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Vol. 27（2022），151 ～ 159

Effect of Seawater Temperatures on the Growth and Biochemical Profile of the TropicalGreen Seaweed, Caulerpa lentillifera J. Agardh
NurFatin Solehah Husin*1,2, Hui-Yin Yeong*2, and Siew-Moi Phang*1,3*1 Institute of Ocean and Earth Sciences (IOES), Universiti Malaya
50603 Kuala Lumpur, Malaysia*2 Institute for Advanced Studies, Universiti Malaya
50603 Kuala Lumpur, Malaysia*3 Faculty of Applied Sciences, UCSI University
Cheras 56000 Kuala Lumpur, Malaysia

Abstract
Seaweeds are marine algae that are used as a food source, a means of producing hydrocolloids forindustrial uses, as well as a source of medication, dietary supplements, fertilizer, and animal feed, particularly
in maritime nations. Caulerpa lentillifera J. Agardh is an edible green seaweed that has been used as a fresh
vegetable and has gained a lot of attention recently due to its nutritional content and beneficial composition forhuman consumption. C. lentillifera, which grows naturally in tropical locations, was first commerciallycultivated in the Philippines, then by Japan in the late 1980s, and more recently by Taiwan, China. Numerous
studies have revealed that C. lentillifera is high in protein, carbohydrates, minerals, vitamins, dietary fibres,
polyunsaturated fatty acids, including omega-3 fatty acids, and bioactive compounds such as phenolic
compounds, polysaccharides, and siphonaxanthin, which are reported to have high antioxidant, anticancer,
anti-diabetic, and immunomodulation properties and potential health benefits. In our current study, theinfluence of temperature on the growth of C. lentillifera was investigated. The experiment was conducted inenriched seawater containing 320μmol/L NO3⁻ and 10.6μmol/L PO43⁻ for 30 days at three different
temperatures: 20, 24, and 28 ºC. Their growth, photosynthetic pigments, and biochemical compositions,
including the contents of protein, carbohydrates, lipids and fatty acids profiling, were determined to investigate
the effects of temperatures and nutrients on C. lentillifera. In comparison to 24 ºC and 20ºC, our results
showed that the highest growth rate and chlorophyll a content were observed at 28 ºC, with 8.77±0.01 % d⁻1
and 3397.02±5.36 μg g⁻1 respectively. C. lentillifera's total protein (38.72±1.40% DW⁻1) and carbohydratecontent (61.56±6.73% DW⁻1) was also found to be highest at 28ºC in comparison to other temperatures.
However, the lipid content of C. lentillifera was shown to be higher at 20ºC with 8.70 ±0.10 % DW⁻1. In
conclusion, lower temperatures impeded the photosynthetic processes, reduced the amount of soluble protein,
and restricted the development rate of C. lentillifera, but they also promoted the accumulation of othersubstances like lipids, fatty acids, and carotenoids. This study is part of an ongoing project to optimize theculture of C. lentillifera in the cold, nutrient-rich effluent generated from an ocean thermal energy conversion(OTEC) system.
Key words : Caulerpa lentillifera, Seaweeds, Temperature, Biochemical composition

1. Introduction
Seaweeds are non-flowering photosynthetic marine flora; function as the primary producers in the marine
ecosystem. Seaweeds are widely distributed in the intertidal, tidal and subtidal zones of tropical to polar regions. In
2020, the world's annual seaweed production had reached 36,165,326 tonnes FW, of which 96.81% (35,013,088 tonnes
Date of Manuscript Acceptance 2023.1.11
E-mail of corresponding author: phang@um.edu.my

*

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NurFatin Solehah Husin, Hui-Yin Yeong and Siew-Moi Phang

FW) was produced by the seaweed aquaculture farming industry, and worth an estimated annual value of USD
16,444,926,480. (FAO, 2022a). It is an important marine resource for phycocolloid industries, serving as food for direct
human consumption, components for medical, pharmaceutical, nutraceuticals, and cosmetics industries as well asbioresources for aquaculture and agriculture industries (García-Poza et al, 2020). C. lentillifera also contains highamount of carbohydrates (phycocolloids such as alginates, agar, carrageenan); proteins, minerals and is also found to bea rich source of important bioactive components.
Tropical seaweed aquaculture farming harvested 99.08 % (15,061,383 tonnes FW) of tropical seaweed production
in 2020, worth USD 5,681,861.48. (FAO, 2022b). In 2020, Caulerpa aquaculture farming produced a total of 1,021.36
tonnes FW with an annual value of USD 623.34. (FAO, 2022a) Caulerpa lentillifera or commonly known as sea
grapes, is widely consumed as a fresh vegetable in Asia, specifically in Southeast Asia.
The most abundant contents in C. lentillifera are carbohydrates (~43 -55%), and dietary fibre (Insoluble dietary
fibres ~ 15.75 - 28.98%). Protein (~ 13-25%), essential minerals (Na, K, Ca, and Mg), vitamins (A and C) are alsopresent (Syakilla et al., 2022). 25 major fatty acids species were identified in C. lentillifera, with high omega-3 fatty
acids, and polyunsaturated fatty acids, which act as strong antioxidants. Bioactive compounds, such as phenolic
compounds, polysaccharides, and siphonaxanthin also found in C. lentillifera where these compounds are reported tohave high antioxidant, anticancer, anti-diabetic, and immunomodulation properties. Reported health benefits
contributed by C. lentillifera included cardioprotective properties such as anti-hypertensive and hypolipidemic,
antibacterial, anticancer, anti-coagulant, anti-hyperglycemic, anti-diabetic, anti-inflammatory, antioxidative,
anti-pyretic, chelating agent, and immunostimulatory (- innate immune system) (Syakilla et al., 2022)
Ocean thermal energy conversion (OTEC) is an emerging technology for renewable energy production. The DSW
that is drawn up, generate energy through the difference in temperature between the cold DSW and the warm surface
water. The cold deep seawater applications from the OTEC effluents can be used in different applications and industries
including seaweed cultivation. It is very useful for extending the range of marine organisms that can be cultivated andtemperate species of algae can now be cultivated in the tropics, using these effluents. The cultivation of seaweeds in theeffluents arising from an OTEC system and the valorisation of the seaweed biomass in the form of high-value products,
would contribute to the sustainability of the energy production system. Furthermore, integrating the OTEC andseaweed cultivation may pave the way for the development of new algal products and the expansion of the algal
industry.
Integration of C. lentillifera cultivation in OTEC industries may provide more avenues for research and productdevelopment, as C. lentillifera can be considered a promising and valuable alternative product with high nutritional
value for the future. For example, a lower temperature of OTEC effluent may increase lipid and fatty acid production
because desaturation of saturated fatty acids is reduced and production of unsaturated fatty acids, consisting of MUFAs
and PUFAs, is increased, which is important in the medicine, cosmetics, and biotechnology industries. Hence, culturing
C. lentillifera in cold OTEC effluent will greatly contribute to the production of valuable products and eventually
improve its market value.

2. Research Objective and Scope of studies
The objectives of this project are to investigate the effect of seawater temperature and nutrients (Nitrate (NO3⁻) &phosphate (PO43⁻) on the growth and biochemical profiles of selected commercial seaweed, C. lentillifera. The purposeof these studies is to provide a baseline for the land-based cultivation and utilization of cold deep seawater sources suchas effluent from the OTEC system. This can serve as a sustainable method to produce more biomass and higher desired
quality products from the seaweed. Besides, the environmental factors such as light, nutrients, temperature and salinity
can be controlled and manipulated throughout the cultivation period compared to an open cultivation system like anoffshore/ near-shore cultivation system. These can help to improve and holds the potential of a year-round production
especially for targeted specific valuable molecules. In addition, a cleaner and marketable biomass

Effect of Seawater Temperatures on the Growth and Biochemical Profile of the Tropical Green
Seaweed, Caulerpa lentillifera J. Agardh

153

3. Research methodology
3.1 Seaweed collection & maintenance
Caulerpa lentillifera were collected from natural seaweed beds in Port Dickson (02˚26’38” N; 101˚51’21” E). The
collected seaweeds were transported in a healthy state in a cool Styrofoam box. The stocks were rinsed with seawater
while epiphytes, dirt and plants parts with any signs of de-pigmentation and necrosis were removed before being usedin the experiments. Healthy cleansed seaweeds were cultured in tanks of aerated seawater (30ppt) and maintained at
28ºC, 25μmol m⁻2 s⁻1 with 12 h light and 12 h dark cycles.

3.2 Experimental Design
Growth studies of Caulerpa lentillifera were conducted in a 30 L tank in a growth incubator with 50 μmol m⁻2 s⁻1
illuminated with cool-white LED tubes and 12h L: 12h D cycles for 30 days. The experiments were carried out in five
replicates in 20 litters of 30 ppt artificial seawater (Instant Ocean® Synthetic Sea Salt) enriched with 320 μmol L⁻1
NO3⁻ and 10.6 μmol L⁻1 PO4⁻, with a starting inoculum of 100 g per replicate for each of the three temperature
treatments (20, 24, and 28 ºC). Samples were harvested from each tank on day 0, 6, 12, 18, 24 & 30 for growth &biomass determination, and biochemical (carbohydrate, protein, lipid and pigment) extraction and determination.

3.3 Biomass and relative growth rate (RGR):
The fresh weight (g) and relative growth rate (%d⁻1) of C. lentillifera were measured at six-day intervals for 30
days. The relative growth rate (RGR) regarding day 0 (RGRi) and previous samples (RGRt-1) were calculated using thefollowing equations (Phang, 1997):(1)
Where Wi is the initial fresh weight, Wt is the fresh weight on day t, Δt is the time interval(2)
Wt is the fresh weight on day t and Wt-1 is the fresh weight on the previous sampling day, Δt is the time interval

3.4 Extraction of chlorophyll-a & carotenoid content:
The determination of chlorophyll-a content and carotenoid was done using the colorimetric method by (Strickland
and Parson, 1972). The supernatant was measured at OD665nm, OD645nm, and OD630nm for chlorophyll –a content andOD452nm for carotenoid content.

3.5 Biochemical composition extraction of Caulerpa lentillifera:
For the biochemical composition assay, the C. lentillifera extracted based on the standard method such as Protein(Bradford, 1976), carbohydrates (Dubois et al., 1956), lipid (Bligh and Dyer 1959) and fatty acid transesterification(Christie et al., 1989) were determined on the day 0,12,18, 24 and 30 of the experiments. The protein, carbohydrates
and lipid content of C. lentillifera were analyzed and expressed in percentages dry weight (% DW-1). The analysis offatty acids was conducted using gas chromatography for the determination of fatty acid composition (Christie et al.,1989).

3.6 Statistical Analyses
Tukey’s – HSD comparisons were applied to determine statistically significant differences (p<0.05) among time
and treatments following ANOVA. One-way ANOVA was used for growth and biochemical analyses of C. lentillifera
at the end of the experiments. A significance level of 95% (p<0.05) was set for all the tests. Data are presented as mean
± SD (the number (n) of replicates is presented in each figure caption). The statistical analyses were carried out using

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NurFatin Solehah Husin, Hui-Yin Yeong and Siew-Moi Phang

Statistica software version 8.0

4. Results and Discussion
4.1 The growth and biomass production
The highest biomass production of C. lentillifera was obtained at 28 ºC with 199.39±3.90 g FW on day 24
compared with 20 ºC & 24 ºC. However, the highest biomass productivity of C. lentillifera was also obtained at 28 ºC
but on day 6 of 141.48±0.23 g m⁻2 d⁻1 with the relative growth rate of 8.77±0.01% d⁻1. On contrary, to the biomass
production under lower temperature of 20 ºC has it is dropped from 100.66±0.18 g FW on day 0 to 63.68±4.41 g FW
on day 6. The low growth rate at 20 ºC could be a result of inadequate acclimatization, which has led to the culture
becoming stagnant. Besides, the temperature shock due to inadequate acclimatization also led to the disintegration of
the algal thallus and later reduced their biomass production. Hence, it is shown that the growth rate and biomass
production of C. lentillifera cultivated at the lower temperatures of 20 ºC and 24 ºC exerted a stronger influence to theseaweed production.

Fig. 1

Biomass production. FW (g) for Caulerpa lentillifera at different temperature of 20 ºC, 24 ºC and 28 ºC
in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5).
The significantly difference was labelled with an asterisk (*), (p<0.05).

Table 1 Biomass productivity (g m⁻2 d⁻1) and relative growth rate (% d⁻1) for Caulerpa lentillifera at different
temperature of 24 ºC and 28 ºC in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates(n = 5). The significantly difference was labelled with an asterisk (*), (p<0.05).
Temperature
Biomass productivity (g m⁻2 d⁻1) Day 6
Biomass productivity (g m⁻2 d⁻1) Day 18
Biomass productivity (g m⁻2 d⁻1) Day 24
Relative growth rate (% d⁻1) Day 6
Relative growth rate (% d⁻1) Day 18
Relative growth rate (% d⁻1) Day 24

24 ºC
10.33±0.24
29.30±2.55
37.13±3.17

28 ºC
141.38±0.23*
79.60±1.8665.73±0.52

0.64±0.01

8.77±0.01
4.94±0.11
4.04±0.10

2.16±0.57
2.56±0.26

4.2 Pigments production
The higher chlorophyll-a content was observed at 28 ºC with 3397.02±5.36 μg g⁻1 and 24 ºC with 3137.27±109.69
μg g⁻1. On the contrary, at the lower temperature of 20 ºC, the chlorophyll-a content decreased under longer exposure to
the lower temperature, as lower temperatures might suppress the enzyme activity in chlorophyll synthesis (Guo et al.,2015) and disrupt the algal cell component for photosynthesis (Allen et al., 2001). Lowering the temperature generallyEffect of Seawater Temperatures on the Growth and Biochemical Profile of the Tropical Green
Seaweed, Caulerpa lentillifera J. Agardh

155

reduces the reaction rates and therefore limits the sinks for the absorbed excitation energy (light), particularly in CO2
fixation and photorespiration. Smaller sinks for absorbed excitation energy increase the potential for oxidative damage
to PSII (Allen et al., 2001) hence affects chlorophyll production and photosynthesis. As C. lentillifera grown at 20ºC
became feebler, the branches and the fronds had lost their pigmentation, became perforated and fragile. which later led
to the disintegration of the algal cells (Andersen et al., 2013).
However, the highest carotenoid content was found at 20 ºC with 2399.40±36.40 μg g⁻1. Carotenoid is an auxiliary
pigment that aids in the maintenance the photosynthetic stability, especially in adverse situations. Hence, moreaccumulation of carotenoid was observed at 20 ºC than its ambient temperature to protect its photosynthetic
mechanism.

Fig. 2Chlorophyll-a content (μg g⁻1) for Caulerpa lentillifera at different temperature of 20ºC, 24ºC and 28ºC
in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5).
The significantly difference was labelled with an asterisk (*), (p<0.05).

Fig. 3

Carotenoid content (μg g⁻1) for Caulerpa lentillifera at different temperature of 20 ºC, 24 ºC and 28 ºC
in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5).
The significantly difference was labelled with an asterisk (*), (p<0.05).

4.3 Biochemical composition (carbohydrates, protein and lipid) production
The highest total carbohydrate content was obtained in Caulerpa lentillifera at 28 ºC with 61.56±6.73% DW⁻1
followed by 24 ºC with 59.24±6.92% DW⁻1 both on day 18. The carbohydrate accumulation of compatible solutes
occurred at a lower temperature to maintain their cell osmolality and to regulate their photosynthetic mechanism. Thisis important to maintain the seaweed’s energy production and consumption in the algae cells. (Klähn and Hagemann2011; Ras et al., 2013; Barati et al., 2019)
As for protein content, the higher accumulation was also observed at 28 ºC with 38.72±1.40% DW⁻1 and at 24 ºC
with 35.35±0.27% DW⁻1 respectively. Studies conducted by Settamongkol et al., 2015, also showed similar protein

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NurFatin Solehah Husin, Hui-Yin Yeong and Siew-Moi Phang

accumulation compared to this study, where the protein content found in the green algae species such as C. taxifolia
and C. linum had 33.83±0.21% DW⁻1 and 30.70±0.70% DW⁻1 respectively. This showed that temperature changes willaffect the stability of cellular components, protein’s membrane and lessen the protein synthesis through protein

Fig. 5 Total carbohydrates content (% DW⁻1) for Caulerpa lentillifera at different temperature of 20 ºC, 24 ºC and 28ºC in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5). The significantly
difference was labelled with an asterisk (*), (p<0.05).

Fig. 6 Total protein content (% DW⁻1) for Caulerpa lentillifera at different temperature of 20 ºC, 24 ºC and 28 ºC in
320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5). The significantly
difference was labelled with an asterisk (*), (p<0.05).

Fig. 7 Total lipid content (% DW⁻1) for Caulerpa lentillifera at different temperature of 20 ºC, 24 ºC and 28 ºC in 320μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5). The significantly difference
was labelled with an asterisk (*), (p<0.05).

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Effect of Seawater Temperatures on the Growth and Biochemical Profile of the Tropical Green
Seaweed, Caulerpa lentillifera J. Agardh

modulation and modify their environment in the algal cells (Eggert 2012; Wagenen et al., 2012; Schroda et al., 2015;
Barati et al., 2019).
The lipid content of C. lentillifera obtained at 20 ºC ranged from 4.84 ±0.13 % DW⁻1 to 8.70 ±0.10 % DW⁻1,
followed by C. lentillifera cultivated at 28ºC with a similar range of 4.58 ±0.29 % DW⁻1 to 8. 57 ±0.23 % DW⁻1. The
higher lipid accumulation at the lower temperature of 20 ºC showed that, temperature has significant effects on thesynthesis and accumulation of lipids and fatty acids. This is because, under stress management and repair strategies,
seaweed modifies its lipid biosynthesis pathways to produce and promote accumulations of neutral lipids in the form of
triacylglycerol. It offers a carbon and energy storage function in the cell that allows algal to tolerate unfavorable
environmental conditions (Hu et al., 2008; Chen et al., 2011; Feng et al., 2011; Mairet et al., 2011; Fakry et al., 2015).

4.4 Fatty Acid distribution
About 25 major fatty acid species were identified in C. lentillifera cultured under different temperature treatmentsof 20 ºC, 24 ºC, and 28 ºC. Among these, C16:0 was the most abundant fatty acid (29.62±0.42% to 40.50±0.16%),
followed by C18:0 (9.09±0.34% to 17.61±1.16%), and C20:1n9 (3.95±0.37% to 14.39±0.19%) for all the treatments.
The extent of unsaturated fatty acids increased for cultivated C. lentillifera at 20 ºC and in other words, theamounts of polyunsaturated fatty acids (PUFA) were highest in C. lentillifera at 20 ºC (18.40±0.35%), followed by 24ºC (13.07±0.61%), and 20 ºC (12.43±0.51%). In particular, C20:1n9-EPA contributed the highest value in total PUFA
species (up to 14.39±0.19%) in C. lentillifera cultured at 20 ºC.
The results indicated that, when C. lentillifera was exposed to a lower temperature, the PUFA amount increased
rapidly to maintain the cell membrane fluidity. This is because, the desaturation of existing C18:0 fatty acid chains orTable 2 Total fatty acid distribution percentages (%) for Caulerpa lentillifera at different temperature of 20 ºC, 24 ºC
and 28 ºC in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates (n = 5).
The significantly difference was labelled with an asterisk (*), (p<0.05).
% Fatty acid distribution
SFA
MUFA
PUFA

20 ºC
60.85±0.80
20.75±0.72
18.40±0.35*

Temperature
24 ºC
68.12±0.69*
18.81±0.91
13.07±0.61

28 ºC
68.77±0.34*
18.80±0.46
12.43±0.51

Table 3 The major species distributed in fatty acid distributions (%) for Caulerpa lentillifera at different temperatureof 20 ºC, 24 ºC and 28 ºC in 320 μmol L⁻1 NO3⁻ and 10.6 μmol L⁻1 PO43⁻. Data are means ± SD of five replicates(n = 5). The significantly difference was labelled with an asterisk (*), (p<0.05).
Temperature
20 ˚C
24 ˚C
28 ˚C
20 ˚C
24 ˚C
28 ˚C
20 ˚C
24 ˚C
28 ˚C
20 ˚C
24 ˚C
28 ˚C

Day
0
12

18

24

30

SFA
C16:0
C18:0
31.22±0.66
11.30±0.11
34.04±1.93
10.99±0.90
38.59±0.73
10.93±0.16
40.50±0.16*
9.09±0.34
34.28±0.23
9.81±0.26
39.25±0.31
11.37±0.2136.35±1.01
9.21±1.03
32.81±0.67
11.24±0.52
31.55±1.94
11.69±0.75
36.87±0.50
14.56±1.20
29.65±1.77
15.06±0.55
29.62±0.4217.61±1.16*
33.58±0.64
17.11±1.45

MUFA
C16:1
C18:2n6c
4.85±0.21
6.04±0.82
5.25±0.12
6.50±0.57
5.53±0.23
5.10±0.08
4.46±0.24
5.85±0.26
5.75±0.28
4.94±0.30
5.20±0.26
4.74±0.55
5.42±0.26
5.51±0.32
6.59±0.36
4.84±0.15
3.54±1.02
3.88±0.39
5.60±0.53
3.81±0.06
3.44±0.30
3.65±0.10
4.52±0.33
2.08±0.44
3.63±0.34
2.46±0.24

PUFA
C20:1n910.10±0.4210.64±0.26
10.22±0.22
8.42±0.29
14.39±0.19*
9.22±0.18
7.47±0.46
5.13±0.45
4.62±0.13
5.03±0.40
5.50±0.54
4.33±0.35
3.95±0.37

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NurFatin Solehah Husin, Hui-Yin Yeong and Siew-Moi Phangof de novo synthesized fatty acids increases as the enzymes encoding the desaturation of this fatty acids such as Δ9 andΔ12 desaturases, are elevated under cold stress conditions (Cao et al., 2017). Hence, temperatures showed a positive
relationship with a higher degree of fatty acid desaturation in both cold and freezing environments (Zhang et al., 2012;
Barati et al., 2019). Fatty acid distribution varied with the changes in the natural environment, like temperature stress,
to maintain their membrane integrity and act as one of the central mechanisms for seaweeds. The mechanism included
fatty acid composition, unsaturation of the cells, and re-distribution of fatty acid species in membrane lipids (Cao et al.,2017).

5. Conclusion
Caulerpa lentillifera can be considered a promising and valuable alternative product for the future with highnutritional value. A marine producer with a unique structure and biochemical composition could be exploited for its
various properties, such as food, energy, medicine, cosmetics, and biotechnology. Lower temperatures may inducechanges in seaweed growth and their biochemical composition, especially for tropical C. lentillifera. Under low
temperatures, saturated fatty acids reduced and increased the production of unsaturated fatty acids, consist of MUFAs
and PUFAs and other seaweed compositions such as lipid and carotenoid productions. This will greatly contribute to
the production of valuable lipids and fatty acids in C. lentilifera, improving its market value. As a result, cultivating C.
lentillifera at low temperatures may open up new avenues for research as well as product development in a variety offields for new sources and the production of high-value products. For example, implementation in OTEC industries
application.

6. Acknowledgement
The research was funded by the Ministry of Higher Education Malaysia under the Malaysia Partnership andAlliances in Research (MyPAiR) Programme - Science and Technology Research Partnership for Sustainable
Development (SATREPS)-OTEC Project 6: H-OTEC – Spin-off- Growth of various new floras that use cold deep
seawater – Seaweeds, Umi Budo (IF045-2019); Higher Institution Centre of Excellence (HICoE) Programme(IOES-2014F). We thank the Algae Research Laboratory of Universiti Malaya and Institute for Advanced Studies (IAS)
together with the Institute of Ocean and Earth Sciences (IOES), Universiti Malaya.

References
Andersen, G. S., Pedersen, M. F., and Nielsen, S. L. (2013). Temperature acclimation and heat tolerance ofphotosynthesis in Norwegian Saccharina latissima (Laminariales, Phaeophyceae). Journal of phycology, 49(4),689-700.
Allen, D. J., and Ort, D. R. (2001). Impacts of chilling temperatures on photosynthesis in warm-climate plants. Trends
in plant science, 6(1), 36-42.
Barati, B., Gan, S. Y., Lim, P. E., Beardall, J., and Phang, S. M. (2019). Green algal molecular responses to temperaturestress. Acta physiologiae plantarum, 41(2), 26.
Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing
the principle of protein-dye binding. Analytical biochemistry, 72(1-2), 248-254.
Bligh, E.G., and Dyer, W.J. (1959) A rapid method of total lipid extraction and purification.Can. J. Biochem. Physiol
37:912-917
Cao, M., Wang, D., Mao, Y., Kong, F., Bi, G., Xing, Q., and Weng, Z. (2017). Integrating transcriptomics andmetabolomics to characterize the regulation of EPA biosynthesis in response to cold stress in seaweed Bangia
fuscopurpurea. PloS one, 12(12), e0186986.
Christie, W. W. (1989). Gas chromatography and lipids. Oily.
Chen, M., Tang, H., Ma, H., Holland, T. C., Ng, K. S., and Salley, S. O. (2011). Effect of nutrients on growth and lipid
accumulation in the green algae Dunaliella tertiolecta. Bioresource technology, 102(2), 1649-1655.
Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. T., and Smith, F. (1956). Colorimetric method for determinationof sugars and related substances. Analytical chemistry, 28(3), 350-356.

Effect of Seawater Temperatures on the Growth and Biochemical Profile of the Tropical Green
Seaweed, Caulerpa lentillifera J. Agardh

159

Eggert A. (2012). Seaweed responses to temperature. In Wiencke C. & Bischof K. (Eds.), Seaweed biology. Ecologicalstudies (analysis and synthesis) (pp. 47-66). Berlin, Heidelberg: Springer
FAO, 2022a. Fishery and Aquaculture Statistics. Global aquaculture production 1950-2020 (FishStatJ). In: FAO
Fisheries
andAquaculture
Division[online].
Rome.
Updated2022.
www.fao.org/fishery/statistics/software/fishstatj/en
FAO. 2022b. Fishery & Aquaculture Statistics. Global production by production source 1950-2020 (FishStatJ). In: FAO
Fisheries
andAquaculture
Division[online].
Rome.
Updated2022.
www.fao.org/fishery/statistics/software/fishstatj/en
Fakhry, E. M., and El Maghraby, D. M. (2015). Lipid accumulation in response to nitrogen limitation and variation oftemperature in Nannochloropsis salina. Botanical studies, 56(1), 1-8.
Feng, D., Chen, Z., Xue, S., and Zhang, W. (2011). Increased lipid production of the marine oleaginous microalgae
Isochrysis zhangjiangensis (Chrysophyta) by nitrogen supplement. Bioresource Technology, 102(12), 6710-6716.
García-Poza, S., Leandro, A., Cotas, C., Cotas, J., Marques, J. C., Pereira, L., and Gonçalves, A. M. (2020). The
evolution road of seaweed aquaculture: Cultivation technologies and the industry 4.0. International Journal ofEnvironmental Research and Public Health, 17(18), 6528.
Guo, H., Yao, J., Sun, Z., and Duan, D. (2015). Effect of temperature, irradiance on the growth of the green alga
Caulerpa lentillifera (Bryopsidophyceae, Chlorophyta). Journal of Applied Phycology, 27(2), 879-885.
Hu, Q., Sommerfeld, M., Jarvis, E., Ghirardi, M., Posewitz, M., Seibert, M., and Darzins, A. (2008). Microalgal
triacylglycerols as feedstocks for biofuel production: perspectives and advances. The plant journal, 54(4),621-639.
Klähn S., and Hagemann, M. (2011). Compatible solute biosynthesis in cyanobacteria. Environmental microbiology,
13(3), 551-562.
Mairet, F., Bernard, O., Masci, P., Lacour, T., and Sciandra, A. (2011). Modelling neutral lipid production by the
microalga Isochrysis aff. galbana under nitrogen limitation. Bioresource technology, 102(1), 142-149.
Phang, S. M. (1997). Sargassum stolonifolium et Yoshida sp. nov. from Penang Island, Peninsular Malaysia. Taxonomyof economic seaweeds, 61-73.
Ras, M., Steyer, J. P., and Bernard, O. (2013). Temperature effect on microalgae: a crucial factor for outdoor
production. Reviews in Environmental Science and Bio/Technology, 12(2), 153-164.
Syakilla, N., George, R., Chye, F. Y., Pindi, W., Mantihal, S., Wahab, N. A., Fadzwi, F. M., et al. (2022). A Review on
Nutrients, Phytochemicals, and Health Benefits of Green Seaweed, Caulerpa lentillifera. Foods, 11(18), 2832.
MDPI AG.
Strickland, J. D. H., and Parsons, T.R., (1972). A practical handbook of seawater analysis. Bulletin Fisheries Research
Board of Canada, 167, 81-86.
Setthamongkol, P., Tunkijjanukij, S., Satapornvanit, K., and Salaenoi, J. (2015). Growth and nutrients analysis inmarine macroalgae. Agriculture and Natural Resources, 49(2), 211-218.
Schroda M., Hemme D., and Muhlhaus T. (2015). The Chlamydomonas heat stress response. Plant J 82(3):466-480
Wagenen, V. J., Miller, T. W., Hobbs, S., Hook, P., Crowe, B., and Huesemann, M. (2012). Effects of light andtemperature on fatty acid production in Nannochloropsis salina. Energies, 5(3), 731-740.
Zhang, H., Li, W., Li, J., Fu, W., Yao, J., and Duan, D. (2012). Characterization and expression analysis of hsp70 genefrom Ulva prolifera J. Agardh (Chlorophycophyta, Chlorophyceae). Marine genomics, 5, 53-58.