Search Results (21)

Smoltification is stressful for salmonids, and cortisol is one of the central endocrine regulators for seawater adaptation. It has been established that cortisol plays both mineralocorticoid and glucocorticoid functions by MR and GR, respectively, since the aldosterone hormone is absent. Recently, investigations have proposed that the 11-deoxycorticosterone (DOC) mineralocorticoid precursor might support cortisol effects, but this mechanism remains unclear. Hence, we assessed the early effects of DOC on rainbow trout pre-smolts, the key smoltification stage, via metabolic and transcriptomic approaches. Thirty-six juveniles (~120 g) were treated for 3 h with DOC (1 mg/kg) and/or mineralocorticoid (eplerenone) or glucocorticoid (mifepristone) receptor antagonists (n = 6 for each group). DOC decreased plasma glucose and pyruvate and increased phosphate and liver glycogen. DOC also downregulated carbohydrate metabolism-related genes in the liver. Finally, gill RNA-seq analysis presented 1660 differentially expressed transcripts in DOC versus vehicle, 1022 for eplerenone + DOC versus DOC and 3324 for mifepristone + DOC versus DOC. The enrichment analysis mainly revealed the upregulation of ion transmembrane transport and carbohydrate metabolism and the downregulation of stress and innate immune responses. This suggests a significant role of DOC in liver carbohydrate metabolism and gill osmoregulation of pre-smolts through both receptors. Hence, this could contribute to improving animal welfare monitoring during smoltification by featuring novel and potential biomarkers. Full article

Microplastic (MP) contamination in aquafeed poses a significant risk to fish health and safety. This study evaluated the effectiveness of a microencapsulated natural antioxidant, astaxanthin (AX), in mitigating the adverse effects of dietary MPs in rainbow trout fry. The microcapsules were composed of an organic wall matrix designed to preserve AX while limiting MP absorption in the intestine. During a 60-day feeding trial, fish were fed diets containing amino formaldehyde polymer fluorescent MP microbeads (1–5 µm; 50 mg/kg), either alone or in combination with microencapsulated AX. MP localization in tissues was assessed via confocal microscopy, and quantification was performed following chemical tissue digestion. Fish welfare was evaluated using histological and molecular analyses. No significant effects on growth or gut morphology were observed across experimental groups. However, MPs were mainly translocated to the liver, where they induced oxidative stress, as evidenced by the upregulation of sod1, sod2, and cat gene expression. The inclusion of microencapsulated AX significantly mitigated the oxidative stress response, and the microcapsules facilitated MP coagulation in the gut, reducing intestinal absorption. These findings highlight the potential of microencapsulated antioxidants to counteract MP-induced oxidative stress and reduce MP bioavailability in aquaculture species, contributing to improved fish welfare and product quality. Full article

Sustained exercise in aquaculture is known to improve the health and growth of finfish. Implementing exercise regimes has become an increasing focus in aquaculture practice. This study examined the relationship between the preferred swimming speed (U_pref) and the optimal swimming speed (U_opt) in rainbow trout (Oncorhynchus mykiss) under non-migratory conditions typical of aquaculture environments. Using a circular raceway, rainbow trout were allowed to swim voluntarily to determine U_pref. U_opt was measured using a forced-swimming test in a swim tunnel respirometer. Experiments were conducted at three temperatures (10 °C, 15 °C, and 20 °C). The results revealed a significant difference between U_pref (1.18 ± 0.14, 1.17 ± 0.19, and 1.24 ± 0.15 BL s⁻¹, respectively) and U_opt (1.4 ± 0.19, 1.5 ± 0.15, and 1.6 ± 0.24 BL s⁻¹, respectively) across all temperatures. Aerobic scope was greatest at 15 °C (3.8), consistent with the species’ thermal range. Notably, swimming at U_pref required 18–22% less energy than U_opt, suggesting that U_pref is more suitable for aquaculture systems. This study introduces a minimally invasive and stress-free method for determining U_pref and provides insights that can optimize flow regimes in aquaculture tanks, improving both energy efficiency and fish welfare. Full article

Monitoring stress in captive fish is crucial for their welfare, but continuous physiological measures in unrestrained animals are challenging. Rainbow trout (Oncorhynchus mykiss) exhibit divergent personalities, ranging from bold to shy, which correlate with cortisol-mediated stress responses. To determine whether personality affects the sympathetic nervous system, heart rate was measured during three potentially stressful events as a proxy for sympathetic nervous system responses. Firstly, trout were classified as bold or shy, using a novel object test. Subsequently, trout were implanted with biologgers to record heart rate in vivo at rest during and after the behavioral tests. Following recovery, the fish underwent a second novel object test, a confinement test, a pair-wise contest, and a final novel object test to explore the degree of boldness over the experimental period, which remained consistent. Heart rate was relatively higher in both bold and shy animals during the confinement test and the pair-wise contest compared with the novel object test, which indicated that heart rate monitoring was a valid gauge of the valence of the experience. Heart rate responses did not differ between bold and shy trout, indicating that behavioral phenotype did not influence the autonomic stress response. Thus, heart rate is a reliable indicator of stress without the need to account for intra-specific behavioral variations. Full article

Reared rainbow trout are vulnerable to environmental stressors, in particular seasonal water warming, which affects fish welfare and growth and induces a temperature response, which involves modifications in tissue lipid profiles. Dietary supplements of plant origin, including the studied mix of a flavonoid, dihydroquercetin and a polysaccharide, arabinogalactan (25 and 50 mg per 1 kg of feed, respectively), extracted from larch wood waste, were shown to facilitate stress tolerance in fish and also to be beneficial for the safety of natural ecosystems and the sustainability of aquaculture production. This four-month feeding trial aimed to determine the effects of the supplement on liver and muscle lipid accumulation and the composition in rainbow trout reared under environmental variables. During periods of environmental optimum for trout, a consistent increase in energy lipid stores, particularly triacylglycerols (2.18 vs. 1.49-fold over a growing season), and an overall increase in lipid saturation due to lower levels of PUFAs, such as eicosapentaenoic (20:5n-3), docosahexaenoic (22:6n-3) and arachidonic (20:4n-6) acids, were observed in both control and supplement-fed fish, respectively. However, in fish stressed by an increase in ambient temperature, dietary supplementation with dihydroquercetin and arabinogalactan reduced mortality (3.65 in control vs. 2.88% in supplement-fed fish, p < 0.05) and alleviated the high-temperature-induced inhibition of lipid accumulation. It also stabilised the membrane phospholipid ratio and moderated the fatty acid composition of fish muscle and liver, resulting in higher levels of n-3 PUFAs and their precursors. Thus, the natural compounds tested are beneficial in accelerating fish tolerance to environmental stressors, reducing mortality and thermal response, and moderately improving fillet quality attributes by increasing the protein/lipid ratio and the abundance of fatty acids essential for human nutrition. Full article

Viral infections are still a major concern for the aquaculture industry. For salmonid fish, even though breeding strategies and vaccine development have reduced disease outbreaks, viral diseases remain among the main challenges having a negative impact on the welfare of fish and causing massive economic losses for the industry. The main entry port for viruses into the fish is through mucosal surfaces including that of the gastrointestinal tract. The contradictory functions of this surface, both creating a barrier towards the external environment and at the same time being responsible for the uptake of nutrients and ion/water regulation make it particularly vulnerable. The connection between dietary components and viral infections in fish has been poorly investigated and until now, a fish intestinal in vitro model to investigate virus–host interactions has been lacking. Here, we established the permissiveness of the rainbow trout intestinal cell line RTgutGC towards the important salmonid viruses—infectious pancreatic necrosis virus (IPNV), salmonid alphavirus (subtype 3, SAV3) and infectious salmon anemia virus (ISAV)—and explored the infection mechanisms of the three different viruses in these cells at different virus to cell ratios. Cytopathic effect (CPE), virus replication in the RTgutGC cells, antiviral cell responses and viral effects on the barrier permeability of polarized cells were investigated. We found that all virus species infected and replicated in RTgutGC cells, although with different replication kinetics and ability to induce CPE and host responses. The onset and progression of CPE was more rapid at high multiplicity of infection (MOI) for IPNV and SAV3 while the opposite was true of ISAV. A positive correlation between the MOI used and the induction of antiviral responses was observed for IPNV while a negative correlation was detected for SAV3. Viral infections compromised barrier integrity at early time points prior to observations of CPE microscopically. Further, the replication of IPNV and ISAV had a more pronounced effect on barrier function than SAV3. The in vitro infection model established herein can thus provide a novel tool to generate knowledge about the infection pathways and mechanisms used to surpass the intestinal epithelium in salmonid fish, and to study how a virus can potentially compromise gut epithelial barrier functions. Full article

Physical enrichment can improve the welfare of captive fish. Previous research has shown that fish often show preference for enriched environments, which can also result in improvements in growth performance. However, effects of enrichment are not always positive and the design and extent of the enrichment needs to be carefully considered. In this regard, information in real aquaculture scenarios is limited. The aim of this study was to serve as a proof of concept to test the feasibility of using simple PVC immersed shelters as a tool for better welfare in an organic rainbow trout farm. Our shelters induced little extra work in farm routines and had no negative effects on fish performance, health or mortality. The behavioral assessment pointed to a preference for sheltered areas in undisturbed conditions. However, no benefits were observed in terms of stress responses during standardized stress tests, and fish showed no obvious shelter-seeking behavior after disturbance. The results in terms of shelter-seeking behavior were probably limited by the short duration of the experiment, which was due to the farm’s routines and needs. It is recommended that strategies for enrichment in real scenarios should be tested covering a relevant part of the life cycle of the fish in captivity, to fully account for their potential to improve welfare in aquaculture. Full article

The rainbow trout (Oncorhynchus mykiss) is one of the most commercially sought-after freshwater fish species and one of the most farmed in the world. On the other hand, aquaculture breeding frequently results in outbreaks of infectious diseases and pests, and compromises the production and welfare of fish. Arthrospira platensis (known as “Spirulina”) has been used as a supplement in diets to enhance fish welfare in recent years because of its beneficial properties. This study aimed to assess the possible protective effects of Arthrospira platensis on rainbow trout specimens exposed to three different doses of the toxicant CdCl₂. The experiment was carried out using five experimental treatments of 40 individuals each: control group; group II (0.2 mg CdCl₂ per kg of commercial fish feed); group III (0.2 mg Kg⁻¹ of CdCl₂ plus 2.5 g per kg of A. platensis); group IV (0.2 mg Kg⁻¹ of CdCl₂ plus 5 g per kg of A. platensis); group V (0.2 mg Kg⁻¹ of CdCl₂ plus 10 g per kg of A. platensis). During the experiment, dietary supplementation of A. platensis normalized all serum and blood parameters altered by the presence of CdCl₂. A. platensis also had a protective effect on markers of oxidative stress. Full article

Background: Although Flavobacterium psychrophilum is recognized as the causative pathogen of rainbow trout fry syndrome (RTFS), often resulting in high fry mortality, it is also responsible for bacterial cold water disease (BCWD) in large and older rainbow trout (Oncorhynchus mykiss). These older fish do not experience high mortality, but sustain, through the shedding of bacteria, a constant infection pressure at farm level, which exposes fry to an unnecessary infection risk. We have produced and assessed the immunogenicity of an experimental injection BCWD vaccine, which may be used to decrease the shedding of bacteria from older fish. Methods: A total of 800 fish were i.p.-injected: 200 fish received the bacterin with adjuvant, 200 fish received the bacterin alone, 200 fish received adjuvant alone and 200 fish were injected with physiological saline. Blood samples were taken at day 0 and at three different time points (4, 8 and 14 weeks) post-vaccination. Plasma antibody levels were measured by ELISA for reactivity against both the homologous F. psychrophilum vaccine strain (serotype Fd) and heterologous strains (serotype Th). Results: Significantly elevated antibody titers were found against all serotypes in vaccinated fish. Welfare parameters associated with the vaccination process were evaluated by analyzing trout plasma samples for six different biochemical parameters, but no adverse effects associated with injection were indicated. Conclusions: The study suggests that an injection vaccine containing formalin-inactivated whole cells of F. psychrophilum (serotype Fd), adjuvanted with FIA, may also induce protection against heterologous strains. We advocate for, as the next step, the performance of field trials evaluating if the vaccination of older rainbow trout will (1) reduce the infection pressure in farms, (2) elevate the general health level in all groups and (3) minimize F. psychrophilum infection in fry at farm level. This may reduce the need for the administration of antibiotics in all age classes. Full article

In this study, the effects of photo-electrocatalysis (PEC) were evaluated as an innovative application of conventional photocatalysis (PC) to remediate water in a recirculating system for rainbow trout (Oncorhynchus mykiss) culture, in relation to fish welfare and health, with a multidisciplinary approach. Three tanks were employed, equipped with conventional biological filters as a control system, and three tanks equipped with the PEC purification system. The concentrations of ammonia, nitrite and nitrate ions in water were monitored, and the fish’s oxidative damage and stress response were evaluated in parallel. The water of the PEC-treated experimental group showed lower ammonia (TAN) and nitrite concentrations and higher nitrate concentration, possibly deriving from TAN oxidation through PEC, also leading to gaseous N₂. Histological analysis did not reveal any pathological alteration in the gills and liver of both groups. The superoxide dismutase (sod1), glutathione reductase (GR), glutathione peroxidase (GPx1), and Tumor necrosis factor (TNFα) gene expressions were significantly higher in the control group than in the PEC-treated group, while the Heat shock protein 70 (Hsp70) expression did not show any difference in the two groups. These results indicate that the use of PEC filters has a positive effect on water quality, compared to the use of conventional biological filters, inducing a high level of welfare in O. mykiss. Full article

The mission of veterinary clinical pathology is to support the diagnostic process by using tests to measure different blood biomarkers to support decision making about farmed fish health and welfare. The objective of this study is to provide reference intervals (RIs) for 44 key hematological, blood biochemistry, blood gasometry and hormones biomarkers for the three most economically important farmed salmonid species in Chile (Atlantic salmon, coho salmon and rainbow trout) during the freshwater (presmolt and smolt age range) and seawater stages (post-smolt and adult age range). Our results confirmed that the concentration or activity of most blood biomarkers depend on the salmonid species, age range and/or the interaction between them, and they are often biologically related to each other. Erythogram and leukogram profiles revealed a similar distribution in rainbow trout and coho salmon, but those in Atlantic salmon were significantly different. While the activity of the most clinically important plasma enzymes demonstrated a similar profile in Atlantic salmon and rainbow trout, coho salmon demonstrated a significantly different distribution. Plasma electrolyte and mineral profiles showed significant differences between salmonid species, especially for rainbow trout, while Atlantic salmon and coho salmon demonstrated a high degree of similarity. Furthermore, electrolytes, minerals and blood gasometry biomarkers were significantly different between age ranges, suggesting a considerably different distribution between freshwater and seawater-farmed fish. The RIs of clinically healthy fish described in this study take into account the high biological variation of farmed fish in Chile, as the 3.059 individuals came from 78 different fish farms, both freshwater and seawater, and blood samples were collected using the same pre-analytical protocol. Likewise, our study provides the Chilean salmon farming industry with standardized protocols that can be used routinely and provides valuable information to improve the preventive vision of aquamedicine through the application of blood biomarkers to support and optimize health, welfare and husbandry management in the salmon farming industry. Full article

This study compared the nutrient-energy retention, digestive function, growth performance, and welfare of rainbow trout (ibw 54 g) fed isoproteic (42%), isolipidic (24%), fishmeal-free diets (CV) over 13 weeks. The diets consisted of plant-protein replacement with graded levels (10, 30, 60%) of protein from poultry by-product (PBM) and black soldier fly H. illucens pupae (BSFM) meals, either singly or in combination. A fishmeal-based diet was also tested (CF). Nitrogen retention improved with moderate or high levels of dietary PBM and BSFM relative to CV (p < 0.05). Gut brush border enzyme activity was poorly affected by the diets. Gastric chitinase was up-regulated after high BSFM feeding (p < 0.05). The gut peptide and amino acid transport genes were differently regulated by protein source and level. Serum cortisol was unaffected, and the changes in metabolites stayed within the physiological range. High PBM and high BSFM lowered the leukocyte respiratory burst activity and increased the lysozyme activity compared to CV (p < 0.05). The BSFM and PBM both significantly changed the relative percentage of lymphocytes and monocytes (p < 0.05). In conclusion, moderate to high PBM and BSFM inclusions in fishmeal-free diets, either singly or in combination, improved gut function and nutrient retention, resulting in better growth performance and the good welfare of the rainbow trout. Full article

Skin mucus is a non-lethal and low-invasive matrix appropriate to assess fish welfare as it contributes to their defence against external aggressions and reflects changes in fish health status. However, more information on the response of this matrix to specific stressors is needed. In this study, rainbow trout (Oncorhynchus mykiss) specimens were subjected to an acute stress by air exposure and sampled after 1, 6, and 24 h post-stress. Blood and skin mucus were collected, and a battery of biochemical biomarkers were measured in both matrices. Cortisol and glucose values showed the expected classical stress response in plasma, increasing after the acute stress. The same pattern was observed in skin mucus, corroborating previous data in fish, and allowing us to confirm that skin mucus can be a useful complementary matrix for stress assessment in fish. The results showed sensitivity to hypoxic stress in skin mucus for cortisol, glucose, alkaline phosphatase (ALP), aspartate transaminase (AST), alanine aminotransferase (ALT), creatinine kinase (CK), and calcium. From the 15 parameters evaluated, 12 did not show statistically significant changes between plasma and mucus; therefore, using skin mucus cannot replace the use of plasma. Finally, the principal component analysis in skin mucus revealed a complete separation between the two experimental groups, being ALP, AST, glucose, cortisol, and CK, the biomarkers that contributed the most to this separation. Full article

The antimicrobial and immunostimulant proprieties of aromatic plant extracts have been widely described, but their effects on serum blood biochemistry in fish have not. For this study, we assessed the changes in serum blood biochemical parameters in rainbow trout fed with a fish diet supplemented with a basil supercritical extract (F1-BEO). Our hypothesis was that treatment and time would be associated with changes in 10 serum blood biochemical parameters. F1-BEO was added to a commercial feed (0.5, 1, 2, 3% w/w). The fish were fed for 30 days, and the blood samples were collected at 2 time points (15 and 30 days). A two-way ANOVA showed a significant effect of treatment, time, and interaction treatment × time on creatinine, urea, total protein, albumin, magnesium, and phosphorus (p < 0.05), a significant effect of both time and interaction treatment × time on cholesterol (CHOL), glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) levels, and a significant effect of time on triglycerides (p < 0.05). While changes in several of the parameters were observed, the levels remained within the normal range for rainbow trout. Notably, after 30 days there was a significant decrease in CHOL in fish treated with 0.5% and 1% w/w F1-BEO (p < 0.05). The statistically non-significant increase in GOT and GPT in the fish fed with F1-BEO up to 3% w/w indicated a positive effect of basil on liver health. Our findings suggest a potential use for basil extracts (for example., F1-BEO) in fish feed to reduce antibiotic use and improve fish welfare. Full article

This study evaluated a diverse range of markers of feeding stress to obtain a more precise assessment of the welfare of rainbow trout in relation to inadequate husbandry conditions. A feeding stress model based on dietary soybean meal was employed to identify suitable minimally invasive “classical” stress markers, together with molecular signatures. In a 56-day feeding experiment, rainbow trout were fed diets containing different levels of soybean meal. The impact of these different soybean meal diets on rainbow trout was assessed by water quality analyses, clinical health observations, classic growth and performance parameters, gut histopathology, blood-parameter measurements and multigene-expression profiling in RNA from whole blood. Soybean meal-induced enteritis was manifested phenotypically by an inflammatory reaction in the posterior section of the intestine and by diarrhoea in some trout. These inflammatory changes were associated with decreased supranuclear vacuolation. The haematocrit values and the levels of plasma cortisol and circulating lymphocytes in the blood were increased in trout that had consumed high amounts of SBM. Notably, the increased haematocrit depended significantly on the bodyweight of the individual trout. The transcript levels of certain genes (e.g., MAP3K1, LYG, NOD1, STAT1 and HSP90AB) emerged as potentially useful indicators in the blood of rainbow trout providing valuable information about inadequate nutrition. The expression-profiling findings provide a basis for improved, minimally invasive monitoring of feeding regimens in trout farming and may stimulate the development of practical detection devices for innovative aquaculture operations. Full article