Effects of clove oil and MS-222 on blood hormone profiles in rainbow trout Oncorhynchus mykiss, Walbaum

Clove oil has been demonstrated to be an effective, inexpensive anaesthetic and euthanizing agent for a number of fish species, including rainbow trout, used in aquaculture and fisheries research. However, the potential for clove oil to cause perturbations in important plasma hormone concentrations has not been investigated. The effect of anaesthesia and euthanasia in trout with eugenol (the active ingredient in clove oil) on plasma cortisol, glucose, growth hormone (GH) and two thyroid hormones [tri‐iodothyronine (T₃) and thyroxine (T₄)] was compared with tricaine methanesulfonate (MS‐222) anaesthesia, and stunning by cranial concussion in two experiments. Effects on blood chemistry were different when comparing the particular anaesthetic method being used. Stunning fish significantly increased plasma cortisol and glucose levels (both P<0.05), while euthanizing fish using either clove oil or MS‐222 had no effect on these hormone levels. In contrast, the levels of GH, T₃ and T₄ hormones were unaffected regardless of whether fish were euthanized by stunning, MS‐222 or clove oil. Variation in effects between hormones were observed using clove oil eugenol. In fish sampled 10 min after anaesthetizing with 150 mg L^?1 of eugenol, cortisol levels were significantly decreased (P<0.03), while there were no differences in either glucose or GH levels. Tri‐iodothyronine and T₄ also showed significantly elevated levels (P<0.05) after 10‐min exposure to eugenol. These results highlight the importance of investigating the potential effects of any new anaesthetic or euthanizing compounds on blood plasma parameters, prior to using them in a research setting, or when comparing results to other studies which have utilized alternative anaesthetic compounds.     

Effects of fasting on metabolic and immunological responses of common carp (Cyprinus carpio) to a further acute stress

Common carp were fasted for 0, 3 and 14 days prior to exposure to a 2‐hr crowding stress. Serum cortisol, glucose, lactate, _{T4, T3}, serum alternative complement (ACH50), lysozyme, total immunoglobulin (Ig) and blood WBC were determined before and 0, 6 and 24 hr after the crowding stress. Three days of fasting led to a significant decrease in serum levels of lactate and thyroid hormones. Fourteen days of fasting led to initiation of physiological process to maintain serum glucose by increase in serum cortisol level and lactate utilization. These fish had lower levels of thyroid hormones suggesting suppression of basal metabolism. The fish also had lower WBC and serum lysozyme and total Ig levels indicating immunosuppression. After the crowding stress, the fish fasted for 3 days had significantly lower lactate, thyroid hormones and total Ig levels. The fish fasted for 14 days had higher cortisol but lower glucose and thyroid hormones levels suggesting impairment in metabolic responses to the crowding stress. Immunological and health indicators including WBC, and ACH50, lysozyme and total Ig of these fish were significantly lower than the other groups after the crowding stress. Overall, the results show that a 14‐day fasting leads to stress responses, reduction in metabolism and immunosuppression. Moreover, 14 days of fasting lead to greater cortisol response to a further crowding stress, which induces marked immunosuppression and fallen welfare.     

Effects of anaesthesia with MS-222, clove oil and CO2 on feed intake and plasma cortisol in steelhead trout (Oncorhynchus mykiss)

We examined the effects tricaine methanesulfonate (MS-222), clove oil and CO₂ on feed intake and cortisol response in steelhead trout, Oncorhynchus mykiss. Even though a body of literature exists about the effects of different anaesthetics on fish, no comparative information seems to be available about their effects on feed intake after anaesthesia, which would be important to know especially in aquaculture research. We anaesthetised juvenile steelhead trout with these three anaesthetics, and then sampled them 4, 24 and 48 h later. Fish in all groups ate relatively well already 4 h after anaesthesia. However, feed intake in fish treated with clove oil or MS-222 was lower than in the controls. There were no differences in feed intake among anaesthetised groups. Plasma cortisol concentrations were elevated 48 h after anaesthetisation, but the treatment means were equal throughout the experiment. Our results support previous findings that clove oil is a reasonable alternative to MS-222.     

Effects of density,PVC‐tubes and feeding time on growth,stress and aggression in African catfish (Clarias gariepinus)

Successive exposure to aquaculture‐related stressors may compromise the allostatic capacity of African catfish and lead to allostatic overload and poor welfare. Therefore, we tested the effect of (i) feeding during the light or dark phase, (ii) density (51 fish per 140 L versus 51 fish per 43 L) and (iii) altered available resting space/shelter on stress handling were studied in African catfish (Clarias gariepinus). Available resting space/shelter was manipulated by providing PVC‐tubes in the tanks. Growth, baseline stress load (cortisol and osmolality), metabolism (glucose, lactate and non‐esterified fatty acids) and aggression (measured by skin scar incidence) were assessed. Upon completion of the experimental period, we determined the adaptive capacity of the fish by the response to air exposure. We show that night‐feeding enhances growth and lowers feed conversion ratio compared to day‐feeding. No effects of density were found for baseline stress hormones, basal metabolism or aggression. Low density seems to stimulate aggressive behaviour after air exposure. Unexpectedly, providing PVC‐tubes under high‐density conditions increased aggression, raised baseline cortisol levels and decreased basal metabolism as well as growth; also the endocrine response to air exposure was stronger. We argue that these effects relate to a reduction in available space, i.e. a further increase in density, or to an increased competition for shelter. Overall, our data suggest that night‐feeding optimizes growth and that care should be taken in using objects as shelter/enrichment for African catfish.     

Effects of thyroid hormone supplementation of canola meal-based diets on growth,and interrenal and thyroid gland physiology of rainbow trout (Salmo gairdneri)

Rainbow trout fed a 26% canola meal-based (CM) diet for 12 weeks at 15°C exhibited reduced growth, lower feed conversion, enlarged thyroid glands and lower plasma thyroid hormone (TH) levels than comparable fish fed equinitrogenous, equicaloric soybean meal-based (SB) diets. Supplementation of the SB diets with either T₄ (20 mg/kg) or T₃ (10 or 20 mg/kg) had no effect on the growth rate, feed conversion and thyroid histology of the trout. However, plasma T₄ levels weredepressed in trout fed the T₄- and high T₃-supplemented SB diets. In trout fed T₄- and T3-supplemented CM diets the growth rates and feed conversion were not significantly different from those of the SB-fed groups. Moreover, in the T₄-supplemented group, plasma T₄ levels were in the normal range. However, thyroid enlargement was evident in all the CM-fed fish, and plasma T₃ levels were markedly elevated in groups fed the T₃-supplemented CM diets. The data suggest that antithyroid components in the CM diets inhibited TH synthesis (but not their release), and impaired TH clearance from the circulation. There were no significant differences in plasma cortisol levels in the 8 treatment groups, nor were there differences in the histological appearance of the interrenal gland. However, when the data from SB- and CM-fed fish were pooled, plasma cortisol levels in the SB-fed fish were significantly lower than in the CM-fed animals. Glucosinolates at a level of 164 mg/kg diet were toxic to young trout, but the effect was ameliorated by dietary TH supplementation.     

Does oral 3,5,3 ′-triiodo-l-thyronine affect dietary glucose utilization and plasma insulin levels in rainbow trout (Salmo gairdneri)?

A factorial experiment was conducted to determine the effect and interaction of dietary carbohydrate level and triiodo-L-thyronine (T₃) supplementation on the growth, physiological response and plasma insulin and cortisol levels of rainbow trout. The oral administration of T₃ significantly increased the growth, protein efficiency ratio and feed efficiency of trout, indicating an increased protein and perhaps energy utilization in these fish. However, T, administration did not significantly increase the utilization of dietary glucose as an energy source by the trout. Similarly, the administration of T₃ did not significantly affect plasma insulin levels in either the fed or the fasted trout. Plasma insulin levels were significantly higher in fed trout reared on the non-T₃ supplemented high carbohydrate diet in comparison to trout reared on the low carbohydrate diets. This indicates that increased dietary carbohydrate stimulates increased insulin secretion in the trout. Therefore, although rainbow trout are not insulin-deficient, they can still be considered a diabetic-like animal due to their poor glucose tolerance. Plasma cortisol levels were not affected by diet composition and altered plasma glucose levels.     

Iodine enrichment of Artemia and enhanced levels of iodine in Atlantic halibut larvae (Hippoglossus hippoglossus L.) fed the enriched Artemia

Flatfish metamorphosis is initiated by the actions of thyroid hormones (TH) and iodine is an essential part of these hormones. Hence, an iodine deficiency may lead to insufficient levels of TH and incomplete metamorphosis. In this study, different iodine sources for enrichment of Artemia were evaluated and the levels of iodine obtained in Artemia were within the range of 60–350 μg g^?1 found in copepods. Larval Atlantic halibut was fed Artemia enriched with either normal DC‐DHA Selco or DC‐DHA Selco (commercial enrichments) supplemented with iodine from days 9 to 60 postfirst feeding. There was no significant difference in growth, mortality or metamorphic development between the groups. The analyses showed that we were able to enrich Artemia with iodine. Further, the larvae‐fed iodine‐enriched Artemia had higher whole body iodine concentration compared to larvae‐fed Artemia without iodine enrichment.     

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.     

Leucocyte profile and growth rates as indicators of crowding stress in pejerrey fingerlings (Odontesthes bonariensis)

Crowding is one of the most common stressors found in intensive aquaculture, compromising growth rates and immune function. Plasmatic cortisol is a classic stress biomarker for fish, but its quantification is expensive and demands blood volumes that small individuals do not provide, constraining the usage of this technique to assess stress in fingerlings. The leucocyte profile is an alternative methodology to quantify stress with reduced costs and volumes of blood. Stress conditions promote neutrophilia and lymphocytopenia as response to elevated glucocorticoids levels. Considering the difficulties to assess stress imposed by intensive fish farming using measurement of glucocorticoid hormones, this study aimed to evaluate the stress‐induced changes in leucocyte profiles and growth rates imposed by crowding in fingerlings of Odontesthes bonariensis, a promising South‐American candidate for freshwater aquaculture. To meet these objectives, fingerlings (initial weight 0.05 ± 0.06 g and length 1.68 ± 0.13 cm) were reared for 45 days under three rearing densities (1, 5 and 10 fingerlings L^?1). At the end of this period, fish were anaesthetized and euthanized to obtain the leucocyte profile, neutrophil:lymphocyte ratio (N:L) and growth rate. Increasing density promoted: significant reduction in growth (final length, weight and specific growth rate); neutrophilia and lymphocytopenia; and increased N:L ratio. Concluding, the tested rearing densities imposed distinct levels of stress characterized by different N:L ratio, demonstrating that the leucocyte profile is a reliable alternative to measure stress levels in O. bonariensis fingerlings and probably in other fish species.     

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.     

Puerarin Promotes Overall Health Through Blood Flow and Immune Response within Chinook Salmon (Oncorhynchus tshawytscha) Fingerlings

An 8-week study was performed to determine the effects of supplementing puerarin, the most biologically active ingredient present in the roots of kudzu (Puerarin lobata), in diets of farmed chinook salmon (Oncorhynchus tshawytscha) fingerlings. Puerarin was given to study fish through supplementation to commercial fish feed. Administration of puerarin reduced cortisol, proteins, and hematocrit levels while improving spleen somatic indices and respiratory burst response compared to fish fed a control diet without affecting growth, and thereby counteracts the negative consequences of stress induced through handling and experienced by fish reared in aquaculture.     

Quantifying risks of volitional consumption of New Zealand Mudsnails by Steelhead and Rainbow Trout

To assess the risk of transferring alien, invasive New Zealand mudsnails (NZMS) Potamopyrgus antipodarum with shipments of live rainbow or steelhead trout Oncorhynchus mykiss, we conducted laboratory trials to quantify and determine the volitional ingestion of snails from the bottom of laboratory tanks. Approximately, 2000 snails were placed on the bottom of a test tank and groups of 10 fish were added to each tank. After 48 h, the fish were removed, euthanized and the snails in the gastrointestinal tract were counted, and the proportion of snails remaining in each tank was measured. We found that both rainbow trout and steelhead consumed NZMS, but rainbow trout consumed nearly twice the number of snails that were consumed by steelhead trout. Feeding fish a maintenance diet increased the total consumption of snails by fish to an average of 64.5 snails per rainbow trout. Our study suggests that management strategies that depurate fish without feed for 48 h to reduce the risk of transfer of NZMS are not viable in preventing the transport of snails if rearing waters contain snails.     

The Efficacy of a Novel Microbial 6‐Phytase Expressed in Aspergillus oryzae on the Performance and Phosphorus Utilization of Cold‐ and Warm‐Water Fish: Rainbow Trout,Oncorhynchus mykiss,and Nile Tilapia,Oreochromis niloticus

The efficacy and tolerance of a novel microbial 6‐phytase were investigated in rainbow trout, Oncorhynchus mykiss, and Nile tilapia, Oreochromis niloticus. Reference diets were sufficient in available phosphorus (P). The test diet limiting in available P was supplemented with phytase at 500, 1000, or 2000 phytase units/kg feed. The enzyme was effective in increasing total P apparent digestibility coefficient in relation to increasing the dose of phytase in rainbow trout and Nile tilapia. Zinc apparent digestibility improved in relation to phytase supplementation in rainbow trout. P release due to phytase supplementation ranged from 0.06 to 0.18% P/kg feed in rainbow trout and from 0.13 to 0.26% P/kg feed in Nile tilapia. A 58‐d performance trial was conducted to evaluate tolerance of fish to phytase supplementation. Dietary treatments consisted of a basal diet without phytase or supplemented with 2000 and 200,000 phytase units/kg feed. Results indicate that this novel microbial 6‐phytase is well tolerated by fish. Significant improvements for growth as well as feed conversion ratio were observed when the phytase was fed at 2000 phytase units/kg feed. This phytase is proven efficient in releasing P from phytate and could be added when plants are used for fish meal replacement in diets for salmonid and omnivorous fish.     

Stress response of juvenile rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)to chemical cues released from stressed conspecifics

The objective of this study was to determine whether exposure of rainbow trout (Oncorhynchus mykiss) to water containing a stressed trout or skin extract from stressed and non-stressed trout would elicit a stress response in conspecifics. Juvenile rainbow trout were exposed for 1 hour to water containing a stressed fish, homogenized skin extracts from a non-stressed fish, skin extract from a stressed fish and water with none of these factors. The stress response was measured over a 24-h period (1, 6, 12, 24 h after exposure). Plasma cortisol levels increased at 12 h in fish exposed to water from a stressed fish and skin extract from a stressed fish. Plasma glucose and hepatic hsp70 levels were not affected by treatments. The results suggest that rainbow trout elicit a stress response when exposed to stress-related alarm cues released from conspecifics.     

Dietary nucleotide supplements influence the growth,haemato‐immunological parameters and stress responses in endangered Caspian brown trout (Salmo trutta caspius Kessler, 1877)

A feeding trial was conducted to evaluate the effects of Optimûn, a commercial nucleotide (NT) product, on the growth, haemato‐immunological and serum biochemical parameters and stress responses of Caspian brown trout fingerlings (average initial weight of 12.26 g). A basal diet was supplemented with levels of 0 (control), 1.5, 2.5, 3.5 and 5 g NT kg^?1 to formulate five experimental diets. After 8 weeks of feeding trial, fish fed diet with 2.5 g NT kg^?1 had the highest final weight compared with other treatments. The fish fed diets with 2.5 g NT kg^?1 had higher blood protein, albumin, albumin/globulin ratio, red blood cells, white blood cells and lymphocyte content and lower alkaline phosphatase. The lysozyme activity in serum was found to be significantly (P < 0.05) greater in fish fed diet with 2.5 g NT kg^?1. Concerning both stressors (confinement and salinity stress), fish fed diet with 2.5 g NT kg^?1 had lower plasma cortisol and glucose levels. Thus, administration of 2.5 g kg^?1 of the Optimûn dietary nucleotide formula is recommended to promote growth and immunity as well as to enhance stress responses of Caspian brown trout.     

Selection for improved stress tolerance in rainbow trout (Oncorhynchus mykiss) leads to reduced feed waste

The magnitude of the cortisol response to a standardised stressor is a heritable trait in salmonid fishes, and selection for stress responsiveness induces differences in both behaviour and neuroendocrine function. For instance, in laboratory studies, fish with a high cortisol response show a greater propensity for stress induced anorexia. Some authors have, however, commented that controlled studies encompassing relatively small groups of fish hold little or no relevance to practical aquaculture. This notion may be flawed, since understanding the mechanisms behind the behaviour of individuals is a proviso to predict behaviour in groups even with the caveat that some behaviors may be modified by group size. As an example, optimal feeding regimes should be easier to predict in a population consisting of individuals whose appetite is relatively less affected by external stressors. In a fluctuating and potentially stressful environment, such a population should also generate less feed waste, if kept on fixed rations. In the present experiment, we tested this hypothesis by monitoring feed waste and feed conversion efficiency in lines of rainbow trout selected for a low (LR) or high (HR) cortisol response to stress. The study was carried out after fish had been transported between rearing sites in the United Kingdom and Norway. There was significantly more feed waste from rearing units containing HR fish, and these fish also showed lower feed efficiency (growth per unit feed consumed). The difference in feed waste became more apparent with increasing time after transport, as rations increased. Simultaneously, size was more variable and growth was slower in HR rearing units. These results suggest that there are several potential benefits of selection for low stress responsiveness in aquaculture.     

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.     

Cortisol and testosterone accumulation in a low pH recirculating aquaculture system for rainbow trout (Oncorhynchus mykiss)

Steroids accumulate in recirculating aquaculture system (RAS), although explanatory factors for such accumulation are still poorly explored. This study investigated the effect of water exchange rate and pH in six replicated RAS on the concentration of the stress hormone cortisol in rainbow trout blood plasma and in the holding water and of the sex steroids testosterone, 11‐ketotestosterone (11‐KT) and 17,20β‐dihydroxypregn‐4‐en‐3‐one (17,20β‐P) over a 70‐day experimental period. Three combinations of water exchange rate and pH were used each treatment, with two replications: (i) high water exchange rate (±1700 L kg^?1 feed) and neutral pH (±7.3), (ii) low water exchange rate (±500 L kg^?1 feed) and neutral pH (±7.3) and (iii) low water exchange rate (±500 L kg^?1 feed) and low pH (±5.8). Plasma cortisol concentrations at day 70 were higher (24.4 ± 9.5 ng mL^?1) for fish kept at low pH when compared to fish kept at neutral pH (12.0 ± 0.1 and 8.7 ± 0.2 ng mL^?1). Water cortisol and testosterone concentrations at day 35 were higher at low pH than at neutral pH, whereas water 11‐KT and 17,20β‐P did not differ among treatments. At day 70, there were no significant differences between low and high pH. These results demonstrate that low pH contributes to increased plasma cortisol concentrations and to its accumulation in water, possibly indicating a stress response to low pH. The higher concentration of testosterone but not of the other sex hormones point to unspecified reproductive effects that need further investigation.     

Alternative matrices for cortisol measurement in fish

Plasma cortisol is the most commonly used indicator of stress in fish but, as the blood sampling procedure itself can be a source of stress, it would be helpful to measure cortisol using less invasive matrices. It is also necessary to find alternative matrices as stress indicators in dead fish in which blood sampling is impossible. In the present study, we investigated transport stress in three aquaculture species, European sea bass (Dicentrarchus labrax L.), common carp (Cyprinus carpio L.) and rainbow trout (Oncorhynchus mykiss Walbaum), by cortisol determination (radioimmunoassay) in plasma and other matrices (skin mucus, gut content, lateral muscle and caudal fin). Cortisol significantly increased after transport in all species and matrices, except in the sea bass gut content, where it remained unchanged. The three species responded to transport stress by producing different cortisol levels. In conclusion, the significant correlation found between plasma cortisol and most of the other matrices opens up the possibility of using them to evaluate stress in fish: mucus sampling is a less invasive method than blood sampling, and in addition to muscle and fin sampling, it can be used in postmortem fish.     

Reproductive performance of fairy shrimp Branchinecta orientalis (G. O. Sars 1901) (Crustacea: Anostraca), fed with effluent of rainbow trout Oncorhynchus mykiss (Walbaum 1792) ponds

Aquaculture production is predicted to increase sharply. In this regard, live feed plays a crucial role in the larval phase of many aquaculture organisms. Hence, a persistent concern in aquaculture is to find low‐cost and eco‐friendly feed sources to culture live feed organisms. Branchinecta orientalis (G. O. Sars 1901), a fresh/brackish water fairy shrimp, was reared using effluent from rainbow trout Oncorhynchus mykiss (Walbaum 1792) ponds, either fresh but supplemented with two species of microalgae, Scenedesmus sp. and Haematococcus sp., or non‐supplemented but after “ageing” of the culture medium. The feeding experiment was designed at a density of 100 individuals L^?1 in 2‐L vessels. The results indicated that differences between final length, survival and most reproductive parameters of the treatment with aged medium and the treatment using fresh medium supplemented with Scenedesmus sp. were non‐significant (p > .05). Better results were obtained for a number of reproductive parameters in the treatment supplemented with Haematococcus sp. Thus, for intensive resting egg production of B. orientalis, microalgae can be replaced by aged non‐supplemented effluent from trout ponds as a nutrient‐rich feed source. This consequently can reduce drainage of nutrients into the environment and thus decrease aquatic pollution.