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。     

Pigmentation, carotenoids, lipid peroxides and lipid composition of skin of red porgy (Pagrus pagrus) fed diets supplemented with different astaxanthin sources

A feeding experiment was carried out to determine the efficiency of different commercial sources, chemical forms and levels, of dietary astaxanthin, to appropriately pigment the red porgy (Pagrus pagrus) skin. According to this, total carotenoid content, profiles and chemical forms present in the skin were determined. In order to establish the potential for antioxidant protecting role of astaxanthin supplemented diets, peroxide levels and lipid composition of skin were also determined.

Red porgy alevins were fed six dietary treatments in triplicate; a basal diet (B) without carotenoids; two diets (N25 and N50) formulated to supply either 25 or 50 mg kg^− 1 of an esterified source of astaxanthin (Haematococcus pluvialis, NatuRose™); two diets (CP25 and CP50) with either 25 or 50 mg kg^− 1 of unesterified astaxanthin (Carophyll® Pink); and a positive control diet (B + S) proved as a successful pigmenting-diet in previous experiences (B + S, 88% basal diet:12% frozen shrimp) [Cejas, J., Almansa, E., Tejera, N., Jerez, S., Bolaños, A., Lorenzo, A., 2003. Effect of dietary supplementation with shrimp on skin pigmentation and lipid composition of red porgy (P. pagrus) alevins. Aquaculture 218, 457–469].

All fish fed carotenoid supplemented diets displayed a pink-coloured skin after 4 months of feeding in contrast to the greyish appearance displayed by fish fed the basal diet not supplemented with carotenoids (B). Furthermore, astaxanthin diesters were the major carotenoid in the skin of pink fish. A second carotenoid, tentatively identified as tunaxanthin diester, was also detected. The best results in terms of skin natural reddish hue, total carotenoid and astaxanthin contents were found by using the esterified forms of dietary astaxanthin (N25, N50 and B + S). Interestingly, the lowest levels of lipid peroxides were found in the fish fed these three treatments. However, no effect of treatment on lipid composition was found. In conclusion, red porgy alevins are able to efficiently utilise dietary natural or synthetic astaxanthin, and deposit this pigment in its esterified form to acquire an acceptable pink-coloured skin compared to that of the wild fish.     

Effect of lipid source and bile salts in diet of Atlantic salmon, Salmo salar L., on astaxanthin blood levels

A study was conducted to determine the effects of dietary lipid and bile acids on astaxanthin absorption in Atlantic salmon (Salmo salar L.). Fish with an average weight of 1500 g were fitted with a dorsal aorta cannula and fed diets containing herring oil, soybean lecithin, lard, or herring oil supplemented with taurocholic acid (2.5 g/kg diet). Each fish was fed all of the experimental diets in successive order to minimize the effect of individual variation. At a given time following the feeding of each diet, blood was collected and analyzed for astaxanthin. Soybean lecithin significantly lowered the absorption of astaxanthin compared to fish fed herring oil. A 20% (p < 0.12) increase in blood astaxanthin was observed when the fish were fed the diet supplemented with taurocholic acid. Feeding lard significantly increased the blood astaxanthin level compared to the control group. It appears that altering the micellar structure by stimulating micellar (taurocholic acid) or mixed micellar (lecithin) systems did not increase the apparent absorption of astaxanthin. However, increasing the phospholipid level may have actually decreased the absorption possibly by lowering the astaxanthin solubility in the micelles. The increased apparent absorption of astaxanthin with lard is possibly linked to the increased content of 16:0, 18:1n − 9 or 18:2n − 6 fatty acids in this diet, or a reduction in very long chain monoenes (20:1n − 9 and 22:1n − 9). This suggests that the solubility of astaxanthin is higher in diets containing higher levels of 16:0 or 18:1n − 1, or alternatively, that reductions in longer chain monoenes (20:1n − 9 and 22:1n − 9) increase the micellar solubility of this pigment.     

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。     

虾青素合成关键酶基因bkt和CrtR-B双价植物表达载体的构建及对花生的遗传转化

虾青素是一种具有极强抗氧化活性的类胡萝卜素,具有广泛的应用价值。β-胡萝卜素酮化酶(BKT)和β-胡萝卜素羟化酶(CrtR-B)是虾青素的生物合成途径中的两个关键酶。采用LaTaqDNA聚合酶用PCR的方法分别从pET-28a(+)-bkt,pET-28a(+)-CrtR中扩增得到bkt和CrtR-B基因,用bkt基因替换pBI221中的GUS基因形成含有CaMV 35S启动子和NOS终止子的bkt基因表达盒,然后插入植物表达载体pCAMBIA1301的多克隆位点,再用CrtR-B基因替换pCAMBIA1301中的另一个GUS基因,形成CrtR-B基因表达盒,最终获得带有抗性基因的双价植物表达载体pCAMBIA1301-bkt-CrtR。通过农杆菌EHA105介导将其转入花生(花育23),获得了经卡那霉素筛选的抗性植株,通过PCR扩增验证,花生基因组中含有这两个基因。     

Effects of dietary supplementation of carotenoids on survival, growth, pigmentation, and antioxidant capacity of characins, Hyphessobrycon callistus

This study aimed to find out if dietary carotenoid (CD) supplement could make differences in survival, growth, pigmentation, and antioxidant capacity of characins Hyphessobrycon callistus, an ornamental fish. Two types of CD and its combination (AX — astaxanthin, BC — β-carotene, MX — 1:1 combination of AX and BC) at three concentrations (10, 20, and 40 mg/kg) were used resulting in nine pigmented diets. A diet without CD supplement served as control. No differences in growth and survival of the fish among treatments were found after 8 weeks rearing. Disregarding the types of dietary CD, AX dominated (> 98%) the body CD, indicating that this fish converted most dietary BC into body AX for storage. Body AX and BC content increased with increasing dietary CD concentration. Body AX in BC-fed fish was lower than that in both AX- and MX-fed fish. No difference in body AX was found between AX- and MX-fed fish, and in body BC in all pigmented fish. Serum total antioxidant status [TAS], serum antioxidant enzymes (superoxide dismutase [SOD], glutathione peroxidases [GPx]) and serum transaminases (alanine aminotransferase [ALT], aspartate aminotransferase [AST]) were chosen as indices of fish antioxidant capacity. Antioxidant activities changed with dietary CD type and concentration. Pigmented fish had lower SOD, GPx and ALT than control fish; dietary CD types only affected SOD and ALT in fish. AX-fed fish had the lowest SOD. Dietary AX had more numbers of negative correlations with antioxidant parameters in fish than BC.     

Replacing fish oil and astaxanthin by microalgal sources produced different metabolic responses in juvenile rainbow trout fed 2 types of practical diets

Dietary fish oil supplementation provides n-3 long-chained polyunsaturated fatty acids for supporting fish growth and metabolism and enriching fillet with eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; c22:6n-3). Two experiments were performed as a 3 × 2 factorial arrangement of dietary treatments for 16 wk to determine effects and mechanisms of replacing 0%, 50%, and 100% fish oil with DHA-rich microalgae in combination with synthetic vs. microalgal source of astaxanthin in plant protein meal (PM)- or fishmeal (FM)- based diets for juvenile rainbow trout (Oncorhynchus mykiss). Fish (22 ± 0.26 g) were stocked at 17/tank and 3 tanks/diet. The 100% fish oil replacement impaired (P < 0.0001) growth performance, dietary protein and energy utilization, body indices, and tissue accumulation of DHA and EPA in both diet series. The impairments were associated (P < 0.05) with upregulation of hepatic gene expression related to growth (ghr1and igf1) and biosynthesis of DHA and EPA (fads6 and evol5) that was more dramatic in the FM than PM diet-fed fish, and more pronounced on tissue EPA than DHA concentrations. The source of astaxanthin exerted interaction effects with the fish oil replacement on several measures including muscle total cholesterol concentrations. In conclusion, replacing fish oil by the DHA-rich microalgae produced more negative metabolic responses than the substitution of synthetic astaxanthin by the microalgal source in juvenile rainbow trout fed 2 types of practical diets.     

Rapid compositional analysis of Atlantic salmon (Salmo salar) using visible‐near infrared reflectance spectroscopy

Rapid measurement of salmon flesh quality parameters (>400 samples day^?1) was demonstrated in the laboratory and remotely at industrial sites. Visible‐near infrared spectroscopy (VNIRS) was applied to predict astaxanthin (AX) and fat content in farmed Atlantic salmon. Fish were sampled from thirteen batches (1–6 kg whole weight, containing 2.3–16.3% fat and 1.2–12.5 μg g^?1 AX), and models validated on small (average ± SD: 1.4 ± 0.4 kg) and large fish (4.2 ± 0.9 kg). Both constituents were well predicted in minced Norwegian Quality Cutlet (NQC) samples (r² ≥ 0.86; standard error of prediction (SEP) ≤0.7% for fat and ≤0.7 μg g^?1 for AX). Comparable metrics were observed for AX prediction in whole NQCs (r² = 0.80–0.88; SEP 0.7 μg g^?1). Fat was better predicted in small fish than large fish for whole NQCs (r² = 0.82, SEP 1.0% cf r² = 0.59, SEP = 0.59%) and non‐destructive scanning through the skin of whole, gutted fish (r² = 0.77, SEP = 1.2% cf r² = 0.49, SEP = 1.5%). Models were also developed for screening polyunsaturated fatty acid (PUFA) concentrations, e.g. in NQCs for docosahexaenoic acid (r² = 0.73) and n‐3:n‐6 PUFA (r² = 0.89).     

豹纹鳃棘鲈(Plectropomus leopardus)早期形态与色素变化及添加剂对其体色的影响

为研究豹纹鳃棘鲈色素变化过程以及添加剂对其产生的影响,本研究在室内工厂化养殖条件下,对豹纹鳃棘鲈的早期色素积累、转变过程进行描述,并采用螺旋藻粉和虾青素两种添加剂对其仔、稚、幼鱼的生长及体色变化的影响进行观察。结果显示,从2 d仔鱼开始,眼点色素增多,并由透明转为黑色,摄食明显;3 d仔鱼其背部鳍褶上的树枝状黑色素扩大,脊椎下方出现1列黑色素细胞丛;22 d的鱼体脊椎上方出现1排黑色素斑点,下方黑色素斑点数量减少,此时口、各鳍基部及沿脊椎两侧黄色素增加;28 d的稚鱼体表黄色素与红色素进一步增多,各鳍沿鳍条均有红色素斑点分布,鱼体呈橘红色,颜色鲜艳;30–33 d的幼鱼体表两侧布满黑色与橘红色斑点,脊椎下方斑点消失,仅在尾椎下方存有1块黑色素斑点。增色剂实验结果显示,实验15 d时,螺旋藻粉组与虾青素组全长、体重差异不显著,但均显著高于对照组(P0.05),各实验组内梯度之间差异不显著(P0.05)。30 d时,3%和6%螺旋藻粉组较对照组的体重值高,但差异不显著(P0.05),9%螺旋藻粉组显著低于对照组(P0.05);虾青素组均高于对照组,除与0.6%组差异不显著(P0.05)外,其他组均差异显著(P0.05)。0.1%实验组体重高于对照组和螺旋藻粉组,并高于组内其他梯度组。经过15 d添加虾青素的幼鱼体色红色素有明显增加,与对照组差异显著;30 d时,螺旋藻组没有达到增色效果。结果表明,螺旋藻粉对幼鱼生长初期有促进作用,随着添加浓度与养殖时间的增加其效果不明显,出现负效应;添加浓度为0.1%的虾青素可使全长、体重增长率最高并具有明显增色效果。     

Comparison of Physical,Thermal and Chemical Methods to Measure Protein Denaturation in Frozen Pacific Whiting (Merluccius productus)

The potential of using the torsion test and differential scanning calorimetry (DSC) to determine the effect of frozen storage on protein denaturation in fish fillets was investigated. Pacific whiting fillets were stored for 12 weeks at three temperature conditions: -20¦C, -8¦C, and at a level varying between 0 and -8¦C. Salt soluble protein (SSP) extractability and Ca++-ATPase activity were used to evaluate the torsion test and DSC. The shear strain value of the torsion test provided a good correlation with SSP extractability, Ca++-ATPase activity, and myosin transition enthalpy as measured by DSC. Therefore, shear strain can be considered as a useful tool for the determination of protein denaturation in Pacific whiting during periods of frozen storage. Because Ca++-ATPase activity, shear stress and shear strain, and myosin transition enthalpy all decreased within one week, protein deterioration in frozen Pacific whiting appears to be rapid at the temperatures tested.