Rainbow Trout Research Articles

First case of ichthyophonosis in farmed rainbow trout Oncorhynchus mykiss in Serbia.

Ichthyophonosis is a disease caused by the mesomycetozoean parasite Ichthyophonus hoferi that affects a variety of fish species, including rainbow trout Oncorhynchus mykiss Walbaum. This disease is characterized by granulomatous lesions and necrosis in various organs, which can have severe impacts on the health and welfare of the fish. Ichthyophonosis has been found in several parts of the world, including Europe, and is a significant concern in the aquaculture industry and for populations of wild marine fishes. The rainbow trout is a widely cultured salmonid species in many countries, including Serbia. Although the presence of I. hoferi in rainbow trout has been reported in several countries, it has never been documented in Serbia. In this article, we report the first case of ichthyophonosis in rainbow trout in Serbia.

Development and validation of reverse-transcription cross-priming amplification-based lateral flow assay for the detection of infectious hematopoietic necrosis virus

Infectious hematopoietic necrosis virus (IHNV) severely and lethally infects salmonid fish, including Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) worldwide. Rapid and accurate viral detection is crucial for preventing pathogen spread and minimizing damage. Although several IHNV detection assays have been developed, their analytical and diagnostic performances have not been evaluated and field usability assessments have not been completely validated. Here, we developed a reverse-transcription cross-priming amplification-based lateral flow assay (RT-CPA-LFA) and validated its diagnostic performance. To detect the IHNV, primers were designed based on the consensus sequence of the nucleocapsid (N) gene. Notably, when combined with a lateral flow dipstick, it could visualize the IHNV amplification products within 5 min and the detection limit of the developed RT-CPA-LFA was 3.28×105 copies/μL. The diagnostic sensitivity and specificity in fish samples (n=140) were 98.88 % and 96.08 %, respectively. Moreover, the IHNV detection rate by RT-CPA-LFA in dead rainbow trout artificially injected with the virus was 100 %, consistent with to the results obtained from second conventional and real-time PCR, indicating its applicability for rapid IHNV detection and presumptive IHN diagnosis during the endemic period.

Ribosome Profiling and RNA Sequencing Reveal Translation and Transcription Regulation under Acute Heat Stress in Rainbow Trout (Oncorhynchus mykiss, Walbaum, 1792) Liver.

Rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) is an important economic cold-water fish that is susceptible to heat stress. To date, the heat stress response in rainbow trout is more widely understood at the transcriptional level, while little research has been conducted at the translational level. To reveal the translational regulation of heat stress in rainbow trout, in this study, we performed a ribosome profiling assay of rainbow trout liver under normal and heat stress conditions. Comparative analysis of the RNA-seq data with the ribosome profiling data showed that the folding changes in gene expression at the transcriptional level are moderately correlated with those at the translational level. In total, 1213 genes were significantly altered at the translational level. However, only 32.8% of the genes were common between both levels, demonstrating that heat stress is coordinated across both transcriptional and translational levels. Moreover, 809 genes exhibited significant differences in translational efficiency (TE), with the TE of these genes being considerably affected by factors such as the GC content, coding sequence length, and upstream open reading frame (uORF) presence. In addition, 3468 potential uORFs in 2676 genes were identified, which can potentially affect the TE of the main open reading frames. In this study, Ribo-seq and RNA-seq were used for the first time to elucidate the coordinated regulation of transcription and translation in rainbow trout under heat stress. These findings are expected to contribute novel data and theoretical insights to the international literature on the thermal stress response in fish.

Improvement of the immunological and antioxidant capacity of rainbow trout (Oncorhynchus mykiss) by using the paraprobiotic Bacillus subtilis subsp. spizizenii

The aim of this study was to produce Bacillus subtilis subsp. spizizenii as a paraprobiotic for the nutrition of rainbow trout (Oncorhynchus mykiss). Fifty-four trout (74.51 ± 3.15 g) were transferred to nine 300-liter tanks and fed PRO (live freeze-dried form), PAR (irradiated freeze-dried form) and control feed (n=3) for 8 weeks. Growth, hematological, immunological and enzymatic liver functions, as well as antioxidant capacity were then analyzed. Kidney, liver, and intestinal tissue as well as the expression of cytokines (TNF-1, TNF-2, IL-1β1, and IL-1β2) in spleen, and kidney were analyzed. The growth indices, white, and red blood cells, neutrophils, monocytes and granulocytes, total protein and globulin improved in the PAR and PRO groups compared to the control group (p<0.05). The results showed that the PAR and PRO groups resulted in modulation of ALT, AST and ALP compared to the control group (p<0.05). Antioxidant indices were highest in the PAR group compared to the PRO and control groups (p<0.05). Intestinal histology results showed that the height of intestinal folds and goblet cells increased in fish fed PRO and PAR compared to the control group (p<0.05). Although melatonin increased in the PAR and PRO groups, it decreased in the liver of PAR compared to PRO (p<0.05). Immunological indices improved in the PAR and PRO groups compared to the control group (p<0.05), with no differences between PAR and PRO (p<0.05). The results of this study show that the paraprobiotic produced in this study is effective under culture conditions and can therefore be used as a supplement in trout farming.

Protective effects of Allium jesdianum essential oil on rainbow trout (Oncorhynchus mykiss) exposed to sub-lethal toxicity of cypermethrin

This study aimed to investigate the protective effects of Allium jesdianum essential oil (AJEO) in decreasing cypermethrin toxicity for rainbow trout. First, the safety of the 0%, 0.5%, 1%, and 1.5% AJEO supplements was assayed after 60 days. Then, the protective effects of AJEO were studied on fish exposed to 12.5% 96h LC50 cypermethrin after 14 days. Results showed that 1 and 1.5% AJEO administration enhanced protease and lipase activities in the intestine and improved growth performance. Moreover, feeding fish with 1 and 1.5% AJEO increased catalase (CAT) and superoxide dismutase activities (SOD) and decreased malondialdehyde (MDA). Also, AJEO increased glutathione peroxidase (GPx) activity in serum. However, exposure to cypermethrin significantly decreased these enzyme activities and increased MDA. The oxidative biomarkers remained normal in fish fed with AJEO after exposure to cypermethrin. The administration of 1 and 1.5% AJEO significantly decreased cortisol and glucose levels, alkaline phosphatase (ALP), lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase activities. Although exposure to cypermethrin significantly increased these biochemical biomarkers, AJEO could adjust them. A significant effect of 1% AJEO on total protein and globulin was observed before and after exposure to cypermethrin. Exposure to cypermethrin decreased all immunological parameters in the serum and mucus. However, administration of 1% AJEO increased protease, lysozyme (LYS) activities, total immunoglobulin (Ig), complement C3 and C4, and nitroblue tetrazolium (NBT) in the serum and ALP, LYS, protease activities and Ig in mucus. In conclusion, results showed that AJEO could potentially decrease the toxicity effects of cypermethrin in fish.

Comparative in vitro hepatic clearances of commonly used antidepressants, antipsychotics, and anti-inflammatory agents in rainbow trout liver S9 fractions

Residues of human pharmaceuticals are widely detected in surface waters and can be taken up by and bioaccumulate in aquatic organisms, especially fish. One of the key challenges in assessing the bioaccumulation potential of ionizable organic compounds, such as the pharmaceuticals, is the lack of empirical data for biotransformation. In the present study, we assessed the in vitro intrinsic clearances (CLINT) of twelve pharmaceuticals, individually and some additionally as mixtures, in rainbow trout (Oncorhynchus mykiss) liver S9 fractions (RT-S9) adhering to the OECD test guidance 319B. The test substances included four anti-inflammatory agents (diclofenac, ibuprofen, ketoprofen, naproxen), seven antidepressants/antipsychotics (citalopram, haloperidol, levomepromazine, mirtazapine, risperidone, sertraline, venlafaxine) and the O-desmethyl metabolite of venlafaxine. Quantifiable intrinsic clearances were detected for diclofenac, ibuprofen, naproxen, levomepromazine, and sertraline. Apart from diclofenac, the in vitro clearances of the other four pharmaceuticals were shown to be critically dependent on the cytochrome P450 (CYP) metabolism. Therefore, we also determined the half-maximal inhibitory concentrations (IC50) of the same twelve pharmaceuticals toward CYP1A-like (7-ethoxyresorufin-O-deethylation, EROD) and CYP3A-like (benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylation, BFCOD) activities in RT-S9 using IC50 shift assay. As a result, levomepromazine and sertraline were identified as the most potent inhibitors of both EROD and BFCOD activity (unbound IC50 < 10 µM each), followed by citalopram and haloperidol (10 µM < IC50 < 100 µM). Additionally, mirtazapine was a selective EROD inhibitor (IC50 ∼ 30 µM). The inhibitory impacts of haloperidol and sertraline were indicatively time dependent. Finally, we carried out intrinsic clearance assays with mixtures of diclofenac, ibuprofen, naproxen, levomepromazine, and sertraline to examine the impacts of EROD and BFCOD inhibitions on their in vitro CLINT in RT-S9. Our in vitro data suggests that the intrinsic clearances of ibuprofen, levomepromazine, and sertraline in rainbow trout can be significantly reduced as the result of P450 inhibition by pharmaceutical mixtures, whereas the clearances of diclofenac and naproxen are less impacted.

Resection of the adipose fin from rainbow trout acutely alters the cerebral transcriptome and respiratory frequency

As many as 100 million salmonid fish are tagged annually by removing the adipose fin, usually for recovery statistics and the fishing industry. In a multi-parametrical approach, we evaluated the response to resection of the adipose fin in rainbow trout Oncorhynchus mykiss over the course of 72 h. Both the concentrations of lactate and the cerebral transcriptomes differed in rainbow trout early after adipose fin clipping compared to the matching intact controls. The number of differentially expressed genes significantly increased, first in the midbrain and later in the forebrain, in fin-clipped compared to intact trout. Several of these genes were predicted to be involved in pain-induced and synaptogenesis-related pathways, suggesting that adipose fin resection may affect the architecture of neuronal networks. In an experimental setup applying swimming tunnels, we tested whether removal of the adipose fin had any consequences on the behavior of fin-clipped versus intact trout exposed to a regularly recurring water flow. Concomitant with increased and prevailing breath frequencies after repeated exposure to water flows, we observed a marked transcriptional response in the forebrain of fin-clipped trout compared with intact unchallenged controls at later time points after the resection. The resulting list of differentially expressed genes was assigned to a broad range of signaling pathways involved in trans-synaptic signaling, neuronal interaction dynamics, and neuroplasticity. Taken together, our findings suggest that resection of the potentially flow-sensory adipose fin alters pathways related to pain perception and likely enhances the physiological response to the current, probably indicating increased physical effort. We also deduce from the transcriptomic data that amputation of the adipose fin leads to reorganization of the high-level integration system in the brain. The short-term effects observed in the present study require further validation to estimate the prevailing stress for the organism. Nevertheless, we propose considering alternative tagging methods for salmonid fish that do not conflict with fish welfare, as the amputation of the adipose fin may have more far-reaching physiological effects than previously assumed.

Assessment of Single-Dose Pharmacokinetics of Oxolinic Acid in Rainbow Trout and Determination of InVitro Antibacterial Activity Against Pathogenic Bacteria From Diseased Fish.

In response to the heightened risk of bacterial diseases in fish farms caused by increased demand for fish consumption and subsequent overcrowding, researchers are currently investigating the efficacy and residue management of oxolinic acid (OA) as a treatment for bacterial infections in fish. This research is crucial for gaining a comprehensive understanding of the pharmacokinetics of OA. The present study investigates pharmacokinetics of OA in juvenile rainbow trout. The fish were given a 12 mg kg-1 dose of OA through their feed, and tissue samples were collected of the liver, kidney, gill, intestine, muscle, and plasma for analysis using LC-MS/MS. The highest concentrations of the drug were found in the gill (4096.55 μg kg-1) and intestine (11592.98 μg kg-1), with significant absorption also seen in the liver (0.36 L/h) and gill (0.07 L/h) (p < 0.05). The liver (0.21 L/h) and kidney (0.03 L/h) were found to be the most efficient (p < 0.05) at eliminating the drug. The study also confirmed the drug antimicrobial effectiveness against several bacterial pathogens, including Shewanella xiamenensis (0.25 μg mL-1), Lactococcus garvieae (1 μg mL-1), and Chryseobacterium aquaticum (4 μg mL-1). The study concludes significant variations among different fish tissues, with higher concentrations and longer half-lives observed in the kidney and intestine. The lowest MIC value recorded against major bacterial pathogens demonstrated its therapeutic potential in aquaculture. It also emphasizes the importance of understanding OA pharmacokinetics to optimize antimicrobial therapy in aquaculture.

Involvement of ahr-dependent Cyp1a detoxification activity, oxidative stress and inflammatory regulation in response to graphene oxide exposure in rainbow trout (Oncorhynchus mykiss)

Graphene oxide (GO) is a very attractive material for use in a vast number of applications. However, before its widespread use, it is important to consider potential issues related to environmental safety to support its safe application. The aim of this study was to investigate effects on fish (rainbow trout) following GO exposure. Using both an in vitro approach with the RTL W1 rainbow trout liver cell line, and in vivo exposures, following OECD TG 203, disturbances at the cellular level as well as in the gills and liver tissue of juvenile trout were assessed. In RTL W1 cells, a time and concentration-dependent loss in cell viability, specifically plasma membrane integrity and lysosomal function, was observed after 96 h of exposure to GO at concentrations ≥18.75 mg/L. Additionally, increased reactive oxygen species (ROS) levels were evidenced at concentrations ≥18.75 mg/L, and an enhancement of metabolic activity was noted with concentrations ≥4.68 mg/L. In vivo exposures to GO did not provoke mortality in rainbow trout juveniles following 96 h exposure but led to histological alterations in gills and liver tissues, induction of enzymatic detoxification activities in the liver, as well as aryl hydrocarbon receptor (ahr)–cytochrome P450 1a (cyp1a) gene expression downregulation, and upregulation of pro-inflammatory cytokines il1b and il8 at GO concentrations ≥9.89 mg/L.

Enhanced immunogenic response of salmonids to Tenacibaculum maritimum formalin-killed vaccine at different water temperatures

Tenacibaculum maritimum is the main organism responsible for yellow mouth (YM) and the primary driver for the use of antibiotics in Canadian Atlantic salmon culture. This creates a potential for the generation of antibiotic-resistant strains and brings significant costs to aquaculture producers. YM only occurs in Atlantic salmon recently transferred to saltwater, wherein affected fish die rapidly with few clinical signs apart from small yellow plaques around the mouth. Interestingly, if fish survive the first three to four months following saltwater transfer, they appear to have long-lasting protection against YM suggesting that immune memory may play a role. As a result, this costly predicament might be solved with an intervention that induces proper immune stimulation prior to the transfer of smolts to saltwater. To explore this, the current study developed an indirect ELISA that was specific for both Atlantic salmon and rainbow trout serum-IgM production towards a North American strain of T. maritimum. This assay was then used to confirm that Atlantic salmon routinely cultured in sea pens develop T. maritimum specific serum IgM and that this antibody development significantly increases over 220 days. To determine whether this antibody production could be induced and possibly optimized, a formalin inactivated T. maritimum vaccine was developed and used for intraperitoneal vaccinations of naïve rainbow trout while being maintained at low (11 °C) and high (15 °C) water temperatures. Analysis of leukocyte recruitment to the injection site revealed that the vaccine significantly enhanced total leukocyte and specifically lymphocyte recruitment when fish were held at the higher water temperature. Pathogen-specific antibody development was also significantly enhanced by the higher water temperature 21 days after receiving the vaccine. However, when a booster was given at day 21, any temperature-related differences in T. maritimum antibody levels between vaccinated groups was eliminated. A better understanding of salmonid antibody development towards T. maritimum could provide valuable insight into the development of an effective vaccine to prevent YM. Such a vaccine intervention could replace currently used antibiotic treatments and drastically improve the sustainability of North American aquaculture.

Ivermectin toxicokinetics in rainbow trout (Oncorhynchus mykiss) following P-glycoprotein inhibition.

Changes to ivermectin (IVM [22,23-dihydro avermectin B1a + 22,23-dihydro avermectin B1b]) toxicokinetics (TK) with and without P-glycoprotein (P-gp) inhibition by cyclosporin A (CsA) were examined in rainbow trout (Oncorhynchus mykiss). Rainbow trout were injected with 175μg/kg 3H-IVM (8.6μCi/mg IVM) with or without co-administration of 480μg/kg CsA into the caudal vasculature. Fish were sacrificed at various time points (0.25, 0.5, 1, 3, 24, 48, 96, and 168h) for organ and tissue sampling (blood, liver, kidney, gill, intestines, brain [5 regions], eye, gonad, and fat) which were analyzed for IVM-derived radioactivity. The IVM concentration decreased over time in blood, liver, kidney, and gill, while concentrations in other tissues remained constant. The highest maximum IVM concentration (Cmax) was found in kidney, followed by liver; the lowest Cmax was found in eye, followed by brain and adipose tissue. The highest % of the administered dose was found in the blood 15min post-IVM administration, followed by the intestine at 60min post-IVM administration. P-gp inhibition by CsA did not significantly affect calculated TK parameters (AUC [7.33 ± 0.73 - 11.5 ± 2.5mg•h/kg], mean residence time [84.7 ± 21 - 125 ± 55h], T1/2 [58.7 ± 15 - 86.8 ± 38h], clearance rate [0.0152 ± 0.0033 - 0.0239 ± 0.0024 L/kg•h], or volume of distribution [1.91 ± 0.47 - 2.02 ± 0.33 L/kg]), but resulted in small but significant changes in the % administered dose found in blood and medulla. These results suggest that P-gp plays a limited role in overall IVM TK, and that its role in xenobiotic protection may be much less robust in fish than it is in mammals.

Toxic effects of the emerging Alexandrium pseudogonyaulax (Dinophyceae) on multiple trophic levels of the pelagic food web

The dinoflagellate Alexandrium pseudogonyaulax, a harmful algal bloom species, is currently appearing in increasing frequency and abundance across Northern European waters, displacing other Alexandrium species. This mixotrophic alga produces goniodomins (GDs) and bioactive extracellular substances (BECs) that may pose a threat to coastal ecosystems and other marine resources. This study demonstrated the adverse effects of A. pseudogonyaulax on four marine trophic levels, including microalgae (Rhodomonas salina), microzooplankton (Polykrikos kofoidii) and mesozooplankton (Acartia tonsa), as well as fish gill cells (RTgill-W1, Oncorhynchus mykiss), ultimately leading to enhanced mortality and cell lysis. Furthermore, cell-free supernatants collected from A. pseudogonyaulax cultures caused complete loss of metabolic activity in the RTgill-W1 cell line, indicating ichthyotoxic properties, while all tested GDs were much less toxic. In addition, cell-free supernatants of A. pseudogonyaulax led to cell lysis of R. salina, while all tested GDs were non-lytic. Finally, reduced egg hatching rates of A. tonsa eggs exposed to cell-free supernatants of A. pseudogonyaulax and impaired mobility of P. kofoidii and A. tonsa exposed to A. pseudogonyaulax were also observed. Altogether, bioassay results suggest that the toxicity of A. pseudogonyaulax is mainly driven by BECs and not by GDs, although further research into factors modulating the lytic activity of Alexandrium spp. are needed.

Effects of electrical and percussive stunning on neural, ventilatory and cardiac responses of rainbow trout

From an ethical standpoint, it is imperative that rainbow trout (Oncorhynchus mykiss) are humanely slaughtered, which entails that they are rendered unconscious immediately by a stunning method and remain so until death. The efficacy of electrical stunning following dewatering (i.e., in-air electrical stunning at intensities of 50 to 920 mA and durations of 5 to 30 s) and percussive stunning, both advocated as humane stunning and/or killing methods, are evaluated here for this species via the presence or absence of visually evoked responses (VERs). In addition, ventilatory and cardiac responses were evaluated to elucidate the physiological basis for the lethality of both methods. While the present study was unable to determine the capability of in-air electrical stunning to induce immediate unconsciousness, our findings revealed that irreversible stuns were induced by both in-air electrical stunning (i.e., ∼25 to 70% of individuals did not recover VERs across the various combinations of stunning intensities and/or durations) and percussive stunning (i.e., ∼100% of individuals did not recover VERs). The efficacy of in-air electrical stunning for permanently abolishing VERs was marginally, but significantly, impacted by stun intensity (i.e., explained 8% of the variation). Furthermore, due to substantial inter-specific variability and a limited sample size, significant impacts of stun intensity and/or duration on the recovery of VERs in reversibly stunned individuals were not detected in the present study (i.e., VERs recovered between <0.5 to 28.8 min). Further investigation is therefore necessary before in-air electrical stunning can be endorsed as a standalone humane slaughter method for rainbow trout. This includes determining its capacity to induce immediate unconsciousness, as well as to identify additional factors that could be modified or enhanced to improve its efficacy. Furthermore, since death following in-air electrical stunning likely entails a prolonged process involving ventilatory failure, hypoxemia, and subsequent vital organ malfunction, rather than immediate cardiac arrest or central nervous system failure, the sequential use of methods such as percussive stunning is recommended to safeguard the welfare of rainbow trout during slaughter.

Probiotic, fructooligosaccharide and yeast extract mixture improves gut health in rainbow trout, Oncorhynchus mykiss.

The present study investigated the effects of a feed additive (FA) containing a probiotic consortium, fructooligosaccharide and yeast extract on growth performance, humoral immune responses, hepatic antioxidant parametersand intestine digestive enzymes, morphology and transcripts in rainbow trout, Oncorhynchus mykiss. The fish were reared for 8 weeks, feeding on diets containing 0 (CNT), 0.3 (0.3FA), 1 (1FA)and 2 (2FA) g/kg FA. The results showed that fish growth parameters were significantly and quadratically related to FA levelsand FA treatments had better growth performance than CNT treatment. Intestinal amylase activity significantly increased in 2FA, whereas, intestinal protease activity increased in all FA treatments. Intestinal villus length and muscular layer thickness significantly increased in 0.3FA treatment. Blood leucocyte and lymphocyte counts, plasma lysozyme activityand hepatic glutathione content significantly increased in 0.3FA and 1FA treatments; whereas hepatic malondialdehyde significantly decreased in these treatments. Blood neutrophil and monocyte counts significantly increased in 0.3FA treatment, while plasma alternative complement activity significantly increased in 1FA treatments. Plasma bactericidal activities against Aeromonas hydrophila, Yersinia ruckeriand Streptococcus iniae, and intestinal expression of heat shock protein 70 and beta-defensin significantly increased in all FA treatments. The abundance of A. hydrophila, Y. ruckeriand S. iniae in fish gut significantly decreased in 0.3FA treatment; these bacteria were absent in the intestines of 1FA and 2FA treatments. The present results suggest that dietary 0.3-1 g/kg of FA can significantly improve growth performance, immune response, intestinal healthand hepatic antioxidant capacity in rainbow trout.

Presence and Prevalence of Motile Aeromonas Species Isolated from Rainbow Trout (Oncorhynchus Mykıss) Offered for Consumption in Kars

This study aimed to determine the presence and prevalence of motile Aeromonas species in rainbow trout offered for consumption in Kars province. Muscle and skin tissues were examined from 50 raw trout, and out of 100 total samples, 42 were found positive for Aeromonas spp. While 20 (40.0 %) of the muscle samples were positive for motile Aeromonas spp., 22 (44.0 %) of the skin samples were also positive. Among these 42 positive samples, 24 % were identified as A. hydrophila, 38 % as A. caviae, 16 % as A. sobria, and 14 % contained both A. hydrophila and A. caviae. Consequently, it was determined that raw rainbow trout offered for sale in Kars is at risk with regard to motile Aeromonas spp. It is important to take the necessary hygiene and cooking precautions to minimize the possible danger for this situation that threatens public health.

Viral Diseases in Trout and Treatment Methods

Viral diseases cause significant economic losses in aquaculture enterprises in Turkey and around the world. In this review, information is given about the symptoms, diagnosis and treatment of infectious pancreatic necrosis (IPN), infectious hematopoietic necrosis (IHN), Viral hemorrhagic septicemia (VHS) diseases in rainbow trout (Oncorhynchus mykis) in different periods.

Novel insights on microbiome dynamics during a gill disease outbreak in farmed rainbow trout (Oncorhynchus mykiss)

The generic term “Gill disease” refers to a wide range of disorders that affect the gills and severely impact salmonid aquaculture systems worldwide. In rainbow trout freshwater aquaculture, various etiological agents causing gill diseases have been described, particularly Flavobacterium and Amoeba species, but research studies suggest a more complex and multifactorial aetiology. Here, a cohort of rainbow trout affected by gill disease is monitored both through standard laboratory techniques and 16S rRNA Next-Generation Sequencing (NGS) analysis during a natural disease outbreak and subsequent antibiotic treatment with Oxytetracycline. NGS results show a clear clustering of the samples between pre- and post-treatment based on the microbial community of the gills. Interestingly, the three main pathogenic bacteria species in rainbow trout (Yersinia ruckeri, Flavobacterium psychrophilum, and Flavobacterium branchiophilum) appear to be weak descriptors of the diversity between pre-treatment and post-treatment groups. In this study, the dynamics of the gill microbiome during the outbreak and subsequent treatment are far more complex than previously reported in the literature, and environmental factors seem of the utmost importance in determining gill disease. These findings present a potential novel perspective on the diagnosis and management of gill diseases, showing the limitations of conventional laboratory methodologies in elucidating the complexity of this disease in rainbow trout. To the authors’ knowledge, this work is the first to describe the microbiome of rainbow trout gills during a natural outbreak and subsequent antibiotic treatment. The results of this study suggest that NGS can play a critical role in the analysis and comprehension of gill pathology. Using NGS in future research is highly recommended to gain deeper insights into such diseases correlating gill’s microbiome with other possible cofactors and establish strong prevention guidelines.

Activin Signaling Pathway Specialization During Embryonic and Skeletal Muscle Development in Rainbow Trout (Oncorhynchus mykiss).

Activin signaling is essential for proper embryonic, skeletal muscle, and reproductive development. Duplication of the pathway in teleost fish has enabled diversification of gene function across the pathway but how gene duplication influences the function of activin signaling in non-mammalian species is poorly understood. Full characterization of activin receptor signaling pathway expression was performed across embryonic development and during early skeletal muscle growth in rainbow trout (RBT, Oncorhynchus mykiss). Rainbow trout are a model salmonid species that have undergone two additional rounds of whole genome duplication. A small number of genes were expressed early in development and most genes increased expression throughout development. There was limited expression of activin Ab in RBT embryos despite these genes exhibiting significantly elevated expression in post-hatch skeletal muscle. CRISPR editing of the activin Aa1 ohnolog and subsequent production of meiotic gynogenetic offspring revealed that biallelic disruption of activin Aa1 did not result in developmental defects, as occurs with knockout of activin A in mammals. The majority of gynogenetic offspring exhibited homozygous activin Aa1 genotypes (wild type, in-frame, or frameshift) derived from the mosaic founder female. The research identifies mechanisms of specialization among the duplicated activin ohnologs across embryonic development and during periods of high muscle growth in larval and juvenile fish. The knowledge gained provides insights into potential viable gene-targeting approaches for engineering the activin receptor signaling pathway and establishes the feasibility of employing meiotic gynogenesis as a tool for producing homozygous F1 genome-edited fish for species with long-generation times, such as salmonids.

Dopaminergic and anti-estrogenic responses in juvenile steelhead (Oncorhynchus mykiss) exposed to bifenthrin

The frequency of detection and concentrations of bifenthrin, a pyrethroid insecticide, in the waterways inhabited by the endangered species, steelhead trout (Oncorhynchus mykiss), has become a significant concern for regulatory agencies. Endocrine disruption has been observed with estrogenic and anti-estrogenic responses in fish species at different life stages. Since several studies have indicated alterations in dopaminergic signaling associated with endocrine responses, juvenile steelhead were exposed to environmentally relevant concentrations of 60 or 120 ng/L bifenthrin for two weeks. Fish brains were assessed for dopamine levels and the expression of genes involved in dopaminergic and estrogenic processes, such as catechol-o-methyltransferase (comt) and monoamine oxidase (mao). Vitellogenin (vtg) and estrogenic receptors (ERα1, ERβ1, and ERβ2) were also evaluated in livers of the animals. Dopamine concentrations were significantly higher in fish brains following bifenthrin exposure. Consistent with a reduction in dopamine clearance, there was a significant decrease in the mRNA expression of comt with increased bifenthrin concentration. Hepatic expression of ERα1 and ERβ2 mRNA was significantly decreased with increased bifenthrin concentration. These data support the possible mechanism of bifenthrin altering the dopaminergic pathway at low ng/L concentrations, in juvenile steelhead, which could interfere with endocrine feedback loops. These findings support the need for and importance of identifying species and life stage differences in pesticide modes of action to reduce uncertainties in risk assessments.

Body length determines flow refuging for rainbow trout (Oncorhynchus mykiss) behind wing dams.

Complex hydrodynamics abound in natural streams, yet the selective pressures these impose upon different size classes of fish are not well understood. Attached vortices are produced by relatively large objects that block freestream flow, which fish routinely utilize for flow refuging. To test how flow refuging and the potential harvesting of energy (as seen in Kármán gaiting) vary across size classes in rainbow trout (Oncorhynchus mykiss; fingerling, 8 cm; parr, 14 cm; adult, 22 cm; n=4 per size class), we used a water flume (4100 l; freestream flow at 65 cm s-1) and created vortices using 45deg wing dams of varying size (small, 15 cm; medium, 31 cm; large, 48 cm). We monitored microhabitat selection and swimming kinematics of individual trout and measured the flow field in the wake of wing dams using time-resolved particle image velocimetry (PIV). Trout of each size class preferentially swam in vortices rather than the freestream, but the capacity to flow refuge varied according to the ratio of vortex width to fish length (WV:LF). Consistent refuging behavior was exhibited when WV:LF≥1.5. All size classes exhibited increased wavelength and Strouhal number and decreased tailbeat frequency within vortices compared with freestream, suggesting that swimming in vortices requires less power output. In 17% of the trials, fish preferentially swam in a manner that suggests energy harvesting from the shear layer. Our results can inform efforts toward riparian restoration and fishway design to improve salmonid conservation.