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.     

Influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum)

The influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied experimentally in the laboratory and at a fish farm. In experiment I, the effect of parasitic infection on columnaris disease was studied using F. columnare carrier fish. The fish were exposed to Diplostomum spathaceum cercariae and a set of other stressors in order to induce clinical columnaris infection. Parasitic infection and other stressors failed to induce the disease. Disease occurred when the fish were challenged with F. columnare, but D. spathaceum infection did not enhance the severity of the infection. In experiment II, the influence of rearing density and water temperature was studied. Overall mortality was highest in fish at normal rearing density with high temperature (+23 degrees C). At low temperature (+18 degrees C) mortality was not affected by rearing density, but the transmission of columnaris disease was faster at normal rearing density at both temperatures. This supports the view that reduction of fish density could be used in prevention of columnaris disease especially if water temperature is high. Because the lower rearing density can also decrease the transmission of ectoparasites and penetrating endoparasites, it could be an efficient tool in ecological disease management.     

Moderate stocking density does not influence the behavioural and physiological responses of rainbow trout (Oncorhynchus mykiss) in organic aquaculture

Welfare in farmed fish got particular attention during the last decades from both governmental and public sides. In aquaculture context, welfare concerns are mainly related to handling procedures, water quality and stoking densities. In Europe, authorities had to clarify the threshold limits of stocking densities to maintain fish good welfare, including for organics aquaculture through the EC regulation 710/2009. However, effects of stocking density on fish welfare are complex and sometimes contradictory. Moreover, there is a lack of knowledge about the impact of density on fish welfare in organic aquaculture. Thus, the aim of the study is to asses welfare state of rainbow trout (Oncorhynchus mykiss) at two initial stocking densities (low density, LD: 12 kg/m³ and high density, HD: 17 kg/m³) fed using organic feed by combining the monitoring of growth performances, behaviour (swimming activity) and physiological indicators (i.e. cortisol, glucose, lactate, hematocrit, red blood cellule count and lysozyme). At the end of experiment, the stocking density reached 21 kg/m³ and 30 kg/m³ for the LD and HD respectively. Overall, growth performances, swimming activity and level of physiological indicators of stress and welfare were similar between HD and LD over the experiment duration. To conclude, we observed no alteration of fish welfare between the two stocking densities monitored. This study suggests that a final stocking density of 30 kg/m³ can be considered for organic aquaculture of rainbow trout respecting welfare.     

Maintenance of steelhead trout (Oncorhynchus mykiss) sperm at different in vitro oxygen tensions alters ATP levels and cell functional characteristics

Adenosine triphosphate (ATP) levels in sperm from steelhead trout (Oncorhynchus mykiss) were found to be 12.0 ± 1.4 pmol ATP per 10⁶ sperm cells. Sperm were stored at 0–2 °C for up to 72 h under 100 and 21% O₂, and 100% N₂. The sperm ATP content of samples maintained under 100 and 21% O₂ both decreased to about 70 and 50% of initial values after 24 and 72 h, respectively. ATP levels of sperm stored under 100% N₂ decreased to 20% of initial values after only 4 h and to less than 10% of initial values after 24 h; they remained unchanged throughout the following 48 h. These low levels, however, were reversible. Following 4 h incubation under 100% N₂, sperm ATP levels rose upon exposure to and maintenance under 100% O₂, increasing to 50% of initial values after a total of 72 h. ATP levels in sperm maintained with cyanide for 4 h decreased to less than 5% of initial values. The magnitude of the change in sperm ATP levels after 24 h in samples maintained under 100% O₂ correlated positively with the initial sperm ATP levels. Sperm ATP levels, motility, and fertility were all correlated positively in a subset of samples examined. The initial percent of living sperm (sperm viability) was 97.5 ± 0.5% and was unaltered through 24 h under all O₂ tensions. Sperm viability remained unchanged through 72 h under 21% O₂ and decreased by 10 and 30% by 72 h under 100% O₂ and 100% N₂, respectively. Therefore, reductions in sperm ATP levels could not be attributed to cell death as viability decreased only modestly over these durations. Spermatocrit values were unaltered throughout the 72 h incubation for samples maintained under 100 and 21% O₂, however, a 95% increase in spermatocrit occurred after 72 h in samples maintained under 100% N₂. These data indicate that intracellular ATP in steelhead trout sperm can be modulated by ambient oxygen without significant loss of viability and that this ATP has a probable role in cell motility and fertility, and sperm packing and cell flexibility.     

Plasma growth hormone levels in fed and fasted rainbow trout (Oncorhynchus mykiss) are decreased following handling stress

Plasma growth hormone concentrations of rainbow trout,Oncorhynchus mykiss, fasted for six weeks, were significantly (p < 0.01) higher than in comparable fed animals; in the fasted fish, the levels fell progressively following acute stress (by displacing the fish within their home aquarium), with significant differences from pre-stressed fish evident between one and thirty-two hours after application of the stressor. Plasma growth hormone concentrations also fell significantly in the fed group, but differences were evident only between two and eight hours after stressor application.Plasma cortisol concentrations in pre- and post-stressed fed and fasted fish were similar. There was a bimodal response to stressor application in both groups, with significantly higher values relative to the pre-stressed sample evident one and eight hours after disturbance, but not after two, four or thirty-two hours. The changes in plasma cortisol levels between the initial (09:00h) sample and the sample taken eight hours later resembles the diet pattern seen in trout given access to self-demand feeders.Plasma glucose concentrations in pre-stressed fed animals were higher than in pre-stressed fasted fish. This relationship was also evident between one and four hours and thirty-two hours after stressor application. The post-stress rise in plasma glucose concentration was evident between one and four hours in the fed group, and between four and eight hours in fasted fish.The diel changes in plasma growth hormone and glucose concentrations could not be attributed to normal circadian patterns, and there was no apparent correlation between changes in plasma growth hormone and cortisol concentrations. There was a significant inverse correlation between plasma glucose and growth hormone concentrations when the total data set were analyzed, but these correlations were not apparent when the treatment groups were analyzed separately.     

The effect of different dietary inclusion levels of rutin (a flavonoid) on some liver enzyme activities and oxidative stress indices in rainbow trout,Oncorhynchus mykiss (Walbaum) exposed to Oxytetracycline

The study was conducted using a completely randomized design with five treatments each with three replicates. In each treatment, 30 fish (10 per replicate) with an average body weight of 30 ± 0.6 g were randomly allocated to 60‐L tanks. Treatments consisted of a control (a diet without rutin and oxytetracycline (OTC)), an OTC treatment (a diet free from rutin and containing 75 mg kg^?1 body weight OTC) and the (OTC + rutin) treatments including diets with different levels of rutin, 500 (R‐500 + OTC), 1000 (R‐1000 + OTC) and 2000 (R‐2000 + OTC) ppm and 75 mg kg^?1 body weight per day OTC which were fed to fish for 28 days. The aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the blood serum after the OTC treatment were significantly (P < 0.05) higher than in the control. After the three (OTC+rutin) treatments, the AST and ALT activities were not significantly different from the control. The level of glucose in the blood serum was significantly lower (P < 0.05) in all treatments as compared to the OTC treatment. Superoxide dismutase (SOD) and catalase (CAT) activities in red erythrocyte lysates were significantly (P < 0.05) reduced after OTC treatment in comparison with the control. After the three (OTC+rutin) treatments, the values of these indices were significantly (P < 0.05) higher than the OTC treatment, in particular for the R‐1000 + OTC treatment. It seems that rutin, as a non‐enzymatic inhibitor, is capable of preventing damage to the liver tissue and reducing oxidative stress.     

Use of electromyogram telemetry to assess the behavioural and energetic responses of rainbow trout, Oncorhynchus mykiss (Walbaum) to transportation stress

Behavioural and energetic responses of domesticated rainbow trout Oncorhynchus mykiss (Walbaum) (mean fork length=440±45 mm) to a brief transportation episode were investigated. Fish implanted with radio transmitters measuring muscle activity (electromyogram; EMGi) were transported in a standard commercial shipping tank for 50 min by truck, and then allowed to recuperate for 48 h in stationary culture tanks. The EMGi telemetry data indicated that vigorous swimming activity occurred during transportation. Telemetry recordings also indicated that the fish's swimming activity returned to baseline levels within the 48 h period after transport. However, even beyond the 48 h resting period, the swimming performance (measured as critical speed and endurance) of transported fish was still impaired relative to non‐transported controls (P<0.05). Respirometry measurements of fish taken after transportation indicated that oxygen consumption (V_o2) was significantly elevated. The rise in V_o2 of post‐transport fish could be attributed to handling procedures, as well as the intense swimming behaviour observed during transportation. Therefore, the behavioural responses of fish during transportation produced physiological consequences that persisted long after the transportation event. This study demonstrates the potential for utilizing behavioural measures, in concert with biotelemetry technologies, as tools to assess the impacts of routine aquacultural procedures on the health and welfare of captive fish.     

Hemoglobin and subunit multiplicity in the rainbow trout (Oncorhynchus mykiss) hemoglobin system

Hemoglobin (Hb) multiplicity is a commonly used index of phylogenetic differentiation and molecular adaptation in fish. Despite improvements in technologies for characterizing protein structure, our knowledge of the component composition of the Hb system in rainbow trout, Oncorhychus mykiss, has remained almost unchanged through three decades. The Hb is considered to consist of four components, of which two (HbI and HbIV) have been extensively characterised with regard to structure and function. Using a variety of molecular techniques we demonstrate the presence of at least nine different Hb fractions composed of nine (five α and four β globin) chains, and that HbI and HbIV consist of two and five individual components, respectively. These findings indicate that the published data on trout HbI and HbIV refer to mixtures of isoHbs and need to be reappraised. This revised version was published online in August 2006 with corrections to the Cover Date.     

Cellular components of probiotics control Yersinia ruckeri infection in rainbow trout, Oncorhynchus mykiss (Walbaum)

Subcellular components of the probiotics Aeromonas sobria GC2 and Bacillus subtilis JB-1, when administered to rainbow trout, Oncorhynchus mykiss , conferred protection against a new biogroup of Yersinia ruckeri . Thus, intraperitoneal or intramuscular injection of rainbow trout with cell wall proteins (CWPs), outer membrane proteins (OMPs), lipopolysaccharides (LPS), whole cell proteins (WCPs) and live cells followed by challenge on day 8 with Y. ruckeri led to 80–100% survival compared with 10% survival in the controls. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles of WCPs and OMPs from GC2 had 10 and 5 variable protein bands in comparison to 11 and 5 bands in the WCPs and CWPs from JB-1. Proteomic analyses were employed following SDS-PAGE to categorize one dominant protein of 104.7 kDa from the CWPs of JB-1 and equated it with ' Bacillus spp. endoglucanase' with a Mascot score >69. These results point to the potential of using cellular components of probiotics for protection of fish against bacterial diseases.     

Nutritional responses in rainbow trout (Oncorhynchus mykiss) fed diets with different physical qualities at stable or variable environmental conditions

Rainbow trout with initial body weight 1144 g were fed two diets with high (feed A) or low (feed B) water stability for 6 weeks. During the last 2 weeks either stable or fluctuating oxygen saturation, salinity and temperature was introduced. High water stability of the feed was associated with harder pellets, less dust formation and less broken pellets compared to feed with low water stability. During the first 4 weeks, feed intake was 23% higher in trout fed diet B than in those fed diet A. Fluctuating environment resulted in a severe drop in feed intake for both dietary groups. At termination of the trial, stomachs of trout fed feed B contained mashed pellets and free water and oil. Stomachs of trout fed feed A contained more intact pellets and little fluid. The apparent digestibility of protein, starch, dry matter and energy was highest in feed A. The apparent digestibility of P and Zn was highest at stable environment, and the absorption of P was highest from feed A. In conclusion, physical quality affected the nutritional value of the feeds, and differences in nutritional value at stable or fluctuating environment appeared to be related to feed intake.     

急性高温胁迫对虹鳟和硬头鳟幼鱼抗氧化酶活性的影响

虹鳟(Oncorhynchus mykiss)与硬头鳟(O.mykiss)为同一种的不同生态型,都属冷水性鱼类.为探讨急性高温胁迫对虹鳟和硬头鳟抗氧化酶活性的影响,选取虹鳟[体重(22.76±2.89)g]和硬头鳟[体重(23.2±1.22)g]幼鱼,分别在不同温度梯度(16℃、20℃、22℃和24℃;16℃、19℃、...     

Immunization of rainbow trout Oncorhynchus mykiss (Walbaum) with a crude lipopolysaccharide extract from Flavobacterium psychrophilum

Control methods for Flavobacterium psychrophilum are limited and oftentimes ineffective; hence, research efforts have focused on vaccine development. This study tested the hypothesis that a crude lipopolysaccharide (LPS) extract from F. psychrophilum will elicit a protective immune response in rainbow trout Oncorhynchus mykiss (Walbaum) against F. psychrophilum challenge. Rainbow trout (mean weight, 3 g) were immunized intraperitoneally with the following treatment and control preparations: 10 μg of crude LPS with or without Freund's complete adjuvant (FCA), 25 μg of crude LPS with or without FCA and saline with or without FCA. Immunization of fish with 10 or 25 μg of crude LPS/FCA resulted in significant antibody responses against F. psychrophilum using ELISA with a whole‐cell lysate as the coating antigen, but only minimal levels of protection were conferred following F. psychrophilum challenge at 14 weeks post immunization. Western blot analyses demonstrated that fish exhibited antibodies specific for low‐molecular mass proteins present in the crude LPS extract, but did not exhibit antibodies specific for F. psychrophilum LPS. The results indicate that higher immunization doses and/or the use of an alternative extraction method that yields larger LPS molecules (23–70 kDa) may be necessary to elicit specific antibody responses against F. psychrophilum LPS.     

Responses of circulating arginine vasotocin, isotocin, and melatonin to osmotic and disturbance stress in rainbow trout (Oncorhynchus mykiss)

In teleost fish exposed to stressors AVT has been implicated in activation of the hypothalamo-pituitary-interrenal axis. Mel has been shown to counteract several behavioural and endocrine consequences of stress in mammals. These studies were undertaken to investigate the effects of either disturbance or osmotic stress, applied either separately or simultaneously, on plasma AVT, IT and melatonin in rainbow trout (Oncorhynchus mykiss). Hormones were determined in plasma by high-performance liquid chromatography preceded by solid-phase extraction. The results showed that both forms of stress caused significant increase in plasma AVT concentration, although more pronounced elevation was observed in physically disturbed fish. Conversely, neither osmotic nor disturbance stress affected plasma IT concentration. The apparent difference in response to stress by the two close related neurohypophysial nonapeptides suggests independent mechanisms controlling their synthesis and/or release and supports the idea that only AVT plays a role in physiological response to stress. Plasma Mel level was depressed in fish subjected to disturbance stress and to both stresses applied simultaneously, an effect possibly associated with the elevation of plasma AVT concentration. Results are discussed in relation to physiological interactions between hormones.     

Mineral availability from barley low phytic acid grains in rainbow trout (Oncorhynchus mykiss) diets

Phosphorus storage within plant seeds occurs mainly as phytic acid, which has profound implications on the use of seeds as food material. Phytic acid phosphorus is unavailable to non‐ruminants, leaches ionic minerals during digestion, and is excreted at elevated levels as a waste product. This presents a problem in nonruminant livestock production including current efforts to develop renewable grain and legume products for use in fish feeds. The development of lines of several cereal grain species that have reduced seed phytic acid concentration provides a novel approach to dietary and environmental problems associated with seed phytic acid. Utilizing four isogenic strains of barley (one normal for seed phytic acid, and three low phytic acid lines that produce seeds with approximately 50, 70 and 95% reductions in phytic acid) the apparent digestibility of nutrients in formulated diets containing these barleys at a level of 30% was measured using rainbow trout (Oncorhynchus mykiss). Also examined was the apparent availability of several minerals including phytate phosphorus and total phosphorus. Results from these studies corresponded well with the results of other animal studies that evaluated low phytic acid cereal grains. With increasing reductions in seed phytic acid, seed available phosphorus increased and faecal phosphorus was reduced by up to 50%. Calcium availability increased, copper and sulphur decreased, and the other tested minerals demonstrated either increased or decreased availability in a manner uncorrelated to grain phytic acid concentration.     

Low temperature acclimation decreases rates of protein synthesis in rainbow trout (Oncorhynchus mykiss) heart

Protein synthesis was assessed in rainbow trout (Oncorhynchus mykiss) hearts perfused with medium containing ³H phenylalanine. Isolated hearts from fish acclimated to 5° and 15°C were used as the model system, and were perfused at variable test temperatures and pH. Protein synthesis expressed as nmol PHE mg protein^–1 h^–1 was two fold higher in hearts from fish acclimated to 15°C and tested at 15°C and extracellular pH 7.6 than in hearts from fish acclimated to 5°C and tested at 5°C and extracellular pH 8.0. The prime determinant of the decreased rate of protein synthesis was thermal history. Fish acclimated to 5°C had lower levels of RNA mg protein^–1 than fish held at 15°C. There was a direct linear relationship between the rate of protein synthesis in nmol PHE mg protein^–1 h^–1 and RNA content. RNA activity (nmol PHE g RNA^–1 h^–1 remained constant regardless of thermal history or perfusion condition. Elevated pH resulted in only a marginal decrease in protein synthesis. Test temperature had no effect on in vitro rates of protein synthesis.