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

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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.     

Total lipid and fatty acid composition of eight strains of marine diatoms

Fatty acid composition and total lipid content of 8 strains of marine diatoms (Nitzschia frustrula, Nitzschia closterium, Nitzschia incerta, Navicula pelliculosa, Phaeodactylum tricornutum, Synedra fragilaroides) were examined. The microalgae were grown under defined conditions and harvested at the late exponential phase. The major fatty acids in most strains were 14∶0 (1.0%–6.3%), 16∶0 (13.5–26.4%), 16∶1n−7 (21.1%–46.3%) and 20∶5n−3 (6.5%–19.5%). The polyunsaturated fatty acids 16∶2n−4, 16∶3n−4, 16∶4n−1 and 20∶4n−6 also comprised a significant proportion of the total fatty acids in some strains. The characteristic fatty acid composition of diatoms is readily distinguishable from those of other microalgal groups. Significant concentration of the polyunsaturated fatty acid 20∶5n−3 (eicosapentaenoic acid) was present in each strain, with the highest proportion in B222 (19.5%). Project supported by the Hi-Tech “863” Programs of the China Ministry of Science and Technology (863-819-02-01).     

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).     

PHYSIOLOGICAL INHIBITORY EFFECT OF OCS IN ARACHIDONIC ACID-RICH PARIETOCHLORIS INCISA （TREBOUXIOPHYCEAE, CHLOROPHYTA）

Parietochloris incisa is an arachidonic acid-rich snow green alga. The main physiological profiles, such as ash free dry weight (AFDW), chlorophyll, carotenoid, protein and total fatty acids (TFA), in this alga exposed to old culture supernatant (OCS) at the decline phase or its crude ethyl acetate extracts (CEAE) were investigated by using tubular photobioreactors of different diameters. Results showed that both OCS and CEAE had strong inhibitory effect on the above physiological parameters. The longer the culture was exposed to OCS and the more CEAE were added into the algal culture, the more the above physiological properties were inhibited. Arachidonic acid (AA), the dominant component of fatty acids in this alga, was also seriously inhibited with respect to total TFA, AFDW of cell mass, or culture volume, due to a prebable reduction of enzymes activities catalyzing chain elongation from C18:1ω9 to AA. These results incontestably evidenced that some CEAE dissolving substances existing in OCS, like auto-inhibitors, inhibited P. incisa growth through feedback. Hence, any efficient removal of auto-inhibitors from algal culture to decrease their bioactivity could be good for maximal production of desired products like AA.     

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.     

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.     

罗氏沼虾三群体间肌肉营养品质的比较

对饲养在相似环境下的罗氏沼虾(Macrobrachium rosenbergii)三群体肌肉营养成分进行了分析和比较,3个群体分别为浙江湖州的人工养殖群体(A)、缅甸引进种子二代群体(B)和“南太湖1号”杂交群体(B-+)。结果表明,3个群体粗蛋白含量的高低顺序依次为:B-+群体(17.57%)、B群体(16.65%)、A群体(15.88%),但三群体之间没有显著性差异(P>0.05);而A群体的粗脂肪含量为2.04%,显著高于B群体的1.24%(P<0.05)。3个群体中,B-+群体的总氨基酸含量、必需氨基酸含量、鲜味氨基酸和脂肪酸中的EPA+DHA含量均为最高,且必需氨基酸指数最高,为68.06;相比之下,A群体的上述各项指标值均较低。可见,以上三群体中杂交种群(B-+)的肌肉营养品质较优,而人工养殖群体(A)的肌肉品质相对较差。     

Biochemical Composition and Nutritional Value of Different Shell Color Strains of Pacific Oyster <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

The shell color of Pacific oyster (Crassostrea gigas) is a desirable trait; but the nutritional studies on C. gigas with different shell colors have not been conducted. Through successive selective breeding, five shell color strains of black (B), purple (P), orange (O), golden (G) and white (W) C. gigas have been developed. The aim of this study was to evaluate the chemical composition and nutritional value of five shell color strains and one commercial population with a common color. The biochemical composition including moisture, total protein, glycogen, ash, total fat, fatty acids (FA), amino acids and minerals was detected. The results indicated that the protein (50.76%–56.57%) was the major component. The content of glycogen showed a significant difference between orange shell and golden shell strains, as well as between commercial population and golden shell strain. In addition, all shell color strains contained a large amount of essential amino acids (12.20–14.15 g (100 g)^?1), of them leucine (2.81–3.29 g (100 g)^?1) and lysine (2.79–3.28 g (100 g)^?1) were predominant. The oysters were rich in polyunsaturated fatty acids (42.26%–45.24% of total fatty acid) with high levels of DHA (18.53%–21.16% of total fatty acid) and EPA (17.23%–18.68% of total fatty acid). Significant differences of mineral contents (Mg, Zn, Fe and Cu) were identified among the six populations. These results indicated that C. gigas with different shell colors presented rich nutritional value with high protein, glycogen, essential amino acids and polyunsaturated fatty acids. The biochemical composition obtained in this study is useful for selective breeding of C. gigas with different shell colors.     

A Fast and Indriect Fluorescent Antibody Assay for the Vibro in Large Yeelow Croaker Pseudosciaena Crocea (Richardson)

A fast and indirect fluorescnet antibody assay for the Vibrio alginolyticus and V.Parahaemolyticus infecting the large yellow croaker has been developed.The specific antisera for the two strains of vibrio were prepared with New Zealand rabbit and the antiserum and cross-reactive efficacy was tested by coagulation in tube.It showed that the goat anti-rabbit IgG had been labeled by fuorescence isothiocyanate(FITC).The results showed that positive reactions were 100% for the large yellow croaker Pseudosciaena crocea with typical symptom of vibrio infection,while the positive reaction to the pathogen in healthy yellow croakers reached 40%,but seemed negative for aquaculter water.The results demonstrated that this fast and indirect fluorescent antibody assay can be used not only to test the vibrio pathogen in diseased yellow croaker but also in infected animals with no symptom.     

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.     

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.     

A comparative analysis of fatty acid composition and fucoxanthin content in six <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> strains from diff erent origins

Phaeodactylum tricornutum is a potential livestock for the combined production of eicosapentaenoic acid (EPA) and fucoxanthin. In this study, six marine diatom strains identified as P. tricornutum were cultured and their total lipid, fatty acid composition and major photosynthetic pigments determined. It was found that the cell dry weight concentration and mean growth rate ranged between 0.24–0.36 g/L and 0.31–0.33/d, respectively. Among the strains, SCSIO771 presented the highest total lipid content, followed by SCSIO828, and the prominent fatty acids in all strains were C16:0, C16:1, C18:1, and C20:5 (EPA). Polyunsaturated fatty acids, including C16:2, C18:2, and EPA, comprised a significant proportion of the total fatty acids. EPA was markedly high in all strains, with the highest in SCSIO828 at 25.65% of total fatty acids. Fucoxanthin was the most abundant pigment in all strains, with the highest in SCSIO828 as well, at 5.50 mg/g. The collective results suggested that strain SCSIO828 could be considered a good candidate for the concurrent production of EPA and fucoxanthin.     

Three-dimensional fluorescence characteristics of dissolved organic matter produced by <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis> Lu

Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrum donghaiense at the exponential growth,stationary and decline stages into <0.45 μm filtrate,100 kDa-0.45 μm,10-100 kDa and 1-10 kDa retentate and <1 kDa ultrafiltrate fractions.The fluorescence properties of different molecular weights of dissolved organic matter(DOM) were measured by excitation-emission matrix spectra.Protein-like and humic-like fluorophores were observed in the DOM produced by P.donghai...     

Changes of Protein and Lipid Contents,Amino Acid and Fatty Acid Compositions in Eggs and Yolk-Sac Larvae of American Shad(Alosa sapidissima)

To investigate the changes of the biochemical composition of American shad(Alosa sapidissima) eggs and larvae at embryonic and early larval stages, samples were collected at different development stages from artificial fertilization to the end of yolk absorption including 2 h, 12 h and 30 h after fertilization and newly hatched larvae including 1 and 3 days after hatching. The composition of lipid, fatty acids, protein and amino acids were analyzed. The content of total protein exhibited a decreasing trend during embryogenesis and larval development, and a significant reduction was detected after hatching(P 0.05). The total lipid content remained relative stable. A significant reduction was detected in almost all amino acids after hatching except for glycine(P 0.05), while a significant decrease was found in the content of cysteine, proline, tyrosine, valine, isoleucine, leucine and phenylalanine during the yolk-sac phase(P 0.05). On the other hand, all the groups of fatty acids remained stable during the period of embryogenesis. But after hatching, a significant decrease was found in the content of C18:2n-6, C18:3n-6, SFA and ratio of EPA/ARA(P 0.05), while a significant increase was found in the content of C18:3n-3, C20:4n-6, C22:6n-3 and ratio of n-3/n-6(P 0.05). In conclusion, the combined data suggested that American shad utilizes the protein content as preferential energy substrates during embryonic and early larval developments with some specificity in the consumption of different amino acids.     

Seasonal variation in imposex intensity of Thais clavigera

Imposex, specifically caused by TBT pollution, refers to the superimposition of male sexual characteristics in gastropod females. Seasonal variation of imposex intensity inThais clavigera from both slightly and severely contaminated sites in Hong Kong waters was studied from 1988 to 1999. The male penis length showed significant difference between both sites and seasons. It was shortest during late autumn and early winter (October to December) and longest during spring and early summer (February to June). Female penis length also showed significant difference between sites. It did not change seasonally, however. The RPS (Relative Penis Size) index was the highest during autumn and early winter, and the lowest during spring and early summer. The VDS (Vas Deferens Sequence) index remained stable throughout the sampling period. This study showed that VDS index is a better indicator when we compare relative intensity of imposex. The comparison can only be meaningful provided the samples from different locations are taken during the same season. Supported by the NSFC (40006007).     

养殖海鲡肌肉的生化组成

对海鲡肌肉的营养成分含量及氨基酸、脂肪酸、无机质组成进行了研究，结果表明，海鲡的蛋白质和脂肪含量分别为21.5%和5.5%，必需氨基酸和鲜味氨基酸分别占氨基酸总量的40.9%和37.8%，不饱和脂肪酸占脂肪酸总量的65.2%，其中C205和C226的含量分别为4.5%和12.0%，3.4kg阶段的养殖海鲡肌肉的必需氨基酸和呈味氨基酸的比率最高。