The effects of elevated winter temperature and sub-lethal pollutants (low pH, elevated ammonia) on protein turnover in the gill and liver of rainbow trout (Oncorhynchus mykiss)

Appetite, growth, and protein turnover (synthesis, growth and degradation) of liver and gills were measured in juvenile rainbow trout (Oncorhynchus mykiss) fed to satiation, and exposed for 90 days to elevated winter temperatures (+2 °C above ambient) and either low pH (5.2) in softwater or 70 M total ammonia (TAmm) in hardwater. All fish increased in weight during the experiments, but those exposed to +2°C grew significantly more than those at ambient temperature due to a stimulation of appetite. During the relatively constant temperature of the first 75 days, +2 °C caused a significant increase in the rates of protein synthesis and degradation in the liver of hardwater-acclimated fish, as a result of an increase in RNA translational efficiency (KRNA). The elevated temperature also induced an increase in gill protein synthesis in softwater-acclimated fish but in this case the underlying mechanism was an increase in Cs, the capacity for protein synthesis (RNA:protein) rather than in KRNA. The addition of 70 M TAmm had no effect on protein turnover in either liver or gills of hardwater-acclimated fish. Low pH inhibited protein growth in the liver of softwater-acclimated fish at day 90 under both temperature regimes. This inhibition was effected via a decrease in protein synthesis at control temperature but via an increase in protein degradation when the fish were exposed to both low pH and +2 °C. From these results we conclude that a simulated global warming scenario has potentially beneficial rather than detrimental effects on protein turnover and growth of freshwater fish during winter.     

Effects of prolonged epinephrine infusion on blood respiratory and acid-base states in the rainbow trout: Alpha and beta effects

Rainbow trout were infused continuously for 12h with epinephrine in the presence or absence of alpha-and/or beta-adrenergic blockade to characterize the specific adrenergic mechanisms involved in the control of blood acid-base status and oxygen transport capacity. Infusion of epinephrine, alone, produced a transient respiratory acidosis, as indicated by an increase in carbon dioxide tension and a decrease in whole blood pH, yet arterial oxygen tension was elevated. Red blood cell pH increased by approximately 0.2 pH units during epinephrine infusion and this increase as well as the increase in oxygen tension were prevented by pretreatment with propranolol (a beta-adrenergic antagonist). Epinephrine infusion during alpha-adrenergic blockade caused a prolonged elevation of blood carbon dioxide tension and abolished the increases in hematocrit and hemoglobin concentrations observed during epinephrine infusion alone. Infusion of the alpha-adrenergic agonists phenylephrine (an alpha₁ agonist) or clonidine (an alpha₂ agonist) caused respiratory acidosis (decreased pH, increased CO₂ tension) and a reduction in oxygen tension. Infusion of isoprenaline (a non-specific beta agonist) caused delayed increases in carbon dioxide and oxygen tensions. We speculate that the increased carbon dioxide tension observed during epinephrine infusion is a result of beta-adrenoceptor mediated inhibition of red blood cell bicarbonate dehydration and not branchial convective or diffusive adjustments. The effects of epinephrine on blood O₂ tension, content and carrying capacity are discussed with reference to the participation of alpha- and beta-adrenergic mechanisms at the gill, spleen and red blood cell.     

Evidence of concealment behavior by adult rainbow trout and brook trout in winter

Abstract – There has been little investigation of the winter ecology of adult trout during winter, especially in regard to concealment behavior. We compared day vs night underwater counts of adult rainbow trout and brook trout from four streams. At water temperatures between 1°C and 9°C, daytime counts accounted for 44% and 16% of nighttime snorkeling counts for rainbow trout and brook trout adults, respectively. As winter progressed, nighttime counts declined more so for brook trout than rainbow trout, but the decline was not significant for either species. Nocturnalism of both species was higher in streams with colder water temperatures. We observed few fish within concealment structure; however, by electrofishing concealment habitat during the day, we captured 10 times more adult trout than we counted immediately beforehand by snorkeling. Adult trout were concealed in cobble-boulder substrate and woody debris during the day. ^Note     

A critical analysis of gas transferin vivo and in a blood-perfused trout head preparation

Gas transfer and blood acid-base status in the blood-perfused trout head preparation (in vitro) were compared with intact trout (in vivo) fitted with oral membranes, dorsal aortic, ventral aortic, and opercular cannulae. Gas transfer rates were calculated from both arterial-venous blood differences and inspired-expired water differences using the Fick method. Oxygen uptake and carbon dioxide excretion were lower, while ammonia excretion was higher, in the blood-perfused head relative toin vivo rates. Gas transfer rates calculated from water were consistently greater than rates calculated from blood, the difference being greaterin vitro compared toin vivo. We conclude that the inadequacy of O₂ and CO₂ transfer in the blood-perfused head was not due to abnormal gill diffusive conductance, but was more likely related to the reduced magnitude of the blood-to-water O₂ and CO₂ diffusion gradients, low hematocrit, and decreased perfusion flow rate . Under the conditions of the present study, the blood-perfused trout head is not a suitable preparation for the study of oxygen transfer. We conclude this preparation is useful for evaluating branchial carbon dioxide or ammonia transfer only when comparable measurements or manipulations cannot be made on intact fish.     

Competitive exclusion of brown trout Salmo trutta L., by rainbow trout Oncorhynchus mykiss Walbaum, in lake tributaries, New Zealand

Spawning runs of brown trout Salmo trutta L., and rainbow trout, Oncorhynchus mykiss Walbaum, were compared in three tributaries of two lakes for timing, numbers and redd characteristics. Redd characteristics for the two species in four other streams were examined. No significant differences in redd characteristics between the two species were observed, but in all lake tributaries the possibilities of superimposition of spring spawning rainbow trout on redds of brown trout were examined. A common feature was the high ratio of numbers of spawners to the area of preferred spawning gravel, and it was concluded that in the lake tributaries, superimposition resulted in increased mortality of brown trout eggs, and in one case elimination of the brown trout population. Management possibilities for control of the species proportions in comparable situations are discussed.     

Response of resident brown trout, Salmo trutta L., and rainbow trout, Oncorhynchus mykiss (Walbaum), to the stocking of hatchery-reared brown trout

Two strains of hatchery-reared adult brown trout, Salmo trutta L., [208–334 mm total length (TL); n =  591] were individually marked and released into a limestone stream. The estimated survival after one month (86%; n =  508) was comparable to that for resident brown trout and rainbow trout, Oncorhynchus mykiss (Walbaum), (89%; n =  771), but declined to 14% ( n =  83) after 8 months compared with 52% ( n =  451) for resident trout. The movement of resident trout out of stocked stretches was higher (14%) than from control sites (5%), but the population size in both individual sites and the overall study area were unaffected. The growth of resident brown trout was unaffected by stocking, but rainbow trout showed lower growth rates in stocked versus unstocked stretches both one and 8 months after stocking ( P <  0.002).     

Temporal segregation in spawning between native Yellowstone cutthroat trout and introduced rainbow trout

Hybridisation with introduced taxa poses a threat to native fish populations. Mechanisms of reproductive isolation can limit or prevent hybridisation between closely related species. Understanding how these mechanisms interact between the same species across geographically distinct occurrences of secondary contact, and how regional factors influence them, can inform our understanding of hybridisation as a threat and management actions to mitigate this threat. We used data collected on adult fish migration timing and approximate emergence timing of subsequent juvenile fish paired with genomic data to assess whether temporal isolation in the timing of spawning exists between Yellowstone cutthroat trout, rainbow trout and hybrids in the North Fork Shoshone River drainage in northwest Wyoming. We found evidence that Yellowstone cutthroat trout spawn, on average, two to four weeks later than rainbow trout and hybrids and two environmental covariates related to water temperature and discharge were associated with differences in spawning migration timing. Despite statistical support for Yellowstone cutthroat trout spawning later, disproportionately high numbers of rainbow trout and hybrids, paired with extended spawning seasons, lead to substantial overlap between all genotypes. Our results provide further evidence of temporal segregation in the timing of spawning as a mechanism of reproductive isolation between closely related species, but substantial spawning overlap suggests temporal segregation alone will not be enough to curtail hybridisation in conservation populations.     

不同硒源对虹鳟生长性能及抗氧化能力的影响

在以鱼粉为主要蛋白源的基础饲料中分别添加亚硒酸钠和硒酵母,使饲料中硒的含量达到1．5％、2．0％、2．5％、3．0％。基础饲料设为对照组,不添加外源硒,对照组的硒含量为1．0％。在水温10℃～15℃、溶解氧5．6mg／L的条件下,对初始体重150g的虹鳟进行为期50d的生长对比试验。实验结束后取鱼的肝脏及血清,分别测定其超氧化物岐化酶（SoD）、谷胱甘肽过氧化物酶（GSH—Px）活力及总抗氧化能力（T—AOC）、丙二醛（MDA）的值。结果表明：虹鳟鱼的增重率、饲料利用率各组之间差异不显著。各组之间的SOD、GSH—Px、T—AOC、MDA没有显著差异。因此在以鱼粉为主要蛋白源的虹鳟鱼饲料中添加外源硒,对虹鳟的生长及抗氧化能力没有显著影响。     

盐度和pH对缢蛏耗氧率及排氨率的影响

缢蛏(Sinonovaculaconstricta)分为壳长约4 0、5 0、6 0cm3组。通过实验生态学方法研究缢蛏在不同盐度和pH下的耗氧率和排氨率变化规律。结果表明,盐度对缢蛏的耗氧率和排氨率有极显著影响(F>F0.01),且缢蛏单位体重耗氧率、排氨率随其个体的增大而变小;当盐度为6～30时,缢蛏的耗氧率和排氨率呈1个峰值变化,在盐度22时均达到最大值。pH对缢蛏的耗氧率无显著影响(FF0.01),此时,缢蛏的排氨率出现1个峰值变化,当pH为8时达到最大值。     

Effect of protein source and nutrient density on growth efficiency, histology and plasma amino acid concentration of rainbow trout (Oncorhynchus mykiss Walbaum)

The effects of protein source and nutrient density on growth efficiency, nutrient digestibility and plasma amino acid concentrations of rainbow trout were evaluated. A 3 by 2 factorial treatment design with three protein sources, fish meal–barley (F–B), plant concentrates (PC) and plant meals (PM), and two nutrient densities were used. A commercial reference diet was also fed. Triplicate tanks of 30 fish (initial wt. 28 g) were fed each diet, and the final weight averaged 240 g fish⁻¹. Protein source and nutrient density affected feed intake, weight gain and feed conversion ratio. Weight gain of trout fed the PC and PM diets was approximately 10% less than fish fed the F–B diets. Protein retention was affected by protein source, but not nutrient density, and was the highest for the fish fed diets containing fish meal and the lowest for the fish fed PM diets. Apparent digestibility coefficients and apparent amino acid availabilities of the diets corresponded with differences in weight gain. This study provides further evidence that growth rates of trout fed fish meal‐free diets, using conventional and concentrated plant protein ingredients, are good but some limitation to growth exists in the fish meal‐free diets.     

Effects of sulfamerazine on selected haematological and immunological parameters in rainbow trout (Onchorhynchus mykiss, Walbaum, 1792)

In the present study, effects of sulfamerazine on some haematological and immunological parameters of rainbow trout ( Oncorhynchus mykiss ) were examined. Four groups of rainbow trout were fed experimental diets containing either no sulfamerazine (control) or supplemented with sulfamerazine at 100 mg kg⁻¹ (Exp-A), 200 mg kg⁻¹ (Exp-B) or 400 mg kg⁻¹ (Exp-C) for 21 days. Blood samples were taken for the haematological and immunological parameters from fish on the third, seventh, 14th and 21st days of feeding. Haematological parameters [haematocrit, leucocrit, numbers of erythrocytes (RBC) and leucocytes (WBC), haemoglobin (Hb), mean corpuscular volume (MCV), mean haemoglobin concentration (MHC) and mean corpuscular haemoglobin concentration (MCHC)] and immunological parameters [phagocytic ratio (PR) and index (PI), glass-adherent NBT-positive cell activation, total plasma protein and total immunoglobulin (Ig)] were evaluated during the experimental trial. It has been observed that MCV ( P <0.05), PR and PI ( P >0.05) were increased and haematocrit and leucocrit value, numbers of erythrocytes and leucocyte, haemoglobin, MHC and MCHC values and total plasma protein and total Ig levels were decreased in rainbow trout after application of three different doses of sulfamerazine.     

Effects of egg aging on the metabolites of ovarian fluid in rainbow trout,Oncorhynchus mykiss

Nuclear magnetic resonance ‐based metabolomics was applied to study effects of egg aging on ovarian fluid metabolites in rainbow trout, Oncorhynchus mykiss. The eggs of three females were pooled and then assigned to three plastic vials for 18 days in vitro storage at 4°C. Ovarian fluid samples were taken 0, 6, 12 and 18 days after storage. Three groups of metabolites including amino acids, osmolytes and energy metabolites were found to change during storage period. The glucose levels of ovarian fluid showed significant decreases on days 12 and 18 after storage (p < .05). For acetoacetate and acetate, significant increases were observed, respectively, on days 12 and 18 after storage (p < .05). The creatine levels of ovarian fluid increased significantly on days 12 and 18 after storage (p < .05). Lactate levels in ovarian fluid elevated during storage period (p < .05). Glycerol levels showed a significant increase on days 12 and 18 after storage (p < .05). The values of osmolytes in ovarian fluid (e.g., betaine, taurine, trimethylamine‐N‐oxide, N,N‐dimethylglycine) showed a decreasing trend on day 12 which continued until the end of storage on day 18 (p < .05). Almost all amino acids elevated on days 12 and 18 after storage (p < .05). After an apparent elevation in isoleucine levels on days 6 and 12, this amino acid decreased on day 18 after storage (p < .05). The osmolytes might act as antioxidant against free‐radicals produced as a result of over‐ripening. Glucose can be used as energy resource for eggs and bacteria during ova storage. Also, change pattern of amino acids indicate hydrolysis of proteins as the time of storage increases.     

Nutritional regulation of somatostatin expression in rainbow trout, Oncorhynchus mykiss

Previously we characterized three DNAs from the endocrine pancreas (Brockmann body) of rainbow trout (Oncorhynchus mykiss) that encode for distinct preprosomatostatin (PPSS) molecules: one containing somatostatin-14 and its C-terminus (PPSS I) and two containing [Tyr⁷, Gly¹⁰]-somatostatin-14 at their C-termini (PPSS II′ and PPSS II″). In the present study, the regulation of PPSS expression was studied in rainbow trout placed on varying nutritional regimes (fed continuously, fasted, fasted then refed). Fish that were fasted for one week displayed reduced growth compared to their fed counterparts, but no alteration in pancreatic PPSS expression was noted between the two groups. Fish fasted for 4 and 6 weeks also were growth retarded and displayed increased levels of PPSS I mRNA and PPSS II″ mRNA compared to fed animals; PPSS II′ mRNA levels were not affected by food deprivation. Refeeding fish for two weeks following 4 weeks of food deprivation restored growth and reduced PPSS I and PPSS II″ mRNA expression to levels similar to those displayed by continuously fed fish. Changes in PPSS expression were correlated with increases in plasma SS. These results indicate that nutritional state modulates differential expression of PPSSs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of unilateral ovariectomy on recruitment and growth of follicles in the rainbow trout,Oncorhynchus mykiss

Virgin female rainbow trout, Oncorhynchus mykiss, were unilaterally ovariectomised at various stages of ovarian development to investigate the effect of the removal of one ovary on subsequent recruitment and growth of follicles in the remaining ovary. The right ovary was removed from groups of 12–15 fish, 12, 7 and 4 months before they were due to ovulate, and the gonadosomatic index and follicle number and size determined just prior to ovulation. There were no differences in fecundity or follicle size in fish unilaterally ovariectomised at 12 and 7 months prior to ovulation compared to the controls. However, in the females unilaterally ovariectomised 4 months prior to ovulation, the remaining ovary either had the normal number of follicles for a single ovary but of a significantly larger size than follicles in the controls, or alternatively had almost 70% more than the normal number of vitellogenic follicles but comprising two distinctly different size populations. Differences in plasma oestradiol-17β concentrations at the final sample were seen only in females unilaterally ovariectomised 4 months prior to ovulation, where the levels were significantly lower than both the sham operated and control fish (p < 0.05). These data show that in the rainbow trout, complete compensatory ovarian hypertrophy following unilateral ovariectomy can occur throughout a major part of ovarian development, but that follicle recruitment is limited to stages up to (and therefore fecundity is determined by) mid-vitellogenesis.     

A study of gross,histological and blood biochemical changes in rainbow trout,Oncorhynchus mykiss (Walbaum), with rainbow trout gastroenteritis (RTGE)

The mechanisms behind the pathogenesis of rainbow trout gastroenteritis (RTGE) are still unknown. This study examined the macroscopic and microscopic changes in trout with RTGE (RTGE+), as well as the blood chemistry. A total of 464 rainbow trout were sampled from 11 sites in the UK, comprising 152 RTGE+ fish and 330 random, apparently healthy fish. A case definition for RTGE was assessed by the analysis of its agreement with three laboratory tests: histopathology, packed cell volume and kidney bacteriology. Cluster analysis indicated the presence of three distinct presentations within the population of RTGE+ fish. Cluster A included gross signs associated with moribund RTGE+ fish, and clusters B and C identified gross signs consistent with concurrent diseases, notably furunculosis, enteric redmouth and proliferative kidney disease. The information gained was used to select RTGE+ fish without concurrent disease for the analysis of RTGE pathogenesis with blood biochemistry. This analysis revealed a severe osmotic imbalance and a reduced albumin/globulin ratio as indicatives of selective loss of albumin. These findings are compatible with a protein losing enteropathy.     

Cholesterolaemia and triacylglycerolaemia in farmed rainbow trout,Oncorhynchus mykiss

Physiological (reference) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for blood plasma cholesterol (CHOL) and triacylglycerol (TGL) in farmed rainbow trout, Oncorhynchus mykiss, in raceway culture. The fish were given dry pelleted diets that contained 37–47% crude protein and 7–14% crude fat and were kept at a stocking density of 50 kg m^?3 in tanks provided with running freshwater at an ambient temperature of 3–16°C and dissolved oxygen, 8.4–11.5 mg L^?1. Blood was sampled between September and November at a photoperiod of 9:00–13:00 hours: 11:00–15:00 hours (light:dark). Cholesterol levels were significantly (P = 0.0001) greater in males (4.7–12.1 mmol L^?1, n = 34, mean weight 406 ± 138 g) than immature females (3.2–9.7 mmol L^?1, n = 386, mean weight 416 ± 147 g). Physiological range for TGL in immature females and males was 2.4–14.4 mmol L^?1 (n = 249, mean weight 418 ± 149 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for CHOL in males and in females, but not for TGL, in which a sinistral asymmetry was found. Correlation and regression analyses indicated significant (P = 0.0000) dependence between the CHOL and total protein (r² = 76.2%), CHOL and Fulton's condition factor (r² = 43.3%) and CHOL and absolute weight of liver (r² = 45.5%). Fluctuation in cholesterolaemia and triacylglycerolaemia, depending on nutrition and the aquaculture method is discussed below.     

Probiotic applications for rainbow trout (Oncorhynchus mykiss Walbaum) I. Effects on growth performance,feed utilization,intestinal microbiota and related health criteria

The effect of dietary probiotics (Bacillus subtilis, Bacillus licheniformis and Enterococcus faecium) used singularly and synergistically on the growth performance, intestinal microbiota and health status of rainbow trout (Oncorhynchus mykiss Walbaum) were assessed after 10 weeks feeding on supplemented diets. No significant improvements of weight gain or specific growth rate were observed in the probiotic fed groups. However, a significant improvement of feed conversion ratio was observed in the group fed E. faecium. High levels of probiotic species were observed in the posterior gastrointestinal tract as transient digesta‐associated populations and potentially resident mucosal populations. Bacillus subtilis and B. licheniformis levels accounted for 36% of the total culturable microbial population adhered to the mucosa and 62% in the digesta. E. faecium levels accounted for 45% of the mucosal population and 89% of the population in the digesta. An increase of serum lysozyme activity was observed in the fish fed diets containing the Bacillus probionts and elevated leukocyte levels were observed in fish fed diets containing Bacillus + E. faecium synergistically. The results of the current study demonstrate potential for B. subtilis, B. licheniformis and E. faecium to improve feed utilization, modulate intestinal microbiota and the health status of rainbow trout.     

Total calcium and inorganic phosphate in the blood plasma of farmed rainbow trout, Oncorhynchus mykiss

Reference (physiological) value intervals determined by the lower 2.5% and upper 97.5% quantiles were calculated for total calcium (Ca _t ) and inorganic phosphate (P) in farmed rainbow trout, Oncorhynchus mykiss , at an age of 10–12 months in raceway culture. Total calcium levels were significantly ( P ≤0.0009) greater in males (2.63–4 mmol L⁻¹, n =47, mean weight 359±142 g) than in immature females (2.48–3.97 mmol L⁻¹, n =410, mean weight 400±145 g). The reference range for P in immature females and males was 3.09–9.32 mmol L⁻¹ ( n =467, mean weight 401±144 g). The distribution and density of the quantiles in the tested reference group were made possible by the use of histograms, which showed normal distributions for Ca _t in males but not for Ca _t and P in immature females and males, in which a sinistral asymmetry was found.     

Insulin stimulates hepatic lipogenesis in rainbow trout,Oncorhynchus mykiss

The effects of the pancreatic hormones, insulin and glucagon, on rates of lipid biosynthesis in liver removed from rainbow trout, Oncorhynchus mykiss, were evaluated in vitro. Livers were removed from animals fasted for 30–36h, cut into ca. 1 mm³ pieces, and incubated in the presence of various concentrations of salmon insulin (sINS), bovine insulin (bINS), or a combination of BINS and bovine/porcine glucagon (GLU). Lipid synthesis was evaluated by total lipid concentration, ³H₂O incorporation into total lipid, and by fatty acid synthetase activity. Both mammalian and sINS tended to increase tissue total lipid concentration in hepatic tissue incubated for 5h. Insulin also stimulated ³H₂O incorporation into total lipid in a dose-dependent manner. Bovine INS (2 × 10^?6 M) stimulated de novo synthesis nearly 6-fold over control rates; sINS (2 × 10^?6 M) stimulated label incorporation more than 7-fold over control rates. Glucagon inhibited INS-stimulated ³H₂O incorporation; whereas, GLU alone had no effect on lipid synthesis in liver pieces incubated 5h. Lipid class analysis indicated that bINS significantly stimulated ³H₂O incorporation into phospholipids, fatty acids, and triacylglycerols. The greatest accumulation of label was in the triacylglycerol fraction, where incorporation was stimulated 17-fold over control levels. Hepatic enzymatic analysis indicated that bINS also significantly stimulated lipogenic enzyme activity 9-fold above control levels. These results indicate that INS is an important regulator of lipid synthesis in the liver of trout.