二丁基羟基甲苯对雨生红球藻虾青素和油脂积累的影响

以雨生红球藻(Haematococcus pluvialis)为对象，研究高光照、缺氮条件下，外源添加不同浓度的二丁基羟基甲苯(Butylated hydroxytoluene, BHT)对雨生红球藻生长、虾青素积累、油脂合成、脂肪酸组成、碳水化合物、蛋白含量以及虾青素和脂肪酸合成相关酶基因的影响。结果显示，添加不同浓度的BHT后，2 mg/L BHT添加组虾青素积累量为最高，显著高于其他实验组和对照组(P<0.05)，达到31.66 mg/g，是对照组的1.87倍。油脂含量达45.56%，高于同期对照组(39.06%)，脂肪酸组成变化不显著。在此条件下，虾青素合成关键酶基因dxs和bkt表达水平分别是对照组的5.19倍和2.04倍；脂肪酸合成关键酶基因kas和acp表达水平较对照组显著提高(P<0.05)，分别是对照组的4.56倍和3.02倍。与对照组相比，2 mg/L BHT添加组的碳水化合物和蛋白含量均呈下降趋势。研究表明，在胁迫条件下，外源添加适量浓度的BHT能有效促进雨生红球藻中虾青素的积累，同时提高了藻细胞内的油脂含量。     

Effects of dietary supplementation of Haematococcus pluvialis powder on gonadal development,coloration and antioxidant capacity of adult male Chinese mitten crab (Eriocheir sinensis)

The green algae Haematococcus pluvialis is an important source of natural astaxanthin as feed additive. This study was conducted to investigate the effects of dietary supplementation of H. pluvialis powder on gonadal development, coloration and antioxidant capacity of adult male Chinese mitten crab Eriocheir sinensis, and four experimental diets were formulated to contain 0, 0.2%, 0.4% and 0.6% of H. pluvialis powder. There were four treatments (defined as D1~D4) in this study and each treatment had three replicates. Dietary H. pluvialis contents had no significant effects on survival, body weight gain rate and gonadal development of male E. sinensis. For colour parameters, the total carotenoids content in carapace and hepatopancreas as well as hepatopancreatic lightness (L*) and carapace redness (a*) increased significantly with increasing dietary H. pluvialis (p < 0.05). For the antioxidant indices in the serum, D4 had the lowest activities of superoxide dismutase (SOD) and peroxidase (POD), but the highest glutathione peroxidase (GSH‐Px) and lactic dehydrogenase (LDH), while the malondialdehyde (MDA) in serum and hepatopancreas decreased significantly with the rising content of dietary H. pluvialis (p < 0.05); D1 had the highest levels of SOD, POD and GSH‐Px in hepatopancreas. For the non‐specific immune indices, the highest activities of acid phosphatase (ACP) and γ‐glutamyl transpeptidase (γ‐GT) were found on the serum of D3 and D4 (p < 0.05). D1 had the highest levels of ACP and alkaline phosphatase (ALP) in hepatopancreas, while D2 and D3 had the lowest levels of ALP and ACP respectively. These results suggested the optimal dietary natural astaxanthin level was around 40 mg/kg diets.     

Dietary carotenoids and skin melanin content influence the coloration of farmed red porgy (Pagrus pagrus)

The present study investigates the effects of dietary carotenoid sources on the coloration of the red porgy, Pagrus pagrus. Red porgies (131.9 ± 16.2 g; mean ± SD) were fed for 12 weeks on five different diets supplemented with red carotenoids (mainly astaxanthin esters) supplied from Haematococcus pluvialis algae and yellow carotenoids (mainly β‐carotene, lutein, and zeaxanthin) supplied from Alfalfa, Medicago sativa L. and Spirulina, Spirulina pacifica. The carotenoid‐supplemented diets did not have any marked effect on the growth rate, the feed conversion ratio, the daily feeding rate, or the hepatosomatic index of red porgy. The biochemical indices measured in plasma including cholesterol, total proteins, glucose, lactate, phospholipids, non‐esterified fatty acids, triglycerides, and thyroid hormones did not differ significantly among groups. Diet did not affect significantly the melanophore‐area coverage, the melanin skin concentration and skin lightness. Carotenoid‐supplemented diets affected significantly the carotenoid deposition in the skin, the presence and distribution of erythorphores and xanthophores, and skin hue and chroma. Overall, data have shown the efficacy of Haematococcus algae in promoting a reddish coloration in red porgy.     

Astaxanthin from Haematococcus pluvialis Prevents Oxidative Stress on Human Endothelial Cells without Toxicity

Astaxanthin, a powerful antioxidant, is a good candidate for the prevention of intracellular oxidative stress. The aim of the study was to compare the antioxidant activity of astaxanthin present in two natural extracts from Haematococcus pluvialis, a microalgae strain, with that of synthetic astaxanthin. Natural extracts were obtained either by solvent or supercritical extraction methods. UV, HPLC-DAD and (HPLC-(atmospheric pressure chemical ionization (APCI)+)/ion trap-MS) characterizations of both natural extracts showed similar compositions of carotenoids, but different percentages in free astaxanthin and its ester derivatives. The Trolox equivalent antioxidant capacity (TEAC) assay showed that natural extracts containing esters displayed stronger antioxidant activities than free astaxanthin. Their antioxidant capacities to inhibit intracellular oxidative stress were then evaluated on HUVEC cells. The intracellular antioxidant activity in natural extracts was approximately 90-times higher than synthetic astaxanthin (5 µM). No modification, neither in the morphology nor in the viability, of vascular human cells was observed by in vitro biocompatibility study up to 10 µM astaxanthin concentrations. Therefore, these results revealed the therapeutic potential of the natural extracts in vascular human cell protection against oxidative stress without toxicity, which could be exploited in prevention and/or treatment of cardiovascular diseases.     

Colouring ornamental fish (Cyprinus carpio and Carassius auratus) with microalgal biomass

Koi carp and goldfish value increases with intensity of skin colour, which is an important quality criterion. Fish cannot fully synthesize their own carotenoid colourings and these must therefore be included in their diet. Two trials were undertaken to investigate skin colour enhancement in ornamental species (i.e. three chromatic varieties of koi carp (Cyprinus carpio), namely Kawari (red), Showa (black and red) and Bekko (black and white) and goldfish (Carassius auratus)) by feeding a dietary carotenoid supplement of freshwater microalgal biomass [Chlorella vulgaris, Haematococcus pluvialis, and also the cyanobacterium Arthrospira maxima (Spirulina)], using a diet containing synthetic astaxanthin and a control diet with no colouring added for comparison. In the first trial, five homogeneous duplicate groups of 25 juvenile koi carp (C. carpio) (initial mean body weight 24.6 ± 0.7 g) were fed, for 10 weeks, one of the four diets containing 80 mg colouring/kg diet. In the second trial, this procedure was repeated for five homogeneous duplicate groups of 25 goldfish (C. auratus) (initial mean body weight of 0.9 ± 0.1 g). Initial and final samples of skin along the dorsal fin were withdrawn, from five fish per group, for subsequent analysis of total carotenoid content (spectrophotometric analysis), and red hue (colorimetric analysis, CIE (1976) L* a* b* colour system). Growth and feed efficiency were not significantly different between groups administered by the various dietary treatments. In both trials, dietary carotenoid supplementation increased total skin carotenoid content. The more efficient colouring for koi carps was found to be C. vulgaris biomass, providing both maximum total carotenoid deposition and red hue for the three chromatic koi carp varieties studied, and particularly for the kawari variety. For goldfish the best colouring obtained, as ascertained by total carotenoid content, was also achieved using C. vulgaris biomass, and red hue was maximum when using H. pluvialis biomass.     

硝酸钠浓度对雨生红球藻生长及虾青素积累的影响

研究了0.15、0.30、0.75、1.50 g/L 4种不同硝酸钠营养浓度条件对雨生红球藻生长状况的影响,同时探究了该试验条件对雨生红球藻中虾青素起始积累期和后期积累量的影响,以寻找既能使雨生红球藻在氮源用尽后细胞密度达到或接近最大,同时又能最快直接进入虾青素积累阶段的硝酸钠营养添加浓度。试验结果表明,硝酸钠浓度为0.15 g/L时雨生红球藻细胞生长状况最好,细胞浓度最高可以达到5.19×105个/ml。此条件下细胞尚未停止生长即已有细胞开始积累虾青素,变红(培养的第23天)。使用有机溶剂萃取法以丙酮提取虾青素,硝酸钠浓度为0.15 g/L时,最终虾青素浓度达到19.136 mg/L,是其他硝酸钠浓度下的虾青素含量最大值。     

雨生红球藻规模化培养工艺的构建与应用

针对雨生红球藻规模化培养中的工艺流程、生物污染、温度控制、溶氧解析和pH值稳定等问题,于2003年到2005年,在山东省潍坊地区,开展了雨生红球藻中试规模的论证研究。建立了可用于雨生红球藻规模化生产的3种不同类型的光生物反应器(柱式、管道、生物幕)和人工调节太阳能的温控车间,并构建了一套二步规模化培养工艺流程。结果表明,研究构建的培养模式降低了培养过程中的潜在风险,并大大提高了雨生红球藻虾青素含量。与传统培养方式相比,年培养天数大约提高了3倍。在每个批次培养中,细胞生长培养效率提高了至少1.75倍,虾青素累积培养所需时间缩短了40%,虾青素含量提高了4倍。总体来看,研究构建的新工艺比传统开放模式的生产效率有很大提高,全年共计提高了35倍。     

Fe2+,醋酸盐和双氧水对雨生红球藻积累虾青素的影响

研究了亚铁离子、醋酸盐和双氧水三种化学因子对雨生红球藻积累虾青素的交叉影响和虾青素积累过程中藻体总可溶性蛋白的变化.结果表明,在营养盐缺乏的前提下,添加45 mM醋酸盐和450 μM亚铁离子,25 ℃,24 h连续光照,6 000 lx光强下进行逆境胁迫诱导,能够大大缩短雨生红球藻虾青素积累周期,比空白对照提前一个月使藻细胞完全变红（显微观察）.但诱导过程中会造成部分藻细胞白化,自溶或破壁死亡,虾青素产量比空白对照下降了7.4%;在雨生红球藻虾青素积累过程中,藻体总可溶性蛋白含量逐渐降低,说明雨生红球藻中虾青素的积累可能以蛋白质的消耗为基础.     

雨生红球藻虾青素粉对锦鲤组织色素沉积的影响

本试验的目的是探讨锦鲤饵料中雨生红球藻虾青素粉的最适添加量及锦鲤的着色规律。试验将平均体重为10g的420尾锦鲤分成7组,每组3个重复。在基础饵料中分别添加0,0．5,1．5,2．5,3．5,4．5,5．5g／kg雨生红球藻虾青素粉,试验结果表明：在相同饲养条件下饲养60d,锦鲤头和鳍中色素的沉积量各组差异不显著（P〉0．05）。而体组织中色素沉积量各组间差异显著（P〈0．05）,其中,雨生红球藻虾青素粉添加量为3．5g／kg的组着色最显著。     

普通小球藻和雨生血球藻之间的化感效应

为了探讨经济微藻之间是否存在化感互利效应的问题,进行了普通小球藻和雨生血球藻之间的化感作用效应试验。试验设置了氮、磷营养限制和正常营养2个水平,普通小球藻和雨生血球藻分别作为受体被培养在相对应的供体藻滤液中的处理,测定了藻细胞密度、生物量和叶绿素a含量等指标。结果表明：①普通小球藻滤液对雨生血球藻的生长具有化感促进效应,雨生血球藻滤液对普通小球藻同时存在化感促进或抑制效应。②在正常营养条件下,2种微藻的细胞密度均呈现较明显的互利促进效应,普通小球藻滤液对雨生血球藻细胞密度的增加量为5.5%-15.8%;雨生血球藻滤液对普通小球藻细胞密度的促进量为2.0%-16.8%。③在营养限制条件下,普通小球藻滤液对雨生血球藻具化感促进作用,细胞密度的促进量为2.3%-13.0%;而雨生血球藻滤液对普通小球藻却产生化感抑制效应,细胞密度的抑制率为10.0%-14.1%。