Effects of E/Z Isomers and Coating Materials of Astaxanthin Products on the Pigmentation and Antioxidation of Rainbow Trout,Oncorhynchus mykiss

The objective of this study was to compare the effects of four astaxanthin preparations with different ratio of E/Z (trans/cis) isomers and different coating materials on the pigmentation and antioxidation properties of rainbow trout. Five diets were designed as basal diet (without astaxanthin supplementation) and four astaxanthin diets (100 mg/kg diet) supplemented with four astaxanthin products, BASF (79% all‐E, gelatin coated), Wisdom‐B (85% all‐E, gelatin coated), Wisdom‐C (94% all‐E, carrageenan coated), and Wisdom‐D (94% all‐E, gelatin coated). After 4 wk feeding, the flesh astaxanthin content, redness, and yellowness of astaxanthin‐supplemented groups were higher than those of control group at the second and fourth week (P < 0.05). Among these astaxanthin‐supplemented groups, the Wisdom‐B group had the highest flesh redness and astaxanthin content. The astaxanthin‐supplemented groups also had the lower flesh frozen loss and malondialdehyde level than control group at the fourth week (P < 0.05). The results indicated that dietary 100 mg/kg astaxanthin could improve the flesh redness and antioxidation abilities of rainbow trout. The ration of E/Z isomers and properties of coating materials of astaxanthin preparations influenced the pigmentation and antioxidation properties, and Wisdom‐B had a better pigmentation effect on the flesh than other astaxanthin preparations.     

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.     

Pigmentation and carotenoid content of shrimp fed with Haematococcus pluvialis and soy lecithin

The aim of this work was to evaluate the effects of Haematococcus pluvialis (H. pluvialis) (carotenoid source) and H. pluvialis plus soy lecithin on development, carotenoid content, and pigmentation of shrimp (Litopenaeus vannamei). One hundred and eighty shrimps (7.8 g) were divided in six tanks (n = 30) and fed with control food, H. pluvialis, and H. pluvialis plus soy lecithin for 2 weeks. Carotenoids were extracted with acetone and quantified by UV–vis spectrophotometry, and astaxanthin was determined by high‐performance liquid chromatography. Colour was analysed by colorimetry. Lecithin/H. pluvialis group presented higher survival rate (100%) when compared to control group (93.3%). Haematococcus pluvialis and lecithin/H. pluvialis groups presented higher red‐like colour (a* 16.4 and 19.9) than control (a* 20.6). Lecithin/H. pluvialis group presented higher carotenoids content (8.2 mg kg^?1 muscle, 26.8 mg kg^?1 exoskeleton) and astaxanthin (8.5 mg kg^?1 muscle, 23.3 mg kg^?1 exoskeleton) than control (carotenoids: 4.2 mg kg^?1 muscle, 12.3 mg kg^?1 exoskeleton; astaxanthin: 3.2 mg kg^?1 muscle, 8.1 mg kg^?1 exoskeleton). Feeding with 60 ppm carotenoids (from H. pluvialis) during 2 weeks was sufficient for favouring red‐like pigmentation in shrimp, and lecithin increased astaxanthin content only in exoskeleton.     

Production of astaxanthin from Haematococcus in open pond by two-stage growth one-step process

We have developed a two-stage growth one-step process for cultivation of Haematococcus using a self-designed system that mimics an open pond in the natural environment. The characteristics of this process are green vegetative cell growth and cysts transformation and pigment accumulation that proceed spontaneously and successively in one open photobioreactor. Four strains of Haematococcus (H. pluvialis 26; H. pluvialis 30; H. pluvialis 34; H. pluvialis WZ) were cultured in this imitation system for a duration of 12 days. The changes in cell density and medium pH were closely monitored, and the astaxanthin content and yield of the four Haematococcus strains were measured at the end of 12 days of cultivation. Two of the strains, H. pluvialis 26 and H. pluvialis WZ, were selected as strains suitable for mass culture, resulting in the astaxanthin yield of 51.06 and 40.25 mg L^− 1 which are equivalent to 2.79 and 2.50% of their dry biomass respectively. Based on the laboratory work, 6 batch cultures of H. pluvialis WZ were conducted successfully to produce astaxanthin in two 100 m² open race-way pond by two-stage growth one-step process. The astaxanthin content ranged from 1.61 to 2.48 g 100 g^− 1dry wt., with average astaxanthin content of 2.10 g 100 g^− 1dry wt. Compared with the one-stage production of astaxanthin based on continuous culture, the superiority of our process is that it can accumulate much more astaxanthin in red cysts. Compared with two-stage production of astaxanthin, the advantage of our process is that it does not need to divide the production process into two parts using two bioreactors. The presented work demonstrates the feasibility for producing astaxanthin from Haematococcus using a two-stage growth one-step process in open pond, culture systems that have been successfully used for Spirulina and Chlorella mass culture. The future of Haematococcus astaxanthin production has been also discussed.     

The degree of carotenoid esterification influences the absorption of astaxanthin in rainbow trout, Oncorhynchus mykiss (Walbaum)

The absorption of astaxanthin from diets (30 mg kg^?1 inclusion) supplemented with either unesterified astaxanthin; isolated astaxanthin monoesters, diesters or a cell‐free carotenoid extract from Haematococcus pluvialis were studied in rainbow trout (>200 g). No significant differences (P > 0.05) were recorded in the apparent digestibility coefficients (ADC) (≈60–65%) between astaxanthin sources. However, following consumption of a single meal, peak serum astaxanthin levels at 32 h (≈1.0–1.6 μg mL^?1) were significantly higher (P < 0.05) in fish fed unesterified astaxanthin and astaxanthin monoester, compared to fish fed astaxanthin diester and the cell free extract. However, no significant differences (P > 0.05) were recorded in serum astaxanthin uptake rates between sources of astaxanthin. Results suggest that the extent of carotenoid esterification negatively influences the peak serum levels of astaxanthin in rainbow trout.     

Effect of dietary bile extracts on serum response of astaxanthin in rainbow trout (Oncorhynchus mykiss): a preliminary study

Effects of porcine bile extracts added at three different dietary concentrations 0, 10 and 20 g kg^?1 were studied on astaxanthin serum concentration in rainbow trout (mean weight 200 ± 7 g). Astaxanthin from micro‐algae Haematococcus pluvialis and synthetic astaxanthin (CAROPHYLL® pink) were incorporated in diets of rainbow trout at a rate of 100 mg astaxanthin kg^?1 of feed. Fish were hand fed twice a day. After 5 days of feeding there was a significant effect of the pigment source on the ratio (total blood astaxanthin per unit body weight to cumulative astaxanthin intake per unit body weight). Trout receiving synthetic astaxanthin showed a significantly (P < 0.05) higher ratio than trout fed algal astaxanthin. Increasing dietary bile extract did not lead to produce any effect on this ratio. The power of the statistical analysis is discussed. Therefore, the interaction (pigment source × dietary bile concentration) showed no more effect.     

Effects of dietary astaxanthin on the growth,innate immunity and antioxidant defence system of Paramisgurnus dabryanus

The effects of astaxanthin supplementationon the growth, innate immunity and antioxidant defence system of loach (Paramisgurnus dabryanus) were investigated in this study. A total of 450 fish (initial average weights of 3.00 ± 0.10 g) were fed five diets with graded levels of astaxanthin (0.00, 50.00, 100.00, 150.00 and 200.00 mg/kg) for 56 days. The results showed that astaxanthin supplementation significantly stimulated the growth (FBW, WG, FER, HSI), innate immunity (AKP activity in the hepatopancreas, intestinal tract, muscle and skin and LZM activity in the hepatopancreas, intestinal tract and skin), antioxidant ability (T‐SOD, GSH‐PX and CAT activities and T‐AOC, GSH and MDA contents in the hepatopancreas, intestinal tract, muscle and skin) of loach. Moreover, dietary astaxanthin supplementation significantly up‐regulated the relative mRNA levels of Nrf2 and Maf, and down‐regulated the relative mRNA level of Keap1 in the hepatopancreas when the supplementation levels of astaxanthin were 50–200 mg/kg. In conclusion, the diets with 100–151.06 mg/kg astaxanthin supplementation were optimal for loach, which based on the growth, immunity and antioxidant‐related indicators, and the astaxanthin supplementation regulated the antioxidant ability partially referring to Keap1‐Nrf2 signallings.     

Carotenoid composition of a New Zealand (Evechinus chloroticus) and an Australian (Heliocidaris erythrogramma) sea urchin in relation to gonad colour

Sea urchins are commercially harvested for their edible gonads, also known as roe or uni. Most species, particularly those from the Northern hemisphere are reported to exhibit limited variation in gonad colour, which is proposed to be due to carotenoid pigments. The commercially harvested species from New Zealand, Evechinus chloroticus, and from Australia, Heliocidaris erythrogramma, however, exhibit considerable gonad colour variation, ranging from a culinary desirable light yellow/orange to an undesirable brown. Gonads from E. chloroticus and H. erythrogramma were characterized spectrophotometrically and the carotenoid content extracted and analysed by reversed‐phase high‐performance liquid chromatography. The main carotenoid found in the gonads of both species was 9′‐cis‐echinenone, with lower levels of all‐trans‐echinenone, lutein and isozeaxanthin. Carotenoid composition contributed to the yellow/orange colour components of the gonads, but was not correlated with the darker roe coloration observed in some E. chloroticus individuals. Our results highlight important considerations relating sea urchin gonad coloration to carotenoid content and as such will provide useful information for the future aquaculture and processing of these two species.     

Effect of dietary conjugated linoleic acid levels on growth performance,muscle fatty acid profile,hepatic intermediary metabolism and antioxidant responses in genetically improved farmed Tilapia strain of Nile tilapia Oreochromis niloticus

An 8‐week feeding trial was conducted to determine the effect of dietary conjugated linoleic acid (CLA) on growth performance, muscle fatty acid profile, hepatic intermediary metabolism and antioxidant responses in genetically improved farmed Tilapia (GIFT) strain of Oreochromis niloticus (initial body weight: 42.6 ± 0.4 g, mean ± standard deviation). Three replicated groups of GIFT strain of Nile tilapia were hand‐fed to satiation, twice a day, with the diets in which CLA oil, containing mainly the bioactive cis‐9, trans‐11 and trans‐10, cis‐12 isomers, was included at 0 (control), 0.5%, 1%, 1.5%, 2% and 2.5%, respectively, at the expense of fish oil to maintain the constant lipid and energy levels. Growth performance and feed utilization showed no significant differences among the treatments (P > 0.05). The dietary inclusion of CLA modified total percentages of the main groups of fatty acids. Increasing saturated fatty acid content and reduced mono‐unsaturated fatty acid contents in muscle were observed with increasing dietary CLA inclusion (P < 0.05). Total n‐3 fatty acids and total polyunsaturated fatty acids tended to decline with increasing dietary CLA levels (P < 0.05), but n‐6 fatty acids showed no significant differences among the treatments (P > 0.05). Dietary CLA supplementation resulted in the significant increase in the trans‐10, cis‐12 and cis‐9, trans‐11 CLA isomers in muscle (P < 0.05) and also significantly influenced several hepatic enzymatic activities, such as succinate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, lipoprotein lipase and hepatic lipase activities (P < 0.05). Reduced superoxide dismutase and glutathione peroxidase activities, and the decline in malondialdehyde levels were observed in fish fed the CLA‐supplemented diets (P < 0.05), indicating that dietary CLA supplementation showed a powerful antioxidant effect for this fish species. Our study was the first report involved in the effect of dietary CLA inclusion on hepatic intermediary metabolism and antioxidant responses in fish, which could be used as indicators of nutritional and physiological status of the fish species.     

Effect of esterification on the absorption of astaxanthin in rainbow trout, Oncorhynchus mykiss (Walbaum)

Gastrointestinal and serum absorption of astaxanthin was studied in rainbow trout, Oncorhynchus mykiss (Walbaum) (217 ± 2 g) fed diets supplemented with either esterified astaxanthin (from Haematococcus pluvialis) or free astaxanthin (synthetic, as 8% w/w beadlets) at similar levels (50 mg kg^?1). After 56 days of feeding, there was a significant difference (P = 0.0582) between steady‐state serum astaxanthin concentrations for fish fed free (2.0 ± 0.3 μg mL^?1) or esterified astaxanthin (1.3 ± 0.1 μg mL^?1) at the 90% confidence level. However, following ingestion of a single meal supplemented with free or esterified astaxanthin, the rates of astaxanthin absorption into serum were not significantly different (P > 0.1) (0.8 ± 0.2 µg mL^?1 h^?1 and 1.0 ± 0.4 µg mL^?1 h^?1 respectively). In fish fed both free or esterified astaxanthin, higher absorption (P < 0.05) of astaxanthin by the ileal (0.8 ± 0.14 μg g^?1 and 0.9 ± 0.15 μg g^?1 respectively) compared with the posterior (0.2 ± 0.01 μg g^?1 and 0.3 ± 0.14 μg g^?1 respectively) intestine was recorded. This confirmed the role of the anterior intestine in carotenoid absorption. Non‐detectable levels of esters in digesta taken from the hind intestine suggest the anterior intestine is also the primary region for ester hydrolysis.     

Effects of dietary astaxanthin on growth,blood biochemistry,antioxidant, immune and inflammatory response in lipopolysaccharide‐challenged Channa argus

The present study investigated the effects of dietary astaxanthin on the growth, blood biochemical, antioxidant, immune and inflammatory response in lipopolysaccharide‐challenged Channa argus. A total of 0, 50, 100 and 200 mg/kg of astaxanthin were added to the basal die for 56 days. After the feeding experiment, each group was subjected to a lipopolysaccharide challenge (except for the Control group). The results showed that adding astaxanthin to the diet can significantly increase the weight gain and specific growth rate and decrease the feed conversion ratio of C. argus; the highest weight gain, specific growth rate and minimum feed conversion ratio occurred in the 100 mg/kg group. Furthermore, dietary astaxanthin supplementation can alleviate the negative effects of lipopolysaccharides by increasing the levels of alkaline phosphatase, lysozyme, complement 3, complement 4, total serum protein, albumin, globulin, superoxide dismutase, catalase and glutathione peroxidase and decrease the serum cortisol, aspartate aminotransferase, alanine aminotransferase, glutathione peroxidase, and malondialdehyde. Dietary astaxanthin supplementation also can decrease the relative expression of inflammatory genes (nuclear factor κB, interleukin‐1, interleukin‐8 and tumour necrosis factor‐α) in the liver, spleen, kidney and intestine. To summarise, dietary astaxanthin addition can improve the growth performance and attenuate the negative effects of lipopolysaccharide challenge in C. argus. The optimal amount of astaxanthin is 100 mg/kg.     

Processing of astaxanthin‐rich Haematococcus cells for dietary inclusion and optimal pigmentation in Rainbow trout,Oncorhynchus mykiss L.

A range of physical cell disruption techniques have been evaluated to aid the processing of astaxanthin‐rich haematocysts of Haematococcus pluvialis for inclusion in salmonid feeds. Cell disruption by a scalable pressure treatment system was shown to be effective in breaking open the haematocysts without altering the content or isomeric composition of carotenoids in the algal cells. Storage of disrupted cells was optimal at ?20°C in the dark under nitrogen. Disrupted cells were spray‐dried, incorporated into commercial diets and fed to rainbow trout (Oncorhynchus mykiss L.). A marketable level of pigmentation in fish muscle was achieved after 10‐week dietary supplementation. The geometric and optimal isomer composition of the astaxanthin deposited in the muscle was nearly identical to that seen in Haematococcus. Changes were observed in the chirality of the astaxanthin deposited in the skin in comparison with that isolated from both the white muscle and the alga.     

Effects of astaxanthin on Brachionus manjavacas (Rotifera) population growth

Astaxanthin is a red secondary carotenoid and powerful antioxidant that is used in aquaculture to enhance colour and improve fish health. Brachionid rotifers are often used as a live feed for larval fish, but do not contain endogenous carotenoids. However, they can be enriched with astaxanthin through their diet and transfer it to larval predators. When supplemented with 2 μg/ml astaxanthin oleoresin extracted from the green alga Haematococcus pluvialis, Brachionus manjavacas rotifer cultures reached significantly higher population densities and maintained them for longer. Furthermore, data are presented that exposure to oleoresin or pure astaxanthin enhances rotifer resistance to oxidative stress, a common cause for the collapse of rotifer mass cultures. Astaxanthin can be visualized in the gut of the rotifers, allowing the time course of uptake to be estimated by image analysis. Using this method, it was found that accumulation of astaxanthin in the rotifer gut saturates after 1.5 hr of exposure. The bioactive dose of astaxanthin oleoresin for rotifers was found to be 1–20 μg/ml. Astaxanthin concentration in rotifer tissues was measured using absorbance spectrophotometry. It was found that treating rotifers with 20 μg/ml for 24 hr; the concentration of astaxanthin absorbed into rotifer tissues was 2.6 mg/g. Overall, these experiments indicate that astaxanthin extracted from H. pluvialis can be used to improve the productivity and stability of rotifer mass cultures by increasing oxidative stress resistance and enhance the nutritional content of rotifers for larval fish.     

Blood appearance, metabolic transformation and plasma transport proteins of 14C-astaxanthin in Atlantic salmon (Salmo salar L.)

The time of appearance in blood, and transport of astaxanthin, and catabolic transformation of astaxanthin to idoxanthin were investigated in Atlantic salmon (Salmo salar) that had been force-fed a single dose of ¹⁴C-astaxanthin. In addition to the LPs, a major protein, associated with radiolabeled astaxanthin was detected. The maximum level of radiolabeled carotenoids in blood was attained 30 h after administration of ¹⁴C-astaxanthin. Radioactive idoxanthin (combined 3,4-cis and 3,4- trans glycolic isomers of idoxanthin) appeared after 6 h and a stable level was obtained after 18 h. LPDP and LP, separated by ultracentrifugation, contained on average 89 and 11% of the total radioactivity in plasma, respectively. During the 168 h experiment, maximum radioactivity in LP appeared after 22 h. Separation of plasma by ultracentrifugation on a discontinuous NaCl/KBr-gradient and an iodixanol-gradient confirmed that most of the radiolabeled carotenoids were present in the HDPF that did not contain LPs (58%), whereas HDL and LDL contained 36 and 6% of the radioactivity, respectively. Of the recovered radioactivity, astaxanthin in the HDPF comprised 82%, idoxanthin 5% and unidentified compounds 12%, whereas HDL contained 78% astaxanthin, 22% idoxanthin and no unidentified compounds. Proteins from the fractions with the high density and high radioactivity (iodixanol-gradient) were separated by PAGE under non-denaturing conditions and showed a radioactive band with parallel migration length to BSA and salmon albumin. These results show that astaxanthin is rapidly converted to idoxanthin and that the majority of astaxanthin in the plasma is associated with a protein other than LPs, presumably albumin. The identity of this protein requires verification.     

Diets supplemented with microalgal biomass: effects on growth,survival and colouration of ornamental fish Hyphessobrycon eques (Steindacher 1882)

This study evaluated the effects of the addition of microalgae (Ankistrodesmus gracilis and Haematococcus pluvialis) to the fish diet in improving the growth and optimal pigmentation (red carotenoid) of Hyphessobrycon eques. The basal mixed diets consisted of a formulated diet, supplemented with dried microalgae biomass of A. gracilis (1.5 g kg^?1) and H. pluvialis (1.5 g kg^?1). The live food diets contained zooplankton was cultured in open ponds, associated with microalgae. All the microalgae were cultured in the laboratory. No mortality was observed with any experimental diets. Fish performance results showed significant differences (P < 0.05) between the basal diet (BD) and the live food diet. The higher weight and total length were observed with mixed diets (BD + H. pluvialis and BD + A. gracilis). The mixed diets promoted more intense values of chroma (Cab*), lightness (L*) and redness (a*) to H. eques. Diaphanosoma birgei (Cladocera) represented more than 32% of zooplankton ingested by ornamental fish in live food (zooplankton and zooplankton + microalgae), and Argyrodiaptomus furcatus (Copepoda) was the species most ingested by H. eques in live food dietary treatment zooplankton. The feeding behaviour observed in the laboratory as well as the food preferences of H. eques was dependent on the zooplankton composition present in the used open ponds. This study showed that diets with microalgae and zooplankton were able to enhance the pigmentation of H. eques, being a good tool to benefit the culture management of this species.     

Digestibility,growth and pigmentation of astaxanthin,canthaxanthin or lutein diets in Lake Kurumoi rainbowfish,Melanotaenia parva (Allen) cultured species

New cultured ornamental fish namely Lake Kurumoi rainbowfish Melanotaenia parva (Allen) run into reduced of colour performances when reared in the aquaria, consequently, fish feed must be added with carotenoids as a pigment source. The aim of this study was to evaluate the digestibility, growth and pigmentation of astaxanthin, canthaxanthin and lutein in diet. Apparent digestibility coefficients (ADC) of dry matter, lipid, protein, carotenoids, growth and pigmentation were studied in twenty fish after 14 and 56 days of observation. The single‐dose supplementation of 100 mg/kg of astaxanthin, canthaxanthin, or lutein diets on fish was fed by apparent satiation. The basal diet without carotenoids was used as control. The result showed that the ADC of carotenoids of test diets was higher compared to control. Fish fed astaxanthin diet had higher survival rate (96.67 ± 2.89%), colour measurements of lightness (57.60 ± 7.46%), a*‐values (4.66 ± 1.20), total carotenoids content in skin (33.75 ± 5.02 mg/kg) and muscle (2.16 ± 0.74 mg/kg). Astaxanthin also increased the growth after 14 days (2.00% ± 0.19%/days) but there was no significantly different at the end of experiment. The yellowish‐orange colour performance was more rapidly achieved by fish fed astaxanthin diet after 28 days experimentation. These values suggested that dietary carotenoids were required and astaxanthin diet was superior to other diets for skin pigmentation of Lake Kurumoi rainbowfish.     

Effect of combination of dietary Bacillus subtilis and trans‐cinnamic acid on innate immune responses and resistance of rainbow trout,Oncorhynchus mykiss to Yersinia ruckeri

The present study investigated the effects of combination of dietary Bacillus subtilis and trans‐cinnamic acid on serum biochemical parameters, innate immune responses and resistance of rainbow trout, Oncorhynchus mykiss to Yersinia ruckeri. Six experimental groups of fish with mean weights of 20.58 ± 0.35 g were used in the study. Five experimental groups of fish were fed diets containing Bacillus subtilis (10⁷ per gram) or a mix of the Bacillus subtilis (BS) and trans‐cinnamic acid (25 mg/kg‐25trcBS, 50 mg/kg‐50trcBS, 75 mg/kg‐75 trcBS, 150 mg/kg‐150 trcBS), whereas an additive‐free basal diet served as the control (Cont). In this study, an increase was observed in granulocyte percentage, respiratory burst activity, phagocytic activity, phagocytic index, myeloperoxidase activity and total antiprotease activity especially in fish fed with mix of the BS and trans‐cinnamic acid‐supplemented diets (p < .05). Moreover, at the end of the 20‐day challenge period the survival rates and antibody titre (p < .05), and relative per cent survival were higher in the BS group and all trcBS groups compared with control group. As a conclusion, the results in the present study show that feeding rainbow trout with diets containing a mix of B. subtilis and trans‐cinnamic acid over a 60‐day period might be sufficient for improving fish immune responses and disease resistance against Y. ruckeri.     

Astaxanthin profiles and corresponding colour properties in Australian farmed black tiger prawn (Penaeus monodon) during frozen storage

The colour of commercial cooked black tiger prawns (Penaeus monodon) is a key quality requirement to ensure product is not rejected in wholesale markets. The colour, due to the carotenoid astaxanthin, can be impacted by frozen storage, but changes in colour or astaxanthin profile, during frozen storage, have not been studied in detail. Subsequently in this study, the aims were to define the astaxanthin (as cis, trans, mono‐ester and di‐ester forms) content, together with the colour properties, in both pleopods (legs) and abdominal segments. Changes in astaxanthin content and colour properties were further determined during frozen storage (?20°C). Total astaxanthin content was seen to decrease in all samples over time, with the rate of degradation being significantly greater (P < 0.05) in pleopods than abdomen. In both pleopods and abdomen, rate of degradation of esterified forms was significantly greater (P < 0.05) than non‐esterified forms. Hue angle (increase), a* value (decrease) and L value (increase) were all seen to significantly change (P < 0.05) during storage, with changes being more prevalent in the pleopods. The pleopods are the key indicator of astaxanthin and colour loss in cooked black tiger prawns and preservation strategies are required to preserve astaxanthin and colour during frozen storage.