Effects of constant light on haematological parameters of cultured rainbow trout (Oncorhynchus mykiss) in the Southern Hemisphere

The effects of artificial photoperiod regimes on reproductive patterns have been studied in several species, as have haematological parameters. However, information on how artificial photoperiods may affect blood components is scarce, especially under field conditions. We have assessed the effects of constant light [long day (LD) photoperiod: 24 h (light):0 h (dark)] on haematological parameters of cultured rainbow trout in Chile (Southern Hemisphere). In the initial stage (March up to June), two groups of fish were maintained under similar conditions and under the ambient (natural) photoperiod (NP). One group was then placed under the LD photoperiod regime for 2 months (June/July), following which it was returned to␣the NP (August–January); the control group remained under the NP throughout the experiment (March–January). All fish were assessed for haematological parameters and growth performance at various times during the experiment. During the initial stage, no differences were found between groups. However, at the end of the LD 24:0 period, the LD fish had increased haematocrit values and erythrocyte numbers and diminished mean corpuscular haemoglobin. After the LD fish had been returned to the NP, they developed secondary sexual characteristics and had a 40–44% decrease in the number of leukocytes.     

Some changes in the haematological parameters of rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) exposed to three artificial photoperiod regimes

Haematological parameters are often used as health status and stress indicators in fish. However, information on the effects of artificial photoperiods on these parameters is scarce and ambiguous. The consequences of three different artificial photoperiod regimes [light/day 12 h:12 h (LD 12:12) for 150 days as controls; LD 14:10 and LD 24:0, both for an initial 60 days, and then LD 12:12 for the remaining 90 days] were evaluated in trout. In all fish the haemogram and the erythrocyte production index (EPI) were assessed at days 7, 14, 30 and 60 (phase 1) and at days 90, 120 and 150 (phase 2). At the end of phase 1, photoperiods LD 14:10 and 24:0 induced higher production of immature (late basophilic) erythrocytes (0.06–0.08 × 10¹² cells/l, P < 0.05) and elevated EPI (young erythrocytes = 1.6 in LD 14:10, 2.25 in LD 24:0; late basophilic erythrocytes = 4.1–4.9 in LD 14:10, 3.3 in LD 24:0) than in controls (late basophilic erythrocytes = 0.02–0.04 × 10¹² cells/l; EPI young erythrocytes = 0.2–1.0; EPI late basophilic erythrocytes = 1.4–2.7). During phase 2, only reduced numbers of lymphocytes and thrombocytes were observed at day 150 in the LD 14:00 group. An increase in the EPI (1.9) of control young erythrocytes was found at day 150.     

Effects of skeleton photoperiods on melatonin secretion in the rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract Plasma melatonin levels were measured at hourly intervals in rainbow trout, Oncorhynchus mykiss (Walbaum), maitained under three different skeleton photoperiods: 8L:2D:2L:12D;8L:7D:2L:7D; and 8L:12D:2L:2D. Blood samples were taken before, during and after the 2-h light pulses. Melatonin levels increased rapidly to mean scotophase (dark period) values of 261 ± 7 pg/ml after the first light-dark transition and had returned to mean photophase (light period) values of 51 ± 2 pg/ml 30min after the end of the scotophase. Light pulses during the early, mid- and late scotophase elicited similar reductions in melatonin levels, followed by rapid increases after the light pulse, reaching previous scotophase values within 90 min. The inability of any of the light pulses to modulate or truncate melatonin secretion under a LD 8:16 photoperiod provides further support for the hypothesis that melatonin secretion in the rainbow trout is a direct response to darkness, and is not under endogenous circadian control as in other vertebrates examined.     

Effects of canola meal on physiological and biochemical parameters in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (initial body weight 4.16 ± 0.25 g) were fed diets [crude protein 420 g kg^?1; gross energy 18.7 MJ kg^?1 dry matter (DM); crude fat 110 g kg^?1] containing graded levels of either a canola meal (crude protein 350 g kg^?1 DM) supplemented with DL‐methionine as partial fish meal protein. A growth trial was conducted over 16 weeks at a water temperature of 12 ± 1 °C. At the end of the growth trial, in addition to body composition analyses, plasma tri‐iodothyronine (T₃) and thyroxine (T₄), cholesterol and liver fatty acid composition were measured. Replacement of fish meal with canola meal (100–570 g kg^?1 replacement) did not affect on growth performance. At 16th week, plasma cholesterol levels were reduced in fish fed all diets in comparison with 8th week. Plasma T₄ levels were significantly higher in the canola meal‐fed fish sampled after 16 weeks, but no significant differences in T₃ levels were obtained (P > 0.05). Proximate compositions were affected by dietary treatments. The liver fatty acid composition reflected that of the diet with a higher level of polyunsaturated (n‐6) fatty acids in fish fed diet canola meal and a higher content in n‐3/n‐6 ratio in fish fed diet without canola meal. These studies show that canola meal has potential to replace substantial levels of fish meal in diets for carnivorous fish without compromising performance.     

Edwardsiellosis in farmed rainbow trout (Oncorhynchus mykiss)

Edwardsiella tarda was isolated from naturally infected rainbow trout (Oncorhynchus mykiss) in raceway culture on a commercial fish farm where the fish were kept at a density of 110 kg m^?3 and at a water temperature of 14°C and with a photoperiod of 13 h light:11 h dark. The clinical signs of diseased fish (150 ± 20 mm standard length) were anorexia and lethargy. The most striking lesions in the fish were in the liver. There were hyperaemia and haemorrhages; on histopathological examination, the liver displayed inflammatory infiltrate in portal area, focal necrosis, dilatation of blood sinus and activation of sinusoidal cells. Infection experiments, performed 2 years after isolation of the original culture of E. tarda, were carried out under laboratory conditions at water temperatures of 15, 18 or 24°C. All experimental fish (common carp, Prussian carp, tench), intraperitoneally injected with 8 × 10⁶ cells, demonstrated a total resistance to E. tarda.     

Effect of lecithin on the haematological and condition indices of the rainbow trout Oncorhynchus mykiss (Walbaum)

Rainbow trout Oncorhynchus mykiss (Walbaum) were studied for 98 days for their response to the administration of a 1% rapeseed lecithin supplement in dry pellets. Lecithin tended to stimulate erythropoiesis, which manifested itself as a higher erythrocyte count, a higher haematocrit level and higher haemoglobin concentration during the course of the trial, compared with the control group. The significant differences that were found in the haematological profile, in the parameters of nitrogen metabolism (total protein, blood nitrogen urea, uric acid, creatinine) and mineral metabolism (inorganic phosphate, total calcium) and in the activity of catalytic enzymes (alanine aminotransferase), in certain periods of the trial do not suggest any adverse effects on the state of health of the fish; the levels of haematological parameters remained within the range of reference values. The catalytic activity of the aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase and α‐hydroxybutyryl dehydrogenase as well as the hepatosomatic index and condition factors calculated after Fulton and after Clark were unchanged. Lecithin supplement to the diet caused no clinical changes in the state of health of the fish, nor were any changes ascribable to the administration of lecithin observed on the basis of the patho‐anatomic and histological examination. Considering this and the good results in terms of the supporting antioxidant properties and the effect on growth capacity, recorded in previous experiments, rapeseed lecithin at the tested concentration level can be recommended as a supplement to dry pellets for rainbow trout.     

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.     

不同倍性雌性虹鳟(Oncorhynchus mykiss)性体指数与肝体指数的关系

文章研究了二倍体和三倍体雌性虹鳟（Oncorhynchus mykiss）在13月龄、17月龄、21月龄和35月龄时的性体指数（GSI）和肝体指数（HSI）及其相关性。二倍体雌性虹鳟的GSI在13月龄时最小（0．083％）,此后逐渐上升,至35月龄达到高峰（8．918％）,变化幅度极大;HSI在13-31月龄呈上升趋势,至31月龄时达最大值（1．444％）,然后下降。三倍体雌性虹鳟的GSI变化幅度很小,17月龄时达到最大值（0．103％）,35月龄时最小（0．032％）;HSI在17月龄达到高峰（1．179％）,而后下降,31月龄最低（1．067％）。试验结果表明,二倍体雌性虹鳟GSI与HSI的变化密切相关,可间接反映鱼类的生殖和生长状况;三倍体雌性虹鳟GSI与HSI无显著相关性,说明其肝脏与卵巢之间没有大规模的营养物质与能量的转移,有利于商品鱼的生产。     

Metabolism of exogenous histamine in rainbow trout (Oncorhynchus mykiss)

Information on the metabolism of exogenous histamine in fish is of much concern regarding the effect of dietary histamine on fish. Histamine catabolic enzymes, diamine oxidase and histamine N-methyl transferase were measured in the tissues of rainbow trout. Diamine oxidase was detected in the stomach, pylorus caeca and intestine. Histamine N-methyl transferase was detected only in the liver.A change in the contents of histamine and its metabolites was observed in the tissues of rainbow trout after oral administration of histamine. A large amount of imidazole acetic acid was observed in the serum, kidney, liver and muscle. On the other hand, 1-methyl histamine was detected only in the liver. Histamine and its metabolites, imidazole acetic acid and 1-methyl histamine were metabolized and diminished within 48 hr in all tissues.These results showed that histamine was metabolized by two metabolic routes in rainbow trout. One is the main pathway producing imidazole acetic acid by intestinal diamine oxidase and the other is the complementary pathway producing 1-methyl histamine by liver N-methyl-transferase.     

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.     

Effects of steroid hormones on immunoglobulin M (IgM) in rainbow trout, Oncorhynchus mykiss

The immunosuppressive effects of steroid hormones were evaluated as the response against implanted steroid hormones, cortisol (F), testosterone (T), estradiol-17 (E2), and 11- ketotestosterone (11-KT), in juvenile rainbow trout. In long term experiments (5 weeks), fish were given a single intraperitoneal implant of F or T. A clear suppressive effect of plasma IgM levels with F and T was not necessarily obtained, although mucus IgM levels were reduced corresponding to the elevated plasma steroid hormone levels. In short term experiments (1 week), intraperitoneal implantation of T, 11-KT and E2 suppressed plasma and mucus IgM levels, although the effects were not dose-dependent. When administered through diet, F and T caused a suppression of plasma IgM levels; F administration at both high and low dosages caused a significant decrease in plasma IgM levels, while only a high dose of T caused the suppression. These results suggest that sex steroid hormones, as well as F, have immunosuppressive functions in rainbow trout.     

Effects of dietary ascorbic acid levels on cholesterol metabolism in rainbow trout (Oncorhynchus mykiss)

An 80‐day feeding trial was conducted to evaluate the effects of dietary ascorbic acid (AA) supplementation at different levels (0, 0.2 and 0.4 g/kg l ‐ascorbate‐2‐polyphosphate; 7.6, 77.2 and 146.7 mg/kg AA, respectively) on cholesterol metabolism in rainbow trout (Oncorhynchus mykiss). Dietary AA supplementation regardless of inclusion level increased the serum total cholesterol and low‐density lipoprotein (LDL) cholesterol levels and LDL cholesterol/high‐density lipoprotein cholesterol ratio. No significant differences were observed in the serum triiodothyronine and thyroxine levels, faecal cholesterol content, hepatic activity and mRNA expression of acyl‐coenzyme A:cholesterol acyltransferase among the dietary treatments. Dietary AA inclusion increased the faecal bile acid content, hepatic activity and mRNA expression of 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase and hepatic cholesterol 7α‐hydroxylase mRNA expression, but decreased the hepatic LDL receptor content. High level of AA supplementation (0.4 g/kg) depressed the serum cortisol levels. These results suggest that dietary supplementation with 0.2?0.4 g/kg l ‐ascorbate‐2‐polyphosphate may increase the serum total cholesterol level in rainbow trout. The cholesterol‐raising effect of AA may be due to the increased hepatic cholesterol production and the depressed cholesterol clearance from serum. In addition, dietary AA inclusion also facilitates the hepatic conversion of cholesterol to bile acids.     

Effects of pH on contractility of rainbow trout (Oncorhynchus mykiss) intestinal muscle in vitro

Extreme acid-base imbalances in salmonids have been shown to impact on gastrointestinal motility. Changes in pH are known to affect cardiac and skeletal muscle in fish and mammalian intestinal smooth muscle preparations. This study was conducted to determine if the contractility of rainbow trout intestinal muscle was sensitive to pH fluctuations and to begin the investigation into the mechanisms by which pH affected contractility. Isolated duplicate or triplicate segments of rainbow trout intestine were suspended in organ baths containing modified Krebs-Henseleit solution. The pH was adjusted by varying the concentration of CO₂ aerating the solution and with HCl or lactic acid. Contractility was determined as response to the administration of 5-hydroxytryptamine (5-HT, serotonin), KCl, and transmural stimulation within the pH range of 6.3 to 8.5. Optimum pH for proximal segments was 7.85 while the range was much wider for distal segments. Decreasing the pH with CO₂ resulted in a greater inhibition of smooth muscle contractility than when the pH was decreased in 100% O₂ with either HCl or lactic acid, particularly when the tissues are stimulated electrically. This effect was ameliorated as the fish attained sexual maturity, although the mechanisms involved were not clear. The Cl^-/HCO ₃ ^- exchanger appeared to be involved in recovery from acidotic stress, particularly at pH levels below 6.5. The role played by the Na⁺/H⁺ exchanger is still unclear. Although contractility was inhibited in the presence of the amiloride analog EIPA (5-(N-ethyl-N-isopropyl)amiloride), whether the inhibition was due to blockage of ion transport or to cytosolic effects is yet to be determined.     

Embryonic expression of UCP2 in rainbow trout (Oncorhynchus mykiss)

Uncoupling proteins are mitochondrial anions transporters that dissociate respiration from ATP synthesis through proton leaks. Uncoupling protein 2 reportedly plays a role in several physiological processes such as energy partitioning, nutrition, and fatty acid metabolism. The mRNA expression of rainbow trout UCP2 genes (UCP2A and UCP2B) was monitored during embryogenesis and early larval development. Both genes were recruited early and displayed similar steadily decreasing patterns from fertilization until hatching. The expression of UCP2A and UCP2B appeared significantly differentiated after hatching and during the yolk sac absorption, with UCP2A displaying higher expression. We suggest that UCP2 expression profiles in the rainbow trout embryo could be associated with the utilization of lipids as a source of energy during development.     

Reproductive performance of female rainbow trout Oncorhynchus mykiss (Walbaum) kept under water temperatures and photoperiods of 13° and 51°N latitude

At Doi Inthanon Fisheries Research Unit (DIFRU), Thailand (13°N), rainbow trout were exposed to natural (13°N) and artificial (51°N) photoperiods, and natural water (NW) temperatures and cooled water (CW) 8 months before first spawning. In group I (51°N, CW), water temperatures of 18°C were never reached. In group II (51°N, NW) and group III (13°N, NW), the mean water temperatures in May exceeded 20°C, and 19°C in June and July. Eggs from 94% of all females in group I were obtained before January. This percentage diminished to 84% and 68% in groups II and III. The weight of the spawners and the size of the eggs were significantly lower in group III than in the other groups. No significant differences were observed for egg number per kg body weight of spawners between the groups. The mean fertilization rate of eggs was the highest, with 71%, in group I, and the lowest, with 50%, in group II. For hatching rates, on average 27%, 24% and 30% in groups I, II and III, respectively, differences were not significant. In group III, 37% of all batches reached fertilization rates above 80% and 16% of egg batches showed hatching rates of more than 60%.     

Isolation and partial characterization of rainbow trout (Oncorhynchus mykiss) gill mucin

Gill mucin from rainbow trout was isolated utilizing two rounds of cesium chloride density ultracentrifugation followed by gel filtration on Sepharose CL-2B. Neither density ultracentrifugation nor gel filtration alone was sufficient for purification of the mucin. Isolated gill mucin had a density of 1.5 g/ml and eluted at the void volume of the Sepharose CL-2B column. Silver-stained reducing 6% polyacrylamide gel electrophoresis of gill mucin produced a band at the origin with a smear entering the separating gel. There was no evidence of a link protein in gill mucin on reducing 12% polyacrylamide gel electrophoresis. Gill mucin had an amino acid profile similar to that of mucins in other species. Specifically, 35.1% of the total amino acids were represented by threonine and serine, while another 27.5% were alanine and proline. Gill mucin contained galactose (26.7 ± 3.2%), galactosamine (22.5 ± 4.4%), glucose (16.6 ± 8.7%), fucose (16.1 ± 1.5%), glucosamine (12.0 ± 1.9%) and mannose (5.1 ± 4.4%). Uronic acid levels from purified mucin were very low (0.7 ± 0.1%). Sialic acid was also present (0.06 g/g of mucin protein). The periodic acid-Schiff assay routinely utilized for detection of mucins was relatively insensitive for detection of gill mucin (6 × less sensitive than for pig gastric mucin) so a rabbit antiserum was raised. The antiserum produced profiles similar to the periodic acid-Schiff assay of fractions following gel filtration. Immunofluorescence of formalin-fixed rainbow trout gill tissue sections showed that the antiserum detected mucin within branchial goblet cells.     

Effects of dietary nucleotides supplementation on rainbow trout (Oncorhynchus mykiss) performance and acute stress response

This experiment was conducted to examine the effect of dietary nucleotides (NT) on fish performance and acute stress response on fingerling rainbow trout (23 g ± 0.01, mean weight ± SEM). Five experimental diets according to different levels of supplemented nucleotides (0, 0.05, 0.1, 0.15, and 0.2%) were assayed on experimental fish for 8 weeks. Growth, hematological parameters (hematocrit, hemoglobin, erythrocyte, lymphocyte, and neutrophil count), serum proteins (globulin, albumin), and plasma enzymatic activity (alkaline phosphatase, ALP; aspartate transaminase, AST; lactate dehydrogenase, LDH; alanine transaminase, ALT) were assayed. At the end of feeding trial, fish fed the control and 0.2% diets were subjected to handling and crowding stress. Modulatory effects of nucleotides on acute stress response (cortisol and glucose) and plasma electrolytes (Na⁺, Cl⁻, K⁺, and Ca²⁺) were studied. The percentage of body weight gain (WG) and feed efficiency (FE) of fish were better when the fish were fed 0.15–0.2% diets. Fish fed the nucleotide-supplemented diets tended to have lower levels of serum enzymes including ALP, AST, LDH, and ALT. Plasma cortisol levels of fish fed on 0.2% diet under handling and crowding stress were significantly lower than fish fed the control diet at all post-stress time intervals. In our study, fish fed nucleotide-supplemented diet had significantly lower concentrations of glucose compared to those fed the basal diet. The concentrations of sodium, chloride, calcium, and potassium of fish fed the control diet were significantly lower than in fish fed nucleotide-supplemented diet. Dietary nucleotides administration seems to promote growth and to enhance resistance against handling and crowding stress in fingerling rainbow trout.