4x3 2x298x311

Transactions on Science and Technology Vol. 4, No.
3-2, 298 - 311, 2017
Growth and Biomass Production of Native

Microalgae Chlorella sp., Chlamydomonas sp.
and Scenedesmus sp. Cultivated in Palm Oil
Mill Effluent (POME) at Different Cultivation
Conditions
Harizah Bajunaid Hariz1#, Mohd Sobri Takriff2
TRANSACTIONS ON SCIENCE AND TECHNOLOGY
1 Faculty of Engineering and Built Environment, Chemical and Process Engineering, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, MALAYSIA.
2 Research Center for Sustainable Process Technology (CESPRO), Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, MALAYSIA.
# Corresponding author. E-Mail: [email protected]
ABSTRACT
The main objective of this research was to study native microalgae Chlorella sp., Clamydomonas sp. and Scenedesmus
sp. growth and biomass productivity at different source of nutrient, inoculum concentration and aeration factor.
Agricultural wastewater, palm oil mill effluent (POME) was used as a nutrient source which was compared with Bold’s
Basal Medium (BBM) by culturing the microalgae at different inoculum concentration in 1-Litre transparent vessel with
700ml of working volume. Fixed aeration rate, 1L/min was set for all with 14000 lux of light intensity at 25±2°C cultivation
temperature. The results were also compared with the non-aerated cultures which only mixing was being provided for the
microalgae growth. Based on the growth rate, 30% of inoculum shows the highest growth rate for all these three
microalgae species Chlorella sp. (µmax =0.2712), Chamydomonas sp. (µmax =0.2547) and Scenedesmus sp. (µmax
=0.1867) cultured in POME with air being supplied as the aeration factor. However, in BBM 30% of inoculum for
Chlamydomonas sp. (µmax =0.5082) and Scenedesmus sp. (µmax =0.3402) manifest highest growth rate while
Chlorella sp. does not exhibit significant effect at different inoculum concentration. The non-aerated culture condition
shows 20% of inoculum for Chlamydomonas sp. (µmax =0.125) and Chlorella sp. (µmax =0.2052) cultures give the
highest growth rate with POME. Similar results were obtained for Chlamydomonas sp. (µmax =0.1673) and Chlorella sp.
(µmax =0.1855) cultured in BBM at non-aerated condition. Scenedesmus sp. shows the highest growth rate with 10% of
inoculum at the non-aerated condition for both POME (µmax =0.1900) and BBM (µmax =0.1975). From the growth
curves, the adaptation (lag phase) and exponential period of microalgae species were determined. Based on the results,
all 3-species cultured in POME required 6 days for the lag phase and 14 days for the exponential phase which is doubled
the period taken for BBM as the nutrient medium.
KEYWORDS: Microalgae, Wastewater, Palm Oil Mill Effluent (POME), Culture Condition, Nutrient Source
Data With Detail Description - Biological sciences
Received 11 January 2017 Revised 1 August 2017 Accepted 2 November 2017 Online 3 November 2017
© Transactions on Science and Technology 2017
INTRODUCTION
Microalgae capable of consuming nutrients for growth and absorb toxin in suspension. Apart
from that, microalgae can be used in CO2 fixation as it undergoes photosynthesis process where CO2
will be consumed for growth and O2 will be released as the respiration product. The simple cell
structure of microalgae with a large surface area per unit volume of microalgae cell explained the
rapid growth and cell division of microalgae. This special characteristic of microalgae allows them to
consume nutrients at a high rate of consumption (Khan et al., 2009). Microalgae cultivation under
optimum growth conditions allows microalgae optimum cell division for every 3-4 hours. However,
most species will take 1-2 days for the population to increase (Williams & Laurens, 2010). Microalgae
metabolic mode can be divided into a few categories; heterotopic, autotrophic, mixotrophic,
photoheterotrophic, and photoautotrophic (Brennan & Owende, 2010; Liang et al., 2009). Some
microalgae species can practise more than one metabolic mode at the same time. This phenomenon
Hariz & Takriff, 2017. Transactions on Science and Technology. 4(3-2), 298 - 311
Hariz & Takriff, 2017. Transactions on Science and Technology. 4(3-2), 298 - 311 299
happens to several microalgae species that has been studied such as Chlorella sp. and Scenedesmus sp.
that are able to switch from autotrophic to heterotrophic mode depending on the source of nutrients
available for utilization. Other studies reported that Chlamydomonas sp. also practises more than one
metabolism mode when it is cultivated in POME and this multimode attribute is known as
mixotrophic metabolism (Ding et al., 2016).
Medium design and composition influence the growth and performance of microalgae.
Insufficient nutrients provided during microalgae cultivation will slow down the growth rate and
productivity of microalgae. There are growth and productivity requirements that need to be fulfilled
in microalgae cultivation such as optimal supply of macronutrients (carbon, nitrogen, phosphorus),
micronutrients (Ca, Mn, Fe, Zn,Cu), growth factors (vitamins, amino acids), functional nutrient for
growth and product formation (non-growth associated and growth associated), physical factors
(temperature, light, water activity, gas exchange) as well as the addition of additives (protectant,
chelating and neutralizing agent). Commonly, commercial medium is used by most microalgae
industries for cultivation. The cost for the medium sometimes exceed the gain from the biomass
production. This has proven that commercialized medium is not feasible for the industries. Thus, the
high cost of commercial medium for microalgae cultivation triggers the idea of using an available
nutrient source from wastewater as an alternative way of microalgae cultivation. The high content of
organic materials in wastewater such as POME provides low cost microalgae cultivation method
which will benefit the industries for a longer-term period.
Palm oil mill effluent (POME) is one of the wastes coming out from palm oil production
industry. POME is an unpleasant, non-toxic suspension waste generated from palm oil milling
process where a large amount of water and steam are being utilized and resulted in wastewater
discharged. Native microalgae strains isolated from the local environment such as palm oil mill
effluent and rubber waste water are used in current research to avoid biosafety issue and enhance
adaptability of microalgae. This native microalgae strains will have high tolerance as it originates
from the same environmental condition. The main objective of current research is to evaluate native
microalgae growth and biomass productivity at different source of nutrient, inoculum concentration
and aeration factor. Initial inoculum concentration is one of microalgae growth factors which need to
be optimized. Higher inoculum concentration support growth and lessen the adaptation period of
microalgae (Lau et al., 1995). Study done by (Khalid et al., 2016) shown the increment of inoculum
concentration promoted better growth of the Characium sp. grown in POME. Other study has came
out with the finding that higher inoculum concentration resulted in better biomass production (Ma
et al., 2012). However, too high inoculum concentration might cause internal shading phenomenon
and stunted the growth (He et al., 2015). Thus optimization of initial microalgae seed concentration
is important to counteract this issue. In this experiment, microalgae growth was totally depended on
the atmospheric air. The ambient CO2 percentage is 0.03% v/v which is below the microalgae growth
requirement (Becker,1994). Mixing is important to provide sufficient turbulence to the medium and
promote gas exchange between medium and the surrounding which will also overcome extreme pH
condition (Gross, 2003). In addition, turbulence can enhance the microalgae productivity by
ensuring the well distributed nutrient reaching microalgae cell for microalgae utilization and
accumulation of organic matters can be avoided (Hariz et al., 2017). Thick boundary layers between
microalgae cell and the nutrient medium will be reduced and improve microalgae nutrient uptake
rate (Mostert & Grobbelaar , 1987; Borowitzka ,1998). Sufficient turbulence for optimum mass
transfer rate of nutrient and gas, promote light penetration by dispersing the accumulated
particulate matters (Gross, 2003) at the same time to avoid energy wastage.
ISSN 2289-8786. http://transectscience.org/

METHODOLOGY
Microorganism and Growth Medium

Native microalgae Chlorella sp. and Chlamydomonas sp. were isolated from palm oil wastewater
while Scenedesmus sp. was isolated from rubber wastewater and the cultures were maintained in
Bold’s Basal Medium (BBM) at the temperature 25ºC. BBM is the commercial nutrient medium that
consists of nutrient as stated in Table 1.
Table 1. Nutrient elements contain in BBM and Anaerobic POME

BBM Nutrient Concentration Anaerobic POME Nutrient Concentration
Element (mol/L) Element (mg/L)
NaNO3 2.94×10−3 M Ammoniacal-Nitrogen 387
(NH3-N)
CaCl2 2H2O 1.70×10−4 M Ammonium (NH4) 498
MgSO4 7H2O 3.04×10−4 M Phosphorus (P) 75
K2HPO4 3H2O 4.31×10−4 M Phosphate (PO4-3) 147
KH2PO4 1.29×10−3 M Chemical Oxygen Demand 705
(COD)
NaCl 4.28×10−4 M Biological Oxygen 388
Demand (BOD)
EDTA 1.71×10−4 M
KOH 5.53×10−4 M
FeSO4 7H2O1.79×10−5 M
H3BO3 1.85×10−4 M
Trace elements ZnSO4·7H2O3.07×10−5 M,
MnCl2·4H2O7.28×10−6 M,
MoO3 4.93×10−6 M,
CuSO4·5H2O6.29×10−6M,
Co(NO3)2·6H2O1.68×10−6 M
Other than BBM, anaerobic palm oil mill effluent (POME) was used as nutrient growth medium.
Anaerobic POME was collected from the palm oil mill and stored at 4OC. The POME sample was
then centrifuged at 8000rpm for 10 minutes before being used for the experiment. This to ensure the
suspended solid is removed from the POME sample before being introduced for microalgae
cultivation. The characteristic of anaerobic POME used were showed in Table 1.
Cultivation Conditions and Analytical Determinations

The experiments were carried out in 1000 ml Erlenmeyer flasks containing growth medium,
POME and BBM at different ratio of inoculated microalgae as stated in Table 2 and Table 3. The
working volume for the experiment was 700ml for each flask and the remaining space was for the
headspace to promote gas exchange. The runs were carried out in triplicates to ensure the
consistency of the result. The same ratio of growth medium and inoculum concentration was used
for all the three microalgae strains with different nutrient growth medium POME and BBM. For the
non-aerated experiment set up, non-absorbance cotton wool was used as a stopper to allow gas
exchange between the atmosphere air and the cultivated microalgae. The cultures were mixed at a
fixed speed of 100 rpm by using the orbital shaker (SHO-1D Wise Shake). For the aerated experiment
set up, fixed aeration rate at 1 Liter/min of air was supplied into the flasks by using the air pump
(Big Boy BB-10200). The temperature for both experiments were maintained at 25±2°C with light
exposure of 14000 lux of fluorescent lamp (Hitachi, Malaysia). The sample was taken daily, and the
microalgae biomass was determined by dry cell weight (DCW) analysis. The activity was done by
using 0.45 µM micro fiberglass filter paper (GF/C) to filter the biomass and dry in 105°C drying oven
for 24 hours. 10ml of samples were collected from the flasks by using measuring string and the
samples were then centrifuged at 8000rpm for 10 minutes by using the centrifuge (Centrifuge 5804,
Eppendorf) to separate the biomass from the supernatant. The supernatant was discarded, and
distilled water was added to the remaining pellet of microalgae biomass and the liquid mixture was
centrifuge for the second time with the same centrifugation speed. This to ensure the dissolved solid
is being removed out of the sample before filtration process. The control experiment set up was
prepared without microalgae being introduced into the growth medium for each experiment
condition and it was ran together with the other samples. This is important for the analysis purpose
as the filtered pellet from this control set up is used to determine the amount of suspended solid
contained. For the biomass calculation, the following is the formula:
DCW = (weight of filtered microalgae biomass – weight of blank filter paper)

Volume of sample collected (1)
(a) (b)
Figure 1. Experiment set up for microalgae cultivation in BBM a) Cultivation of microalgae with air
supply from the air pump; b) Cultivation of microalgae with mixing by using shaker
Table 2. Microalgae cultivation in BBM at different inoculum concentration
Inoculum Concentration 10% 20% 30%

Volume of Inoculum (ml) 70 140 210
Volume of BBM (ml) 630 560 490

(a) (b)
Figure 2. Experiment set up for microalgae cultivation in POME a) Cultivation of microalgae with
air supply from the air pump; b) Cultivation of microalgae with mixing by using shaker
Table 3. Microalgae Cultivation in POME at different inoculum concentration
Inoculum Concentration 10% 20% 30%

Volume of Inoculum (ml) 70 140 210
Volume of POME(ml) 630 560 490
RESULT AND DISCUSSION

Based on the results from dry cell weight analysis, growth curve can be constructed from the
logistic model by using MATLAB. The biomass productivity, ρ and specific growth rate, µ can be
obtained from this logistic model (Yang et al., 2011).
X = X0 Xmax е µmax t / ((Xmax-X0) + X0 е µmaxt) (2)

x = microalgae concentration in the medium.
Table 4. Growth rate of microalgae cultivated with aerated growth condition
Nutrient Growth
Bold Basal Medium (BBM) Palm Oil Mill Effluent (POME)
Medium
Inoculum
10% 20% 30% 10% 20% 30%
Concentration (v/v)
Native Microalgae Growth Rate (μ )

Strain max
Chlorella sp.
0.3231 0.3636 0.3537 0.2270 0.1517 0.2712
UKM2
Chlamydomonas sp.
0.3076 0.2574 0.5082 0.0968 0.1260 0.2547
UKM6
Scenedesmus sp.
0.2037 0.2369 0.3402 0.1557 0.1732 0.1867
UKM9

Table 5. Growth rate of microalgae cultivated with non-aerated growth condition
Nutrient Growth
Bold Basal Medium (BBM) Palm Oil Mill Effluent (POME)
Medium
Inoculum
Concentration 10% 20% 30% 10% 20% 30%
(v/v)
Native Microalgae Growth Rate (μ )

Strain max
Chlorella sp.
0.1635 0.1855 0.1445 0.1401 0.2052 0.0970
UKM2
Chlamydomonas
0.1172 0.1673 0.1562 0.0981 0.125 0.0980
sp. UKM6
Scenedesmus sp.
0.1975 0.0925 0.1352 0.1900 0.1629 0.0797
UKM9
Cultivated microalgae strains with air supply as the aeration source showed higher growth rate
compare to microalgae growth which mixing is provided as shown in Table 4 and Table 5. The
reason this happened might be because of the turbulence factor in aerated condition enhance the
growth of the microalgae strain in both BBM and POME. Turbulence as a result of air flux,
producing different movement regimes of the cells within the media under an equal light intensity,
could be an important factor. Adequate turbulence is important in this case of microalgae cultivation
condition as it enhances mass transfer rate of nutrient to microalgae cell, promote gases exchange
that can overcome photo-oxidative effect and thick boundary layer can be minimized (Mostert &
Grobbelaar, 1987; Borowitzka, 1998; Gross, 2003; Hariz et al., 2017). Other than that, efficiency of
light utilization by microalgae can be improved by having optimum turbulence in cultivation
condition. In addition, during photosynthesis process, microalgae consume carbon dioxide and
gives out oxygen. Hence, the oxygen level in the cultivation system increases. However, an excessive
amount of oxygen in the system can cause photo-oxidative and damages microalgae chlorophyll
which results in the disruption of the photosynthesis process and reduces the productivity of
microalgae. Aeration promotes mass transfer which allows the removal of excess oxygen released
out from the medium during photosynthesis process by microalgae that can inhibit microalgae
growth (Woertz et al., 2009). Moreover, nutrient uptake rate can be enhanced by minimizing the
boundary layer between microalgae cell and the surrounding nutrient concentration. It is proven
that turbulence influences the microalgae productivity as an optimum turbulence enhances mass
transfer rate and overcomes the thick boundary layer of unstirred suspension (Mostert &
Grobbelaar, 1987; Borowitzka, 1999). Microalgae reproduction is also enhanced in a sufficient
turbulence system compared to a still medium which can cause biomass accumulation.

Table 4 shows the summary of the specific growth rate of the three microalgae species UKM2,
UKM6 and UKM9 in aerated cultivation condition in comparison to their differences of nutrient
source from different medium used. As the result obtained, growth of these three strains showed
higher biomass productivity in BBM compare to POME. This can be explained based on the nutrient
contain in BBM is complete for the growth requirement of microalgae in comparison to POME.
Nutrient is one of the most critical factors for microalgae productivity (Stephens et al., 2010). The
limitation of nutrient components in the effluent can cause a low growth rate of microalgae. In
POME the level of nutrient content may fluctuated from time to time depending of palm oil mill
activities. Sometimes, the level of organic compound is too high that might cause inhibition towards
microalgae and affect the nutrient uptake performance. The tolerance towards the organic
compound is totally depending on microalgae species as different species shows different tolerance
towards highly concentrated medium such as POME. A strong relationship is shown between
microalgae growth and the availability of nutrients for microalgae consumption (Khalid et al., 2016).
This happened as microalgae assimilate the nutrient components from wastewater, the biomass
increases and resulted in positive growth of microalgae at the same time reduce nutrient or known
as pollutant in the wastewater. The chemical composition of microalgae biomass, C106H263O110N16
represents the fraction of elements contained in the biomass cell and it proves that the elements in
the biomass are the keys of microalgae requirement and elements involved (Stumm & Morgan,
1993). A research has proven that highly concentrated POME results in low growth rate and biomass
yield of Chlamydomonas sp. which is grown in 100% v/v and 50% v/v of POME dilution (Ding et al.,
2016).
In terms of microalgae strains sensitivity, BBM shows the least growth limitation compared to
other medium (Milling et al., 1988). The extremely high concentration of nutrient such as nitrogen
and phosphorus can inhibit microalgae growth. The agricultural wastewater is an example of
wastewater with high nitrogen and phosphorus concentration (Wilkie & Mulbry, 2002) which
sometimes might not be suitable for microalgae growth. POME is loaded with abundance of nutrient
that might not be suitable to certain microalgae species especially the low resistance towards
concentrated wastewater. Microalgae might take a long time to adapt to the environment with a
high nutrient load and low light penetration condition that limits its growth and performance in
nutrient removal (Ding et al., 2016). However, different microalgae species show differences in their
tolerance towards high nutrient concentration.
Meanwhile, light penetration is another factor that is important and causing low growth and
productivity when light is limited. In POME microalgae cultivation, light penetration is poor due to
the high turbidity of POME which is about 460 NTU and it appears in extremely dark brown in
colour which is limiting the light entering the cultivation system. The high amount of suspended
solid that cause the internal shading and prevent certain amount of light from penetrating through
the medium for microalgae utilization. The light penetrated the cultivation column might be
unevenly distributed and limits the growth. It is important to reduce the suspended solid in POME
by having the pre-treatment process such as flocculation to have effective microalgae cultivation
environment. Optimum mixing speed or aeration rate can also overcome this internal shading
problem as it prevents the accumulation of particulate matters that block light penetration.

Aeration source and nutrient growth medium are not the only parameters being considered in
this experiment, the microalgae inoculum concentration is the other factor being tested as it is
important and might influence the growth pattern of all these three strains. Based on the result in
Table 4, 30% of inoculum concentration of UKM6 and UKM9 shows the highest growth rate in BBM
compare to the 20% and 10% of the inoculum strain. While UKM2 does not give any significance
difference at 10%, 20% and 30% initial inoculum concentration. However, in POME all the three
strains have highest growth rate at 30% inoculum concentration with air supply as the aeration
source. The data proved that high inoculum concentration required to cultivate microalgae in POME
as it is important for the adaptation of microalgae to the environment as higher initial inoculum
concentration will enhance the microalgae adaptability to the growth environment. The result of
specific growth rate from Table 5 showed that 20% of inoculum concentration resulted in highest
growth for UKM2 and UKM6 in both BBM and POME in non-aerated condition while 10% of
inoculum concentration gave the highest growth for UKM9 for both BBM and POME. In BBM,
microalgae do not show significance effect on adaptation period compare to the adaptation period in
POME as it might take longer time to adapt due to the nutrient availability factor and other
particulate matters present in POME. The inoculum concentration also influences the exponential
period of microalgae growth as higher inoculum concentration shows exponential phase that start at
early stage of growth curve compare to lower inoculum concentration. This is important to optimize
the initial inoculum concentration as it able to reduce the lag phase and enhance the adaptability of
microalgae in the system especially for wastewater treatment where treatment period will be faster.
According to Figure 3-6, the comparison of microalgae growth pattern can be made at different
nutrient medium used. Microalgae growth in POME showed longer lag phase of growth about 6
days which is doubled the lag phase of microalgae in the commercial medium, BBM about 3 days at
average. The same pattern showed for the exponential phase where microalgae took up to 14 days in
POME compare to BBM which is at average of 7 days. The microalgae growth pattern study is
important as it is related to the hydraulic retention time (HRT) for microalgae based treatment
process. It is important to have optimum HRT for the system as too short HRT will resulted in
insufficient time for the microalgae to adapt and efficiently utilize the nutrient in the effluent. If the
HRT is too long, the thick boundary of the suspension liquid will start to form and cause limitation
of light to penetrate through for microalgae cultivation system. It is proven that with an optimum
HRT, the competition between species that might lead to nutrient and CO2 limitations can be
avoided.

1.6
a)
1.4 10% ukm2
1.2 20% ukm2

30% ukm2
1
DCW (g/L) 0.8
0.6
0.4
0.2
0
0 2 4 6 8 10 12 14 16
Cultivation Period (Day)
b)
1.2
10% ukm6
1
20% ukm6
0.8
DCW (g/L)
30% ukm6
0.6
0.4
0.2
0
0 2 4 6 8 10 12 14 16
c)
1.4
10% ukm9
1.2
20% ukm9
1
30% ukm9
DCW (g/L)
0.8
0.6
0.4
0.2
0
0 2 4 6 8 10 12 14 16
Figure 3. Dry cell weight (DCW) growth curve in Bold’s Basal Medium (BBM) with air supply as the
aeration source a) Growth curve of Chlorella sp. (UKM2), b) Growth curve of Chlamydomonas sp.
(UKM6), c) Growth curve of Scenedesmus sp. (UKM9)

a)
1.2
10% ukm2
1
20% ukm2
0.8
DCW (g/L) 30% ukm2
0.6
0.4
0.2
0
0 5 10 15 20 25 30
-0.2
b)
1.2
1 10% ukm6
20% ukm6
0.8
30% ukm6
DCW (g/L)
0.6
0.4
0.2
0
0 5 10 15 20 25 30
-0.2
c)
1.4
10% ukm9
1.2
20% ukm9
1 30% ukm9
0.8
DCW (g/L)
0.6
0.4
0.2
0
0 5 10 15 20 25 30
-0.2
Figure 4. Dry cell weight (DCW) growth curve in Palm Oil Mill Effluent (POME) with air supply as
the aeration source a) Growth curve of Chlorella sp. (UKM2), b) Growth curve of Chlamydomonas sp.
(UKM6), c) Growth curve of Scenedesmus sp. (UKM9)

a)
0.7
10% ukm2
0.6
20% ukm2
0.5
30% ukm2
DCW (g/L) 0.4
0.3
0.2
0.1
0
0 5 10 15 20 25 30
-0.1
b)
0.45
0.4 10% ukm6
0.35
20% ukm6
0.3
30% ukm6
DCW (g/L)
0.25
0.2
0.15
0.1
0.05
0
0 5 10 15 20 25 30
c)
0.45
0.4 10% ukm9
0.35 20% ukm9
0.3 30% ukm9
0.25
DCW (g/L)
0.2
0.15
0.1
0.05
0
-0.05 0 5 10 15 20 25 30
Figure 5. Dry cell weight (DCW) growth curve in Bold’s Basal medium (BBM) with agitation by
mixing a) Growth curve of Chlorella sp. (UKM2), b) Growth curve of Chlamydomonas sp. (UKM6), c)
Growth curve of Scenedesmus sp. (UKM9)

a)
1.4
10% ukm2
1.2 20% ukm2
1 30% ukm2
0.8
DCW (g/L)
0.6
0.4
0.2
0
0 5 10 15 20 25 30
-0.2
b)
1
0.9
0.8 10% ukm6
0.7 20% ukm6
DCW (g/L)
0.6 30% ukm6

0.5
0.4
0.3
0.2
0.1
0
0 5 10 15 20 25 30
c)
0.8
10% ukm9
0.7
20% ukm9
0.6
30% ukm9
0.5
DCW (g/L)
0.4
0.3
0.2
0.1
0
0 5 10 15 20 25 30
Figure 6. Dry cell weight (DCW) growth curve in Palm Oil Mill Effluent (POME) with agitation by
mixing a) Growth curve of Chlorella sp. (UKM2), b) Growth curve of Chlamydomonas sp. (UKM6), c)
Growth curve of Scenedesmus sp. (UKM9)

CONCLUSION
The main objective of this research was to study native microalgae Chlorella sp., Clamydomonas
sp., and Scenedesmus sp. growth and biomass productivity at different source of nutrient, inoculum
concentration and aeration factor. The findings proved that turbulence, nutrient availability, initial
inoculum concentration influence the microalgae growth and productivity. The results show that
POME can be the alternative of the commercial microalgae nutrient medium and promotes the cost-
effective strategies of microalgae cultivation system towards a better environment.
ACKNOWLEDGEMENTS
The authors gratefully acknowledge Yayasan Sime Darby for the scholarship and UKM-YSD
Research Grant for the financial support.
REFERENCES
[1] Becker, E. W. (1994). Microalgae: Biotechnology and Microbiology (Vol. 10). Cambridge University
Press.
[2] Borowitzka, M. A. (1998). Limits to growth. In: Wong, Y-S & Tam, N. F. Y. (eds). Wastewater
Treatment With Algae. Springer Berlin Heidelberg.
[3] Borowitzka, M. A. (1999). Commercial production of microalgae: ponds, tanks, tubes and
fermenters. Journal of Biotechnology, 70(1-3), 313–321.
[4] Brennan, L. & Owende, P. (2010). Biofuels from microalgae-A review of technologies for
production, processing, and extractions of biofuels and co-products. Renewable and Sustainable
Energy Reviews, 14(2), 557–577.
[5] Ding, G. T., Yaakob, Z., Takriff, M. S., Salihon, J. & Abd Rahaman, M. S. (2016). Biomass
production and nutrients removal by a newly-isolated microalgal strain Chlamydomonas sp in
palm oil mill effluent (POME). International Journal of Hydrogen Energy, 41(8), 4888-4895.
[6] Gross, E. M. (2003). Allelopathy of aquatic autotrophs. Critical Reviews in Plant Sciences, 22(3-4),
313-339.
[7] Hariz, H. B. & Takriff, M. S. (2017). Palm oil mill effluent treatment and CO2 sequestration by
using microalgae-sustainable strategies for environmental protection. Environmental Science and
Pollution Research, PMID: 28791508, DOI: 10.1007/s11356-017-9742-6 1-32.
[8] He, Q., Yang, H. & Hu, C. (2015). Optimizing light regimes on growth and lipidaccumulation
in Ankistrodesmus fusiformis H1 for biodiesel production. Bioresource Technology, 198, 876–
883.
[9] Khalid, A.A.H., Yaakob Z., Sheikh Abdullaha, S. R. & Takriff M. S. (2016). Enhanced growth
and nutrients removal efficiency of Characium sp. cultured in agricultural wastewater via
acclimatized inoculum and effluent recycling. Journal of Environmental Chemical Engineering 4,
3426–3432
[10] Khan, S. A., Hussain, M. Z., Prasad, S. & Banerjee, U. C. (2009). Prospects of biodiesel
production from microalgae in India. Renewable and Sustainable Energy Reviews, 13(9), 2361–
2372.
[11] Lau P.S., Tam N. F. Y. & Wong Y. S. (1995). Effect of algal density on nutrient removal
fromprimary settled wastewater. Environmental Pollution, 89, 59–66.
[12] Liang, Y., Sarkany, N. & Cui, Y. (2009). Biomass and lipid productivities of Chlorella vulgaris
under autotrophic, heterotrophic and mixotrophic growth conditions. Biotechnology Letters,
31(7), 1043–1049.

[13] Ma, Q., Wang, J., Lu, S., Lv, Y. & Yuan Y., (2012). Quantitative proteomic profiling
revealsphotosynthesis responsible for inoculum size dependent variation in
Chlorellasorokiniana. Biotechnology and Bioengineering, 110 (2013) 773–784.
[14] Mostert, E. S. & Grobbelaar, J. U. (1987). The influence of nitrogen and phosphorus on algal
growth and quality in outdoor mass algal cultures. Biomass, 13(4), 219–233.
[15] Stephens, E., Ross, I. L., Mussgnug, J. H., Wagner, L. D., Borowitzka, M. A., Posten, C. &
Hankamer, B. (2010). Future prospects of microalgal biofuel production systems. Trends in
Plant Science, 15(10), 554-564.
[16] Stumm, W. & Morgan, J. J. (1993). Aquatic Chemistry: Chemical Equilibria and Rates in Natural
Waters (3rd ed). New York : Wiley-Wiley-Interscience.
[17] Wilkie, A. C. & Mulbry, W. W. (2002). Recovery of dairy manure nutrients by benthic
freshwater algae. Bioresource Technology, 84(1), 81-91.
[18] Williams, P. J. L. B. & Laurens, L. M. L. (2010). Microalgae as biodiesel & biomass feedstocks:
Review & analysis of the biochemistry, energetics & economics. Energy & Environmental
Science, 3(5), 554 - 590.
[19] Woertz, I., Feffer, A., Lundquist, T. & Nelson, Y. (2009). Algae Grown on Dairy and Municipal
Wastewater for Simultaneous Nutrient Removal and Lipid Production for Biofuel Feedstock.
Journal of Environmental Engineering, 135(11), 1115–1122.
[20] Yang, J., Rasa, E., Tantayotai, P., Scow, K. M., Yuan, H. & Hristova K. R. (2011). Mathematical
model of Chlorella minutissima UTEX2341 growth and lipid production under
photoheterotrophic fermentation conditions. Bioresource Technology, 102, 3077–3082.

4x3 2x298x311

Uploaded by

Copyright:

Available Formats

4x3 2x298x311

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

4x3 2x298x311

Uploaded by

Copyright:

Available Formats

Transactions on Science and Technology Vol. 4, No.

3-2, 298 - 311, 2017

Growth and Biomass Production of Native

ISSN 2289-8786. http://transectscience.org/

Microorganism and Growth Medium

Table 1. Nutrient elements contain in BBM and Anaerobic POME

Cultivation Conditions and Analytical Determinations

DCW = (weight of filtered microalgae biomass – weight of blank filter paper)

Table 2. Microalgae cultivation in BBM at different inoculum concentration

Inoculum Concentration 10% 20% 30%

ISSN 2289-8786. http://transectscience.org/

Table 3. Microalgae Cultivation in POME at different inoculum concentration

Inoculum Concentration 10% 20% 30%

RESULT AND DISCUSSION

X = X0 Xmax е µmax t / ((Xmax-X0) + X0 е µmaxt) (2)

Table 4. Growth rate of microalgae cultivated with aerated growth condition

Native Microalgae Growth Rate (μ )

ISSN 2289-8786. http://transectscience.org/

Table 5. Growth rate of microalgae cultivated with non-aerated growth condition

Native Microalgae Growth Rate (μ )

ISSN 2289-8786. http://transectscience.org/

ISSN 2289-8786. http://transectscience.org/

ISSN 2289-8786. http://transectscience.org/

1.2 20% ukm2

ISSN 2289-8786. http://transectscience.org/

ISSN 2289-8786. http://transectscience.org/

Cultivation Period (Day)

ISSN 2289-8786. http://transectscience.org/

0.6 30% ukm6

ISSN 2289-8786. http://transectscience.org/

ISSN 2289-8786. http://transectscience.org/

ISSN 2289-8786. http://transectscience.org/

You might also like