EFFECT OF LIGHT INTENSITY ON THE TOTAL LIPID AND FATTY ACID COMPOSITION OF SIX STRAINS OF MARINE DIATOMS

The effect of light intensity ( 1500 Ix and 5000 Ix) on the total lipid and fatty acid composition of six strains of marine diatoms Cylindrotheca fusiformis (B211 ), Phaeodactylum tricornutum (Bl14, Bl18 and B221) Nitzschia closterium (B222) and Chaetoceros gracilis (B13) was investigated.The total lipids of B13, Bl14, and B211 grown at 5000 Ix were lower than those grown at 1500 Ix. No evident changes were observed in Bl18, B221 and B222. Fatty acid composition changed considerably at different light intensity although no consistent correlation between the relative proportion of a single FA and light intensity. The major fatty acids of the 6 strains were 14:0, 16:0, 16:l(n- 7) and 20:5(n-3) . Cylindrothecafusiformis had high percentage of 20:4n - 6 (9.2 - 10.9% ) . The total polyunsaturated fatty acid /rl all 6 strains decreased with increasing light intensity. The percentage of the highly unsaturated fatty acid eicosapentaenoic acid (EPA) decreased with increasing light intensity in all strains except Chaetoceros gracilis.     

Contents of carbohydrates and lipids in the bodies of two planktonic crustaceans

The carbohydrate and lipid contents in two planktonic crustaceans, i. e.Pseudeuphausia sinica andAcetes chinensis sampled from Xiamen coastal waters were estimated. InP. sinica, the contents of carbohydrates (in dry weight) and lipids varied from 2.19–2.33% and 21.2–21.9% respectively; inA. chinensis from 1.74–2.55% and 14.43–15.10% respectively. The analyses of fatty acids by gas-chromatograph (Model 103) showed that eight fatty acids of 14∶0, 15∶0, 16∶0, 17∶0, 18∶0, 18∶2, 20∶2 and 22∶2 were found in these two animals, that 16∶0, 18∶0, 18∶2, and 20∶2 formed the major constituents with 91.08% (P. sinica) and 74.80% (A. chinensis) of the total fatty acids, and that the values of odd carbon types (15∶0 and 17∶0) were fairly low, 4.47% and 3.36% respectively. Three monosaccharides: glucose, fructose, and galactose detected by high-liquid chromatograph (Waters 208) accounted for more than 60% of the total carbohydrates, especially the glucose, exceeding by 40%.     

Effect of growth phase on the fatty acid compositions of four species of marine diatoms

The fatty acid compositions of four species of marine diatoms (Chaetoceros gracilis MACC/B13, Cylindrotheca fusiformis MACC/B211, Phaeodactylum tricornutum MACC/B221 and Nitzschia closterium MACC/B222), cultivated at 22 ℃±1 ℃ with the salinity of 28 in f/2 medium and harvested in the exponential growth phase, the early stationary phase and the late stationary phase, were determined. The results showed that growth phase has significant effect on most fatty acid contents in the four species of marine diatoms. The proportions of 16:0 and 16: 1n-7 fatty acids increased while those of 16 :3n-4 and eicosapentaenoic acid (EPA) decreased with increasing culture age in all species studied. The subtotal of saturated fatty acids (SFA) increased with the increasing culture age in all species with the exception of B13. The subtotal of monounsaturated fatty acids (MUFA) increased while that of polyunsaturated fatty acids (PUFA) decreased with culture age in the four species of marine diatoms. MUFA reached their lowest value in the exponential growth phase, whereas PUFA reached their highest value in the same phase.     

Effects of temperature, salinity, light intensity, and pH on the eicosapentaenoic acid production of Pinguiococcus pyrenoidosus

The effects of temperature, light intensity, salinity, and initial pH on the growth and fatty acid composition of Pinguiococcus pyrenoidosus 2078 were studied for eicosapentaenoic acid (EPA) production potential. The fatty acid composition was assayed by gas chromatography-mass spectrometry, which indicated that the main fatty acids were C14:0, C16:0 and EPA. The highest EPA percentage 20.83% of total fatty acids was obtained at 20℃ with the temperature being set at 20, 24, and 28℃. Under different salinities and light intensities, the highest percentages of total polyunsaturated fatty acids (PUFAs) and EPA were 17.82% and 31.37% of total fatty acids, respectively, which were achieved at salinity 30 and 100μmol photon m-2s-1 illumination. The highest percentages of total PUFAs and EPA were 38.75% and 23.13% of total fatty acids, respectively, which were reached at an initial pH of 6 with the test range being from 5.0 to 9.0.     

Comparative Analysis of Nutrient Composition of Caulerpa lentillifera from Different Regions

This study investigated the difference of nutrient composition in Caulerpa lentillifera collected from different regions.The nutrient compositions of C.lentillifera from China's Hainan and Shandong provinces were determined and compared with those from Semporna(Malaysia),Sabah(Malaysia),Petchburi(Thailand),and two other species of seaweeds(Malaysia).The results showed that the polysaccharide and protein contents of C.lentillifera from Hainan(44.82%±0.98%and 12.5%±0.70%),Shandong(43.22%±1.42%and 14.7%±0.72%),Petchburi(59.27%±2.31%and 12.49%±0.30%),and Sabah(44.02%±2.01%and 19.38%±1.48%)were higher than those of Eucheuma cottonii(26.49%±3.01%and 9.76%±1.33%)and Sargassum polycystum(33.49%±1.70%and 5.40%±0.07%),respectively.The polyunsaturated fatty acid contents in C.lentillifera from Hainan(29.98%)and Shandong(22.11%)were higher than that in Semporna(16.76%±0.27%),Sabah(9.49%),and S.polycystum(20.34%±0.43%),but lower than that in E.cottonii(51.55%±0.57%).In Caulerpa lentillifera of Hainan and Shandong provinces,the essential amino acid(EAA)/total amino acid(TAA)ratios were 35.22%and 38.78%,respectively;and the EAA/ETAA ratios were 54.36%and 63.35%,respectively.The EAA composition of C.lentillifera was close to the ideal model of Food and Agriculture Organization/World Health Organization.C.lentillifera was rich in mineral elements,including calcium((3315.85±127.55)mg(100 g)^-1 and(3728.35±92.38)mg(100 g)^-1),magnesium((6715.74±82.58)mg(100 g)^-1 and(8128.59±242.72)mg(100 g)^-1),and trace elements,including iron((510.65±5.47)mg(100 g)^-1 and(1972.97±183.35)mg(100 g)^-1),selenium((1.04±0.08)mg(100 g)^-1 and(0.83±0.15)mg(100 g)^-1),and zinc((33.90±0.13)mg(100 g)^-1 and(11.75±2.05)mg(100 g)^-1).These results showed that C.lentillifera was more nutritious than S.polycystum or E.cottonii in terms of polysaccharide,protein,and fatty acid contents.Furthermore,both C.lentillifera species collected from Hainan and Shandong provinces show more basic nutrients.Therefore,C.lentillifera has important development and application prospects.     

Decomposition of algal lipids in clay-enriched marine sediment under oxic and anoxic conditions

A series of laboratory incubation experiments were conducted to examine the decomposition of algal organic matter in clay-enriched marine sediment under oxic and anoxic conditions. During the 245-day incubation period, changes in the concentrations of TOC, major algal fatty acid components (14:0, 16:0, 16:1, 18:1 and 20:5), and n-alkanes (C₁₆–C₂₃) were quantified in the samples. Our results indicate that the organic matters were degraded more rapidly in oxic than anoxic conditions. Adsorption of fatty acids onto clay minerals was a rapid and reversible process. Using a simple G model, we calculated the decomposition rate constants for TOC, n-alkanes and fatty acids which ranged from 0.017–0.024 d⁻¹, 0.049–0.103 d⁻¹ and 0.011 to 0.069 d⁻¹, respectively. Algal organic matter degraded in two stages characterized by a fast and a slow degradation processes. The addition of clay minerals montmorillonite and kaolinite to the sediments showed significant influence affecting the decomposition processes of algal TOC and fatty acids by adsorption and incorporation of the compounds with clay particles. Adsorption/association of fatty acids by clay minerals was rapid but appeared to be a slow reversible process. In addition to the sediment redox and clay influence, the structure of the compounds also played important roles in affecting their degradation dynamic in sediments.     

Monthly Changes of Glycogen, Lipid and Free Amino Acid of Oyster

Monthly difference of the chemical composition of oyster cultured along the eastern coast of Shandong Province was analyzed. The components analyzed included glycogen, fatty acid and free amino acid （FAA）. The content of glycogen was high in January and March （2.89 and 2.82 g（100 g） ^-1 on average, respectively） and low in October （2.07g（100g）^- 1 on average）. The low content of neutral lipids in October reflected a relatively poor nutritional value of oyster （1.42 g（ 100 g）^- 1 on average）. The main fatty acids of oyster were palmitic acid （16：0）, oleic acid （18： 1）, eicosapentaenoic acid （EPA, 20： 5（0 3） and docosahexaenoic acid （DHA, 22：6（0 3）. The major FAAs of oyster were Taurine, Glutamicacid, Glycin, Alanine, Arginine and Proline. Taurine was the most abundant FAA with its content ranging from 603 mg（100 g） ^-1 to 1 139 mg（ 100 g） ^-1. The high contents of glycogen, polyunsaturated fatty acid and FAA showed that oyster cultured along the eastern coast of Shandong Province was nutritionally good in January and March.     

Triploidy induction with heat shocks toPenaeus chinensis and their effects on gonad development

Heat shocks effectively produced triploids inPenaeus chinensis. Fertilized eggs heat shocked (28–32°C) for 8 to 16 minutes, starting from 8 to 20 minutes after spawning, resulted in triploidy induction rates of 39%–75%. Several triploid populations were cultured to 10 cm. In a triploid population, two kinds of ovaries were observed. Histological examination showed apparent differences between these two kinds of ovaries, whereas among male shrimp, there were no such differences. Contribution No. 3465 from the Institute of Oceanology. Chinese Academy of Sciences. The study was supported by the International Foundation for Science, Asian Fisheries Society and Chinese Climbing Project PDB 6–2, and Chinese National 863 Project (No. 863-819-01-05).     

Effects of sodium nitrate and sodium acetate concentrations on the growth and fatty acid composition of <Emphasis Type="Italic">Brachiomonas submarina</Emphasis>

One isolate of Brachiomonas submarina was tested for its ability to grow heterotrophicly on 5 different organic compounds. Sodium acetate and glucose were found to be effective in supporting the growth. Sodium acetate was chosen as the organic nutrient to test the combined effects of organic and inorganic solutions on the growth and fatty acid composition of Brachiomonas submarina. The best growth rates were achieved at 3 mmol L⁻¹ CH₃COONa and 0.88 mmol L⁻¹ NaNO₃ in heterotrophic condition, and 4 mmol L⁻¹ CH₃COONa and 3.52 mmol L⁻¹ NaNO₃ in mixotrophic condition. The differences between fatty acid contents were significant. The total polyunsaturated fatty acids (T. P. U. F. As) varied from 55.79% to 67.72% in heteritrophic growth and from 52.39% to 65.55% in mixotrophic growth. It is concluded that CH₃COONa and NaNO₃ at 3 mmol L⁻¹ and 3.52 mmol L⁻¹ should respectively be used in order to achieve the highest growth rate and fatty acid content.     

Seasonal variation in fatty acid composition of seston and the copepod Calanus sinicus (Brodsky, 1962) in Jiaozhou Bay and its trophic implications

The fatty acid compositions of seston and Calanus sinicus were investigated to study trophic relationships in Jiaozhou Bay. Principal component analysis was carried out to ordinate the fatty acid patterns of seston in stations and months. The results showed that diatoms were most abundant in the phytoplankton at station A5 (located in the northwest of the bay:36°9′N, 120°20′E) and least abundant at station D7 (located outside of the bay: 35°59′N, 120°26′E). By contrast, dinoflagellates were most abundant at station D7 and least abundant at station A5. According to the annual variations of 16:1ω7 and 18:4ω3/ 16:1ω7, diatoms flourished mainly in spring and summer, while dinoflagellates bloomed exclusively in summer. A distinctive feature of the fatty acid composition of C. sinicus was the prevalence of 20:5ω3 and 22:6ω3. The higher content of 16:1ω7 over 18:4ω3 in females indicated that diatoms contributed more than dinoflagellates to the diet of C. sinicus. The feeding intensity of C. sinicus on diatoms was higher in spring and autumn than in other seasons. The herbivorous indicators 20:1 and 22:1 were comparatively low, suggesting that besides phytoplankton, C. sinicus might feed on a wider range of particles including organic detritus, bacteria and small copepods.     

Measurement of trace manganese (II) by the catalytic kinetic spectrophotometric method

A new kinetic spectrophotometric method is developed for the measurement of manganese (II) in water. The method is based on the catalytic effect of manganese (II) with the oxidation of weak acid brilliant blue dye (RAWL) by KIO₄ using the Nitrilo triacetic acid (NTA) as an activation reagent. The optimum conditions obtained are 40 mgL⁻¹ RAWL, 1×10⁻⁴molL⁻¹ KIO₄, 2×10⁻⁴ molL⁻¹ Nitrilo triacetic acid (NTA), pH = 5.8, the reaction time of 3.00 min and the temperature of 20.0 °C. Under the optimum conditions, the proposed method allows the measurement of manganese (II) in a range of 0–50.0 ng mL⁻¹ and with a detection limit of down to 0.158 ng mL⁻¹. The recovery efficiency in measuring the standard manganese (II) solution is in a range of 98.5%–102%, and the RSD is in a range of 0.76%–1.25%. The new method has been successfully applied to the measurement of manganese (II) in both fresh water and seawater samples with satisfying results. Moreover, few cations and anions interfere with the measurement of manganese (II). Compared with other kinetic catalytic methods and instrumental methods, the proposed method shows fairly good selectivity and sensitivity, low cost, cheapness, low detection limit and rapidity. It can be applied on boats easily.     

Effect of n?3 HUFA levels in rotifers and Artemia on growth and survival of larval black sea bream ( Sparus macrocephalus )

Requirement for dietary n−3 HUFA (n−3 highly unsaturated fatty acid) for growth and survival of black sea bream (Sparus macrocephalus) larvae was studied using rotifers andArtemia at various levels of n−3 HUFA. Five treatments with rotifers andArtemia differing in n−3 HUFA were prepared by enriching them with various oil emulsions. Results indicated that dietary n−3 HUFA significantly influence fish n−3 HUFA levels and are essential for growth and survival of black sea bream larvae. The results also indicated that the incorporation of n−3HUFA TG (triacylglycerols) into tissues of larval black sea bream was more effective from natural fish oil in comparison with n−3 HUFA fatty acid ethyl esters from ethyl-esterified oil. Project No. 39770588 supported by the NSFC.     

EFFECTS OF HARVEST STAGE ON THE TOTAL LIPID AND FATTY ACID COMPOSITION OF FOUR CYLINDROTHECA STRAINS

Four strains of Cylindrotheca (B156, B164, B196, and B200) were harvested at three phases in the growth curve and their total lipid and fatty acid composition were determined. Total lipid reached highest values in the late stationary phase in B156 (24.0%), B164 (26.6%) and B200 (17.3%), in the exponential phase in B196 (15.5%). Saturated fatty acids increased with the devel-opment of the culture in B156 and B196, and reached their highest values in the late stationary phase in B156 (41.7%) and B196 (45.1%), in the early stationary phase in B164 (45.4%) and B200 (37.6%). Monounsaturated fatty acids increased with the development of the culture in B196 and B200, and reached their highest values in the late stationary phase in B196 (32.4%) and B200 (32.8%), in the early stationary phase in B164 (31.0%) and in the exponential phase in B156 (29.3%). Polyunsaturat-ed fatty acids decreased in the later phases of the culture in B164, B196 and B200, and peaked in theexponential phase in B164 (29.5%), B196 (42.9%) and B200 (37.3%), and in the early stationary phase in B156 (32.0%).     

Induced nest spawning and artificial hatching of the fertilized eggs of mudskipper, Boleophthalmus pectinirostris

In this study, nest spawning was successfully induced by exogenous hormone injections and seawater flow stimulation, and optimum condition for hatching fertilized eggs of burrow fish mudskipper,Boleophthalmus pectinirostris, was searched. Apart from spawning inside the nests, females also spawned outside the nests. The percentages of spawned nests were 8.0% to 24.2%. Most eggs were observed adhered to the inner wall of the top half of the nest. Fertilization rates of the nest-spawned eggs varied from 17.3% to 80.8%. Females could spawn after being artificially confined inside the nests with males at ratios of 1∶1 or 1∶2, but the spawned eggs were not fertilized. Mean hatching rates of artificially fertilized eggs incubated in round plastic buckets were 32.7%–70.6%, and in the net cages, were 4.2%–20.5%, respectively. Mean hatching rates of nest-fertilized eggs incubated in the round plastic buckets were 33.6%–76.3%, and in the net cages, were 5.9%–25.2%. Results showed that round bucket incubation was the best way for hatching fertilized eggs of mudskipper. Keeping the hatching seawater flowing is an important way for increasing the hatching rates of the mudskipper fertilized eggs. Supported by the NSFC (No. 30170739), and also supported by the Provincial Natural Science Foundation of Fujian Province (No. B0210003).     

DETERMINATION OF TRACE BENZENE HYDROCARBONS IN JIAOZHOU BAY BY ENRICHMENT AND CAPILLARY COLUMN GAS CHROMATOGRAPHY

Trare amounts of benzene hydrocarbons obtained in Jiaozhou Bay (Qindao) were enriched bysorption on a GDX-102 column and eluted by carbon disulfide. The eluted was concenttaled and then de-temened by capillary column gas cbornatognphy.The contents of virious kinds of benzene hydrocarbons in Jiaozhou Bay coastal water were benzene(22.3-141.6)× 10～(-9)g/L, toluate (15.2-94.0) × 10~(-9) g/L, ethyl benzene(11.8-85.1)×10~(-9) g/L, p -xylene(15.2-78.5) ×10(-9) g/L, m-xylene (10.9-79.4) ×10(-9) g/L, o -xylene (12.4-80.1) x ×10(-9)g/L; iso-propyl(8.4- 73.1) x ×10(-9)g/L, n -propyl (6.9-76.4) ×10(-9) g/L, 1, 3, 5-trimethylbenzene (10.9- 35.9)×10(-9) g/L, 1,2, 4-trimethybenzene (10.0- 38.0)×10(-9) g/L, n - butydriare (8. 1 - 34.6) ×10(-9)g/L. The recovery of benzenehydrocarbons was (85.1 -95.6)%.     

STUDY ON EMBRYONIC DEVELOPMENT AND EARLY GROWTH OF TRIPLOID AND GYNOGENETIC DIPLOID LEFI—EYED FLOUNDER, PARALICHTHYS OLIVACEUS （T. et S. ）

The early effects of chromosomal manipulation of eggs and sperm on the yields of trip-loid and gynogenefic diploid larvae of Paralichthys olivaceus were investigated. Triploidy was achieved by cold shocking fen.ilized eggs at 0 - 2℃ for 45 minutes duration 5 minutes after fen.ilization, and the in-duced triploidy rates were 31.2% - 50% and the relative hatching rates were 53.3% - 99%. Gynnge-aetic diploids were obtained when eggs were inseminated with irradiated sperm and cold shocked at 0 -2℃ for 45 minutes duration 5 minutes after fertihzation. The induced gynogenetic diploid rates and the relative hatching rates were 94 % - 96 % and 48.5 % - 68.5 % respectively. The embryonic development of the triploid experimental group and of the gynogenetic diploid experimental group was delayed at first compared with the control group. But from the gastrula stage, it was not delayed anymore. There were no significant differences in the growth of the triploid experimental group larvae and the control group larvae, and in the growth of the gynogenetic diploid experimental group larvae and the control group larvae ac-cording to Student‘s t-test (α = 0.05). The relationship between the early growth of the triploid experi-mental group larvae and that of gynogenetic diploid experimental group larvae was also studied.     

Effect of artificial regulations ofArtemia n−3 HUFA content on growth and survival of black seabream (Sparus macrocephalus) larvae

The requirement for dietary n−3 HUFA (highly unsaturated fatty acid) for growth and survival of black seabream (Sparus macrocephalus) larvae was tested usingArtemia with various levels of n−3 HUFA. Four treatments withArtemia differing in their n−3 HUFA were prepared by feeding them various oil emulsions. The results indicated that dietary n−3 HUFA significantly influences fish n−3 HUFA levels and are necessary for good growth and survival of black seabream larvae. The proper value of n−3 HUFA level and DHA, EPA level inArtemia should be 4.273% and 0.873% (wet weight) for good growth and survival in black seabream larvae respectively. Contribution No. 3044 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39470562 supported by NSFC.     

Studies on culture conditions of benthic diatoms for feeding abalone

Effects of salinity, pH, nitrogenous and phosphate nutrients on the growth rate of four species of benthic diatoms were studied by using the method of in situ optical density measurement. The optimal culture conditions for the four species of diatoms are as follows: forAmphora coffeaeformis, s 35; pH 8.5; NO₃ ⁻−N 1.54, NH₄ ⁺−N, 7.0; CO (NH₂)₂−N, 2.5 (mg/L); PO₄ ³⁻−P 1.13 (mg/L); forCocconeis scutellum varparva, s 30; pH 8.5; NO ₃ ⁻ −N 3.08, NH ₄ ⁺ −N 3.5, CO(NH₂)₂−N 5.0 (mg/L); pO ₄ ³⁻ −P 0.283 (mg/L); forNavicula corymbosa s 25; pH 8.0; NO ₃ ⁻ −N 1.54, NH ₄ ⁺ −N 3.5 CO(NH₂)₂−N 5.0 (mg/L); PO ₄ ³⁻ −P 0.565 (mg/L); forNavicula mollis. s 25; pH 8.0; NO ₃ ⁻ −N 1.54, NH ₄ ⁺ −N 1.75, CO(NH₂)₂−N 1.25 (mg/L); PO ₄ ³⁻ −P 0.141 (mg/L). Part of the results of this paper were exchanged in the Fourth Asian Aquaculture Forum. Oct. 16–20. 1995. Beijing, P. R. China.     

Heritability and genetic correlation of survival in turbot (Scophthalmus maximus)

We analyzed the survival data of the offspring from 21 sires and 42 dams of turbot. The results show that the cumulative survival rates for turbot from 2 to 18 months range from 17.5% to 28.5%; main mortality occurred during months 2–5; and the highest survival rates of families were 97.9%, 98.8%, 99.4%, 99.7% during months 2–5, 5–6, 6–8, 8–11, and 11–18, respectively, and 99.5%, being 53.5%, 23.8%, 19.5%, 14.9%, and 13.2% higher, respectively, than the mean values in each period. In all periods, the estimated heritabilities for survival were very low without significant difference from zero (P≫0.05) (values ranged from 0.06 to 0.12), indicating low additive genetic effects. The genetic correlations of survival among families in different periods were all positive, but low in magnitude (values range from 0.03 to 0.31). Genetic correlations between long-term survival and other periods’ survival had negative values (−0.06 and −0.15) and three positive values (0.16, 0.12 and 0.14). Genetic correlations between survival and weight were all positive, except for survival at months 2–5 and weight at 18 months, which was not significantly negative (−0.18).     

FATTY ACIDS PROFILE IN A HIGH CELL DENSITY CULTURE OF ARACHIDONIC ACID-RICH PARIETOCHLORIS INCISA （TREBOUXIOPHYCEAE, CHLOROPHYTA） EXPOSED TO HIGH PFD

The changes in arachidonic acid (AA) and fatty acids profiles along the growth curve ofParietochloris incisa, a coccoid snow green alga, were studied in a 2.8 cm light-path flat photobiorcactor, exposed to strong photon flux density [PFD, 2400 μEmol/(m²·s)]. Sixteen fatty acids were identified by gas chromatography showing that AA was the dominant fatty acid (33%–41%) followed by linoleic acid (17%–21%). AA content was closely investigated with respect to total fatty acids (TFA), ash free dry weight (AFDW) of cell mass as well as total culture content. These parameters were influenced significantly in a similar manner by culture growth phase, i.e., slightly decreasing in the lag period, gradually increasing in the logarithmic phase, becoming maximal at the early stationary phase, starting to decrease at the late stationary phase, sharply dropping at the decline phase. The increase in AA per culture volume during the logarithmic phase was not only associated with the increase in AFDW but also connected with a corresponding increase in AA/TFA, TFA/AFDW as well as AA/AFDW. The sharp decrease in AA content of the culture during the decline phase was mainly due to the decrease in AA/TFA, TFA/AFDW and AA/AFDW, although AFDW declined only a small extent. Maximal AA concentration, obtained at the early stationary phase, was 900 mg/L culture volume, and the average daily net increase of AA during 9 days logarithmic growth was 1.7 g/(m²·day). Therefore, harvesting prior to the decline phase in a batch culture, or at steady state in continuous culture mode seems best for high AA production. The latter possibility was also further confirmed by continuous culture with 5 gradients of harvesting rate. Contribution No. 4138 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39970575 supported by NSFC and A/2786-2 supported by International Foundation for Sciences (IFS).