Effects of dietary keto-carotenoids (canthaxanthin and astaxanthin) on the reproductive performance of female rainbow trout Oncorhynchus mykiss (Walbaum)

The aim of this study was to clarify further the relationship between dietary keto-carotenoids (canthaxanthin and astaxanthin) and reproductive performance in female rainbow trout. Three experiments were undertaken in three successive breeding seasons. In addition to a control diet, fish were fed a canthaxanthin-supplemented diet (200 mg canthaxanthin kg⁻¹ feed, designated as 6mC200) for 6 months prior to spawning (experiment A); the same diet as in A but for either 3 months (3mC200) or 6 months (6mC200) before spawning (experiment B); diets with one of two levels of astaxanthin supplementation, 50 mg (6mA50) or 100 mg (6mA100) kg⁻¹ feed, or a diet with 100 mg canthaxanthin kg⁻¹ feed (6mC100) fed for 6 months (experiment C).
There was no significant influence of carotenoid supplementation on either the frequency of maturing females or the date of maturation. The number of ova per kg of female body weight averaged 2700 and did not vary significantly among fish fed the different diets. Across experiments A, B and C there was no significant difference in egg and larval survival among fish fed carotenoid-supplemented and control diets. In experiment A, 6mC200 females produced smaller eggs than controls but this result was not confirmed in experiments B and C. In general, eyed egg yield appeared partly dependent upon egg size. Alevin weight was also correlated with egg weight. The growth test conducted on fingerlings from experiment B failed to provide any evidence of an effect of feeding carotenoid supplemented diets to the female parent. Pigment analyses conducted on alevins revealed that canthaxanthin fed to the female parent was transferred into the eggs and therefore to the larvae, although canthaxanthin was metabolized within a few weeks after hatching.     

Pharmacokinetics and bioavailabilities of 14C-keto-carotenoids, astaxanthin and canthaxanthin, in rainbow trout, Oncorhynchus mykiss

The pharmacokinetics and bioavailabilities of ¹⁴C‐astaxanthin and ¹⁴C‐canthaxanthin were studied in the blood of rainbow trout following intra‐arterial (i.a.) and oral (p.o.) administration. Sixteen months old 1 kg trout were cannulated in the dorsal aorta. [6,7,6′,7′‐¹⁴C]‐keto‐carotenoids were administered i.a. and p.o. at a dose of 573.5 kBq kg^?1 fish body weight for astaxanthin and 836.2 kBq kg^?1 fish body weight for canthaxanthin. After i.a. distribution, total body clearance (Cl_tot) was 17.30±20.29 mL kg^?1 of fish h^?1 for ¹⁴C‐canthaxanthin and 3.30±1.50 mL kg^?1 of fish h^?1 for ¹⁴C‐astaxanthin. The volume of distribution at steady‐state (V_ss) was 208.32±124.79 mL kg^?1 of fish and 71.84±64.15 mL kg^?1 of fish for ¹⁴C‐canthaxanthin and ¹⁴C‐astaxanthin respectively. Less than 0.4% of the administered radioactivity was recovered in urine. Radioactivity (expressed as percent of the dose) excreted in the bile of fish that received ¹⁴C‐canthaxanthin by i.a. route was 20‐fold higher than that observed for fish treated p.o. This ratio was lower for ¹⁴C‐astaxanthin (7.6‐fold). The mean keto‐carotenoid bioavailabilities calculated were 10–15% for both compounds. Findings suggest one daily astaxanthin application is preferable, while 12‐h time intervals between applications are preferable for canthaxanthin.     

虾青素和角黄素对虹鳟肌肉着色和肝脏总抗氧化能力的影响

分别在基础饲料(对照组)中添加100 mg/kg的虾青素、角黄素,混合色素(50 mg/kg虾青素+50 mg/kg角黄素)饲喂初始体重为(56.60±0.63) g的虹鳟60 d,考察虾青素和角黄素对虹鳟肌肉着色和肝脏总抗氧化能力的影响。结果显示,饲料中添加了虾青素、角黄素和混合色素后对虹鳟增重率、饲料系数及肌肉常规成分、肌肉失水率、含肉率均无显著影响(P> 0.05)。虾青素组、角黄素组和混合色素组虹鳟肌肉的比色卡得分、红度、虾青素含量和血清总类胡萝卜素含量均比对照组有显著提高(P< 0.05);虾青素组虹鳟肌肉比色卡得分(26.25)和红度值(18.40)显著高于角黄素组(22.38, 14.13)和混合色素组(24.00, 15.70)(P< 0.05);虾青素组虹鳟肌肉虾青素含量为4.75 mg/kg (30 d)和6.45 mg/kg (60 d),均显著高于混合色素组的3.87 mg/kg (30 d)和5.48 mg/kg (60 d)(P< 0.05);在虹鳟血清总类胡萝卜素含量方面,虾青素组 > 混合色素组 > 角黄素组;虾青素组、角黄素组、混合色素组虹鳟肝脏的总抗氧化能力之间无显著差异(P> 0.05),分别为2.39 U/mg,2.25 U/mg,2.39 U/mg,均较对照组(2.03 U/mg)显著提高(P< 0.05)。上述结果表明：饲料中添加100 mg/kg虾青素、角黄素及虾青素+角黄素混合(1∶1)均能有效改善虹鳟肌肉颜色,提高肝脏总抗氧化能力,虾青素、虾青素+角黄素混合(1∶1)对虹鳟肌肉的着色效果优于角黄素。     

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.     

Feeding increasing levels of corn gluten meal induces suboptimal muscle pigmentation of rainbow trout (Oncorhynchus mykiss)

A 24‐week growth trial was conducted to evaluate the effects of feeding levels of corn gluten meal (CGM) on growth performance and pigment deposition in the muscle of rainbow trout (Oncorhynchus mykiss). Three isonitrogenous and isoenergetic (digestible energy basis) experimental diets were formulated to contain increasing levels of CGM (0%, 9% and 18%) and 50 mg kg^?1 of astaxanthin. Each diet was fed in triplicate to groups of 75 fish (initial average body weight = 549 g fish^?1) reared at 8.5°C. The inclusion of CGM did not significantly (P > 0.05) affect final body weight, thermal growth efficiency (TGC) or feed efficiency. Carotenoid concentration determined by liquid chromatography showed a significant (P < 0.05) linear reduction in the concentration of one astaxanthin isomer, all‐trans astaxanthin and all‐trans lutein in the muscle of fish in response to increasing levels of CGM. Tristimulus colour analysis of the muscle showed a significant (P < 0.05) linear reduction in a* (redness) and C*ab (chroma). Salmofan^? score showed a significant (P < 0.05) linear and quadratic reduction in response to increasing levels of CGM. In conclusion, the inclusion of CGM up to 18% does not significantly impact growth performance of rainbow trout. However, the concentration of all‐trans astaxanthin as well as the expression of important colour attributes of the muscle can be negatively affected at levels exceeding 9% of CGM in the diet. More research on this topic is needed to discern the mechanism(s) behind the negative effects of dietary CGM and/or its intrinsic yellow pigments on muscle pigmentation of rainbow trout.     

虾青素在虹鳟体内的沉积与降解研究

为研究着色虹鳟在摄食状态和饥饿状态下的虾青素降解规律,实验选用平均体质量为(101.9±1.3)g的未着色虹鳟和已着色虹鳟,对未着色虹鳟饲喂含虾青素(100 mg/kg)饲料以研究虾青素在机体的沉积规律(虾青素沉积组);对着色虹鳟,饲喂含虾青素(100 mg/kg)饲料(正对照)、不含虾青素饲料(摄食降解组)或予以饥饿处理(饥饿降解组),实验共设4个处理组,每个处理组3个重复.实验共持续4周,每周采样一次,测定肌肉、肝脏和全鱼虾青素含量,血清类胡萝卜素含量,肌肉色差值(L*,a*,b*)和肌肉罗氏比色卡得分.结果表明,虾青素沉积组的虹鳟肌肉、肝脏虾青素含量、肌肉红度值和罗氏比色卡得分随饲养时间延长而增加(P＜0.05),在第3周时达到基本稳定,与第4周测定值无显著差异(P＞0.05);在4周养殖期内,全鱼虾青素含量呈增加趋势,而全鱼虾青素沉积率则呈下降趋势,第1周、第4周时的全鱼虾青素含量和全鱼虾青素沉积率分别为3.71 mg/kg、8.59 mg/kg和21.31％、15.26％;饥饿降解组和摄食降解组虹鳟的全鱼、肌肉、肝脏虾青素含量、肌肉红度值和罗氏比色卡得分随养殖时间延长逐渐降低(P＜0.05),其中饥饿降解组的下降趋势更为明显,第3周后,这种降低趋势减缓.上述研究表明,在虹鳟饲料中添加100 mg/kg虾青素,经过3周饲养即可使肌肉达到着色要求,虾青素的沉积率随时间延长而下降;饥饿和投喂不含虾青素的饲料均可使已着色虹鳟肌肉虾青素含量和红色度显著下降,饥饿使虾青素的减少更加迅速,虾青素在鱼体内的减少量随时间的延长而降低.     

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.     

Digestibility, growth and nutrient retention in rainbow trout (Oncorhynchus mykiss) fed diets extruded at two different temperatures

Digestibility and utilization of a fishmeal‐based diet extruded at 103 or 137 °C were examined. Each of the diets was fed to 0.5‐kg rainbow trout in nine tanks supplied with freshwater. Specific growth rate was significantly (P < 0.05) elevated for trout fed the feed extruded at 137 °C. The apparent digestibilities of protein and energy, feed conversion ratio, retentions of nitrogen and energy were not significantly affected by extrusion temperature. Digestibility of ash was significantly reduced in the diets produced at 137 °C.     

Effect of dietary DP/DE ratio on apparent digestibility, growth and nitrogen and phosphorus retention in rainbow trout, Oncorhynchus Mykiss (Walbaum)

The effect of DP/DE ratio in diets for rainbow trout, Oncorhynchus mykiss (Walbaum), was investigated. To evaluate growth and body composition, groups of trout were fed three experimental diets with a constant level of gross energy (25.4 ± 0.12 MJ kg^?1 dry matter (DM)) and different digestible protein/digestible energy (DP/DE) ratios (diet A, 16. 35; diet B, 17.21; dietC, 18.23 g Mr^?1). Fat, protein and energy digestibility coefficients were not affected by the DP/DE ratio of the diets. Growth and feed utilization improved markedly as dietary DP/DE ratio increased (P < .01). The efficiency of fat, protein and energy utilization tended to increase with increasing DP/DE ratio of the diets. Nitrogen discharge in effluent water per kg of weight gain was not affected by dietary treatments (mean values for: diet A, 29.9; diet B, 29.8; diet C, 29.1 g N kg^?1 weight gain) while phosphorus discharge in effluent water fell using diets with a higher DP/DE ratio (mean values for: diet A, 7.3; diet B, 6.7; diet C, 5.9 g P kg^?1 weight gain).     

Changes in smooth muscle contractility of rainbow trout (Oncorhynchus mykiss Walbaum) intestine during acclimation to altered temperature

The effects of altered water temperature in vivo on in vitro smooth muscle contractility of rainbow trout intestine were investigated. Temperature has a significant effect on receptor-mediated intestinal smooth muscle contractility in the rainbow trout. The efficacy of 5-HT, carbachol, and transmural stimulation increased with temperatures above 10°C, with an optimal increase at 15°C. There was also a modest increase in the potency of 5-HT and carbachol within 2 days of establishing trout at 20°C. By day 8, most of these changes had either stabilized or were returning to control values, suggesting that acclimation changes in membranes and enzyme activities were taking effect. However, the contractile responses to carbachol and transmural stimulation were still increasing at this time. This may imply that the muscarinic receptors are more resistant to membrane acclimation changes and may take longer to adapt. Because these experiments were controlled for handling stress and seasonal changes that affect contractility, we have been able to demonstrate some early changes in smooth muscle contractility that occur during acclimation to altered temperature.     

Utilization of (3S,3'S)-astaxanthin acyl esters in pigmentation of rainbow trout (Oncorhynchus mykiss)

The deposition of natural, optically active, astaxanthin fatty acid esters in rainbow trout ( Oncorhynchus mykiss ) was studied. Mono-esterified and di-esterified (3 S ,3' S ) astaxanthin were purified from the green microalga Haematococcus pluvialis and incorporated into extruded diets and compared with diets containing synthetic racemic astaxanthin (Carophyll Pink) and a total carotenoid extract from the alga. All sources of astaxanthin achieved >4 mg kg⁻¹ in the white muscle after 6 weeks feeding. No significant difference ( P  > 0.05) between the deposition of astaxanthin or total carotenoid for the different diets was observed. Other xanthophylls, namely lutein, zeaxanthin and idoxanthin were found in the white muscle of rainbow trout fed all diets and together accounted for 10–14% of total carotenoid. Astaxanthin was deposited in the white muscle in the stereochemical form administered in the diet, i.e. racemic astaxanthin for Carophyll Pink and ˜100% (3 S ,3' S )-astaxanthin for the algal sources. In contrast, epimerization of (3 S ,3' S ) astaxanthin from the alga was observed for the astaxanthin esters deposited in the skin of rainbow trout, with a ratio close to 1.0:2.0:1.0 (3 S ,3' S :3 R ,3' S :3 R ,3' R ).     

Effect of growth hormone on muscle protein synthesis in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar)

This paper reports on the effect of administration of mammalian growth hormone (GH) on muscle protein synthesis as measured in white muscle using the phenylalanine flooding technique. The effect of exogenous GH was compared with that of insulin and prolactin, and with endogenous GH.The rate of protein synthesis in white muscle of rainbow trout 6 h after the injection of bovine GH or bovine insulin was twice (2.6 and 2.9% d^–1) that of the control saline-injected fish (1.2% d^–1). A metabolic effect of GH, as observed with insulin, is suspected.The rates of change in body weight and body length and the fractional rate of protein synthesis in muscle of rainbow trout were enhanced by mammalian GH administration. The effect of GH on muscle RNA/protein ratios was not significant. An opposite effect of antibodies against salmon GH (Lebailet al. 1989) on growth rate and muscle protein synthesis rate was found in rainbow trout. It is suggested that the effects of exogenous and endogenous GH on capacity and efficiency of muscle protein synthesis were similar.The long-term effects of mammalian GH on presmolt Atlantic salmon was also tested. The same trends were found with ovine prolactin supplementation in Atlantic salmon but not as high as those observed with ovine GH.     

Effect of alternate distribution of astaxanthin on rainbow trout (Oncorhynchus mykiss) muscle pigmentation

The aim of the present study was to determine the effects of the use of astaxanthin alternate feeding on rainbow trout pigmentation in term of astaxanthin serum concentration, muscle colour and astaxanthin muscle retention. Four hundred and fifty rainbow trout were fed the same basal diet supplemented with two different astaxanthin levels, 100, and 200 mg astaxanthin kg^? 1 of diet, hereafter designated as AX100 and AX200, respectively. An additional astaxanthin-free (AX0) diet was used. The experimental treatments were as follows: (1) REF = AX100 diet at each meal each day, served as reference; (2) SD1 = AX100 diet at each meal the first day followed by AX0 diet at each meal every second day; (3) SD2 = AX100 diet and AX0 diet in alternate meals each day; (4) R2 = AX200 diet and AX0 diet in alternate meals each day; (5) R4 = AX200 diet at the first meal the first day followed by AX0 diet at the second meal the first day and at each meal every second day. Fish were fed the experimental feeding schedule for 42 days.At the end of the experiment there were no significant differences among fish fed the different feeding schedules in term of final mean weight, specific growth rate and feed efficiency ratio. SD2 fish group displayed the highest (P < 0.05) astaxanthin serum concentration and the R4 fish group the lowest one. REF and R2 fish groups showed similar astaxanthin serum concentrations. Muscle chroma showed the most pronounced effect. It increased significantly for all fish groups during the experiment. At the end of the experiment REF and R2 fish groups displayed higher values than SD1 and R4 fish groups. Muscle astaxanthin concentrations increased significantly during the experiment whatever the astaxanthin feeding schedule. At the end of the experiment, the highest muscle astaxanthin concentration was recorded for R2 fish group while the lowest was noted for R4 fish group. Except for SD1 and R4 fish groups, muscle astaxanthin retention decreased significantly during the experiment. At the end of the experiment, muscle astaxanthin retention coefficients for SD2 fish group were significantly higher than those for REF fish group. The results reported here provide further evidence of the potential applicability of alternate astaxanthin feeding on rainbow trout pigmentation. Extending the optimisation of the SD2 treatment will therefore be subject for future studies. Its application could result in cost saving in the fish farming industry.     

Qualitative modification of muscle metabolic organization with thermal acclimation of rainbow trout,Oncorhynchus mykiss

Rainbow trout (Oncorhynchus mykiss) were acclimated to 4 and 18°C and fed at rations levels which led to an equal change in mass at these temperatures during the six week acclimation. Thermal acclimation markedly modified the metabolic organization of red and white muscle. Cold-acclimated fish had activities of -hydroxyacyl CoA dehydrogenase in both red and white muscle that were twice those of warm-acclimated fish. By contrast, the activities of cytochrome oxidase were unchanged by thermal acclimation. Thus, the capacity for -oxidation of lipids is specifically enhanced in the muscle of cold-acclimated trout. In white muscle, citrate synthase and phosphofructokinase activities were also enhanced by cold acclimation (increases of 125% and 35% respectively), while cytochrome oxidase levels were unchanged. The non-parallel changes in the activities of mitochondrial enzymes strongly suggest that trout muscle mitochondria undergo qualitative reorganization during cold acclimation. The relative activities of mitochondrial enzymes suggest that mitochondria from red muscle have a threefold greater capacity to oxidize lipids than those from white muscle.     

Seasonal variation of muscle metabolic organization in rainbow trout (Oncorhynchus mykiss)

This study examined how muscle metabolic organization varied during an annual cycle in which rainbow trout (Oncorhynchus mykiss) were held in outdoor holding ponds in which they were exposed to natural changes in temperature (range 0.2 to 15.6°C) and photoperiod. We examined the activities of glycolytic and mitochondrial enzymes in red and white muscle to evaluate whether trout enhance their capacity for lipid and carbohydrate oxidation during cold-acclimization. When assayed at habitat temperature, the enzyme activities generally increased in spring to reach a maximum in summer followed by a decrease in the fall. This led to significantly higher activities at warm than cold periods for all enzymes measured in red muscle and all but one in white muscle. The activities at 10°C provided little evidence for compensatory adjustments of aerobic capacity. Particularly in red muscle, enzyme levels at 10°C were generally lower during cold than warm periods. The variation of enzyme activities throughout the cycle was not due to changes in protein concentration, as the same responses were observed when activities were expressed per g wet mass or per mg protein. Although the aerobic capacity did not increase with cold-acclimatization, the relative capacity for lipid oxidation was higher in winter than in summer trout. In contrast, the relative capacity for aerobic glycolysis was higher in summer than in winter trout. Thus, the metabolic capacities of trout muscle undergo seasonal reorganization.     

The effects of acute temperature change on smooth muscle contractility of rainbow trout (Oncorhynchus mykiss Walbaum) intestine

The effects of altered temperature in vivo on in vitro smooth muscle contractility of rainbow trout intestine were investigated. Initial analysis of the data revealed a seasonal variation in the maximal tension of intestinal smooth muscle attainable with 5-hydroxytryptamine (serotonin), carbachol, KCl, and transmural stimulation in vitro. Peaks occurred in spring and troughs in autumn. There was no seasonal cycling of the potency of the stimulants. All data regarding the efficacy of the stimulants were subsequently corrected for seasonal variation. The response of smooth muscle depends on the temperature of the water in which the fish are placed (2°C−20°C). There was a marked linear increase in efficacy and a slight increase in potency of the stimulants with increasing temperature. Changes in responsiveness of the intestinal smooth muscle occurred within 30 min of moving the fish between tanks. Smooth muscle reactivity returned to pretreatment values by 48h. Any changes in responsiveness with regards to time were unlikely to be as a consequence of water temperature, but may have been a result of handling stress.     

The effect of water-borne magnesium on the dietary magnesium requirement of the rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) (mean initial weight 0.84 g) were fed diets containing graded levels of magnesium (Mg) (78 to 725µg/g) while being exposed to one of several levels in the rearing water (1.4 to 1000 mg/l). Uptake of Mg from the water, in Mg-deficient fish, was linearly related to the water Mg concentrated. It appears that the fish's Mg requirement can be met from either or both the diet or water. Under the experimental conditions, a water-borne concentration of 46 mg/l was calculated to be sufficient to meet the Mg requirement of the fish fed a Mg-free diet.     

Influence of dietary garlic (Allium sativum) on the antioxidative status of rainbow trout (Oncorhynchus mykiss)

Oxidative damage by free radicals has been implicated in the pathogenesis of several diseases. In this study, the antioxidative effect of dietary garlic on rainbow trout was examined. Trout fingerlings were fed on diets containing 10, 20, 30, 40 and 50 g garlic powder kg^?1 diet. Serum lipid peroxides and activities of antioxidant enzymes were measured. Serum thiobarbituric acid reactive substances (TBARS) assay showed that garlic consumption that resulted in a significant decrease in lipid peroxidation. The lowest levels of TBARS were observed in fish fed diet supplemented with 30 g kg^?1 garlic. However, higher doses of garlic (40 and 50 g kg^?1 diet) caused no further reduction in serum TBARS. The results showed a significant increase in superoxide dismutase in all of the garlic‐treated groups compared with the control. Ingestion of 10, 20 and 30 g kg^?1 dietary garlic resulted in a significant reduction in the catalase activity compared with all but the 10 g kg^?1 group. There was no significant difference in glutathione peroxidase activity among the different groups. Serum alanine aminotransferase and aspartate aminotransferase levels increased significantly in trout‐fed diets containing 40 and 50 g kg^?1 garlic powder. These results suggest that dietary garlic may improve the antioxidant status of rainbow trout. However, undesirable effects of higher doses of garlic should be considered.     

Identification of a novel gill-specific calpain from rainbow trout (Oncorhynchus mykiss)

Calpains are calcium-dependent neutral proteases responsible for many cellular functions. The two forms of calpain ubiquitously expressed in mammalian tissues are known as μ-calpain and m-calpain. We report here the identification of a novel calpain that is similar to but distinct from the μ- and m-calpains in rainbow trout. The cDNA of the novel gene is 2623 bp in length with a single open reading frame. The predicted protein (676 amino acids) contains the conserved calpain characteristic domains that include: domain I (pro peptide), II (cysteine catalytic site), III (electrostatic switch), and IV (calmodulin-like) with five Ca²⁺-binding EF hands. Northern blot and RT-PCR analyses demonstrated that the novel calpain gene is predominantly expressed in rainbow trout gills. Comparison of the novel protein with the ubiquitously expressed calpains and several mammalian tissue-specific calpains revealed that the novel calpain is an orthologue of the mammalian digestive tract specific calpain (calpain 9).