Joon-Baek Lee

Potential of Benthic Diatoms Achnanthes longipes, Amphora coffeaeformisand Navicula sp. (Bacillariophyceae) as Antioxidant Sources

Reactive oxygen species (ROS) such as hydrogen peroxide, hydroxyl radicals and superoxide anions, which are produced because of exogenous factors (tobacco smoke, ionizing radiation and certain pollutants) and endogenous factors (normal aerobic respiration) have been implicated in cellular processes such as mutagenesis, carcinogenesis and premature aging. ROS have the ability to react with a large variety of easily oxidisable cellular components, such as proteins, lipids and lipoproteins (Fridovich 1995). Oxidation is one of the major reasons of food deterioration, resulting to the destruction of fat-soluble vitamins and development of off colors and toxicants (Yang et al. 2000; Ukeda et al. 2002). Besides, in the food industry lipid peroxidation is an important deteriorative reaction during processing and storage. Commercial antioxidant supplements such as butylated hydroxyl anisole (BHA), butylated hydroxy tolouene (BHT), α-tocopherol and propyl gallate (PG) have been used in order to reduce oxidative damages in human body (Sherwin 1990; Gulcin et al. 2002). However, it is suspected that those antioxidants are responsible for some side effects such as liver damage and carcinogenesis. Antioxidants are involved with the oxidation mechanism by scavenging free radicals, chelating catalytic metals and by acting as oxygen scavengers (Shahidi and Wanasundara 1992; Buyukokuroglu et al. 2001). As a natural antioxidant source, plants have an ability to absorb the sun’s radiation for generating high levels of oxygen as secondary metabolites of photosynthesis. Oxygen is easily activated by ultra violet (UV) radiation and heat from the sunlight to produce toxic ROS. Therefore, plants produce various antioxidative comAlgae Volume 24(1): 47-55, 2009

Seasonal Cycle of Phytoplankton in Aquaculture Ponds in Bangladesh

A study on the seasonal changes in the phytoplankton community was carried out in four aquaculture ponds of Bangladesh over a period of 16 months from August 2000 to November 2001. Out of 45 phytoplankton species identified, 30 belong to Cyanophyceae, 7 to Chlorophyceae, 5 to Bacillariophyceae and 3 to Euglenophyceae. The highest phytoplankton abundance was observed in spring followed by early autumn, summer, and the lowest was in winter. The annual succession of Cyanophyceae was characterized by spring and early autumn period dominated by Microcystis sp. Anabaena sp. and Planktolymbya sp. with Microcystis sp. as the main blue-green algae represented. Chlorophyceae was characterized by rainy season domination of Chlorella vulgaris, Pediastrum sp. and Scenedesmus denticulatus with maximum abundance of Chlorella vulgaris. Whereas Bacillariophyceae was dominant during the winter period. Navicula angusta and Cyclotella meneghiniana were the most frequently occurring species of Bacillariophyceae throughout the study period. Euglenophyceae was dominant in late autumn and Euglena sp. was the dominant species. The effect of various physicochemical water quality parameters on the seasonal distribution and succession of the above mentioned phytoplankton population as well as the interaction and eutrophication are discussed.

Growth Characteristics, Bio-chemical Composition and Antioxidant Activities of Benthic Diatom Grammatophora marina from Jeju Coast, Korea

Benthic diatoms are known as a good food for shellfish in nature and in commercial hatchery of Jeju Island, Korea. Grammatophora marina is commonly found as dominant benthic micro-algae in Jeju coastal waters throughout the year. To know the best growth conditions of this species, culture was done in terms of three parameters; water temperature, salinity and nutrients. Each parameter was controlled by temperature of 15, 20 and 25°C; salinity of 25, 30 and 35 psu; and nutrient concentrations of 50, 100 and 200%. F/2 media was used with artificial seawater for the culture, which was continued for two weeks with L:D cycle 12:12 by using fluorescent light. Maximum specific growth rate was recorded 1.68 d ‐1 at temperature of 25°C with salinity of 35 psu and nutrient concentration of 200% on 6 th day during the culture period. Maximum biomass was also observed 4.9 × 10 5 cells mL ‐1 in the same condition. This species may belong to the euryhaline and eutrophic habitat with warm condition. For nutritional aspects of this species, protein, lipid and carbohydrate were measured. The value of protein, lipid and carbohydrate was 4.96%, 15.82% and 5.65%, respectively. The antioxidant activities of 80% methanolic extract were 46.7%, 23.7% and 23.8% on DPPH (1,1-Diphenyl-2-picrylydrazy) radical, superoxide anion radical and hydrogen peroxide scavenging, respectively. Percentage metal chelating activity was 81.2%. Enzymatic extracts of Alcalase and Ultraflow showed remarkable scavenging activities on DPPH radical (86.5% and 57.2%, respectively), and superoxide anion scavenging activities were 45.3% and 41.4% from Kojizyme and Viscozyme extracts, respectively. Extract of Protomex revealed 24.8% activity on hydrogen peroxide and Neutase showed 30.8% on hydroxyl radical scavenging effects. Celluclast and Viscozyme extracts showed 33.2% and 32.1% activities on nitric oxide scavenging, respectively, while Alcalase showed 61.5% on metal chelating. This species contains higher lipids among the biochemical compounds and higher metal chelating activities from both 80% methanolic and enzymatic extracts.

Seasonal Characteristics of Phytoplankton Dynamics and Environmental Factors in the Coast of Mara-do and U-do, Jeju Island, Korea

A study on seasonal characteristics of phytoplankton dynamics and environmental factors was carried out at four stations including Mara-do and U-do located in the western and eastern coast of Jeju Island in southern Korea from April 2003 to March 2004. Out of 101 phytoplankton species identified, 84 belong to Bacillariophyceae, 9 Dinophyceae, 6 phytoflagellates and 2 coccolithophorids, and the highest value of species diversity was observed in April. Phytoplankton was more abundant at the western coast than at the eastern coast from March to September and its highest abundance was 49.24 10 cells L at Mara-do in July. The pennate diatoms were more abundant at the western coast than at the eastern coast with the highest abundance of 38.75 10 cells L at Mara-do in July, and during this period Nitzschia longissima contributed 68.5% of the total phytoplankton abundance. Naviculaceae was more abundant at Gosan (western coast) in November when Stauroneis membranacea represented 80.1% of the abundance. Leptocylindrus dances contributed 49.4% of the abundance at U-do in November. Dinophyceae was more abundant at U-do in August. Water temperature and pH fluctuated from 11.7 to 27.1 and from 7.31 to 8.70, respectively. Water temperature of Mara-do was about 1-2 higher than the other stations. Salinity varied from 30.4 to 35.0 psu with the minimum in rainy season and the maximum at the end of winter. The concentration of NH-N, NO-N, NO-N, PO-P and SiO-Si ranged 0.07-6.79, 1.0-62.0, 1.0-8.0, 1.0-7.0 and 7.0-191.0 g-at L, respectively. Chlorophyll a concentrations varied from 0.10 to 1.17 g L. NH-N concentrations were high at U-do from May to December, and at Mara-do from January to February. The high concentrations of NO-N were found at Mara-do from June to September and at U-do from January to May. The effects of various physicochemical parameters on the seasonal distribution and succession of phytoplankton population suggest that there is a classical pattern of phytoplankton dynamics in Jeju coastal waters.

Seasonal Dynamics of Phytoplankton and Environmental Factors around the Chagwi-do off the West Coast of Jeju Island, Korea

The dynamics of phytoplankton abundance with seasonal variation in physicochemical conditions were investigated monthly at 10 stations around the Chagwi-do off the west coast of Jeju Island, Korea, including inshore, middle shore, and offshore in the marine ranching area from September 2004 to November 2005. Water temperature varied from 12.1 to 28.9°C (average 18.8°C), and salinity from 28.9 to 34.9 psu (average 33.7 psu). The chlorophyll a concentration was 0.02-2.05 µg L1 (average 0.70 µg L1), and the maximum concentration occurred in the bottom layer in April. A total of 294 phytoplankton species belonging to 10 families was identified: 182 Bacillariophyceae, 52 Dinophyceae, 9 Chlorophyceae, 12 Cryptophyceae, 6 Chrysophyceae, 4 Dictyophyceae, 13 Euglenophyceae, 6 Prymnesiophyceae, 5 Prasinophyceae, and 5 Raphidophyceae. The standing crop was 2.21-48.69x104 cells L1 (average 9.23x 104 cells L1), and the maximum occurred in the bottom layer in April. Diatoms were most abundant throughout the year, followed by dinoflagellates and phytoflagellates. A phytoplankton bloom occurred twice: once in spring, peaking in April, and once in autumn, peaking in November. The spring bloom was represented by fourChaetoceros species andSkeletonema costatum; each contributed 10–20% of the total phytoplankton abundance. The autumn bloom comprised dinoflagellates, diatoms, and phytoflagellates, of which dinoflagellates were predominant.Gymnodinium conicum, Prorocentrum micans, andP. triestinum each contributed over 10% of the total phytoplankton abundance.

Growth Charateristics of Five Microalgal Species Isolated from Jeju Island and Four Microalgal stock Strans in Hatchery

Five microalgal species isolated from the Jeju coast and four microalgal stock strains in hatchery were cultured in order to investigate their adapation to extreme changes in environmental factors such as salinity, water temperatue, adn nutrients. In case of salinity variation, Nitzschia sp. of Bacillariophyceae, Isochrysis galbana of Haptophyceae and Tetraselmis gracilis of Prasinophyceae showed optimum growth at the low salinity of 20 and 25 psu. Amphora coffeaeformis and Chetoceros simplex of Bacillariophyceae, and Pavlova lutheri of Haptophyceae adapted well at the relatively high salinities of 30 and 35 psu. However Phaeodactylum tricornutum of Bacillariophyceae and Chlorella sp. of Chlorophyceae showed euryhaline property In case of water temperature variation, most of all the species studied wer inhibited at 10℃. C. simplex, Nitzschia sp., p. tricornutum, Chlorella sp. and T. gracilis grew well at above 20℃. A. coffeaeformis, I. galbana and P. lutheri adapted also at the high temperature of 30℃. Each microalgal strain showed different growth rates and its maximum biomass. Generally microalgal populations from the Jeju coast grow well in relatively high salinity and high water temperature. Their growth were inhibited at low water temperature, but not likely affected at low salinity. This study indicates that the microalgal populations could not be affected by abnormally low salinity phenomena, which have happened occasionally around the west Jeju coast in summer and have led macrobenthic animals to mass mortality.

OPTIMAL GROWTH CONDITIONS AND ANTIOXIDATIVE ACTIVITIES OF CYLINDROTHECA CLOSTERIUM (BACILLARIOPHYCEAE)1

We isolated the unialgal strain of Cylindotheca closterium (Ehrenb.) Reimann et J. C. Lewin and produced an axenic strain using an antibiotic cocktail of enriched f/2 artificial seawater medium. The optimal growth conditions were estimated under 27 different combinations of temperature, salinity, and nutrients, and mass culture was performed based on the best specific growth conditions. Its antioxidant activities were determined from the extracts of methanol, water, and enzymes (proteases and carbohydrases). The maximum specific growth rate (μmax) varied from 0.63 to 0.97 · d−1. The maximum cell density was 7.20 × 104 cells · mL−1, while the μmax was 0.82 · d−1 in culture conditions of 20°C, 30 psu (practical salinity unit), and “F” nutrient concentrations on day 10 of the culture period. The scavenging rates for 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) free radical were 72.5% and 69.4% from Viscozyme and methanol extracts, respectively. The enzymatic extracts of C. closterium prepared by the hydrolyses of Amyloglucosidase (AMG) and Viscozyme showed 45.8% and 45.5% nitric‐oxide‐scavenging rates, slightly lower than the activity of alpha‐tocopherol (α‐tocopherol) but similar to butylated hydroxytoluene (BHT). The extract from methanol and water showed 44.8% and 44.4% scavenging rates, statistically similar with BHT. The metal‐chelating activities of the Kojizyme, Alcalase, methanol, Viscozyme, and Neutrase extracts were 67.1, 53.9, 53.2, 52.1, and 50.2 %, respectively, five to six times higher than the commercial antioxidants. The AMG, Viscozyme, and Neutrase extracts showed a remarkable linoleic acid peroxidation inhibition, which was higher than BHT and statistically similar with α‐tocopherol.

Seasonal Abundance of Epiphytic Dinoflagellates around Coastal Waters of Jeju Island, Korea

The seasonal abundance of epiphytic dinoflagellates in the intertidal zone of Jeju Island, Korea was quantitatively estimated by monthly collection of macroalgal samples (Rhodophyta, Phaeophyta, and Chlorophyta) from six sampling locations from July 2012 to June 2013. Ten epiphytic dinoflagellate taxa, including eight potentially toxic species Amphidinium carterae, A. operculatum, Gambierdiscus sp., Ostreopsis ovata, Prorocentrum concavum, P. emarginatum, P. lima, and P. rhathymum, were identified. Two Amphidinium and three Prorocentrum species are newly recorded in Korean coastal waters. A significant change in seasonal abundance was recorded with maximum (751.82 ± 223.12 cells g^(-1) wet weight of algae; cells g^(-1) hereafter) in June (summer), followed by October (autumn) (650.45 ± 225.02 cells g^(-1)) and September (598.02 ± 197.82 cells g^(-1)). O. ovata was the most abundant (338.21 ± 11 cells g^(-1)), reported in October and Gambierdiscus sp. was the least abundant, which was found only in September (6.92 ± 16.97 cells g^(-1)) and October (6.54 ± 6.54 cells g^(-1)) at Hamduk (St 2). Significantly, highest spatial abundance of total dinoflagellates for all sampling stations was found at Hamduk (St 2) (547.91 ± 315 cells g^(-1)), while it was lowest at Hwasun (St 5) (232.59 ± 144.93 cells g^(-1)). Abundance of all dinoflagellate species was significantly correlated with environmental parameters, with some exceptions. During summer and autumn, increasing abundance of dinoflagellate at all sampling stations compared to the other seasons emphasizes environmental and biological interactions of epiphytic dinoflagellate with host macroalgae. Generally, each of the epiphytic dinoflagellates did not show specific preference of macroalgae as host. However, Chlorophytes were less preferred host by most of the dinoflagellates.

Mobility of Amphidinium carterae Hulburt measured by high-frequency ultrasound

The over-growth of phytoplankton causes harmful algal blooms (HABs) in marine ecological environments. Mobility measurement is important in understanding the action of HABs. In this study, the mobility of Amphidinium carterae Hulburt (A. carterae) was investigated using high-frequency ultrasound in the laboratory. Mobility in response to light was illustrated with M-mode images reconstructed from echoed signals. This study suggests that mobility of the swimming speed of A. carterae in response to light can be measured and calculated with M-mode images through high-frequency ultrasound. This finding may be helpful in understanding the fundamental behavior of HABs.

Feeding Stimulants and Feeding Preference of Haliotis discus Reeve (Jeju Island) to Marine Algae

Haliotis discus, a useful abalone of herbivorous gastropod, shows feeding preference to marine algae depending upon their growth stage and recognition of taste. This study was carried out to investigate this abalone’s algal preferences and the presence of feeding stimulants. In single-choice experiments the small (S) group generally preferred Ulva pertusa (Chlorophyta), whereas the medium (M) and large (L) group preferred both Laminaria japonica (Phaeophyta) and Undaria pinnatifida (Phaeophyta). In multi-choice experiments using 4 algal species of L. japonica, U. pertusa, U. pinnatifida and Ecklonia cava (Phaeophyta), the results were same as in the single-choice experiments; the S group preferred U. pertusa the most, while the M and L group preferred both U. pinnatifida and L. japonica. However E. cava was not preferred by any groups. In order to examine the presence of feeding stimulant, chemical compounds from algae used as feed were isolated and identified. The abalone responded to water soluble matters of L. japonica, U. pinnatifida and U. pertusa, but those of E. cava and Sargassum sagamianum (Phaeophyta) were not attractive to them. In feeding stimulant experiments using fat soluble matters, the S group preferred the fat soluble matter of U. pertusa the most, while the M group and the L group preferred those of U. pertusa and U. pinnatifida, and those of L. japonica, respectively. However the fat soluble matter of S. sagamianum was not attractive to the abalone. The results of feeding stimulant experiments were same as those of single-choice or multi-choice experiments, which showed that compound lipids in fat soluble matter might act as feeding-stimulant.