Natural bixin as a potential carotenoid for enhancing pigmentation and colour in goldfish (Carassius auratus)

Discovering natural carotenoids for colour enhancement and health benefits of fish is important to develop new feed formulations. We have purified natural bixin from achiote seeds and evaluated the effect of colour enhancing and pigmentation in goldfish. Varying levels of bixin‐based diets were prepared with 420 g kg^?1 of crude protein and 120 g kg^?1 of lipid content. Our results clearly showed that bixin (0.05, 0.10, 0.20 and 0.60 g kg^?1) based diets significantly (P < 0.05) enhanced the skin and fin colour at 30 and 60 days compared to diet without bixin. Interestingly, diet which contains 0.20 g kg^?1 bixin and commercial feed (with astaxanthin) had similar effect on carotenoid deposition in skin. Moreover, total carotenoid deposition in fin was higher than in skin of all bixin‐containing diets. However, 0.60 g kg^?1 bixin‐containing diet had lower specific growth rate (1.01 ± 0.01) and higher feed conversion ratio (2.05 ± 0.19) compared to the control group. The present results demonstrate that achiote bixin can be successfully used as an alternative natural carotenoid source against synthetic astaxanthin in fish feed. Our data indicate that 0.20 g kg^?1 is a suitable dietary level of bixin to ensure strong pigmentation, acceptable growth and feed utilization in goldfish.     

虾青素对大鳞副泥鳅幼鱼生长、体成分及抗氧化指标的影响

【目的】研究饲料不同虾青素添加水平对大鳞副泥鳅幼鱼生长、体成分及抗氧化指标的影响。【方法】选取450尾初始体质量为(3.00±0.10) g/尾的大鳞副泥鳅幼鱼,随机分成5组,每组3个重复,每个重复30尾鱼,分别饲喂基础饲料(对照组)以及添加50,100,150,200 mg/kg虾青素的试验饲料,在控温养殖系统中进行为期8周的饲养试验。试验结束后,称量鱼体质量并计算生长指标,测定试鱼体成分及肝胰脏抗氧化指标。【结果】试验期间大鳞副泥鳅成活率为100%。大鳞副泥鳅的终末体质量、平均增重率和特定生长率在虾青素添加水平为50～200 mg/kg时显著高于对照组(P0.05),饲料效率和蛋白质效率在虾青素添加水平为100～200 mg/kg时显著高于对照组(P0.05);结合抛物线回归分析结果与方差分析结果可知,大鳞副泥鳅平均增重率最大时的最适虾青素添加水平为100～151.06 mg/kg,饲料效率最大时的虾青素添加水平为100～157.04 mg/kg。当虾青素添加水平为50～200 mg/kg时,大鳞副泥鳅全鱼蛋白质含量显著高于对照组(P0.05)。肝胰脏总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性及谷胱甘肽(GSH)含量在虾青素添加水平为50～200 mg/kg时均显著高于对照组(P0.05);当虾青素添加水平为50～200 mg/kg时,丙二醛(MDA)含量显著低于对照组(P0.05)。【结论】本试验条件下,综合生长、体成分及抗氧化指标,大鳞副泥鳅饲料中虾青素的最适添加水平为100～151.06 mg/kg。     

Antioxidant defence to ammonia stress of characins (Hyphessobrycon eques Steindachner) fed diets supplemented with carotenoids

This study aimed to find out whether dietary carotenoid (CD) supplement could influence the resistance of characins (Hyphessobrycon eques Steindachner) to ammonia stress. Two types of CD and its combination [astaxanthin (AX), β‐carotene (BC), 1 : 1 combination of AX and BC (MX)] at three concentrations (10, 20 and 40 mg kg⁻¹) were used resulting in nine pigmented diets. No differences in growth and survival of the fish among treatments were found after 8‐week rearing. Experimental and control fish were then exposed to 15 mg total ammonia nitrogen L⁻¹ (stress group) and 0.15 mg total ammonia nitrogen L⁻¹ (normal group) for 72 h, and their blood was withdrawn. No mortality resulted under such TAN concentrations. 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 or stress resistance. SOD, GPx and AST were affected by the interactions of dietary CD and ammonia stress. The activities of TAS, SOD, GPx and AST increased under the stress. Dietary CD reduced serum SOD, GPx, ALT and AST activities. In conclusion, dietary CD increased the resistance of characins to ammonia stress.     

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.     

The pigmenting effect of different carotenoids on fancy carp (Cyprinus carpio)

The optimal concentration of a panel of individual and combined carotenoid sources on skin pigmentation in fancy carp was investigated by nine experimental diets that were formulated and supplemented with astaxanthin at 25 mg kg^?1, lutein at 25 and 50 mg kg^?1, β‐carotene at 25, 50 and 75 mg kg^?1, and lutein combined with β‐carotene at 25 : 25 and 50 : 50 mg kg^?1, while a diet without supplemented carotenoid served as a control. The results showed that serum TC of fish fed diets containing supplemented with lutein plus β‐carotene at 25 : 25; 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet were higher than the other treatments (P ≤ 0.05). Serum TC of the respective treatments was 6.2 ± 2.0, 7.8 ± 3.3 and 7.3 ± 1.9 μg mL^?1 serum, respectively. Fish fed diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 diet had serum astaxanthin concentrations similar to fish fed the diet with astaxanthin alone at 25 mg kg^?1. Serum astaxanthin concentrations was 0.7 ± 0.01, 0.9 ± 0.01, 0.4 ± 0.02 and 1.7 ± 0.18 μg mL^?1 serum, respectively. The chromaticity of fish body skin of red and white position was assessed by colourimetry using the CIE L*a*b (CIELAB) system. Pigmentation response of skin redness of fancy carp fed with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were higher than other treatments (P ≤ 0.05) but they were similar to fish fed with 25 mg kg^?1 astaxanthin diet. The redness (a* values) of fish fed diets with diets combined with lutein and β‐carotene at 25 : 25, 50 : 50 mg kg^?1 and lutein 50 mg kg^?1 were 28.3 ± 0.53, 29.9 ± 1.38, 28.8 ± 3.95 and 28.5 ± 2.49, respectively. After 3 weeks of feeding the experimental diets, the fish fed on a diet without carotenoid supplement for one week demonstrated that the same three groups still retained their redness and had an overall tendency to improve skin colouring. Finally, concentrations 50 mg kg^?1 of lutein, or the combination of lutein and β‐carotene at 25 : 25 mg kg^?1 showed the highest efficiency for improving skin pigmentation and redness of skin.     

Larval pigmentation, survival and growth of Penaeus indicus fed the nematode Panagrellus redivivus enriched with astaxanthin and various lipids

The highly unsaturated fatty acid (HUFA) content of the free-living nematode Panagrellus redivivus was successfully increased when three different lipid sources (cod-liver oil, marilla oil and capelin oil) from marine animals were used in the nematode culture medium. The lipid enrichment significantly increased the ω3 HUFA content, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), of the nematodes compared with lipid non-enriched nematodes. Penaeus indicus larvae fed the lipid-enriched nematodes had significantly ( P < 0.05) greater larval survival (69–77%) until metamorphosis than those fed non-enriched nematodes (54%).
This nematode species was also enriched with astaxanthin to determine the effects of this carotenoid on pigmentation, survival and growth during the larval development of P. indicus . The pigment, delivered via nematodes, significantly ( P < 0.05) improved larval coloration and survival (88%) compared with that of placebo-pigmented nematodes (78%). However, there was no strong evidence to show the benefit of either pigment and/or lipid enrichment on larval growth and development of P. indicus .
This study has shown that the nutritional value of the nematodes can be enhanced by the addition of fish oil into the culture medium. Supplementation of EPA and DHA, together with synthetic astaxanthin, allow the nematodes to be used as a sole diet for the larval culture of P. indicus .     

Effects of dietary astaxanthin concentration and feeding period on the skin pigmentation of Australian snapper Pagrus auratus (Bloch & Schneider, 1801)

A single‐factor experiment was conducted to investigate the effects of dietary astaxanthin concentration on the skin colour of snapper. Snapper (mean weight=129 g) were held in white cages and fed one of seven dietary levels of unesterified astaxanthin (0, 13, 26, 39, 52, 65 or 78 mg astaxanthin kg^?1) for 63 days. Treatments comprised four replicate cages, each containing five fish. The skin colour of all fish was quantified using the CIE L^*, a^*, b^* colour scale after 21, 42 and 63 days. In addition, total carotenoid concentrations of the skin of two fish cage^?1 were determined after 63 days. Supplementing diets with astaxanthin strongly affected redness (a^*) and yellowness (b^*) values of the skin at all sampling times. After 21 days, the a^* values increased linearly as the dietary astaxanthin concentration was increased before a plateau was attained between 39 and 78 mg kg^?1. The b^* values similarly increased above basal levels in all astaxanthin diets. By 42 days, a^* and b^* values increased in magnitude while a plateau remained between 39 and 78 mg kg^?1. After 63 days, there were no further increases in measured colour values, suggesting that maximum pigmentation was imparted in the skin of snapper fed diets >39 mg kg^?1 after 42 days. Similarly, there were no differences in total carotenoid concentrations of the skin of snapper fed diets >39 mg kg^?1 after 63 days. The plateaus that occurred in a^* and b^* values, while still increasing in magnitude between 21 and 42 days, indicate that the rate of astaxanthin deposition in snapper is limited and astaxanthin in diets containing >39 mg astaxanthin kg^?1 is not efficiently utilized. Astaxanthin retention after 63 days was greatest from the 13 mg kg^?1 diet; however, skin pigmentation was not adequate. An astaxanthin concentration of 39 mg kg^?1 provided the second greatest retention in the skin while obtaining maximum pigmentation. To efficiently maximize skin pigmentation, snapper growers should commence feeding diets containing a minimum of 39 mg unesterified astaxanthin kg^?1 at least 42 days before sale.     

Effects of dietary highly unsaturated fatty acids and astaxanthin on the fecundity and lipid content of pond-reared Penaeus monodon (Fabricius) broodstock

Five diets that contained fresh squid meat as the basic constituent and were supplemented with different amounts of highly unsaturated fatty acids (HUFA) and astaxanthin were fed to pond‐reared Penaeus monodon broodstock. Diet A was sole squid meat. Diets B and C were supplemented with astaxanthin 50 and 100 mg kg^?1 respectively. Diets D and E were supplemented with HUFA 5 and 10 g kg^?1 and astaxanthin 50 mg kg^?1 respectively. The result showed that the group fed diet E had the best reproductive performance in all experimental groups. It had a higher proportion of spawns (71.5%), spawning rate (0.047), a shorter latency period (7.7±0.3 d), higher absolute fecundity (× 10³) (361.6±5.5) and egg production/female (× 10³) (597.0±18.0) than all the other experimental groups. The fatty acid composition in broodstock diets strongly affected the tissue and fecundity of broodstock. Good correlations between the content of 20:4n‐6 in eggs and the fecundity (r²=0.6109) and egg production (r²=0.9876) of broodstock were found. On the other hand, 22:6n‐3 and DHA/EPA ratio was negatively correlated with the fecundity of broodstock (r²=0.5362, 0.8702 respectively). The result also showed that the balance between n‐3 and n‐6 fatty acid families, total polyunsaturated fatty acids and total saturated fatty acid and 20:5n‐3 (EPA) and 22:6n‐3 (DHA) may play vital roles in maturation and reproductive performance of P. monodon broodstock.     

Calanus oil as a natural source for flesh pigmentation in Atlantic salmon (Salmo salar L.)

The present study was to understand how efficiently the astaxanthin in Calanus oil is utilized for flesh colouration in Atlantic salmon ( Salmo salar ). Postsmolts of the fish (309 g) were held at 7.9 °C and they were fed diets containing 20 or 60 mg astaxanthin per kilogram feed derived from a synthetic source or from Calanus oil for 181 days. Besides growth and feed intake assessments, at day 81 and 181, fish flesh were subjected to colour analysis and astaxanthin determination. Growth and feed performance did not vary between the groups. There were significant differences in the amount of astaxanthin in muscle between almost all groups both at day 81 ( P  < 0.05) and at day 181 ( P  < 0.001). However, a notable similarity between fish receiving 20 mg astaxanthin from the synthetic source and those receiving 60 mg astaxanthin from Calanus oil ( P  > 0.05) at day 181 indicated that comparable amounts were deposited only with the greater level of the natural source. Tristimuli colorimeter a* values support the analytical results at day 181. Although Calanus oil did serve as a natural dietary pigment source for farmed salmon, its inclusion level should provide more than 60 mg astaxanthin per kilogram feed to achieve colouration preferred by the market.     

法夫酵母诱变选育高产虾青素菌株的研究

以法夫酵母(Phaffia rhodozyma)为试验材料,对其先进行紫外线和氯化锂(UV-LiCl)复合诱变处理,获得了一株产虾青素为635．82μg·g~(-1)的菌株UL-40,其中紫外线适宜诱变条件为30 W、20 cm处照射菌株3 min,LiCl筛选剂量为3 g·L~(-1)。对UL-40菌株用N-甲基-N′-硝基-N-亚硝基胍(NTG)和2-脱氧-D-葡萄糖再进行复合诱变处理,获得了1株遗传性状稳定、25℃条件下的产虾青素为894．54μg·g~(-1)菌株ND-25,其中NTG在25℃条件下最适诱变剂量为150μg·mL~(-1),2-脱氧-D-葡萄糖最适筛选剂量为1 g·L~(-1)。