profound, but still a 36 % higher carotenoid timal fertilizer mixture is required to combine high-quality
content was obtained compared to the inorganic fertilizer fruits with satisfactory yields. The importance of nitrate to
treatment. These observations confirm the stimulating effect stimulate plant growth and fruit yield suggests that conven-
of phototrophic biomass on the sugar and carotenoid content tional fertilizers should be employed as the main source of
of fruits (Kobayashi and Kobayashi 1995; Kumari et al. 2011). macronutrients, while the addition of microalgal biomass
Moreover, it is in line with other findings, in which the could improve the market value of the products. Although

Fig. 5 Fresh weight (a), dry weight (b), sugar (c), and carotenoid (d) content of red tomato fruits (n = 36) for the different fertilizer treatments. Error bars
indicate standard deviation. Significant differences among fertilizer strategies are indicated with a different letter
J Appl Phycol

further research is required to assess the optimal amount of The low economic impact of the use of fertilizers therefore
microalgae to be amended to the fertilizers, previous findings allows incorporation of more expensive fertilizers if an
related to the application of phototrophic organisms as economic added value can be delivered through superior
biofertilizers suggest that the microalgal biomass can already fruit quality.
have beneficial effects on crop output at a 10 % concentration On average, the economic value of auctioned organically
(Tripathi et al. 2008; Kumari et al. 2011). Microalgae-based grown tomatoes is 33 % higher than those obtained through
fertilizers can therefore partake in the transition of convention- conventional inorganic cultivation (Auction Hoogstraten
al greenhouse practices toward an integrated plant nutrition SCRL (2015); Provincial center for crop cultivation,
system (IPNS). This integrated cultivation concept combines Kruishoutem, personal communication). If a comparable price
and optimizes the use of inorganic, organic, and biofertilizers would be obtained for tomatoes grown on inorganic and
to sustain desired crop productivity with a minimal impact on microalgae-based fertilizers, microalgae could be blended in
the environment (Chen 2006). The beneficial effects obtained for up to 18 % of the nitrogen demand of the plants while
in this study demonstrate the potential of microalgal fertilizers remaining economically competitive. The amendment of
to give an added value within this sustainability framework. 10 % of the nitrogen demand of the plant as a microalgal
Furthermore, in organic horticulture, conventional organic fer- fertilizer to a conventional inorganic fertilizer system could
tilizers can be amended with microalgal biomass in order to potentially combine the improved plant growth of nitrate-
improve fruit quality while preserving the sustainable organic based inorganic fertilizers with the beneficial effects on fruit
cultivation practices. quality of microalgae. This system would result in a 9 % in-
crease of the production cost, which may be balanced by the
Economic assessment of microalgal fertilizers added market value due to improved taste and pigmentation
for conventional and organic greenhouse horticulture (Fig. 6). This supports the economic viability of microalgae-
based fertilizers.
The economic evaluation of an outdoor raceway pond for This economic evaluation does not take into account the
microalgae cultivation specifies a microalgal production cost additional savings which can be obtained within the green-
of €23 kg−1 biomass or €289 kg−1 fertilizer-N (Table S3). house cultivation system through on-site microalgae produc-
Commercial inorganic NPK fertilizers (14 % N, 7 % P, tion. The production of microalgae allows for the recovery of
15 % K) and organic slow-release fertilizers (4 % N, 2 % P, excess nutrients present in the greenhouse effluents, thereby
5 % K) have a market value of €7.9 and €11 kg−1 N, respec- reducing discharge to the environment and enabling the rec-
tively (Table S5). This indicates that microalgal biomass is not lamation of water for further use. Additionally, the utilization
economically competitive with commercial horticulture fertil- of unused greenhouse space for indoor microalgae cultivation
izers when applied as the main nitrogen fertilizer source. Nev- would stimulate microalgal growth through the availability of
ertheless, a cost assessment of greenhouse tomato cultivation artificial lighting and heating. This would improve production
shows that tomato production using inorganic and organic yields and hence reduce the total production costs. The
fertilizers comes at a total production cost of €0.73 and € recycling of greenhouse wastewaters would thus not only
1.09 kg−1 fruit, respectively (Fig. 6). While labor contributes reduce the dependency of inorganic fertilizers but also gen-
38 % and is the predominant cost factor, fertilizers only erate revenues from unused greenhouse space and waste-
contribute 3 % to the total production cost (Table S4). water treatment facilities.

Fig. 6 Cost of different fertilizer


mixtures of microalgae and
inorganic and organic fertilizers
and the respective production cost
of the tomato fruits. The dotted
line indicates the average auction
price of organically grown
tomatoes in 2013–2014
(€1.16 kg−1; PCG, personal
communication)
J Appl Phycol

Microalgae-based fertilizers also offer advantages within a and mineral composition of self-grafted and grafted tomatoes. Scient
Hortic 149:61–69
larger sustainability framework. In contrast to nutrient-rich
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waste streams such as manure, microalgal biomass can func- by microalgae: status and prospects. Renew Sust Energ Rev 19:360–
tion as a stable, predictable, transportable, and concentrated 369
fertilizer product. This allows it to be introduced in modern Chaves B, De Neve S, Hofman G, Boeckx P, Van Cleemput O (2004)
greenhouse horticulture. Furthermore, the additional plant Nitrogen mineralization of vegetable root residues and green ma-
nures as related to their (bio)chemical composition. Eur J Agron 21:
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vation give an added value compared to the direct application or biofertilizer for crop growth and soil fertility. In: International
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demonstrates that microalgal biomass can be used as an or- affects yield and taste of tomatoes. J Sci Food Agric 85:1405–1414
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Jourquin S, Maertens E, Deuninck J, D’hooghe J (2013) Het
of fruits. Further research is required to determine optimal bedrijfsinkomen van de tomatenteler: resultaten van bedrijven uit
fertilizer mixtures that produce high quality fruits with satis- het landbouwmonitoringsnetwerk (The income of the tomato famer:
factory yields. In this context, our economic evaluation sup- survey results from the agricultural monitoring network).
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Acknowledgments J.C. was supported by a PhD grant from the Insti- Netherlands, pp 1269–1282
tute for the Promotion of Innovation by Science and Technology in Flan- Kondo K, Nakata N, Nishihara E (2010) Effect of the purple non-sulfur
ders (IWT-Vlaanderen, SB-101187). O.G. was supported by the project bacterium (Rhodobacter sphaeroides) on the Brix, titratable acidity,
grant IWT Baekeland mandate 120200. S.V.D.H. was supported by the ascorbic acid, organic acid, lycopene and β-carotene in tomato fruit.
INTERREG IVB NWE program, the Flemish Government, and Province Food Agric Environ 8:743–746
West-Flanders within the EnAlgae project. The authors thank Stephen J. Kumari R, Kaur I, Bhatnagar AK (2011) Effect of aqueous extract
Andersen, Francis Meerburg, Jochen Hanssens, Tom De Swaef, and of Sargassum johnstonii Setchell & Gardner on growth, yield
Alessia Landi for inspiring scientific discussions. and quality of Lycopersicon esculentum Mill. J Appl Phycol
23:623–633
Lachman J, Orsak M, Pivec V, Kratochvilova D (2001) Anthocyanins and
carotenoids-major pigments of roses. Horticultural Science-UZPI,
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