Egyptian Journal of Aquatic Research (2012) 38, 51–57

National Institute of Oceanography and Fisheries

Egyptian Journal of Aquatic Research

http://ees.elsevier.com/ejar
www.sciencedirect.com

FULL LENGTH ARTICLE

Production and nutritive value of Spirulina platensis

in reduced cost media
Fedekar Fadel Madkour b, Abd El-Wahab Kamil a,*
, Hoda Shafik Nasr a

a
National Institute of Oceanography and Fisheries, Inland Waters, Egypt
b
Marine Science Department, Faculty of Science, Port Said University, Egypt

Received 8 July 2012; accepted 6 September 2012

Available online 10 November 2012

KEYWORDS Abstract This study aimed to provide a cost effective medium to large scale production of Spirulina
Spirulina platensis; platensis. This intention was implemented by substituting all the nutrients present in Zarrouk’s med-
Mass production; ium (SM) with cheaper and locally available commercial fertilizers and chemicals. The Reduced Cost
Reduced Cost medium; medium contained single super phosphate (SSP), commercial sodium bicarbonate, Muriate of pot-
Cyanobacteria; ash (MOP) and crude sea-salt, (Syahat salt). Four grades of nitrogen concentrations representing
Nitrogen source 10%, 20%, 30% and 40% of SM nitrogen concentration (29.42 mM-N) were taken from ammonium
nitrate (Treatments 1–4) or urea (Treatments 5–8) respectively, for testing. The alga was grown for
33 days at 30 ± 2 C, pH 9, 30 lEm2 s1 irradiance. The growth characteristics (maximum biomass
Xm, cell productivity Px, specific growth rate lm and chlorophyll concentration), and biochemical
composition (proteins, carbohydrates and lipids) of the alga grown in these media were compared
with that cultivated in SM. Significant differences in the growth parameters and biochemical com-
position were observed for the different nitrogen sources and concentrations. The results revealed
that S. platensis could utilize ammonium nitrate most efficiently and that growth was enhanced with
increasing the concentrations of ammonium nitrate giving maximum biomass at 0.353 g/L
(Treatment 3). Further increasing the concentration limited growth. The growth parameters in urea
showed a significant decrease associated with increasing urea concentrations. The maximum bio-
mass, chlorophyll and protein yield (0.813 ± 0.018 mg/L, 0.0685 ± 0.0024 lg/L and 52.62%,
respectively) were recorded using Treatment 3 which was comparable with that of SM
(0.840 ± 0.008 mg/L, 0.0701 ± 0.0089 lg/L and 52.95%, respectively). The results indicated that

* Corresponding author. Address: Abd Elwahab Abd Elshafi kamil,

National Institute of Oceanography and Fisheries, Inland Waters, 101
Kaser El Aini, Cairo, Egypt. Tel.: +20 42188003/24127815, mobile:
+20 1111615711; fax: +20 42185320/27921341.
E-mail address: abdelwahabkamil@gmail.com (A.El-Wahab Kamil).
Peer review under responsibility of National Institute of Oceanography
and Fisheries.

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52
the newly prepared medium can be used profitably for large-scale mass production of protein-rich
Spirulina and yields similar performance with cost effective to Zarrouk’s medium.
ª 2012 National Institute of Oceanography and Fisheries. Production and hosting by Elsevier B.V. All
Introduction
Spirulina is a multicellular, filamentous cyanobacterium which
can colonize environments that are unsuitable for many other
organisms, forming populations in freshwater and brackish
lakes and some marine environments, mainly alkaline saline
lakes (Vonshak, 1997). Spirulina contains a high content of
protein (up to 70%), along with high amounts of essential fatty
acids, essential amino acids, minerals, vitamins (especially
B12), antioxidant pigments (phycobiliproteins and carotenoids)
and polysaccharides (Belay et al., 1993; Vonshak, 1997). Con-
sequently, the commercial production of Spirulina has gained
worldwide attention for use in human food supplements, ani-
mal feed and pharmaceuticals. In aquaculture, Spirulina is
used as a feed additive to improve growth, feed efficiency, car-
cass quality, and physiological response to disease in several
species of fish (Mustafa et al., 1994). Furthermore, it is the
richest algal source of Gamma-linolenic acid (GLA), a precur-
sor for the biologically-active compound (prostaglandins,
PGE1) (Habib et al., 2008) which is necessary for the enhance-
ment of the immune system in shrimp larvae (Belay et al.,
1996; Yuan-Kun et al., 2003). In addition to its good nutri-
tional value, the evidence for its potential therapeutic applica-
tion is overwhelming (Belay et al., 1993; Belay, 2002).
The growth of Spirulina and the composition of the biomass
produced depend on many factors, the most important of which
are nutrient availability, temperature and light (Cornet et al.,
1992). In addition, Spirulina requires relatively high pH values
between 9.5 and 9.8 (Belkin and Boussiba, 1971), which effec-
tively inhibits contamination by most algae in the culture. Pro-
duction of Spirulina with reduced costs is necessary when
considering large-scale cultivation for industrial purposes. The
cost of nutrients is considered the second major factors influence
the cost of Spirulina biomass production after labor (Vonshak,
1997). Zarrouk’s medium has successfully served as the standard
medium (SM) for Spirulina culture for many years (Zarrouk,
1966). Consequently, many media have been developed using
seawater (Faucher et al., 1979), sewage water (Saxena et al.,
1982) and industrial effluents (Tanticharoen et al., 1993).
Therefore, the present study was undertaken to evaluate the
growth of Spirulina platensis in a medium based on commercial
grade chemicals and fertilizers and to compare the growth
characteristics and biochemical composition of the produced
organism with that cultivated in Zarrouk’s medium. This will
allow the derivation of a new medium to assist in decreasing
the cost of Spirulina production.
Materials and methods
Microorganism
The cyanobacterium S. platensis, strain K-2, used in the pres-
ent study was obtained from the Freshwater Hydrobiology
Laboratory, National Institute of Oceanography and Fisheries
F.F. Madkour et al.
rights reserved.
(NIOF), Cairo, Egypt. The strain was maintained in 500 mL
sterilized Erlenmeyer flasks containing 100 mL Zarrouk’s med-
ium (Zarrouk, 1966) at 30 ± 2 C, pH 9 with continuous illu-
mination using cool white fluorescent tubes (2500 Lux) and
twice daily shaking by hand.
Culture media and experimental design
All constituents of Zarrouk’s medium were autoclaved without
bicarbonate salt, which was sterilized by filtration, and used as
the standard control medium (SM). To prepare the new Re-
duced Cost media, all of the constituents present in Zarrouk’s
medium were substituted with commercial fertilizers and other
cost-effective ingredients. Sodium nitrate was substituted with
one of two nitrogen sources: ammonium nitrate (NH4NO3) or
urea [COÆ(NH2)2]. Four grades of nitrogen concentrations rep-
resenting 10%, 20%, 30% and 40% of SM nitrogen concentra-
tion (29.42 mM-N) were taken from ammonium nitrate
(Treatments 1–4) or urea (Treatments 5–8), respectively for test-
ing. Analytical grade potassium hydrogen phosphate (K2HPO4)
was substituted with single super phosphate fertilizer SSP
[CaÆ(H2PO4)2ÆH2O], to provide a source of phosphorus. Analyt-
ical grades of sodium bicarbonate (NaHCO3), potassium sulfate
(K2SO4) and sodium chloride (NaCl) were substituted with com-
mercial grades of NaHCO3, Muriat of potash (MOP) and crude
sea-salt (Syahat salt, NaCl raw material), respectively, with con-
centrations equivalent to that found in SM. The composition of
SM and the Reduced Cost media are shown in Table 1.
Sterilized Erlenmeyer flasks (500 mL) containing 100 mL of
either SM or the Reduced Cost media, were used as culture
units. Cultures were initially inoculated with S. platensis bio-
mass concentration of 0.06 mg/L dry weight. The experiment
was carried out in triplicates for 33 days at 30 ± 2 C, pH 9,
under 30 lEm2 s1 irradiance using cool white fluorescent
lamps with a photoperiod cycle of 12:12 h light/dark and thrice
daily shaking by hand.
Multiplication characteristics evaluation
Biomass concentration was determined every three days by mea-
suring the optical density at k 560 nm to produce a standard
curve (Leduy and Therien, 1977). This standard curve was sub-
sequently used to calculate the biomass of individual samples
based on their optical density. The dry weight of biomass was
determined by filtration of sample (10 mL) through dried pre-
weighed Gelman GA-6 filter (Ø 47 mm and nominal pore size
0.45 lm). The biomass obtained was washed twice with distilled
water, dried at 80 C for 4 h, cooled in desiccator and the result-
ing dry weight determined (Olguin et al., 2001). Chlorophyll
content was determined by filtering 5 mL of cultivation medium
in late exponential phase through GF/C filters (Ø 47 mm and
nominal pore size 0.45 lm). The material was submitted to
extraction with acetone and chlorophyll a was measured
spectrophotometrically according to American Public Health
Production and nutritive value of Spirulina platensis in reduced cost media 53

Table 1 Chemical composition of Zarrouk’s medium and the new Reduced Cost media.
Zarrouk’s medium Reduced Cost media
Component Concentration Component Concentration
g/L (mM) N Nitrogen sources Treatments g/L (mM) N
NaNO3 2.5 29.42 NH4NO3 types 1 0.118 2.94
K2HPO4 0.5 2 0.235 5.88
K2SO4 1 3 0.353 8.83
NaCl 1 4 0.471 11.77
MgSO4Æ7H2O 0.2 CO (NH2)2 types 5 0.088 2.94
CaCl2Æ2H2O 0.04 6 0.176 5.88
FeSO4Æ7H2O 0.01 7 0.265 8.83
EDTA 0.08 8 0.353 11.77
NaHCO3 16.8 Non-nitrogen sources
Micronutrient* 1 mL Single super phosphate 1.25
Muriate of potash 0.898
Crude sea-salt (Syahat salt) 1
Commercial grade NaHCO3 16.8
Distilled water 1L Filtered, boiled and cool tap water 1L
*
Micronutrient solution: H3BO3, 2.86; MnCl2Æ4H2O, 1.81; ZnSO4Æ4H2O, 0.222; Na2MoO4, 0.0177; CuSO4Æ5H2O, 0.079 (g/L). Note: Non
nitrogen’s sources were added in all treatments.

Association (APHA) (1985). The growth induces of S. platensis,
was evaluated using the following equations:
(1) Cell productivity (Px) according to Ogbonna et al.
(1995)
Px ðmg=L=dayÞ ¼ ðXm  Xi =tm Þ;
where: Xi = initial cell concentration (mg/L), Xm = maxi-
mum cell concentration (mg/L), tm = cultivation time related
to maximum cell concentration (days).
(2) Maximum specific growth rate (lm) and doubling time
(td) according to Levasseur et al. (1993),
lm ðdiv=dayÞ ¼ ln ðX2 =X1 Þ=t2  t1
td ð=dayÞ ¼ ln 2=l ¼ 0:693=l;
where: X1 = cell concentration at time t1, X2 = cell concentra-
tion at the time t2.
Biochemical analysis
Aliquots from culture units in late exponential phase were fil-
tered through GF/C filters (pore size 0.45 lm) and kept frozen
at 80 C for biochemical composition analysis. For protein
analysis, filters were homogenized with 5 mL of 10% trichloro-
acetic acid to precipitate protein (Dortch et al., 1984), and the
protein concentration was determined using the method de-
scribed by Lowry et al. (1951) and modified by Clayton (1988)
after re-dissolving precipitated protein in NaOH (1 M). Total
cellular carbohydrate was determined according to Dubious
et al. (1956) and total lipids were estimated using sulfophosph-
ovanilin method as described by Chobral and Castelleno (1961).
Statistical analysis
Data were statistically analyzed by one-way analysis of vari-
ance (ANOVA) using SPSS version 17 (SPSS Inc., Chicago,
IL, USA). Duncan’s multiple range test was used to compare
differences between treatment means when significant F values
were observed, at (p 6 0.05) level (Duncan, 1955).
Results
The growth rates of S. platensis, expressed as dry weight (mg/
L), in SM and the Reduced Cost media are shown in Fig. 1a
and b. The growth curves lacked a lag phase for both SM
and the Reduced Cost media. A rapid increase in biomass val-
ues was observed in SM giving the maximum biomass values
(0.840 ± 0.008 mg/L on the 15th day). In contrast, a gradual
increase in the dry weight was seen at all concentrations of
the Reduced Cost media but with different rates of increase.
The growth curves for different concentrations of the Reduced
Cost media showed similar growth rate behavior with peak
biomass concentrations being observed between the 27th and
33rd days. In each case the peak biomass value differed.
In cultures supplemented with ammonium nitrate, the in-
crease in the concentration of ammonium nitrate was associ-
ated with increased growth rate in treatments up to number
3, the growth rate was greatly reduced with the addition of a
higher concentration of ammonium nitrate (Treatment 4).
The maximum biomass obtained at different ammonium
nitrate concentrations was 0.591 ± 0.018 and 0.813 ±
0.018 mg/L for Treatments 4 and 3, respectively, with a longer
exponential phase for the Treatment 4 (Fig. 1a). In Contrast,
cultures supplemented with urea showed slower growth rates,
when compared to either SM or ammonium nitrate media,
reaching the end of exponential phase between 27 and 33 days
of cultivation. The growth rate and maximum biomass values
were inversely correlated with the concentration of urea. The
maximum quantity of biomass was lower than that recorded
in SM and ammonium nitrate media, ranging between
0.360 ± 0.003 and 0.640 ± 0.006 mg/L for Treatments 8 and
5, respectively (Fig. 1b).
Table 2 shows the maximum chlorophyll a concentration
(chl am), cell productivity (Px), maximum specific growth rate
54
Biomass concentration (mg/L) 0.90
(a)
0.80
0.70
0.60
0.50
0.40
0.30
0.20
0.10
0.00
3 6 9 12 15 18
0.90
(b)
0.80
Biomass concentration (mg /L)
0.70
0.60
0.50
0.40
0.30
0.20
0.10
0.00
3 6 9 12 15 18 21 24
Growth time (days)
Figure. 1 Changes in biomass concentration of S. platensis (mg/
L) grown on Zarrouk’s medium and the Reduced Cost media
supplemented with different concentrations of (a) ammonium
nitrate (Treatments 1–4), and (b) urea (Treatments 5–8). Each
value is the mean of three samples.
(lm) and doubling time (td) found for the various treatments.
In all variants of the new medium, S. platensis produced lower
values of chl am, Px and lm coupled with an increased doubling
time compared with that observed in SM. The growth param-
eters showed the same trends seen with changes in dry weight:
values of chl am, Px and lm showed a direct increase with
increasing concentration of ammonium nitrate up to Treat-
ment 3 then decreased for Treatment 4; while parameters de-
creased with increasing urea concentration. Values of chl am
for SM and Treatment 3 (0.0701 ± 0.0089 and 0.0685 ±
0.0024 lg/L, respectively) were close although their Px
(0.052 ± 0.0005 and 0.028 ± 0.0006 mg/L/day, respectively)
and lm (0.321 ± 0.010 and 0.233 ± 0.016 div/day, respec-
tively) were significantly different (p < 0.05). Although the
highest values of cell productivity (Px) were recorded on differ-
ent days of incubation for SM and different concentrations of
the Reduced Cost media, the maximum specific growth rate
(lm) for all culture was observed between the 3rd and 6th days.
The percentages of protein (52.95 ± 0.53), carbohydrate
(13.20 ± 0.57) and lipid (7.16 ± 0.68) contents obtained with
SM were comparable with that observed for Treatment 3
(Table 3). Compared to SM, protein content of S. platensis cul-
tivated in all concentrations of urea and Treatment 4 was sig-
nificantly decreased (ranging between 37.79 ± 7.64 and
47.10 ± 6.67), while carbohydrate content was increased
(ranging between 16.01 ± 4.24 and 24.50 ± 2.49). Lipid con-
tent showed different pattern with insignificant lower values
in ammonium nitrate and significant higher values (p < 0.05)
in urea (ranging between 5.64 ± 0.11 and 15.39 ± 1.32).
F.F. Madkour et al.
Discussion
Zarouk
In mass cultures of microalgae, nutrition condition is one of
the key factors that controls their growth and productivity
1
(Vonshak and Richmond, 1988; Faintuch et al., 1991). This
2
investigation was conducted with the basic aim of providing
3
a simple and inexpensive medium to decrease the cost of large
4
scale production of Spirulina. This intention was implemented
by substituting all the nutrients of Zarrouk’s medium with
cheaper and locally available commercial fertilizers and
21 24 27 30 33 chemicals.
Among the nutrients required for the growth of algae,
phosphorus is considered as an important element which plays
an essential role in maintaining high production rates of mic-
roalgae (Mostert and Grobbelaar, 1987). Commercial agricul-
Zarouk
ture fertilizer, SSP, was selected as a source of phosphorus
5
(16% P2O5), and as a supplier of (EDTA) and some essential
6 micronutrients, since it contains these elements as impurities.
7 Raoof et al. (2006) have grown successfully S. platensis in Zar-
ouk’s medium substituted with SSP in place of K2HPO4,
8
EDTA and A5 micronutrient solution. Becker (1994), reported
that the best required phosphate concentration for growth of
27 30 33 microorganisms falls in a wide range (between 0.05 and
20 mg/L), and does not depend on the concentration of other
nutrients. However, the concentration of phosphate in the
present study was adjusted to be the same as SM, which is
above Becker’s concentrations. In addition, SSP contains rea-
sonable amount of calcium (19–25%) and sulphur (8–12%)
which meets the requirement for these nutrients.
Other expensive chemical, such as K2SO4, were substituted
with using commercial grade fertilizer, MOP. The concentra-
tion of MOP was adjusted to supply amount of potassium
equivalent to that provided by K2SO4 in SM. The advantage
of utilizing crude sea-salt (Syahat salt) was that the macro
and micro-nutrients required for S. platensis growth are mainly
provided by this kind of low-cost raw material.
Changes in nitrogen source and amount of cultivated media
limit the intensive growth of microalgae and alter their pig-
ment and biochemical composition (Mostert and Grobbelaar,
1987). Cyanobacteria preferentially use inorganic nitrogen for
growth, particularly nitrates and ammonium but some of them
are able to grow on organic nitrogen (Fogg et al., 1973). In the
present study, different concentrations of two nitrogen sources
were tested to determine their effectiveness on productivity of
S. platensis. Ammonium nitrate and urea were chosen because
they are cheaper than others (e.g. potassium nitrate, sodium
nitrate or ammonium chloride), and contain two nitrogen
atoms (35% and 46% nitrogen, respectively) whereas other ni-
trates have only one (14–16% nitrogen).
When the response of S. platensis grown in SM and the Re-
duced Cost media was monitored, the results showed signifi-
cant differences in the growth parameters for the different
nitrogen sources. There was no lag phase in the growth curves
of SM and the new media because initial inocula were S. plat-
ensis cells that had been harvested in exponential phase, thus
the alga grew immediately, independent of different nitrogen
source. The final biomass yield in ammonium nitrate was com-
parable with that seen using SM but with a longer exponential
phase and greater doubling time (td). Growth in urea was poor
giving significantly lower biomass (p < 0.05) than that of
either SM or ammonium nitrate. In spite of the increased
Production and nutritive value of Spirulina platensis in reduced cost media 55

Table 2 Maximum chlorophyll (Chl am), cell productivity (Px), maximum specific growth rate (lm) and doubling time (td) of S.
platensis grown on SM and the Reduced Cost media.
Media Treatment Chl am (lg/L) Px (mg/L/day) lm (div/day) td (/day)
SM control 0.0701 ± 0.0089a 0.052 ± 0.0005a 0.321 ± 0.010a 2.157 ± 0.066f
NH4NO3 1 0.0580 ± 0.0059b 0.019 ± 0.0003e 0.157 ± 0.012e 4.419 ± 0.329b
2 0.0629 ± 0.0054ab 0.023 ± 0.0003c 0.192 ± 0.008d 3.606 ± 0.150cd
3 0.0685 ± 0.0024a 0.028 ± 0.0006b 0.233 ± 0.016b 2.978 ± 0.200e
4 0.0369 ± 0.0034cd 0.016 ± 0.0005f 0.096 ± 0.006f 7.198 ± 0.437a
Urea 5 0.0389 ± 0.0042c 0.021 ± 0.0002d 0.225 ± 0.020bc 3.088 ± 0.262de
6 0.0340 ± 0.0062cd 0.016 ± 0.0001f 0.209 ± 0.012cd 3.319 ± 0.190cde
7 0.0278 ± 0.0041d 0.014 ± 0.0002g 0.191 ± 0.007d 3.634 ± 0.139c
8 0.0143 ± 0.0005e 0.009 ± 0.0001h 0.099 ± 0.007f 7.015 ± 0.506a
All values are mean ± standard deviation (n = 3). Different superscript letters across a row indicate significant difference between means
(p < 0.05).

Table 3 Protein, carbohydrate and lipid contents (% dry weight) of S. platensis grown on SM and the Reduced Cost media.
Media Treatment Protein Carbohydrate Lipid
a d
SM control 52.95 ± 0.530 13.20 ± 0.575 7.16 ± 0.683d
NH4NO3 1 51.30 ± 1.357ab 16.30 ± 0.431cd 6.50 ± 0.173d
2 51.70 ± 5.962ab 17.15 ± 1.524cd 6.42 ± 0.280d
3 52.62 ± 0.911a 15.00 ± 1.431cd 6.50 ± 0.113d
4 44.07 ± 0.881 bc 24.50 ± 2.486a 5.64 ± 0.113d
Urea 5 47.10 ± 6.678ab 16.01 ± 4.236cd 11.01 ± 1.456c
6 45.71 ± 3.742ab 17.12 ± 3.096cd 11.31 ± 0.926c
7 43.00 ± 3.822cd 19.21 ± 1.761bc 13.01 ± 1.534b
8 37.79 ± 7.642d 22.05 ± 2.323ab 15.39 ± 1.315a
All values are mean ± standard deviation (n = 3). Different superscript letters across a row indicate significant difference between means
(p < 0.05).

length of the exponential phase and the increased td in ammo-
nium nitrate and urea when compared with SM may be due to
adaptation to the different media by S. platensis.
The results revealed that S. platensis could most efficiently
utilize ammonium nitrate as compared to urea. The high val-
ues of specific growth (lm) for ammonium nitrate confirmed
that the metabolic efficiency of the alga was best on ammo-
nium nitrate. Although urea has been recognized as a good
nitrogen source and successfully metabolized by algae (Baldia
et al., 1991), the present results indicated that S. platensis pre-
fers utilizing nitrogen from ammonium nitrate. The preference
of Spirulina for nitrate and ammonium salts as a nitrogen
source was demonstrated by Richmond (1988) who reported
that nitrates and ammonium salts were the main nitrogen
sources assimilated by Spirulina as well as urea in lesser con-
centration. The higher growth rate in ammonium nitrate
may be due to a greater efficiency of ammonia grown cells in
the conversion of photosynthetically generated reducing power
into net growth as mentioned by Rhee and Lederman (1983).
Several studies have shown that nitrate and ammonium ions
appear to be actively transported in cyanobacteria, while it en-
ters into the cells of alkalophilic species such as Spirulina by
passive diffusion (Boussiba, 1990; Rodriguez et al., 1994).
The growth of S. platensis was enhanced by increasing the
concentration of ammonium nitrate giving the maximum bio-
mass in Treatment 3, which was comparable with that of SM
while further increasing in the ammonium nitrate concentration
limited growth. On the other hand, the growth rate of S. platen-
sis in urea showed a significant decrease associated the increase
in urea concentration giving the best biomass production at the
lowest concentration (Treatment 5). This may be due to the
inhibitory effect of urea on Spirulina production at concentra-
tion more than 0.3 g/L as mentioned by Richmond (1990). Filali
et al. (1997) concluded that among inorganic and organic nitro-
gen sources, nitrate is the more convenient because it may be
provided to the culture at high concentrations while ammonium
and urea are toxic at concentrations higher than 2 mM. The
combination between nitrate and ammonium sustained the
same efficiency as nitrate if maintained at lower concentrations,
giving higher growth rates and reduced toxicity. This may ex-
plain our finding that to obtain the maximum biomass of S. plat-
ensis, the concentration of ammonium nitrate or urea should not
be more than 8.83 or 2.94 mM N, respectively.
For a long time, condition of the culture nutrition has been
recognized one of the important factors that plays a major role
in determining the biochemical composition of microalgae
(Mostert and Grobbelaar, 1987). The present results revealed
that the presence of variations in the protein, carbohydrate
and lipid contents produced in respect to nitrogen source type
and concentration. Although these variations in some treat-
ments were not large, significant differences (p < 0.05) were
observed in many treatments. Considering, ammonium nitrate,
the higher concentrations gave higher values for cellular
proteins and carbohydrates. Since nitrogen is required for
56
synthesis of the amino acids, which make up proteins, the in-
crease in nitrogen concentration caused an increase in protein
biosynthesis in Spirulina. With regard to lipids, higher concen-
trations of ammonium nitrate showed no increase in their lev-
els. The same finding was obtained by Piorreck et al. (1984),
who found that for cyanobacteria, total lipid content remained
constant at all nitrogen concentrations studied. On the other
hand, the biochemical composition of Spirulina grown on med-
ia supplemented with urea was changed significantly when
compared with SM giving lower production of protein and
higher production of both carbohydrates and lipids. More-
over, increasing the concentration of urea favored the accumu-
lation of carbohydrate and lipids rather than proteins. The
changes in biochemical composition are considered to be an
adaptation mechanism to toxicity of high urea concentration
causing the accumulation of carbohydrates and lipids content
in the alga at the expense of protein production.
In terms of the costs of local chemical companies’ prices of-
fers, it was found that the preparation of 1000 L of SM would
EGP 480 (US$ 80) compared to EGP 78 (US$13) for 8.83 mM
N as ammonium nitrate media (Treatment 3). This emphasizes
the merits of the new modified medium, not only as a low cost
alternative but also as a highly productive input, which can be
used profitably by the rural population for large-scale biomass
production of protein-rich Spirulina.
Conclusion
Although, sodium nitrate seemed more optimal for S. platensis
growth, ammonium nitrate and urea can serve as alternative
nitrogen sources, since this cyanobacterium shows utilization
of these nitrogen sources at low concentrations. The maximum
dry weight, chlorophyll and protein yields were obtained using
SM and the Reduced Cost medium containing 8.83 mM N as
ammonium nitrate. In these media, the observed yields were
statistically comparable. Any further increase in the ammo-
nium nitrate concentration led to a considerable decrease in
biomass and other parameters. This indicates the critical
importance of the level of nitrogen in large scale production.
The present study clearly indicates that the newly prepared Re-
duced Cost medium supplemented with ammonium nitrate as
nitrogen source is comparable with Zarrouk’s medium regard-
ing growth performance of Spirulina in terms of dry biomass,
chlorophyll and chemical composition. Therefore, cultivation
in 8.83 mM N as ammonium nitrate can be exploited when
the purpose is to produce high biomass density of S. platensis
with good nutritional characteristics. From a practical point
of view, these results draw attention to the importance of
selecting the source and concentration of nitrogen in phyto-
plankton cultures, because it may modify the metabolic activ-
ities and consequently, the composition and nutritional value
of microalgae, which is important to consider in aquaculture
practices.
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