INTERNATIONAL JOURNAL OF AGRICULTURE & BIOLOGY

ISSN Print: 1560–8530; ISSN Online: 1814–9596

10–293/AWB/2011/13–2–297–300
http://www.fspublishers.org

Short Communication  
The Nutritional Value of Edible Freshwater Alga Cladophora sp.
(Chlorophyta) Grown under Different Phosphorus
Concentrations
TAWEESAK KHUANTRAIRONG1 AND SIRIPEN TRAICHAIYAPORN
Algae and Water Quality Research Unit, Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai
50200, Thailand
1
Corresponding author’s e-mail: [email protected]

ABSTRACT

Culture of an edible freshwater alga Cladophora sp. was subjected to phosphorus (P) supply (1.07-14.78 mg L-1 of PO4-P) in
order to determine its proximate compositions, vitamins, minerals and carotenoid compositions. The alga was mass cultured
by using 10% of canteen wastewater with the addition of di-potassium hydrogen orthophosphate at the concentrations of 0, 5,
10, 15 and 20 mg L-1. The results showed that with increased P concentration, protein, vitamin A, P, β-carotene, lutein and
zeaxanthin contents increased but carbohydrate content decreased indicating that P supply could enhance carotenoid
production and some nutritional values of this alga. © 2011 Friends Science Publishers

Key Words: Cladophora; Edible algae; Nutritional value; Carotenoid composition

INTRODUCTION (Menendez et al., 2004), Skeletonema costatum,

Chaetoceros muelleri (Leonardos & Lucas, 2000) and the
Among the major freshwater edible algae of Thailand higher plants lentil (Zeidan, 2007; Togay et al., 2008) and
are Cladophora, Microspora, Nostoc, Nostochopsis and forage maize (Eltelib et al., 2006). This study assessed the
Spirogyra. In Thailand edible freshwater algae, Cladophora nutrition of a macroalga Cladophora sp. culture in different
is known as Kai. It is abundant in the Nan and Mekong phosphorus concentration i.e., proximate compositions,
rivers in the Northern part of Thailand. The local people vitamins, minerals and carotenoid compositions contents.
around these rivers collect it for domestic consumption and
it is sold in the markets and is now cultured by using MATERIALS AND METHODS
canteen wastewater for fish supplement feed
(Traichaiyaporn et al., 2010). Many reports on nutritional Canteen wastewater preparation: Canteen wastewater
value of Cladophora show that in contains significant (1000 L) was collected from canteen wastewater clarifier of
amounts of protein and carotenoids, which are essential for Maejo University and allowed to settle in an open-air
human and fish nutrition (Khuantrairong & Traichaiyaporn, cement pond for 3 weeks for the breakdown of solid organic
2009; Traichaiyaporn et al., 2010). Studies on Cladophora wastes by microorganisms. The water was then filtered
have been reported especially on its taxonomy and through an 80 µm plankton net filter (Promya et al., 2008).
nutritional values (Powtongsook, 2000; Peerapornpisal et The filtrate was analyzed for pH (8.64), total hardness
al., 2006; Khuantrairong & Traichaiyaporn, 2009) but (145.50 mg L-1), orthophosphate phosphorus (4.84 mg L-1),
reported of relationship between its nutrition and ammonia nitrogen (3.05 mg L-1) and nitrate nitrogen (6.44
environmental are few. mg L-1). Then the filtrate was diluted to 10% for algal
Algae are significant source of human food, especially cultivation.
in Asia and cultivated for nutrition and pigments for Culture conditions: Cladophora sp. was obtained from
supplemental use as human food and animal feed (Promya Algae and Water Quality Research Unit, Chiang Mai
et al., 2008). Factor such as cell size and shape, rate of University, Thailand. The alga was bath cultured by
ingestion, digestibility and biochemical composition attachment on plastic nets (60 g m-2) in cement raceway
determine the nutritive quality of algae and their utility as ponds (size 1.2×2.3×0.5 m) using 10% canteen wastewater,
food (Whyte, 1987; Doroudi et al., 2003). with continuous flow of 0.15 m s-1 for 12 weeks
Studies on effect of phosphorus on nutritional values (Khuantrairong & Traichaiyaporn, 2009; Traichaiyaporn et
have been reported in algae Chaetomorpha linum al., 2010). A complete randomize design (CRD) was carried

To cite this paper: Khuantrairong, T., and S. Traichaiyaporn, 2011. The nutritional value of edible freshwater alga Cladophora sp. (Chlorophyta) grown
under different phosphorus concentrations. Int. J. Agric. Biol., 13: 297–300
 KHUANTRAIRONG & TRAICHAIYAPORN / Int. J. Agric. Biol., Vol. 13, No. 2, 2011

out using 4 treatments with 3 replications by addition of 4 vitamin B2 (0.05-0.06); vitamin C (1.89-1.94) and vitamin E
concentrations of di-potassium hydrogen orthophosphate (5.96-6.00). Statistical analysis indicated that increasing the
(K2HPO4) at 5, 10, 15 and 20 mg L-1 (treatment 1, 2, 3 & 4, concentration of phosphorus increased of vitamin A content
respectively) to the cultured media and the control group in Cladophora sp., but had no effect on vitamin B1, B2, C
was 10% canteen wastewater without adding K2HPO4. and E contents (Table I).
Analysis method: Algal samples at 12 weeks of culturing The levels of Ca, Fe, K, Mg, P and Zn contents (in mg
were harvested, washed, air dried and freeze-dried. The 100 g-1) were similar among the treatments and ranged from
samples were analyzed for the purposes: 6585-6863, 29.40-30.87, 2512-2658, 241.60-255.30 and
Proximate composition, vitamin and mineral analysis: 1.87-1.95, respectively. The P contents ranged from 69.17-
The protein, fat, ash, moisture and fiber were determined by 111.79 mg 100 g-1. There was significant difference among
standard AOAC (2000) methods. Carbohydrate was treatments and the highest value was observed in treatment
determined according to the method of Crampton and Harris 4 which addition of 20 mg L-1 K2HPO4 (Table I).
(1969). The vitamin A content was calculated on the basis Carotenoid composition: Fig. 1 shows the HPLC
of β-carotene content according to the method of Tee and chromatogram of carotenoid compositions in the extracted
Lim (1991) as vitamin A (retinol equivalent, RE) = (µg β- sample of Cladophora sp. β-carotene, lutein and zeaxanthin
carotene)/6. The level of four vitamins (B1, B2, C & E) and contents (in µg g-1) of this species culture at different
six minerals (Ca, Fe, K, Mg, P & Zn) were determined by in phosphorus concentrations were observed as 20.01-96.59,
house method base on AOAC (2000). 172.80-437.28 and 24.62-171.75, respectively (Table I). The
Carotenoid compositions analysis: β-carotene, lutein and highest values of these pigments were observed in treatment
zeaxanthin were extracted following the method described 4, which addition of 20 mg L-1 K2HPO4, whereas the lowest
by Yoshii et al. (2004) with some modifications. The values were observed in the control group. Statistical
samples (0.5 g) were extracted on ice in the dark with 25 analysis showed that the levels of β-carotene, lutein and
mL of 100% acetone, stored in the dark at –20oC for 18 h zeaxanthin were significantly different among all the
and the supernatant was filtered. Investigation and treatments (Table I) suggesting that phosphorus effected the
quantification of these pigments were performed by reverse increase in carotenoid compositions production of
phase high liquid chromatography (HPLC) with method Cladophora sp.
modified from Mostaert et al. (1998) and Yoshii et al.
(2004), using a C30 carotenoid column (5 µm; 250 × 4.6 DISCUSSION
mm). The flow rates were 1.0 mL min-1 at 0-8 min and 2.0
mL min-1 at 8-18 min. The mobile phase was methanol: The biochemical composition of algae varies between
chloroform (4:1). Peak responses were determined by a UV- species and according to culture conditions (Leonardos &
Vis at the absorbance of 456 nm. The carotenoids peaks Lucas, 2000; Martínez-Fernández et al., 2006). It is
were identified with compared by pure β-carotene, lutein influenced by growth phase and culture medium
and zeaxanthin standards from Sigma Chemical Company. composition as well as environment factors (Brown et al.,
Statistical analysis: For all analyses, the mean and standard 1993; Valenzuela-Espinosa et al., 2002). In the present
deviation were calculated and reported. Analysis of variance study, proximate composition, vitamin and mineral contents
(ANOVA) and Duncan’s Multiple Range Test (DMRT) of Cladophora sp. i.e., protein, vitamin A and P increased
were adopted for statistical analyses and comparative with increased phosphorus levels in culture media, while
studies of proximate compositions, vitamins, minerals and carbohydrate decreased. The previous studies have been
carotenoids of Cladophora sp. growth under different shown the environmental conditions influence to algal
phosphorus concentrations. nutritional values. Khuantrairong and Traichaiyaporn (2009)
suggested that P content of Cladophora sp. was positively
RESULTS correlated with phosphorus levels in culture medium.
Leonardos and Lucas (2000) reported that phosphorus
Proximate composition: The proximate composition of limitation affected to decrease protein content of a
Cladophora sp. indicated that the alga contained: protein microalga Chaetoceros muelleri.
(10.71-17.69%); lipid (2.04-2.56%); ash (14.71-16.86%); Elenkov et al. (1996) reported that high salinity
moisture (9.87-11.32%); fiber (20.67-26.10%) and affected increasing lipid content of Cladophora vagubunda.
carbohydrate (52.54-60.98%). Protein contents increased In addition, C. siwaschensis living in high salt water was
under high phosphorus (P) concentration, while considerably increased calcium and phosphorus content
carbohydrate contents decreased. Statistical analysis showed (Alfimov & Proshkina-Lavrenko, 1961). Auer and Canale
that protein and carbohydrate contents were significantly (1982) observed that high dissolved P in water induced an
different among the treatments (Table I). increase in tissue P content of Cladophora. Moreover,
The vitamins contents (in mg 100 g-1) of Cladophora enrichment of P in culture media increased tissue P of a
sp. culture at different P concentrations were observed as green macroalga C. linum (Menendez et al., 2004). In the
follows: vitamin A (0.33-1.61); vitamin B1 (0.05-0.06); higher plants, Zeidan (2007) and Togay et al. (2008) stated

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Table I: Nutritional values of Cladophora sp. cultured at different phosphorus concentrations

Nutritional values Amount of nutritional values
Addition of K2HPO4 (mg L-1)
0 5 10 15 20
Proximate (%)
Protein 10.71±1.14a 11.94±0.55a 15.00±0.88b 15.46±0.47b 17.69±1.75c
Lipid 2.04±0.78a 2.56±0.45a 2.29±0.93a 2.09±0.30a 2.46±1.19a
Ash 15.29±1.11a 15.56±0.67a 15.14±1.52a 14.71±0.64a 16.86±0.84a
Moisture 10.98±1.91a 11.11±1.19a 9.87±0.75a 11.32±0.86a 10.45±1.24a
Fiber 23.05±2.63a 26.10±2.72a 25.4±2.93a 20.67±1.28a 22.65±3.78a
Carbohydrate 60.98±1.57c 58.83±1.63b 57.70±1.21b 56.43±0.47b 52.54±2.26a
Vitamin (mg 100g-1)
Vitamin A 0.33±0.23a 1.02±0.15b 1.11±0.21b 1.59±0.40c 1.61±0.08c
Vitamin B1 0.05±0.01a 0.06±0.02a 0.06±0.02a 0.05±0.01a 0.06±0.01a
Vitamin B2 0.05±0.00a 0.06±0.01a 0.06±0.02a 0.06±0.01a 0.05±0.01a
Vitamin C 1.89±0.10a 1.91±0.04a 1.91±0.13a 1.93±0.11a 1.94±0.05a
Vitamin E 5.97±0.13a 6.01±0.01a 5.98±0.20a 5.96±0.08a 6.00±0.21a
Minerals (mg 100g-1)
Calcium (Ca) 6,631±209.03a 6,657±14.14a 6,585±279.31a 6,863±211.42a 6,740±169.00a
Iron (Fe) 29.91±1.93a 29.40±0.88a 29.55±0.64a 30.87±1.93a 29.57±1.08a
Potassium (K) 2,658±200.11a 2,652±137.18a 2,512±149.20a 2,652±136.47a 2,633±43.13a
Magnesium (Mg) 241.60±18.67a 245.45±6.43a 251.55±17.18a 254.00±5.66a 255.30±8.20a
a
Phosphorus (P) 69.17±1.17 71.03±1.40a 87.82±3.24b 94.55±1.42b 111.79±13.07c
Zinc (Zn) 1.91±0.04a 1.94±0.09a 1.91±0.01a 1.95±0.16a 1.87±0.09a
Carotenoid (µg g-1)
β-carotene 20.01±13.99a 61.03±9.26b 66.38±12.83b 95.56±24.11c 96.59±5.10c
Lutein 172.80±48.11a 296.70±16.97b 395.39±18.17c 370.80±1.32c 437.28±26.38d
Zeaxanthin 24.62±12.49a 72.17±3.00b 79.40±2.27b 92.47±7.17b 171.75±46.69c
*Data in the same row with different superscripts are significantly different (p<0.05)
Fig. 1: HPLC chromatogram of carotenoid were observed under P limitation. Leonardos and Geider
compositions extracted from Cladophora sp. Peaks are (2005) stated that P and nitrogen ratio was related to
lutein (1), zeaxanthin (2) and β-carotene (3) carotenoid production in cryptophyte Rhinomonas
reticulata. These reports strongly suggested that phosphorus
affected carotenoids production in Cladophora sp.
In conclusion, P supply had effect on the increase
carotenoids production and some nutritional values of
Cladophora sp. Further study should be investigated effect
of other factors such on nutritional values of this alga.
Acknowledgment: Financial support from the Thailand
Research Fund through the Royal Golden Jubilee Ph.D.
Program (Grant No. PHD/0130/2548) is acknowledged.

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