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ABSTRACT
The aim of this study was to determine the effect of differences addition of culture medium
concentrations on the growth of microalgae type Chlorella sp. Samples were obtained from
the Center for Brackish Water Cultivation Fisheries (BBPBAP), Jepara. This research was
conducted at the Laboratory of Nutrition and Natural Feed Sorong Marine and Fisheries
Polytechnic with an experimental method using four treatments with addition of walne
medium, there were A (adding only at the beginning of cultivation); treatment B (adding
every day); C (adding every two days), and treatment; D (adding every three days). The cell
growth performance was significantly found in treatment C with the addition of nutrients
every two days with a peak cell density of Chlorella sp was 2,628,450 cells / ml. There is a
need for mass culture assay to determine the level of effectiveness and efficiency.
Keywords: Chlorella sp.; growth performance; natural food
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Jurnal Airaha, Vol.10, No.02 (Dec 2021):295 – 301, p-ISSN 2301-7163, e-ISSN 2621-9638
(Chilmawati, 2008). In addition, walne also cultivated on a laboratory scale for seven
contains 83.9 mg/l nitrate and 17.7 mg/l days in an Erlenmeyer with sterile seawater.
phosphates (Ajijah et al., 2020). An inoculum volume was 100 mL were
Several researchers have made efforts added into 200 mL of sterile seawater. Walne
to increase nutrients in Chlorella sp. through dosage was added at the beginning about 2
various media. They ranged from using mL/L of total volume then were kept under
inorganic fertilizers in mass-scale culture to temperature of 23oC, 30 ppt salinity, lighting
grow Chlorella sp. (Rahardini et al., 2018), 1 for 24 hours, and adequate aeration provided
g/L sucrose and 10% inorganic carbon (CO2) by blower.
to increase the number of Chlorella sp. (Lin Research methods
& Wu, 2015), and adding of 0.5 ml/L walne This research was an experimental
fertilizer as a source of microalgae study with a completely randomized design
(Chlorella sp.) nutrition (Purnamawat et al., (CRD) consisting of four treatments and
2014). This study aimed to determine the three replications. The independent variable
effect of differences of culture medium in this study was the difference time in
concentration of microalgae Chlorella sp. nutrient concentration adding of the Walne
fertilizer, while the dependent variable was
METHODS the population of Chlorella sp. Thus, A
Time and Location group was added fertilizer only at the
This study was conducted on January, beginning; B group was added fertilizer in
2019, at the Natural Feed and Nutrition every day; C group was added fertilizer each
Laboratory of the Sorong Marine and two days; and D group was added fertilizer
Fisheries Polytechnic. each three days.
Procedures The number of cells was observed
Materials Preparation. using an electric microscope at 40x
Materials were used in this study was magnification with five fields of view and
500 mL Erlenmeyer flask (Pyrex); Dropper three replications for each view. The
pipette (BRAND) were washed with observations were made from day 0 (t2) until
detergent and rinsed using flow water then day 7 (t1) every 24 hours. The observations
were dried. Seawater were filtered by filter of the number of cells were used to calculate
paper and sterilization by adding 60 ppm of cell density.
chlorine then aerated for 24 hours. Chlorine Data Analysis
on the seawater was neutralized using 20 The observed growth parameters of
ppm Na-Thiosulfate. The 200 mL of Chlorella sp. consisted of peak population,
seawater were prepared before were added specific growth rate, and cell density. The
into 500 mL Erlenmeyer flask and wrapped peak population was seen from the highest
using alumunium foil then were prepared for cell density value during the culture period.
sterilization to avoid contamination by using Microalgae cell density was calculated based
autoclave with temperature of 121oC for at on the formula of (Hutami et al., 2015) as
least 15-20 minutes under 1 psi of preasure. follows:
The seawater were kept at room temperature 1000 x n
N=
(23oC) until use. LP x p
Cultivation of Chlorella sp. Note:
The microalgae seeds used in this study N : the density of Chlorella sp.
were Chlorella seeds which were harvested n : the number of observed Chlorella sp.
after 7 days and then can be used for LP : field of view
reculture. The seeds was pure Chlorella sp. P : the number of field of view
which was obtained from Balai Besar The specific growth rate of microalgae
Perikanan Budidaya Air Payau (BBPBAP) (µ) was calculated using the formula by (Das
Jepara, West Java. Chlorella sp. was et al., 2011) as follows :
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Jurnal Airaha, Vol.10, No.02 (Dec 2021):295 – 301, p-ISSN 2301-7163, e-ISSN 2621-9638
2,000,000 concentrations
1,500,000 A Treatmen Growth parameter
1,000,000 B t -1
500,000 µ (day ) Peak Average N
C
- Populatio (cell/mL)
D n
0 1 2 3 4 5 6 7
(cell/mL)
Culture period (days) A 0.53 ± 2,540,694 1,560,206a
0.08b
The cultivation results of microalgae B 0.61 ± 2,384,996 1,501,365a
Chlorella sp with different nutrient 0.15b
C 0.83 ± 2,628,450 1,814,175a
concentrations are shown in Figure 1. The 0.13a
density cell of Chlorella sp. in each treatment D 0.65 ± 2,587,403 1,667,172a
increased from the beginning of cultivation 0.08b
to the peak of the highest population and then Based on ANOVA, Chlorella sp.
decreased. A group showed increasing showed a different significance (p<0.05) was
microalgae cell continuously from the the C group with the highest specific growth
beginning cultivation to seventh day. B group rate than other groups. The highest cell
showed increasing in cell density from the density (peak population) of C group was
first day to the sixth day cultivation, then 2,628,450 cell/mL with an average 1,814,175
decreased in the seventh day. C group cell/mL (Fig.1). Moreover, another group
showed an increasing in cell density from the showed no significant difference with the
first day to the fifth day, than decreased on lowest specific growth rate being A group,
the sixth day to seventh day. D group but the lowest cell density was B group with
experienced an increase in the number of cell value 2,384,996 cells/mL, in an average cell
density from the first day to the sixth day and density was 1,501,365 cells/mL.
decreased on the seventh day.
Figure 1. Growth rate phase of Chlorella sp. DISCUSSION
with varying nutrient concentrations The nutrient addition of walne fertilizer
Cell density of Chlorella sp. showed an every two days could increase the cell
increase from the first day to the seventh day density of Chlorella sp. The nutrients needed
in almost all treatments. Increased cell by phytoplankton were well utilized and
density due to Chlorella sp. utilized the reached an exponential phase significantly,
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Jurnal Airaha, Vol.10, No.02 (Dec 2021):295 – 301, p-ISSN 2301-7163, e-ISSN 2621-9638
compared to the other treatment groups. the culture media. The greater the number of
Generally, microalgae growth is divided into particles in the culture medium, the longer
4 (four) phases which are lag phase, log or the time required to pass through the
exponential phase, stationary phase and death adaptation phase. These particles can
phase (Moazami et al., 2012). The increase the value of turbidity, BOD and
availability of nutrients in a microalgae COD so that photosynthesis becomes more
growth medium affects the growth rate. inhibited and also slows down the growth
Futhermore, (Sánchez et al., 2000) stated that rate of Chlorella sp. (Dianursanti et al.,
the culture medium greatly affects the 2014). Then the D group showed a lower
biomass yield and protein content of level of cell density as well, this is
microalgae (Chlorella). Based on R. Ramaraj presumably due to the nutrients that can be
et al., (2016) and Rameshprabu Ramaraj et used by Chlorella sp. enough but not
al., (2015), explained that algae are easy to optimal. Nutrient composition plays an
grow and cultivate anywhere with less important role in microalgae cultivation,
energy requirements and few nutrients. excessive or depletion source of nutrient
However, the addition of nutrients to might affect the biomass quality (Yaakob et
Chlorella culture media made a significant al., 2021). Dianursanti et al., (2014) stated
contribution to growth and could even that Chlorella biomass growth using Walne
increase biomass productivity and nutritional medium was more stable than using liquid
value (Kim et al., 2013; Blair et al., 2014). B waste from tofu because Walne media
group showed differences in the level of cell contained micro nutrients such as Fe, Mn,
density compared to the C group, presumably Mg and Cl.
due to the nutrients were given was excesive
that Chlorella sp unable to absorb nutrients CONCLUSION
effectively, leading to the buildup of toxic Statistically, the addition of culture
organic matter that decreased growth medium every two days (C group) showed a
eventually. Excessive nutrients will be toxic significant difference (P<0.05) on the
to microalgae growth, causing growth to be specific growth rate of microalgae Chlorella
suboptimal (Arinta, 2012; Swandewi et al., sp. with a value of 0.83 ± 0.13 µ (day -1).
2017). The specific growth rate can also be However, the difference in nutrient
used as an indicator of the carrying capacity concentration does not have a different effect
of the medium and the availability of energy on the cell density of Chlorella sp. The
for cells to divide (Wahyuni et al., 2019; highest population peak was found in
Musa et al., 2013). The low growth rate may treatment C, with a value of 2,628,450
be caused by nitrate and phosphate, which cells/ml. There is a need for mass culture
cells cannot adequately utilize for growth and assay to determine the level of effectiveness
division. According to Arinti, (2012) and and efficiency.
Swandewi et al., (2017), excess phosphate
concentration will inhibit the process of REFERENCES
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Jurnal Airaha, Vol.10, No.02 (Dec 2021):295 – 301, p-ISSN 2301-7163, e-ISSN 2621-9638
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