Antioxidant and Antimicrobial Activity of <italic>Zostera marina</italic> L. Extract
Antioxidant and Antimicrobial Activity of Zostera marina L. Extract
ALGAE. 2009. Jun, 24(3): 179-184
Copyright ©2009, The Korean Society of Phycology
  • Received : July 14, 2009
  • Published : June 30, 2011
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About the Authors
Han Gil Choi
Faculty of Biological Science and Research Institute for Basic Science and Institute of Biotechnology, Wonkwang University, Iksan, Jeonbuk 570-749, Korea
Ji Hee Lee
Faculty of Biological Science and Research Institute for Basic Science and Institute of Biotechnology, Wonkwang University, Iksan, Jeonbuk 570-749, Korea
Hyang Ha Park
Faculty of Biological Science and Research Institute for Basic Science and Institute of Biotechnology, Wonkwang University, Iksan, Jeonbuk 570-749, Korea
Fotoon A. Q. Sayegh
Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia

Methanol crude extract of the sea grass Zostera marina L. and organic solvent fractions (n-hexane, chloroform, ethyl acetate, n-butanol, and water) were screened for antioxidant activity (total phenolic contents, DPPH scavenging activity, and reducing power) and antimicrobial activity against three human skin pathogens, two bacteria and a yeast; Staphylococcus aureus, Staphylococcus epidermidis , and Candida albicans . Total phenolic contents and 2, 2- diphenyl-1-picrylhydrazyl (DPPH) scavenging activity were highest in the ethyl acetate fraction with 968.50 μg gallic acid equivalent per milligram of extract, and ca. 95% scavenging activity on the DPPH radicals at 10 mg ml ?1 . In antimicrobial activity tests, MICs (Minimum Inhibitory Concentration) of each Zostera marina extract partitioned ranged from 1mg to 8 mg ml ?1 (extract/ 10% DMSO) against all three human skin pathogens. The MICs of the ethyl acetate and n-butanol fractions were the same with 1 mg ml ?1 against S. aureus and C. albicans . The ethyl acetate fraction of Z. marina does protect against free radicals and may be used to inhibit the growth of human skin pathogens.
Antioxidants are very important not only for the prevention of food oxidation but also for the defense of living cells against oxidative damage (Kim et al . 2003). Antioxidants reduce harmful reactive free radicals and reactive oxygen species (ROS) in cells (Anchana et al . 2005), thereby preventing cancer and heart disease (Yan et al . 1999; Qi et al . 2005). Several synthetic antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and tert-butylhydroquinone (TBHQ) are extensively used because of their excellent efficacy and low cost. However, these artificial chemicals have major side-effect issues including toxicity and DNA damage problems (Choi et al . 1993; Sasaki et al . 2002). Therefore, the identification and isolation of new natural antioxidants from aquatic and terrestrial plants are desirable (Nishida et al . 1996).
Zostera marina L. is an important species in coastal ecosystems because it contributes for nutrient cycling and sediment stabilizer, and provides food stuffs and habitat for many marine organisms such as invertebrates and fishes (Harrison 1882; Moore and Short 2006). Z. marina produces many secondary metabolites (e.g. phenolic compounds) in order to protect itself against microorganisms, epiphytes, and predation (Harrison and Chan 1980; Harrison 1982; Vergeer et al . 1995). As halophytes and mangroves growing in stressful environments make phenolic compounds to suppress the growth of yeast and mold (Bandaranayake 2002), Z. marina produces phenolic contents for protecting itself against Labyrinthula zosterae Porter & Muehlstein, a marine slime mould-like protist (Buchsbaum et al . 1990; Vergeer et al . 1995; Vergeer and Develi 1997). In marine plants, bioactive chemicals such as phenolic compound play important roles for survival and growth, and are now being used in the development of new drugs and health foods for human (Baker 1984; Smit 2004; Katalinic et al . 2006).
Harrison and Chan (1980) reported that methanol extracts of Zostera marina inhibit the growth of various invading microalgae (diatoms and flagellates) and a bacterium ( Staphylococcus aureus ). Antioxidant activity for the polysaccharide zosterine isolated from Z. marina has been examined by the activation of free radical peroxide oxidation in mice (Khasina et al . 2003) and the activity was compared with two antioxidant drugs (Kolenchenko et al . 2005). Stirk et al . (2007) found that the amount of bioactive chemicals in seaweeds is changed seasonally and regionally due to fluctuation of environmental conditions. For Z. marina methanol extracts, antioxidant activities using DPPH and reducing activity methods, and antimicrobial activities against two human skin pathogens (Staphylococcus epidermidis and Candida albicans ) have not been tested yet. It is also worth to note the antioxidant and antimicrobial activities of Korean Z. marina for the first time. Thus, the aim of present study was to examine the antioxidant and antimicrobial activities for Korean Z. marina methanol and fractioned extracts to determine which solvent fraction could be a potential source for developing natural antioxidant and antibiotics.
- Antioxidant activities
Preparation of Z. marina extract: Zostera marina was collected at Ihoijin Jangheung, Korea (34° 27’ N, 126° 56’ E), transported within an ice-box to the laboratory, and washed carefully in freshwater to remove sediment and epiphytes. Prior to experiment, samples were freeze dried and stored in a refrigerator below ?15°C. Extractions were made with methanol (MeOH) at room temperature for 24 hrs, filtered (Whatman No. 2), and concentrated by rotary evaporation (RE-111, Switzerland), 35°C below.
The MeOH crude extract (83.40 g) of Z. marina were partitioned into n-hexane, chloroform, ethyl acetate, nbutanol, and water fractions. The partition procedures were as follows; First, MeOH crude extract was dissolved in 200 ml distilled water followed by the addition of 100 ml n-hexane, chloroform, ethyl acetate, and nbutanol, three times respectively (Fig. 1) . Subsequently, the dried solvent fractions were dissolved in methanol and stored at below ?15°C.
Chemicals: Chemicals used were Folin-Ciocalteu phenol reagent, DPPH (α-diphenyl-β-picrylhydrazyl), Gallic acid, EDTANa 2 , trichloroacetic acid, FeCl 3 .6H 2 O, K 3 Fe(CN) 6 , Na 2 CO 3 , FeCl 2 , n-hexane, chloroform, ethyl acetate, methanol, n-butanol, butylated hydroxyanisole (BHA), and ferrozine [3(2pyridyl) 5,6bis (phenyl sulfonic acid) 1,2,4-triazine].
Total phenolic content: Total phenolic content of Z. marina crude and portioned extracts was quantified according to the methods of Yuan et al. (2005) and expressed as GAE (gallic acid equivalents). Each solvent extract (0.1 ml) of Z. marina was mixed with 2.0 ml of 2% Na2CO3 (dissolved in distilled water) in a test tube and kept at room temperature for 2 min. Subsequently, 0.1 ml of Folin-Ciocalteu’s phenol reagent (50%) was added to the above solution which was kept at room temperature for a further 30 min before absorbance was read at 720 nm.
DPPH radical-scavenging activity: To test radicalscavenging activity, reactions with the DPPH radical were carried out according to the method of Wangensteen et al . (2004). Each solvent extract of Z. marina was made in various concentration (0.1, 0.5, 1, 5, 10, and 20 mg ml ?1 ) and mixed with 2.90 ml of a DPPH solution, which was prepared at a concentration of 0.2 mM in MeOH. After standing for 30 min, absorbance was measured by a UV-VIS spectrophotometer at 517 nm (Biochrom, Libsra S22, England), using MeOH as the blank. The DPPH solution was prepared daily, stored in a flask at room temperature, covered with aluminum foil, and all determinations were performed three times. Antioxidant capacity of the test extracts was expressed as the concentration necessary for reduction of DPPH. Percent radical scavenging was calculated as 100× (A start ? A end )/A start , where A start is the absorbance before addition of test extract and A end is the absorbance value after 30 min of reaction time.
Reducing power: Reducing activity of various solvent fractions in Z. marina was determined according to the methods of Yen and Chen (1995). Extracts in phosphate buffer (2.5 ml, 0.2 mol, pH 6.6) were added to potassium ferricyanide (2.5 ml, 10 g L ?1 ) and the mixture was incubated at 50°C for 20 min. Trichloroacetic acid (2.5 ml, 100 g L ?1 ) was added to the mixture and then centrifuged for 10 min at 3,000 rpm. Finally, 2.5 ml of the upper layer was mixed with 2.5 ml of distilled water and 0.5 ml of 0.1% aqueous FeCl 3 , after which absorbance was recorded at 700 nm. Higher optical density of the reaction mixture indicated greater reducing power.
- Antimicrobial activity
The various solvent fractions of Z. marina were used for assessing antimicrobial activity. Antimicrobial activity was examined against three human skin pathogens; two bacteria ( Staphylococcus aureus KCTC 1927, S. epidermidis KCTC 1917) and a yeast ( Candida albicans KCTC 7596), using the MIC (minimum inhibitory concentration) method.
The two bacteria were cultured in nutrient broth (8 g L ?1 , Difco, USA) on agar (15 g L ?1 ), while C. albicans was cultured on Sabouraud dextrose broth (30 g L ?1 , Difco, USA) on agar (15 g L ?1 ). Suspensions of Staphylococcus aureus , S. epidermidis , and C. albicans were adjusted to an optical density (O.D.) of 0.1 at 600 nm. The cell density of test microorganisms containing 0.1 ml of broth solution was 10 8 CFU (colony forming unit).
The solvent fractions of Z. marina were dissolved in 10% dimethylsulfoxide (DMSO). Blank DMSO were also placed in separate wells and served as controls. Two-fold serial dilutions of sample solutions were made and final concentrations ranged from 0.125 to 8 mg ml ?1 . Antimicrobial activity of each solvent fraction against the three skin pathogens was examined by estimating MIC after culture for 24 hrs at 37°C.
- Statistical analyses
Statistical analyses were carried out using STATISTICA version 5.0 software. A one-way ANOVA was used to determine the differences in total phenolic content between the solvent fractions of Zostera marina extract. When significant differences between means were detected, Tukey HSD test was applied (Sokal and Rohlf 1995). Homogeneity of the variance was tested using Cochran’s test (Underwood 1997).
- Antioxidant activity
Total phenolic content: MeOH crude extract of Zostera marina was fractionated with various solvents, from which the following were obtained; 4.97 g in n-hexane, 0.34 g in chloroform, 0.35 g in ethyl acetate, 1.08 g in nbutanol, and 50.42 g (60.45%) in water fraction.
Total phenolic contents were significantly different between solvent fractions of Z. marina (F 5, 12 = 244.75, p < 0.001). Total phenolic content was 968.50 μg per 1 mg of ethyl acetate fraction (Fig. 1) which was significantly greater than the other fractions (Tukey HSD test). Also, phenolic content was significantly lesser at water fraction (50.25 μg) than at crude fraction (204.63 μg) of Z. marina , but no differences in total phenolic contents were found between n-hexane, chloroform, n-butanol, and crude solvent fractions (Tukey HSD test). This indicates that the phenolic compounds of Z. marina are mainly contained in non polar extracts rather than in polar solvent extracts.
DPPH radical scavenging activity: DPPH radical scavenging activity was enhanced with increasing concentration (between 0.1 and 20 mg ml ?1 ) for the five solvent fractions and crude extracts of Z. marina ( Table 1 ). At water fraction, however, DPPH inhibition activities were very low with less than 10% in the tested concentrations. DPPH radical scavenging activity was significantly greater at ethyl acetate fraction with IC 50 values of 0.46 mg ml ?1 than the other solvent fractions including MeOH crude extracts in the range of between 0.1 and 5 mg ml ?1 . At 5 mg ml ?1 of ethyl acetate fraction, DPPH scavenging activity was significantly different between treatments (F 7, 16 = 905.99, p < 0.001), but it was not significantly different compared to crude extract, BHA, and ascorbic acid (Tukey HSD test; Table 1 ). As shown in Table 1 , the ethyl acetate fraction was most effective in DPPH radical scavenging activity with 95.17%, followed by n-butanol fraction (91.27%) and the MeOH crude extract (90.62%) at a concentration of 10 mg ml ?1 . In the present study, DPPH radical scavenging activity was relatively high in the non-polar solvent fractions of ethyl acetate and n-butanol.
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Total phenolic content (mean ± SD, n = 3) in different solvent fractions and crude extract of Zostera marina at 1mg ml?1. Total phenolic content is expressed as gallic acid equivalents (GAE; μg mg?1).
Reducing power: For determination of reducing power, the reduction of Fe 3+ to Fe 2+ was measured in various solvent fractions of Zostera marina . At 0.1 mg ml ?1 , reducing power was significantly different between solvent fractions, crude extract, and BHA (F 6, 14 = 943.96, p < 0.001), and it was significantly greater at n-butanol fraction than the other solvent fractions (Tukey HSD test). However, reducing power was significantly lower at n-butanol compared to the control, BHA at 0.1 mg ml ?1 concentration. At lower concentrations (between 0.1 and 1 mg ml ?1 ), reducing power was stronger in the nbutanol fraction than in the ethyl acetate fraction. However, it was very similar between ethyl acetate and n-butanol fractions at higher concentrations of between 5 and 20 mg ml ?1 (Table 2) . Thus, the reducing power of the ethyl acetate and n-butanol fractions exhibited higher activity than other fractions.
Dose-dependent DPPH inhibition activity (%, mean ± SE, n = 3) ofZostera marinaextracts at 517 nm
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Dose-dependent DPPH inhibition activity (%, mean ± SE, n = 3) of Zostera marina extracts at 517 nm
Dose-response of reducing power (mean ± SE, n = 3) ofZostera marinaextracts at 700 nm
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Dose-response of reducing power (mean ± SE, n = 3) of Zostera marina extracts at 700 nm
- Antimicrobial activity
As the MeOH crude extract (10 g dry wt.) of Zostera marina plants was partitioned, the yields of each solvent fraction varied from 0.03 g (0.32%) in ethyl acetate to 1.19 g (11.87%) in water. Antimicrobial activity of Z. marina fractions was in the range of between 1 and 8 mg ml ?1 concentration against three human skin pathogens but it was not detected at water fraction (Table 3) . Minimum inhibitory concentration for Staphylococcus aureus was found at 1 mg ml ?1 in the ethyl acetate fraction. MICs of S. epidermidis were the same at 8 mg ml ?1 , irrespective of Z. marina solvent fractions. Candida albicans showed different MICs from 1 mg ml ?1 in the n-butanol fraction, 4 mg ml ?1 in chloroform fraction, to 8 mg ml ?1 in the three solvent fractions (Table 3) .
Plant products such as flavonoids, cumarins, phenolic acids and terpenoids are reported to have DPPH scavenging activity (Puertas-Mejia et al . 2002). At the same concentration (3×10 ?5 mol L ?1 ), DPPH quenching activity was stronger in the extract of non-polar solvents (chloroform, ethyl acetate and acetone) than at water, ethanol, or methanol extract in several Rhodomelaceae seaweeds (Yan et al . 1998; Yuan et al . 2005). Jimenez-Escrg et al . (2001) found that DPPH radical scavenging activities are positively correlated with the total polyphenol content in aqueous and organic solvent extracts of brown and red algae. In the present study, phenolic content was approximately 968.50 μg at 1mg of non-polar ethyl acetate fraction of Zostera marina , and DPPH radical scavenging activity (95.55% at 5 mg ml ?1 ) was the highest among various solvent fractions. These results indicate that the presence of compounds with phenolic functional groups in the ethyl acetate fraction.
Minimum inhibitory concentration (MIC, mg ml?1) ofZostera marinafractions against three human skin pathogens
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Minimum inhibitory concentration (MIC, mg ml?1) of Zostera marina fractions against three human skin pathogens
In the present study, 1 mg ml ?1 of n-butanol fraction of Zostera marina appeared to possess 5 times higher reducing activity than at ethyl acetate fraction of Capsicum annuum L. and 35 times greater than at MeOH extract of Palmaria palmata (L.) Kuntze (Kim et al . 2003; Yuan et al . 2005). It is well known that ROS induce oxidative damage to biomolecules like nucleic acids, lipids, proteins, and carbohydrates, and this damage causes cancer, and other disease (Duan et al . 2006). Present results show that ethyl acetate and n-butanol fractions of Z. marina are potential candidates in the development of medicinal compounds relating to prevention of human diseases.
Antimicrobial activity depends on solvent fractions and test organisms. The MICs was the same with 1 mg ml ?1 in the ethyl acetate fraction of Z. marina for Staphylococcus aureus and in the n-butanol fraction against C. albicans . For S. aureus , MICs of ethyl acetate fractions were 1 mg ml ?1 for Z. marina and 8 mg ml ?1 for Neorhodomela aculeata (Perestenko) Musuda (Lee et al . 2006). Harrison and Chan (1980) reported that growth inhibition of several micro-algae and a bacterium, Staphylococcus aureus in Z. marina MeOH extracts. For S. aureus , however, they examined antimicrobial activity with paper disc method and it is impossible to compare the degree of bioactivity between previous and present results. Lee et al . (2006) found that MICs of S. epidermidis were higher (between 16 and 32 mg ml ?1 ) than S. aureus (8-16 mg ml ?1 ) and C. albicans (16 mg ml ?1 ) at four organic solvents (n-hexane, chloroform, ethyl acetate, and methanol) of Neorhodomela aculeata . In the present study, MICs were 8 mg ml ?1 for S. epidermidis and 1-8 mg ml ?1 for S. aureus and C. albicans in all tested solvent fractions. These results indicate that S. epidermidis is less sensitive than two human skin pathogens in both Z. marina and N. aculeata , and various solvent extracts of Z. marina show higher antimicrobial activity than those of N. aculeata against Staphylococcus epidermidis and C. albicans . Vergeer et al . (1995) reported that the production of phenolic compounds in Z. marina increased when infected by Labyrinthula zosterae , indicating that the production of phenolic compounds is an antimicrobial response.
Research studies have already described the positive relationship among total phenolic content, antioxidant activity, and antimicrobial activity (Velioglu et al . 1998; Da Silva et al . 2006). In the present study, antioxidant (DPPH radical scavenging activity and reducing power) and antimicrobial activities against three human skin pathogens were excellent in ethyl acetate fraction of Z. marina compared to other seaweeds such as Palmaria palmata and N. aculeata . Although the chemical compounds of ethyl acetate fractions from Z. marina have not yet been fully identified, they might be related to phenolic contents which were maximal in ethyl acetate fraction. In conclusion, the ethyl acetate and n-butanol fractions of Z. marina might be useful for developing natural antioxidants for many human diseases and new antibiotics against human skin pathogens but more researches on the isolation and identification of the bioactive compounds are required.
We would like to thank three anonymous reviewers for helpful comments which improved the manuscript. This paper was supported by Wonkwang University in 2009.
Anchana C , Aphiwat T , Nuansri R 2005 Screening of antioxidant activity and antioxidant compounds of some edible plants of Thailand Food Chemistry 92 (3) 491 - 497    DOI : 10.1016/j.foodchem.2004.07.035
Baker J.T 1984 Seaweeds in pharmaceutical studies and applications Hydrobiologia 116-117 (1) 29 - 40    DOI : 10.1007/BF00027635
Baker J.T 2002 Bioactivities bioactive compounds and chemical constituents of mangrove plants Wetlands Ecology and Management 10 (6) 421 - 452    DOI : 10.1023/A:1021397624349
Baker J.T 1990 Phenolic-nitrogen interactions in eelgrass Zostera marina L: possible implications for disease resistance Aquatic Botany 37 (3) 291 - 297    DOI : 10.1016/0304-3770(90)90075-V
Choi J.S , Lee J.H , Park H.J , Kim H.G , Young H.S , Mun S.I 1993 Screening for Antioxidant Activity of Plants and Marine Algae and Its Active Principles from Prunus davidiana Korean Journal of Pharmacognosy 24 (4) 299 - 303
Da Silva J.F.M , de Souza M.C , de Andrade S.M.M.R , Vidal F.V.N , Vidal F.V.N. 2006 Correlation analysis between phenolic levels of Brazilian propolis extracts and their antimicrobial and antioxidant activities Food Chemistry 99 (3) 431 - 435    DOI : 10.1016/j.foodchem.2005.07.055
Duan X.J , Zhang W.W , Li X.M , Wang B.G 2006 Evaluation of antioxidant property of extract and fractions obtained from a red alga Polysiphonia urceolata Food Chemistry 95 (1) 37 - 43    DOI : 10.1016/j.foodchem.2004.12.015
Harrison P.G 1982 Control of microbial growth and of amphipod grazing by water-soluble compounds from leaves of Zostera marina Marine Biology 67 (2) 225 - 230    DOI : 10.1007/BF00401288
Harrison P.G , Chan A.T 1980 Inhibition of the growth of micro-algae and bacteria by extracts of eelgrass (Zostera marina) leaves Marine Biology 61 (1) 21 - 26    DOI : 10.1007/BF00410338
Jimenez-Escrg A , Jimenez-Jimenez I , Pulido R , Saura-Calixto Saura-Calixto 2001 Antioxidant activity of fresh and processed edible seaweeds Journal of the Science of Food and Agriculture 81 (5) 530 - 534    DOI : 10.1002/jsfa.842
Katalinic V , Milos M , Kulisic T , Jukic M 2006 Screening of 70 medicinal plant extracts for antioxidant capacity and total phenols Food Chemistry 94 (4) 550 - 557    DOI : 10.1016/j.foodchem.2004.124
Khasina E.I , Kolenchenko E.A , Sgrebneva M.N , Kovalev V.V , Khotimchenko Y.S 2003 Antioxidant Activities of a Low Etherified Pectin from the Seagrass Zostera marina Russian Journal of Marine Biology 29 (4) 259 - 261    DOI : 10.1023/A:1025493128327
Kim J.H , Jeong C.H , Shim K.H 2003 Biological Activities of Solvent Fractions of Capsicum annuum Leaves Korean Journal of Food Preservation 10 (4) 540 - 546
Kolenchenko E.A , Sonina L.N , Khotimchenko Y.S 2005 Comparative in vitro Assessment of Antioxidant Activities of Low-Etherified Pectin from the Eelgrass Zostera marina and Antioxidative Medicines Russian Journal of Marine Biology 31 (5) 331 - 334    DOI : 10.1007/s11179-005-0098-2
Lee J.H , Lee K.H , Yoo H.I , Zhou X.L , Choi H.G , Kim Y.S , Nam K.W 2006 Antimicrobial Activity of Neorhodomela aculeata Extracts Against Human Skin Pathogens Journal of the Korean Fisheries Society 39 (3) 292 - 296
Moore K.A , Short F.T 2006 Zostera: Biology Ecology and Management Springer The Netherlands
Nishida T , Matsukawa R , Masaki K , Dubinsky Z , Karube I 1996 A method for screening potential antioxidant activity Journal of Biotechnology 51 (2) 149 - 155    DOI : 10.1016/0168-1656(96)01611-2
Puertas-mejia M , Hillebr S , Stashenko E , Winterhalter P , Stashenko Elena , Winterhalter Peter 2002 In vitro radical scavenging activity of essential oils from Columbian plants and fractions from oregano (Origanum vulgare L) essential oil Flavour and Fragrance Journal 17 (5) 380 - 384    DOI : 10.1002/ffj.1110
Qi H , Zhao T , Zhang Q , Li Z , Zhao Z , Xing R 2005 Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta) Journal of Applied Phycology 17 (6) 527 - 534    DOI : 10.1007/s10811-005-9003-9
Sasaki Y.F , Kawaguchi S , Kamaya A , Ohshita M , Kabasawa K , Iwama K , Taniguchi K , Tsuda S 2002 The comet assay with 8 mouse organs: Results with 39 currently used food additives Mutation Res Gen Toxicol Mutation Research/Genetic Toxicology and Environmental Mutagenesis 519 (1-2) 103 - 119    DOI : 10.1016/S1383-5718(02)00128-6
Smit A.J 2004 Medicinal and pharmaceutical uses of seaweed natural products: A review Journal of Applied Phycology 16 (4) 245 - 262    DOI : 10.1023/B:JAPH00047783.36600.ef
Sokal R.R. , Rohlf F.J. 1995 Biometry 3rd edition WH Freeman New York
Stirk W.A , Reinecke D.L , van Staden D.L 2007 Seasonal variation in antifungal antibacterial and acetylcholinesterase activity in seven South African seaweeds Journal of Applied Phycology 19 (3) 271 - 276    DOI : 10.1007/s10811-006-9134-7
Underwood A.J 1997 Experiments in Ecology: Their Logical Design and Interpretation Using Analysis of Variance Cambridge University Press Cambridge
Velioglu Y.S , Mazza G , Gao L , Oomah B.D 1998 Antioxidant Activity and Total Phenolics in Selected Fruits Vegetables and Grain Products Journal of Agricultural and Food Chemistry 46 (10) 4113 - 4117    DOI : 10.1021/jf9801973
Vergeer L.H.T , Aarts T.L , de Groot J.D 1995 The 'wasting disease' and the effect of abiotic factors (light intensity temperature salinity) and infection with Labyrinthula zosterae on the phenolic content of Zostera marina shoots Aquatic Botany 52 (1) 35 - 44    DOI : 10.1016/0304-3770(95)00480-N
Vergeer L.H.T , Develi A 1997 Phenolic acids in healthy and infected leaves of Zostera marina and their growth-limiting properties towards Labyrinthula zosterae Aquatic Botany 58 (1) 65 - 72    DOI : 10.1016/S0304-3770(96)01115-1
Yan X.J , Nagata T , Fan X 1998 Antioxidative activities in some common seaweeds Plant Foods for Human Nutrition 52 (3) 253 - 256    DOI : 10.1023/A:1008007014659
Yan X , Chuda Y , Suzuki M , Nagata T 1999 Fucoxanthin as the major antioxidant in Hijikia fusiformis a common edible seaweed Bioscience Biotechnology and Biochemistry 63 (3) 605 - 607    DOI : 10.1271/bbb.63.605
Yen G.C , Chen H.Y 1995 Antioxidant Activity of Various Tea Extracts in Relation to Their Antimutagenicity Journal of Agricultural and Food Chemistry 43 (1) 27 - 32    DOI : 10.1021/jf00049a007
Yuan Y.V , Bone D.E , Carrington M.F 2005 Antioxidant activity of dulse (Palmaria palmata) extract evaluated in vitro Food Chemistry 91 (3) 485 - 494    DOI : 10.1016/j.foodchem.2004.04.039
Wangensteen H , Samuelsen A.B , Malterud K.E 2004 Antioxidant activity in extracts from coriander Food Chemistry 88 (2) 293 - 297    DOI : 10.1016/j.foodchem.2004.01.047