Characteristics of dissolved organic matter in lakes with different eutrophic levels in southeastern Hubei Province,China

Dissolved organic matter(DOM) plays a crucial role in both the carbon cycle and geochemical cycles of other nutrient elements,which is of importance to the management and protection of the aquatic environme nts.To ac hieve a more comprehensive understanding the characteristic s of DOM in the Changjiang(Yangtze) River basin,water samples from four natural lakes(Xiandao,Baoan,Daye,and Qingshan) in southeastern Hubei Province in China with different eutrophication levels were collected and analyzed.The optical characteristics were analyzed using ultraviolet-visible spectrophotometry and excitationemission matrix spectroscopy coupled with parallel factor analysis.The results show that:(1) two humiclike components(C1 and C2) and two protein-like substances(C3 and C4) of DOM were identified in all waterbodies;(2) C3 originated primarily from the degradation of microalgae and contributed substantially to humic-like components during transformation.C4 was widely present in the Changjiang River basin and its formation was related to microbial activity,rather than algal blooms or seasons.Influenced by the water mixing,the protein-like components were more likely to be transformed by microorganism,whereas humiclike components were more easily to be photobleached;(3) the concentration of DOM and the fluorescence intensity of humic-like components gradually increased with rising lake eutrophication levels.With respect to protein-like components,only C3 showed changes along the eutrophication gradients;(4) DOM showed a high affinity with permanganate index(COD_(Mn)) and chlorophyll a(chl a) while the relationship was variable with phosphorus.This study helps us systematically understand the DOM characteristics,microbial activities,and pollutant transfo rmation in the Changjiang River basin and provide s reference to the ecological re storation of aquatic environments.     

Effect of Alexandrium tamarense on three bloom-forming algae

We investigated the allelopathic properties of Alexandrium tamarense (Laboar) Balech on the growth of Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Heterosigma akashiwo (Hada) Hada in a laboratory experiment. We examined the growth of A. tamarense, C. marina, P. donghaiense and H. Akashiwo in co-cultures and the effect of filtrates from A. tamarense cultures in various growth phases, on the three harmful algal bloom (HAB)-forming algae. In co-cultures with A. tamarense, both C. marina and H. akashiwo were dramatically suppressed at high cell densities; in contrast, the growth of P. donghaiense varied in different inoculative ratios of A. tamarense and P. donghaiense. When the ratio was 1:1 (P. donghaiense: A. tamarense), growth of P. donghaiense was inhibited considerably, while the growth of P. donghaiense was almost the same as that of the control when the ratio was 9:1. The growth difference of P. donghaiense, C. marina and H. akashiwo when co-cultured with A. tamarense indicated that the allelopathic effect may be one of the important factors in algal competition and phytoplankton succession involving A. tamarense. In addition, the filtrate from A. tamarense culture had negative impacts on these three HAB algae, and such inhibition varied with different growth phases of A. tamarense in parallel with reported values of PSP toxin content in Alexandrium cells. This implied that PSP toxin was possibly involved in allelopathy of A. tamarense. However, the rapid decomposition and inactivation of PSP toxin above pH 7 weakened this possibility. Further studies on the allelochemicals responsible for the allelopathy of A. tamarense need to be carried out in future.     

Effects of different phosphorus concentrations and N/P ratios on the growth and photosynthetic characteristics of <Emphasis Type="Italic">Skeletonema costatum</Emphasis> and <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis>

The effects of different phosphorus (P) concentrations (0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen (N)/P ratios on the growth and photosynthetic characteristics of Skeletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P (HP) concentration group showed the greatest algal density and highest specific growth rate. Changes in the maximum efficiency of photosystem II (F _v /F _m ) were monitored under the various P and N/P conditions. The largest decrease in F _v /F _m was in the low-P (LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields (Φ _PSII) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinoflagellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _{PS II} values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang (Yangtze) River estuary from a photosynthesis perspective.     

The inhibitory degree between Skeletonema costatum and dinoflagllate Prorocentrum donghaiense at different concentrations of phosphate and nitrate/phosphate ratios

Interactions between Skeletonema costatum (S. costatum) and Prorocentrum donghaiense (P. donghaiiense) were inves-tigated using bi-algal cultures at different concentrations of phosphate (PO4-P) and nitrate/phosphate (N/P) ratios. Experiments were conducted under P-limited conditions and the Lotka-Volterra mathematical model was used to simulate the growth of S. costatum and P. donghaiense in the bi-algal cultures. Both of these two species were inhibited significantly in bi-algal culture. The results of the simulation showed that the inhibitory degree of S. costaum by P. donghaiense was high when the concentration of PO4-P was low (0.1μmolL-1/2 d), but that of P. donghaiense by S. costaum was high with increased PO4-P supply (0.6μmolL-1/2 d). At low concen-tration of PO4-P (0.1μmolL-1/2 d), or high concentration of PO4-P (0.6μmolL-1/2 d) with high N/P ratio (160), the interactions be-tween S. costatum and P. donghaiense were dependent on the initial cell densities of both species. At high concentration of PO4-P (0.6μmolL-1/2 d) with low N/P ratio (25 or 80), S. costatum exhibited a survival strategy superior to that of P. donghaiense. The de-gree of inhibition of P. donghaiense by S. costaum increased with elevated N/P ratio when the medium was supplemented with con-centration 0.1μmolL-1/2 d of PO4-P. The degree of inhibition to P. donghaiense by S. costaum increased with elevated N/P ratio at low concentration of PO4-P (0.1 μmolL-1/2 d). This trend was conversed at high concentration of PO4-P (0.6μmolL-1/2 d). However, the degree of inhibition of S. costaum by P. donghaiense increased with the increased N/P ratio at different PO4-P concentrations (0.1μmolL-1/2 d and 0.6μmolL-1/2 d). These results suggested that both phosphate concentration and N/P ratio affected the competition between S. costaum and P. donghaiense: P. donghaiense is more competitive in environments with low phosphate or high N/P ratio and the influence of N/P ratio on the competition was more significant with lower phosphate concentration.     

Experimental study on the interspecific interactions between the two bloom-forming algal species and the rotifer Brachionus plicatilis

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms (HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×10⁴ cells mL⁻¹ of P. donghaiense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (5, 10, 15 and 20 ×10⁴ cells mL⁻¹), and the increase rate of B. plicatilis population (r) at this algal density was 0.104 ± 0.015 rd⁻¹. Cell densities of P. donghaiense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of the control.     

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide （DMS）, acrylic acid （AA）, and dimethylsulfoniopropionate （DMSP） during growth of three microalgae： Prorocentrum micans, Gephyrocapsa oceanica, and Platymonas subcordiformis. The DMSP, AA, and DMS concentrations in culture media varied significantly among algal growth stages, with the highest concentrations in the late stationary growth stage or the senescent stage. In the stationary growth stage, the average DMSP concentration per cell in P. micans （0.066 5 pmol/cell） was 1.3 times that in G. oceanica （0.049 5 pmol/ cell） and 20.2 times that in P. subcordiformis （0.003 29 pmol/cell）. The average concentrations of AA were 0.044 6, 0.026 9, and 0.003 05 pmol/cell in P. micans, G. oceanica, and P. subcordiformis, respectively, higher than the concentrations of DMS （0.272, 0.497, and 0.086 2 fmol/cell, respectively）. There were significant positive correlations between cell density and AA, DMSP, and DMS concentrations. The ratios of DMS/AA and AA/（DMSP＋AA） in the three algae differed significantly over the growth cycle. In all three microalgae, the DMS/AA ratios were less than 25% during the growth period, suggesting that the enzymatic cleavage pathway, which generates DMS, was not the main DMSP degradation pathway. The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence. In all three microalgae, the AA/（DMSP＋AA） ratio （degradation ratio of DMSP） decreased during the exponential growth phase, and then increased. The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.     

Effects of co-existing microalgae and grazers on the production of hemolytic toxins in Karenia mikimotoi

Karenia mikimotoi (Miyake & Kominami ex Oda) Hansen & Moestrup is associated with harmful algal blooms in temperate and subtropical zones of the world. The hemolytic substances produced by K. mikimotoi are thought to cause mortality in fishes and invertebrates. We evaluated the composition of the hemolytic toxin produced by K. mikimotoi cultured in the laboratory using thin-layer chromatography.In addition, we evaluated the effect of co-occuring algae (Prorocentrum donghaiense and Alexandrium tamarense) and the cladoceran grazer Moina mongolica on hemolytic toxin production in K. mikimotoi. The hemolytic toxins from K. mikimotoi were a mixture of 2 liposaccharides and 1 lipid. Waterborne clues from P. donghaiense and A. tamarense inhibited the growth of K. mikimotoi but increased the production of hemolytic toxins. Conversely, K. mikimotoi strongly inhibited the growth of caged P. donghaiense and A. tamarense. In addition, the ingestion of K. mikimotoi by M. mongolica induced the production of hemolytic toxins in K. mikimotoi. Taken together, our results suggest that the presence of other microalgae and grazers may be as important as environmental factors for controlling the production of hemolytic substances. K. mikimotoi secreted allelochemicals other than unstable fatty acids with hemolytic activity. The production of hemolytic toxins in dinoflagellates was not only dependent on resource availability, but also on the risk of predation. Hemolytic toxins likely play an important role as chemical deterrents secreted by K. mikimotoi.     

Significance of different microalgal species for growth of moon jellyfish ephyrae, <Emphasis Type="Italic">Aurelia</Emphasis> sp.1

The scyphozoan Aurelia aurita (Linnaeus) sp. l., is a cosmopolitan species-complex which blooms seasonally in a variety of coastal and shelf sea environments around the world. The effects of different microalgal species on the growth of newly-released Aurelia sp.1 ephyrae were studied under laboratory conditions. We fed ephyrae with four different microalgal species (diatom, autotrophic dinoflagellate, heterotrophic dinoflagellate, and chlorophyta) plus Artemia nauplii for 12-24 d at 18°C. Results showed that the growth rate diverged significantly for Artemia nauplii compared to other food types. In addition, there was no significant variation between the growth rates for Skeletonema costatum and Prorocentrum donghaiense, and no significant variation was found in the growth rates for N. scintillans and P. subcordiformis. Artemia nauplii could support the energy requirement for the newly-released ephyrae to develop to meduase, and the ephyrae with Artemia nauplii showed a significant average growth rate of 25.85% d^?1. Newly-released ephyrae could grow slightly with some species of microalgae in the earliest development stage. Chain diatom Skeletonema costatum and autotrophic dinoflagellate Prorocentrum donghaiense, could not support the growth of the ephyrae, while heterotrophic dinoflagellate Noctiluca scintillans and chlorophyta Platymonas subcordiformis could support the growth of the ephyrae. However, none of the ephyrae fed with the tested phytoplankton could mature to medusae.     

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 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%).     

Salt response of the freshwater microalga Scenedesmus obliquus (Turp.) Kutz is modulated by the algal growth phase

Growth and biochemical responses of the coenobial green alga Scenedesmus obliquus to salinity stress were monitored across different phases of growth. The alga was cultured on BG11 growth medium and subjected to 0, 30, 100, and 200 mmol/L NaCl for a period of 20 d, during which algal cultures were harvested at 4-d intervals. The salinity-induced inhibition of algal growth was accompanied with prolongation of timing of the different growth phases. The sharp and progressive salinity-induced inhibition of algal growth rate during the early phase of growth points to salt shock but moderation of inhibition at the subsequent stages of growth means algal acclimation to salinity. The concentrations of chlorophylls a and b, soluble sugars, proteins as well as those of K+ and Na+ in the alga exhibited peaks at the initiation of the exponential phase of growth, with increasing magnitude in proportion to the increase in the level of salinity. Nevertheless, whereas soluble sugars of the alga peaked at initiation of the exponential phase, starch concentration progressively increased with culture age, reaching saturation towards the stationary phase. Whereas the salinity-induced increase in soluble sugars was most evident at the early stages of growth the reverse was true for starch. The present results point to fast acclimation of S. obliquus to salt stress post a brief salt shock, utilizing soluble sugars, K+ and Na+ for osmotic adjustment. Increasing salinity from 0 to 200 mmol/L NaCl led to progressive increase in soluble sugars, proteins, K + and Na + concentrations of the algal cells, particularly at the early stages of growth. However, the salinity-induced increase in chlorophyll concentration approached a limit at 100 mmol/L NaCl whereas that in starch concentration was more evident at the later stages of growth.     

Antiangiogenic activity of low-temperature lysozyme from a marine bacterium <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

We extracted marine low-temperature lysozyme (MLTL), a novel lysozyme, from a marine microorganism through fermentation. Our previous study suggested that a low molecular weight (16 kDa) may exert anti-tumor activity through antiangiogenesis. In this study, we extracted a high weight (39 kDa) and investigated its antiangiogenic activity in vivo and in vitro. Using zebrafish embryos as an in vivo study model, we found that treatment with MLTL significantly inhibited the growth of subintestinal vessels (SIVs) in a dose-dependent manner and that 400 μg/ml MLTL was sufficient to block the growth of SIVs. An in vitro study conducted using human umbilical vein endothelial cells (HUVECs) revealed that MLTL suppressed the proliferation, migration and tube formation of HUVECs in a dose-dependent manner. Interestingly, assays by flow cytometry and DNA electrophoresis indicated that MLTL was able to induce apoptosis of HUVECs. Moreover, further study demonstrated that the disruption of intracellular Ca²⁺ homeostasis may play an important role in MLTL induced apoptosis of HUVECs. Taken together, the results of this study demonstrate for the first time that MLTL inhibits angiogenesis through its pleiotropic effects on vascular endothelial cells and induces apoptosis through regulation of cellular Ca²⁺ levels. The results of this study also revealed a possible mechanism underlying the antiangiogenic effect of MLTL and suggested that MLTL may be a promising new antiangiogenic agent for use in cancer therapy.     

湖南锡矿山锑矿区水环境中DOM三维荧光特征及其对锑污染的指示意义

溶解性有机质(dissolved organic matter, 简称DOM)是影响锑迁移转化的重要因素之一。湖南锡矿山锑矿是世界最大的锑矿, 水环境中锑污染情况严重。为查明锡矿山矿区水环境中DOM特征及其影响, 对锡矿山水环境样品进行三维荧光分析, 利用平行因子法提取水环境中天然有机组分, 分析荧光特征, 探究各组分之间与锑的相关关系。分析表明, 矿区水体环境中DOM以低腐殖化、陆源与微生物源混合来源为特点, 多数水样以陆源有机物为主。锡矿山锑矿区水体中包括了3种不同的组分: C1组分为陆源类腐殖质, C2组分为醌类腐殖质, C3组分为类蛋白质(酪氨酸); 水环境中C1组分相对含量最高, 地表水中C3组分相对含量高于地下水。研究认为, 存在以下途径影响地下水环境中锑的释放: ①类蛋白质组分与锑的络合促进锑的溶解释放; ②腐殖质组分与锑的直接络合。低锑地表水中天然有机组分相对含量受稀释作用的影响, 高锑地表水中天然有机组分相对含量对锑的来源有一定的指示作用。      

太湖浮游藻类的后向散射分离及其叶绿素a浓度反演

浮游藻类的后向散射是水体光谱构成的重要组成部分,作为水体辐射传输模型中的重要参数,高精度的藻类后向散射系数对水体叶绿素a浓度的遥感反演精度至关重要。本文以简化的辐射传输模型-生物光学模型为基础,尝试性分离了太湖浮游藻类的后向散射系数。通过藻类后向散射规律分析,建立了浮游藻类吸收、后向散射特征的叶绿素a反演模型,改善了叶绿素a浓度的遥感反演精度。分析表明：藻类颗粒物的后向散射系数与吸收系数之间存在反比关系,且在560 nm、700 nm附近存在明显的散射峰,与叶绿素a浓度之间相关性显著;低密度藻类水体总悬浮颗粒的后向散射以非色素颗粒为主导,适合采用经典的指数模型模拟后向散射系数,而藻类密度较高的富营养化水体,水体总悬浮颗粒的后向散射以藻类颗粒为主导,传统的指数模型已不适用;采用分离藻类后向散射系数的方法,使得叶绿素a浓度的反演值与真实值相关系数从0.66提高到0.98,相对误差从55%降低到38%,均方根误差(RMSE)也由60.95 μg/L降低至13.98 μg/L。其真实性检验表明,与经典指数模型方法相比,利用藻类颗粒后向散射分离方法反演叶绿素a浓度,能够显著改善反演精度。     

Development of a real-time PCR method (Taqman) for rapid identification and quantification of Prorocentrum donghaiense

Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs.     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Amino acid composition and functional properties of giant red sea cucumber (Parastichopus californicus) collagen hydrolysates

Giant red sea cucumber (Parastichopus californicus) is an under-utilized species due to its high tendency to autolysis.The aim of this study was to evaluate the functional properties of collagen hydrolysates from this species.The degree of hydrolysis (DH),amino acid composition,SDS-PAGE,emulsion activity index (EAI),emulsion stability index (ESI),foam expansion (FE),and foam stability (FS) of hydrolysates were investigated.The effects of pH on the EAI,ESI FE and FS of hydrolysates were also inves-tigated.The results indicated that the β and α1 chains of the collagen were effectively hydrolyzed by trypsin at 50℃ with an En-zyme/Substrate (E/S) ration of 1:20 (w:w).The DH of collagen was up to 17.3% after 3 h hydrolysis with trypsin.The hydrolysates had a molecular weight distribution of 1.1 17 kDa,and were abundant in glycine (Gly),proline (Pro),glutamic acid (Glu),alanine (Ala) and hydroxyproline (Hyp) residues.The hydrolysates were fractionated into three fractions (< 3 kDa,3 10 kDa,and > 10 kDa),and the fraction of 3 10 kDa exhibited a higher EAI value than the fraction of > 10 kDa (P<0.05).The fraction of > 10 kDa had higher FE and FS values than other fractions (P<0.05).The pH had an important effect on the EAI,ESI,FE and FS.All the fractions showed undesirable emulsion and forming properties at pH 4.0.Under pH 7.0 and pH 10.0,the 3 10 kDa fraction showed higher EAI value and the fraction of > 10 kDa showed higher FE value,respectively.They are hoped to be utilized as functional ingredients in food and nutraceutical industries.     

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).