Expression of four muscle proteins at different growth stages of Günther's walking catfish Clarias macrocephalus

The complete cDNA sequences of four contractile muscle genes of walking catfish Clarias macrocephalus were characterized by assembling partial EST sequences from a skeletal muscle cDNA library. The four genes were parvalbumin 4 (PV4) (670 bp), troponin C (TnC) (1065 bp), troponin I (TnI) (843 bp) and myosin light chain 3 (MLC3) (953 bp), leading to deduced amino acid sequences of 109, 160, 176 and 150 residues respectively. During the larval stage, TnC mRNA showed the highest levels of expression with a 1.4‐fold increase from day 1 to day 30 post hatch. At 90 days, the relative expression levels of PV4, TnC and MLC3 were the highest, with similar proportions in the skeletal muscle, corresponding to the highest relative growth rate of walking catfish. Expression of the three calcium‐binding proteins remained high in 6‐month‐old fish, with higher levels of PV4. The different proportions of muscle proteins expressed suggested the significance of their contributions to fish growth and appeared to be correlated with the functional properties of muscle cells, which were observed from changes in the swimming activity of the fish.     

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.     

Effect of diets containing laurel seed oil on growth and fatty acid composition of rainbow trout,Oncorhynchus mykiss

The aim of this study was to determine the effects of replacing fish oil (FO) with laurel seed oil (LSO), as an alternative plant lipid source in diets on the growth and fatty acid composition of rainbow trout (Oncorhynchus mykiss; 111.47 ± 0.2 g mean individual weight). At the end of the feeding trial, survival was 100% in all treatments. No significant differences were seen in growth between the dietary groups (P > 0.05). The protein, lipid and ash contents were not significantly different among the groups (P > 0.05); however, there was a significant difference in protein and ash content between the treatment groups and the initial, and between the 50LSO group and the initial group, respectively (P < 0.05). The viscerosomatic index (VSI) and hepatosomatic index (HSI) values were not affected by increasing LSO percentages in the diets. The n‐6 polyunsaturated fatty acid (PUFA) concentration increased with increasing LSO levels in the diets. In contrast, the n‐3 PUFA levels decreased with increasing LSO levels in the diets. The liver and muscle were used for the analysis of fatty acids. The highest level of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) concentrations was recorded in fillet of fish fed the FO diet and the lowest in those fed the 50LSO diet. However, EPA and DHA ratios in the liver of fish fed the 75LSO diet were higher than those in fillet of fish fed the FO and 50LSO diets. No significant differences were seen in fatty acid composition between the dietary groups (P > 0.05). Based on the results of growth performance and fatty acid composition of the experimental fish in this study, it can be concluded that the 75% concentration of laurel seed oil performed best among the diets tested in the experiment.     

Digestive efficiency,free amino acid pools and quality of growth performance in Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis> L.) affected by light regimes and vaccine types

This study comprised the results of three different seawater trials using unique combination of techniques to study protease digestive efficiency and growth performance quality to illustrate the effects of light regimes and vaccine types in Atlantic salmon (Salmo salar L.). Fish with higher growth had higher trypsin (T) and chymotrypsin (C) specific activities with higher T/C ratio or slope T/C ratio [calculated from the regression between trypsin (y) and chymotrypsin (x) specific activities] in the pyloric caeca. The T/C ratios indicated fish growth rates over a period of 1–2 months, while the slope T/C ratios indicated fish growth rates at sampling. Adaptation period for adjustment to the new environment of continuous light was 70 days, indicated by the differences in trypsin specific activities and the crossing of slope T/C ratio regressions following with the changes in growth rate directions between the control and the treated group. Vaccine types affected fish vertebral growth, and additional continuous light enhanced the impact of vaccines on fish growth during springtime, indicated by differences in slope T/C ratios. Continuous light stimulated fish growth during winter to spring, when the natural day length was short, without significantly changing white muscle and oocyte qualities in the fish of about 500 g, except for significantly increased white muscle RNA concentration. Continuous light also reduced fish growth rate later during summer, when the natural day length was long, by precedently decreasing the T/C ratio in late spring. Interestingly, plasma levels of free lysine related to tryptic digestion were correlated with trypsin specific activity levels. Continuous light caused higher levels of most free amino acids (FAA) involved in nitrogen metabolism, higher incorporation of essential FAA for protein synthesis, and higher protein turnover rate (free hydroxyproline levels) in both plasma and white muscle. However, continuous light did not affect higher protein content, intracellular buffering capacity and RNA levels in the white muscle of the fish of about 1 kg, probably due to limitation of FAA available for protein synthesis. It is therefore suggested that enhancing fish growth by continuous light stimulation should be accompanied by increasing availability or content of dietary protein (and probably minerals), which in turn would improve the quality of fish growth performance through increasing fillet protein concentration, strengthening vertebral growth, and delaying oocyte development.     

Cichlasoma dimerus responds to refeeding with a partial compensatory growth associated with an increment of the feed conversion efficiency and a rapid recovery of GH/IGFs axis

Many fish species display compensatory growth (CG), a phenomenon by which fasted fish grow faster during refeeding. However, most studies use a group‐housed fish approach that could be problematic in social fish when interaction between individuals is not considered or eliminated. Additionally, the growth hormone (GH)/insulin‐like growth factors’ (IGF‐1 and IGF‐2) axis is implicated in postnatal growth in vertebrates, but its relevance in CG is not fully understood. Thus, the aim of this work was to determine whether CG occurs in a social fish, Cichlasoma dimerus, using an individually held fish approach and secondly, to evaluate the GH/IGFs expression profile during refeeding by 3 days and 3 weeks. C. dimerus showed partial CG. The feed conversion efficiency (FCE) was higher in three‐day‐refed fish, which presented higher GH plasma and mRNA levels than controls but shown no differences in liver and muscle GH receptors (GHR1 and GHR2) and IGFs mRNA levels. Surprisingly, three‐week‐refed fish exhibited GHR1 and IGF‐2 increments, but a reduction in GHR2 expression in muscle. These results show a strong association between GH levels, growth rate and FCE during refeeding, and a long‐lasting effect of refeeding on muscular expression of GHRs and IGF‐2.     

Effects of supplemental dietary l‐carnitine and ractopamine on the performance of juvenile rainbow trout,Oncorhynchus mykiss

This study was carried out to investigate a possible protein‐sparing action of l ‐carnitine and ractopamine in rainbow trout, Oncorhynchus mykiss. An 8‐week feeding trial was carried out to evaluate the effects of supplementation of three levels of l ‐carnitine (0, 1 and 2 g kg^?1) and two levels of ractopamine (0 and 10 mg kg^?1) on growth performance, fillet fatty acid compositions and blood biochemical parameters in a 3 × 2 factorial experimental design. Ractopamine and 1 g kg^?1 carnitine improved the specific growth rate (1.03% and 1.05% day^?1), feed conversion ratio (FCR, 1.3 and 1.29), protein efficiency ratio (PER, 1.88 and 1.85) of fish and crude protein (73.5 and 73.8) content of fish fillet. l ‐carnitine and ractopamine increased the levels of albumin, total protein and globulin in the serum of fish. Apart from eicosapentaenoic acid and docosahexaenoic acid, other fatty acids of fish fillet were increased by ractopamine, while total saturated fatty acids were almost intact. However, the total n‐3 poly unsaturated fatty acids were reduced by l ‐carnitine supplementation (P<0.05). The present study showed that 1 g kg^?1l ‐carnitine and 10 mg kg^?1 ractopamine each can improve the performance of rainbow trout and their combination in diet could enhance the protein level and change the fatty acids profile in fillet muscle.     

饲料中n-3/n-6多不饱和脂肪酸水平对黄河鲤幼鱼生长性能及生长相关基因mRNA表达的影响

为了分析饲料n-3/n-6多不饱和脂肪酸(PUFAs)水平对黄河鲤幼鱼生长性能和生长相关基因的影响,实验以鱼油和混合植物油(花生油和紫苏籽油)为脂肪源配制5组等氮等能饲料。对照组(T1)以鱼油为唯一脂肪源,18:3 n-3/18:2 n-6为0.97,第二组(T2)以花生油为唯一脂肪源,18:3 n-3/18:2 n-6为0.02,其他3组实验饲料以花生油和紫苏籽油为脂肪源,且n-3/n-6比值分别为0.46(T3)、1.09(T4)和1.53(T5)。10周养殖实验结束后,分析各组鱼体的生长性能、血清中生长激素(GH)含量及不同部位肌肉中生长相关基因的表达水平。结果显示,与对照组相比,n-3/n-6对鱼体的脏体比(VSI)、饲料系数(FCR)、摄食率(FI)无显著影响,增重率(WGR)随n-3/n-6比值的增加先升后降,n-3/n-6比值等于1.09时,WGR达到峰值,为218.53%±24.32%。T2组血清GH含量显著高于其他处理组,随着n-3/n-6比值增大GH含量逐渐降低。背部肌肉中生长相关基因gh、ghr、igf-1和myod均表现为先升后降的趋势,igf-1r为先降后升,mstn无显著差异。红肌中gh、myod含量逐渐降低,ghr、mstn不存在显著差异,igf-1与背部肌肉相似,先升后降,igf-1r先降后升。腹部肌肉中gh、ghr和igf-1表达情况与背部肌肉一致,igf-1r和myod的表达不存在显著差异。T2组和T4组背部肌肉肌纤维数目存在显著差异,且T4组肌纤维较T2组粗。研究表明,n-3/n-6不仅在一定程度上提高鱼体的WGR,且不同比例的n-3/n-6水平会影响血清GH的含量,不同部位肌肉组织中GH/IGF轴生长相关基因的水平也受饲料中n-3/n-6比例的影响。     

Partial replacement of soybean meal with Methylobacterium extorquens single‐cell protein in feeds for rainbow trout (Oncorhynchus mykiss Walbaum)

A feeding trial was conducted with juvenile rainbow trout (15–16 g initial weight) to assess the effects of including single‐cell protein (SCP) produced from Methylobacterium extorquens in trout feeds. Three isonitrogenous and isoenergetic diets were produced: a control diet and two experimental diets containing 5% or 10% bacterial protein meal replacing soybean meal. Triplicate tanks, each containing 35 fish, were fed each diet to apparent satiation in a constant‐temperature (15°C), flow‐through tank system for 12 weeks. No statistically significant differences in final fish weight or other fish growth parameters were observed. Similarly, feed efficiency parameters showed no significant differences among groups. Nutrient retention indices (protein, fat, energy) were relatively high and similar among fish in each dietary treatment group, as were whole body proximate compositions. Fish survival was high, with a small but statistically significant increase for the 10% SCP diet. Overall, results demonstrate that SCP from M. extorquens is a safe and effective alternative protein for rainbow trout diets at the low inclusion levels tested. Slightly lower weight gain in fish fed the 10% SCP diet was largely due to lower feed intake, suggesting that adding palatability‐enhancing ingredients to feeds may allow higher levels of M. extorquens SCP to be used without compromising fish growth.     

Inclusion of macroalgae meal (Macrocystis pyrifera) as feed ingredient for rainbow trout (Oncorhynchus mykiss): effect on flesh fatty acid composition

The use of macroalgae as an additional component in animal feeding has been studied. However, information on how it could influence muscle composition of fish body is scarce. This study evaluates four diets with different macroalgae inclusion levels (0%, 1.5%, 3% and 6%) to test the effect on body fatty acid composition of rainbow trout. Tanks with a volume of 600 L were stocked with 60.6 ± 7.9 g fish at a density of 45 individuals tank^?1 and fed for 124 days. At the end of the experiment there were not significant differences (P<0.05) in muscle proximate composition among fish fed the different treatments. However, it was determined that inclusion of 3% and 6% of macroalgae meal resulted in a significant increase (P<0.05) of polyunsaturated fatty acids (PUFAs) in muscle. In summary, macroalgae meal in rainbow trout diets do not enhance the quantity of protein and lipid contents at muscle level but an addition of 3–6% might contribute to increase the level of PUFAs, specially EPA, DHA and LIN. Thus, use of macroalgae meal might help to increase lipid quality content in the final product due the beneficial effects of PUFAs for human health.     

Spawning induces a shift in energy metabolism from glucose to lipid in rainbow trout white muscle

Enzymatic changes that occur in the white somatic muscle of rainbow trout (Oncorhynchus mykiss) in response to spawning were investigated, and the evenness of their distribution across the ventral-dorsal plane of this muscle was assessed. Four enzymes that are involved in energy metabolism were measured (phosphofructokinase: glycolytic capacity, 3-hydroxyacyl-CoA dehydrogenase: -oxidation, citrate synthase: citric acid cycle, cytochrome oxidase: oxidative capacity). The enzyme activities were followed in different parts of the white muscle of non-spawning female rainbow trout from May, four months after their first spawning, until December, at second spawning. Samples were taken from white epaxial muscle along the lateral line, on the dorsum, and in between. Samples were also taken from red muscle of non-spawning fish. The isoforms of myosin heavy chains (MyHC) were electrophoretically identified on 6% SDS-PAGE gel to study possible changes in contractile properties of the muscle.Transformation from the non-spawning to spawning phase was associated with dramatic changes in the activity of the enzymes studied in white muscle: glycolytic capacity decreased to less than half, whereas oxidative metabolism increased about two- to four-fold in all areas. Significant quantitative differences in enzyme activities were found between the three epaxial muscle areas: in the non-spawning fish lateral line samples differed from those taken in the other two areas, whereas in spawning fish the dorsal sample difered from the other two. No difference in the expression of MyHC-isoforms was found between spawning and non-spawning fish. Co-expression of both slow and fast isoforms was found in single fibres isolated from red muscle.The results show that the energy metabolism in white muscle of domestic rainbow trout is altered during spawning; i.e., the metabolism becomes increasingly aerobic, with an increased capacity for fatty acid utilization, concomitant with phenotypic changes associated with sexual maturation. These changes are especially pronounced in ventral, superficially located fibres.     

Effects of iodized feed on stress modulation in steelhead trout, Oncorhynchus mykiss (Walbaum)

In the aquaculture industry, the physiological systems of fish can be chronically stressed by various biological, chemical and physical factors. Chronic stress leads to a decrease in the overall health and growth of the fish, making them more prone to diseases. Dietary iodine has been shown to reduce this stress response in chickens and increase disease resistance in dairy cattle, but the mechanisms by which iodine affects stress resistance and immunocompetency is not completely understood and has not been extensively studied in fish. This study investigated the effects of iodized feed as a nutritional supplement in relation to stress modulation in steelhead trout (Oncorhynchus mykiss). We investigated effects on primary, secondary and tertiary stress responses by measuring plasma levels of cortisol, glucose and thyroid hormones as well as haematocrit percentage and average growth rate in steelhead trout. Iodine‐supplemented fish, on average, had lower levels of plasma cortisol and glucose and lower packed cell volumes than fish fed with regular commercial feed (P<0.05). Iodine‐supplemented fish also showed higher levels of thyroid hormones and exhibited better growth over the period of the experiment (P<0.05). It was concluded that, used with other husbandry practices, use of iodine‐supplemented feed could lead to better host defence, growth and survival in fish raised in the aquaculture industry.     

Juvenile Atlantic salmon decrease white trunk muscle IGF‐1 expression and reduce muscle and plasma free sulphur amino acids when methionine availability is low while liver sulphur metabolites mostly is unaffected by treatment

We previously reported that juvenile Atlantic salmon with mean initial BW 11.5 g offed a methionine deficient diet had lower weight gain due to a reduced protein accretion, while lipid gain was unaffected. Muscle of the fish fed the methionine deficient diet was depleted for sulphur amino acids, while in liver, the concentration of these metabolites was maintained within narrow limits. We speculated whether this could be due to an increased muscle proteolysis to support a prioritized liver metabolism in fish fed the low methionine diets. In this study, we assessed whether genes associated with muscle proteolysis increased under methionine deficiency. The composition of the diets was similar to those used previously containing 1.6 or 2.1 g Met/16 g N. We confirmed that the fish fed the low methionine diet gained less protein compared to fish fed the DL‐methionine enriched diet (P = 0.014), but growth did not reduce significantly. Also the deficient fish maintained the concentrations of liver sulphur amino acids and reduced muscle free methionine. Several of the other free amino acids within muscle increased. Further, methylation capacity was maintained in liver but reduced in the muscle (P = 0.78 and 0.04, respectively). Gene expression of muscle IGF‐1 was lower (P = 0.008) and myosin light chain 2 tended (MLC2, P = 0.06) to be reduced in fish fed low methionine diet, concurrently the activity of cathepsins B+L increased (P = 0.047) in muscle of fish fed the low methionine diet. Gene expression of the muscle‐specific E3 ubiquitine ligases (Murf and MaFbx) was not affected by treatment. Thus, the lower protein gain observed in fish fed the low methionine diet may be caused by reduced protein synthesis in line with the reduced IGF‐1 gene expression in the white trunk muscle. Thus, to support metabolism, the dietary protein needs to be balanced in amino acids to support metabolism in all compartments of the body and secure maximal protein gain.     

Response of Rainbow trout (Oncorhynchus mykiss) to unrefined peanut oil diets: Effect on growth performance,fish health and fillet fatty acid composition

A 60‐day feeding study with rainbow trout, Oncorhynchus mykiss, was conducted to determine the effects of replacement of fish oil (FO) by unrefined peanut oil (PO) on growth performance, feed utilization, body composition, fatty acid composition and serum biochemical and haematological parameters. Rainbow trouts (51.60 ± 0.75 g) were fed five experimental diets formulated by replacing dietary FO with PO at levels of level 0 (PO₀), 1/4 (PO₂₅), 1/2 (PO₅₀), 3/4 (PO₇₅) and 4/4 (PO₁₀₀), respectively. As a result, the best growth performance was observed in fish fed with PO₀ and PO₅₀ diet. No significant differences were detected among the groups in terms of body compositions. Fatty acid profiles of the fish fillets reflected the fatty acid profiles of the feeds that the fishes were fed with. In this study, the haematological parameters detected that there were no significant differences compared to the control group, whereas the serum biochemical parameters generally worsened as the ratio of peanut oil in the ration exceeded half of fish oil. As a conclusion, the results of the study suggested that the unrefined peanut oil could be used as a replacer of fish oil in diets for rainbow trout.     

Fishmeal alternative from renewable CO2 for rainbow trout feed

The rapid global growth in fish farming and limited supply of fish meal (FM) has consequently reduced FM inclusion levels in compound feeds leading to a higher reliance on alternative protein sources. Sasya is a single cell protein (SCP) product that has a similar amino acid profile as FM. The aim of this study was to evaluate the effects of replacing FM with SCP on in vivo digestibility, growth, feed efficiency, whole‐body proximate/amino acid composition and gene expression levels of various hepatic enzymes in rainbow trout (Oncorhynchus mykiss). Three isonitrogenous (470 g/kg crude protein) and isolipidic (18 g/kg crude lipid) diets were formulated as follows: Diet 1: control (30% FM); Diet 2:24% FM + 6% SCP and Diet 3:18% FM + 12% SCP. Each diet was hand‐fed to triplicate tanks containing 30 rainbow trout fingerlings (4.99 ± 0.20 g) for 9 weeks. Apparent digestibility coefficients of SCP for dry matter, crude protein, lipid and energy were 60, 80, 93 and 74% respectively. Growth performance (final weight: 69–71 g), feed conversion ratios (0.91–0.94) as well as whole‐body protein and amino acid composition were unaffected by diets. However, Diet 3 significantly increased whole‐body crude fat and energy. Fish fed the SCP‐based diets had significantly higher expression for carnitine palmitoyltransferase‐1b (CPT1b), fatty acid delta 6 desaturase (FADS6) and fatty acid elongase 5 compared to the control. Overall, the quality of the SCP was similar as FM. Therefore, this product could enlarge the portfolio of alternative protein sources that can be used in fish diets and thus open a new market opportunity for use of a new feed resource in the feed industry.     

低鱼粉饲料中添加酶解豆粕对凡纳滨对虾生长性能和抗胁迫机能的影响

为研究在养殖过程中降低鱼粉用量的同时保持凡纳滨对虾良好的生长性能和抗逆能力,实验在含10%鱼粉的基础饲料中,分别添加酶解豆粕(PSM) 0%(A)、2.5%(B)、3.5%(C)、4.5%(D)、5.5%(E)制成5组等氮等能饲料,分别投喂初始体质量为(0.45±0.02) g的凡纳滨对虾幼虾8周,检测对虾生长性能及抗胁迫机能。结果显示,8周养殖实验结束后,各实验组对虾的终末体质量为14.65～15.38 g/尾,各组间对虾终末均重、成活率和饲料系数指标均无显著性差异;A组对虾肌肉粗蛋白质含量显著低于其他各实验组;C组、D组和E组对虾肌肉粗脂肪含量显著高于A组;各组间灰分和水分均无显著性差异;D组和E组对虾肝胰腺蛋白酶、淀粉酶、脂肪酶、血清溶菌酶和血清总超氧化物歧化酶活性(T-SOD)均显著高于A组;A组对虾血清丙二醛(MDA)含量显著高于D组和E组。人工急性感染高剂量副溶血性弧菌的胁迫实验中,A组对虾在弧菌感染48和60 h时的累积死亡率均显著高于D组对虾同期的累积死亡率;低剂量副溶血性弧菌人工急性感染后,在凡纳滨对虾鳃组织中检测Toll受体、免疫缺陷(IMD)和溶菌酶3种免疫相关基因的表达量,结果显示,对虾Toll受体、IMD和溶菌酶mRNA表达量最大峰值分别出现在添加酶解豆粕的C组、B组和D组,峰值出现时刻分别为感染后24、42和24 h。研究表明,含10%鱼粉的饲料中添加0%～5.5%酶解豆粕对凡纳滨对虾的生长性能改善效果不显著,酶解豆粕会显著提高凡纳滨对虾肌肉粗蛋白质含量和粗脂肪含量;显著降低对虾血清丙二醛含量;同时也会显著改变凡纳滨对虾对弧菌的抵抗力及其免疫相关基因的时空表达,酶解豆粕添加量达到4.5%时可使养殖的凡纳滨对虾获得最佳的抗弧菌能力。     

Cellular responses to temperature stress in steelhead trout (Onchorynchus mykiss) parr with different rearing histories

This study compared the heat-shock response and metabolic energy status in hatchery-raised and two groups of wild-caught steelhead trout parr collected from the Navarro River watershed, California. Wild parr were from coastal and inland sites with different thermal regimes. Fish were exposed in the laboratory to 25 ± 0.2°C for 2-h and the heat-shock response was assessed in muscle tissue via induction of heat-shock proteins (hsps) 63, 72, 78, and 89. Metabolic measurements included muscle phosphocreatine (PCr), ATP, ADP, and AMP, and hepatic glycogen. Inland and coastal fish overlapped considerably with regard to their hsp responses and energetic endpoints, but hatchery fish were distinct in the biochemical patterns they exhibited. Hsp expression levels after temperature shock were significantly lower in hatchery than in wild fish. Hatchery fish also had significantly lower hepatic glycogen and higher muscle ADP, ATP, and PCr concentrations than wild fish. Coastal and inland steelhead did not differ significantly with regard to peak hsp72 and hsp89 levels or to concentrations of energy metabolites. However, fish from the warm-water, inland site expressed significantly less hsp63, maintained higher basal levels of hsp72, and induced hsp89 more slowly than fish from the cold-water, coastal site. Discriminant function analysis revealed that hatchery fish can be distinguished from wild Navarro River fish with 84.9% certainty using the following function: f(x) = − 43.6 + 0.14(Gly) + 4.1(PCr) + 186.4(AMP) + 80.8(ADP) − 0.14(hsp63) + 0.005(hsp72). This study demonstrates that within a single species, rearing conditions or genetic variation can influence an organism’s disposition and cellular response to thermal stress. Extrapolation of results from laboratory studies on hatchery fish to wild fish may therefore not be possible, and caution must be used when interpreting hsp data obtained for wild fish with different thermal histories.     

The effects of dietary organic or inorganic selenium in rainbow trout (Oncorhynchus mykiss) under crowding conditions

In the present study, the effects of different sources of selenium (Se; sodium selenite or selenomethionine) supplementation on the growth and serum concentrations of oxidative stress markers [malondialdehyde (MDA), 8‐isoprostane, glutathione peroxidase (GSH‐Px) activity] and muscle Se, MDA and heat shock protein 70 (Hsp70) levels in rainbow trouts were evaluated. The fish (n = 360; 0 + years old) with initial average weight of 20 ± 0.8 g were randomly assigned to 12 treatment groups consisting of 3 replicates of 10 fish each in a 2 × 2 × 3 factorial arrangement of treatments (stocking densities, Se sources, Se levels). The fish were kept at low (25 kg m^?3) or high (100 kg m^?3) stocking densities and fed a basal (control) diet or the basal diet supplemented with either 0.15 or 0.30 mg of Se kg^?1 of diet from two different forms: sodium selenite or selenomethionine. High stocking density decreased weight gain, feed intake and feed conversion ratio (FCR) when basal diet was fed (P = 0.001). A linear increase in feed intake and weight gain and improvement in FCR were found in sodium selenite (P = 0.01)‐ or selenomethionine (P = 0.001)‐supplemented fish reared under crowding conditions. Serum and muscle Se levels and serum GSH‐Px activity increased (P = 0.001) linearly, whereas serum and muscle MDA concentrations and serum 8‐isoprostane decreased linearly as dietary sodium selenite (P = 0.01) or selenomethionine (P = 0.001) supplementation increased. Selenomethionine and sodium selenite supplementation decreased Hsp70 in the muscle of fish reared under crowding conditions (P < 0.05). Supplementation with Se improved growth and antioxidant status of fish and the effects of selenomethionine were relatively greater than sodium selenite in the crowded groups. Results suggest that crowding conditions cause significant detrimental effects in rainbow trout indicated by increased oxidative stress, reduced feed intake and body weight gain. ?t also indicates that dietary Se supplementation offers a feasible way of reducing the losses in performance of rainbow trout reared under crowding conditions. Selenomethionine seems to be more effective than sodium selenite and the higer dose in the present study also seems to be more effective than the lower dose.     

Size-fractionated fish hydrolysate as feed ingredient for rainbow trout (Oncorhynchus mykiss) fed high plant protein diets. I: Growth, growth regulation and feed utilization

Six experimental diets were fed to rainbow trout to examine the effect of fish hydrolysate and ultra filtered fish hydrolysate on growth performance, feed utilization and growth regulation using diets low in dietary fish meal inclusion. One diet contained a high level of plant protein sources (90.6% of total dietary protein) and a low level of fish meal (9.4% of dietary protein). Two diets contained different levels of hydrolysate in exchange for the plant protein sources, reducing the plant protein level to 73.9% and 57.2%, respectively. Two further diets were identical in composition except that the hydrolysate was ultra filtered to remove low molecular weight compounds. A moderate level of fish meal was used in the sixth diet which had a dietary plant protein level of 57.0%. All diets were made equal in protein, lipid, energy and lysine. The feeding trial lasted for 90 days and for the fastest growing group, fed moderate level of fish meal, the fish increased in weight from 149 g at start to a final weight of 443 g. All groups showed significant differences in growth and feed utilizations. Specific growth rates were; 0.30% day^− 1 for the plant protein diet, 0.98% day^− 1 for the high hydrolysate diet, 0.72% day^− 1 for the group containing the high level of ultra filtered hydrolysate, and 1.21% day^− 1 for the moderate fish meal diet. Feed efficiencies (g fish weight gain per g feed intake) were found to be 0.57 for plant protein diet, 0.97 for high level of hydrolysate, 0.83 for ultra filtered hydrolysate and 1.03 for the moderate fish meal diet. Half dietary inclusions of hydrolysate and ultra filtered hydrolysate revealed values between the plant protein diets and high levels of these ingredients, respectively. Feed consumption in percentage of average fish weight per day, correlated with the feed efficiency for all groups. PER, PPV and BV correlated with the differences in growth. Protein digestibilities were equal for all groups, while the moderate fish meal diet showed higher lipid and energy digestibilities than the plant protein diets. Although some of the differences may be due to growth inhibitors in plant resources other explanations may be relevant. Plasma growth hormone (GH) levels were significantly higher in fish fed the plant protein diet than fish fed the fish meal or high hydrolysate diet, which is most likely a result of their poor feeding status. Plasma IGF-I levels were not affected by diet. Comparisons of groups with similar inclusion of plant ingredients, and thus equal level of growth inhibitors, show that in removing small molecular weight compounds from fish hydrolysate, the growth and feed efficiency were significantly reduced. Some of these small compounds in fish hydrolysate thus seem to be essential for biological performance. Further, as fish meal revealed the best performance, fish muscle protein is not the only nutrient that makes fish meal an essential ingredient in aqua feed for carnivorous fish. This information is important in the work to find replacement of fish meal in a sustainable growing global aquaculture industry.     

Effects of Acute Handling Stress on Expression of Growth‐Related Genes in Pampus argenteus

In fish, growth and development are mainly regulated by growth hormone/insulin‐like growth factors. Common aquaculture practices subject fish to stress. To investigate the effects of acute stress on growth‐related genes in cultured fish, the expression of growth hormone receptors (ghr1 and ghr2), insulin‐like growth factor binding proteins (igfbp1, igfbp4, and igfbp5), preprosomatostatin I and II (ppss1and ppss2), somatostatin receptors (sstr2 and sstr5), myostatin (mstn1 and mstn2), and glucocorticoid receptors (gr1 and gr2) were examined in Pampus argenteus subjected to handling stress. Plasma growth hormone levels increased significantly and peaked at 12 h and then decreased significantly at 24 h after treatment (P<0.05). Plasma cortisol and glucose concentrations in stressed fish began to increase significantly at 2 and 6 h after treatment, respectively. Real‐time quantitative polymerase chain reaction analysis showed that hepatic ghr2 mRNA levels in liver and muscle decreased sharply in response to the stressor. Igfbp1, 4, and 5 mRNA expressions in muscle also decreased sharply after exposure, while expression of ppss1, sstr2, and mstn2 increased significantly. This study showed that acute handling stress can affect expression of growth‐related genes in P. argenteus. Our findings could be helpful for the further study on response to stress in this species.     

Cellular activity and development of enteric,hepatic and muscle tissues of juvenile pacu Piaractus mesopotamicus (Holmberg 1887) fed with lyophilized bovine colostrum

Bovine colostrum, an alternative food of animal origin provided as partial protein source, may constitute an innovative ingredient in fish feeding. Serum insulin‐like growth factor 1 (IGF‐1) and cellular activity of enteric, hepatic and muscle tissues of juvenile pacu Piaractus mesopotamicus fed with lyophilized bovine colostrum (LBC) were evaluated. Pacu (8.46 ± 0.74 g; 7.75 ± 0.27 cm, n = 18 in triplicate) were stocked into 18 tanks (300 L; 18 fish per tank, in triplicate) and fed with control diet containing 0% of LBC or experimental diet containing 10% and 20% of LBC inclusion for either 30 or 60 days of experiment. IGF‐1 serum concentration was not affected (P > 0.05) by the diet containing LBC and experimental period. The intestinal and hepatic cellular activity was not influenced by LBC (P > 0.05); however, intestine protein/RNA decreased at 60 days (P < 0.05). The muscle cellular activity was affected by LBC, highest in fish fed with diet containing 10% of LBC (P < 0.05). Between the periods DNA and protein/RNA increased at 60 days, while RNA, protein/DNA and RNA/DNA ratio decreased in this same period (P < 0.05). Increased DNA muscle content indicates fish growth by hyperplasia. The results of the present study is related with suitable development of enteric, hepatic and muscle tissues and proper condition of juvenile pacu fed with LBC considering the absence of differences in relation to juveniles that consumed diet without colostrum.