Effects of dietary co-exposure to fungal and herbal functional feed additives on immune parameters and microbial intestinal diversity in rainbow trout (Oncorhynchus mykiss).

Misuse and overuse of antibiotics in aquaculture has proven to be an unsustainable practice leading to increased bacterial resistance. An alternative strategy involves the inclusion of immunostimulants in fish diets, especially fungal and herbal compounds already authorized for human consumption, hence without environmental or public health concerns. In this study, we used a holistic and cross-disciplinary pipeline to assess the immunostimulatory properties of two fungi: Trametes versicolor and Ganoderma lucidum; one herbal supplement, capsaicin in the form of Espelette pepper (Capsicum annuum), and a combination of these fungal and herbal additives on rainbow trout (Oncorhynchus mykiss). We investigated the impact of diet supplementation for 7 weeks on survival, growth performance, cellular, humoral, and molecular immune parameters, as well as the intestinal microbial composition of the fish. Uptake of herbal and fungal compounds influenced the expression of immune related genes, without generating an inflammatory response. Significant differences were detected in the spleen-tlr2 gene expression. Supplementation with herbal additives correlated with structural changes in the fish intestinal microbiota and enhanced overall intestinal microbial diversity. Results demonstrated that the different treatments had no adverse effect on growth performance and survival, suggesting the safety of the different feed additives at the tested concentrations. While the mechanisms and multifactorial interactions remain unclear, this study provides insights not only in regard to nutrition and safety of these compounds, but also how a combined immune and gut microbiota approach can shed light on efficacy of immunostimulant compounds for potential commercial inclusion as feed supplements.

The extensive transgenerational transcriptomic effects of ocean acidification on the olfactory epithelium of a marine fish are associated with a better viral resistance.

BACKGROUND: Progressive CO2-induced ocean acidification (OA) impacts marine life in ways that are difficult to predict but are likely to become exacerbated over generations. Although marine fishes can balance acid-base homeostasis efficiently, indirect ionic regulation that alter neurosensory systems can result in behavioural abnormalities. In marine invertebrates, OA can also affect immune system function, but whether this is the case in marine fishes is not fully understood. Farmed fish are highly susceptible to disease outbreak, yet strategies for overcoming such threats in the wake of OA are wanting. Here, we exposed two generations of the European sea bass (Dicentrarchus labrax) to end-of-century predicted pH levels (IPCC RCP8.5), with parents (F1) being exposed for four years and their offspring (F2) for 18 months. Our design included a transcriptomic analysis of the olfactory rosette (collected from the F2) and a viral challenge (exposing F2 to betanodavirus) where we assessed survival rates. RESULTS: We discovered transcriptomic trade-offs in both sensory and immune systems after long-term transgenerational exposure to OA. Specifically, RNA-Seq analysis of the olfactory rosette, the peripheral olfactory organ, from 18-months-old F2 revealed extensive regulation in genes involved in ion transport and neuronal signalling, including GABAergic signalling. We also detected OA-induced up-regulation of genes associated with odour transduction, synaptic plasticity, neuron excitability and wiring and down-regulation of genes involved in energy metabolism. Furthermore, OA-exposure induced up-regulation of genes involved in innate antiviral immunity (pathogen recognition receptors and interferon-stimulated genes) in combination with down-regulation of the protein biosynthetic machinery. Consistently, OA-exposed F2 challenged with betanodavirus, which causes damage to the nervous system of marine fish, had acquired improved resistance. CONCLUSION: F2 exposed to long-term transgenerational OA acclimation showed superior viral resistance, though as their metabolic and odour transduction programs were altered, odour-mediated behaviours might be consequently impacted. Although it is difficult to unveil how long-term OA impacts propagated between generations, our results reveal that, across generations, trade-offs in plastic responses is a core feature of the olfactory epithelium transcriptome in OA-exposed F2 offspring, and will have important consequences for how cultured and wild fish interacts with its environment.

Changes in defense capacity to infectious hematopoietic necrosis virus (IHNv) in rainbow trout intergenerationally exposed to glyphosate.

Glyphosate, the most widely used herbicide active substance worldwide, has raised many scientific, political and public debates in the context of its recent re-registration in the European Union, highlighting in particular a lack of data concerning its potential generational effects. In this study, we investigated the intergenerational toxicity of this active substance used alone or coformulated in glyphosate-based herbicides (GBHs) on the ability of rainbow trout (Oncorhynchus mykiss) to face a viral challenge. Juvenile trout from parents exposed for eight months to four different chemical exposure conditions (non-exposed control, pure glyphosate, Roundup Innovert®, and Viaglif Jardin® were experimentally infected with the infectious hematopoietic necrosis virus (IHNv). Various enzymatic and hemato-immunological markers were assessed before and after the viral challenge. Chemical contamination with GBHs strongly modulated viral trout susceptibility. Pure glyphosate induced a cumulative mortality of 35.8%, comparable to the control (37.0%), which was significantly reduced with Roundup Innovert® (-9.9%) and increased (+14.8%) with Viaglif Jardin®. No modification was observed for the biomarkers analysed for any conditions. These results demonstrate that the nature of the co-formulants associated to glyphosate in GHBs can modulate the susceptibility of fish to pathogens.

Growth and immune system performance to assess the effect of dispersed oil on juvenile sea bass (Dicentrarchus labrax).

The potential impact of chemically and mechanically dispersed oil was assessed in a model fish of European coastal waters, the sea bass Dicentrarchus labrax. Juvenile sea bass were exposed for 48h to dispersed oil (mechanically and chemically) or dispersants alone. The impact of these exposure conditions was assessed using growth and immunity. The increase observed in polycyclic aromatic hydrocarbon metabolites in bile indicated oil contamination in the fish exposed to chemical and mechanical dispersion of oil without any significant difference between these two groups. After 28 days of exposure, no significant differences were observed in specific growth rate,apparent food conversion efficiency and daily feeding). Following the oil exposure, fish immunity was assessed by a challenge with Viral Nervous Necrosis Virus (VNNV). Fish mortality was observed over a 42 day period. After 12 days post-infection, cumulative mortality was significantly different between the control group (16% p≤0.05) and the group exposed to chemical dispersion of oil (30% p≤0.05). However, at the end of the experiment, no significant difference was recorded in cumulative mortality or in VNNV antibodies secreted in fish in responses to the treatments. These data suggested that in our experimental condition, following the oil exposure, sea bass growth was not affected whereas an impact on immunity was observed during the first days. However, this effect on the immune system did not persist over time.

Innate immunity and antioxidant systems in different tissues of sea bass (Dicentrarchus labrax) exposed to crude oil dispersed mechanically or chemically with Corexit 9500.

The aim of the study was to evaluate effects of chemically dispersed oil by the dispersant Corexit 9500 on innate immunity and redox defenses in a marine model fish. Sea bass (Dicentrarchus labrax) were exposed 48h to four experimental conditions: a control group (C), a group only exposed to the dispersant (D; 3.6mg/L) and two groups exposed to 80mg/L oil mechanically or chemically dispersed (MD; CD). Alternative pathway of complement activity and lysozyme concentration was measured in plasma in order to evaluate the general fish health status. Total glutathione, glutathione peroxidase (GPX) and superoxide dismutase (SOD) were analyzed in gills, liver, brain, intestine and muscle. The chemical dispersion induced a significant reduction of lysozyme concentration when compared to the controls, and the hemolytic activity of the alternative complement pathway was increased in mechanical and chemical dispersion. The analysis of SOD, GPX and total glutathione showed that antioxidant defenses were activated in liver and reduced in intestine and brain. Dispersant was also responsible for an SOD activity inhibition in these two last tissues, demonstrating a direct effect of this dispersant on reactive oxygen species homeostasis that can be interpreted as a signal of tissue toxicity. This result should raise concern about the use of dispersants and show that they can lead to adverse effects on marine species.

Effects of in vivo chronic exposure to pendimethalin on EROD activity and antioxidant defenses in rainbow trout (Oncorhynchus mykiss).

Pendimethalin, an herbicide active substance frequently used in terrestrial systems, has detected in European aquatic ecosystems. Reliable indicators still need to be found in order to properly assess the impact of pesticides in fish. After an in vivo chronic exposure to pendimethalin, the detoxification process and the antioxidant defense system were assessed in 120 adult rainbow trout, Oncorhynchus mykiss. Four nominal exposure conditions were tested: control (C), 500 ng L(-1) (P500), 800 ng L(-1) (P800) and the commercial formulation Prowl(®) at 500 ng L(-1) (Pw500). Fish samples were made after a 28 day exposure period (D28) and after a fifteen day recovery period in clean fresh water (D43). At D28, ethoxyresorufin-O-deethylase (EROD) activity was not activated in liver in spite of the pendimethalin uptake in fish. At D43, EROD activity in fish exposed to the commercial product was lower than in control fish, which may be explained by the high presence of herbicide in fish (613±163 ng g bile(-1)). Furthermore, antioxidant defense responses were set up by trout in gills and liver following chronic exposure to 800 ng L(-1) of pendimethalin concentration. While the glutathione content (GSH) decreased in gills, it increased in liver associated with higher activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD). These disturbances could lead to reactive oxygen species production and oxidative stress in the vital organs in fish. After fifteen days in clean water, while the SOD activity was restored, the GSH content and GPx activity were still significantly disturbed in fish exposed to pendimethalin in comparison with control. These significant differences between treatments in antioxidant defenses parameters measured, attesting to the irreversibility of the effects.

Effect of chronic exposure to pendimethalin on the susceptibility of rainbow trout, Oncorhynchus mykiss L., to viral hemorrhagic septicemia virus (VHSV).

In this study, the in vivo effects of chronic pollution by the active substance (AS) pendimethalin, a dinitroaniline herbicide, on the susceptibility of rainbow trout, Oncorhynchus mykiss L., to an experimental challenge with viral hemorrhagic septicemia virus (VHSV) were assessed. After four weeks of exposure to fresh water (C group) or 500 ng L(-1) of AS (P500 group), the fish were challenged by immersion in water containing 10(4) TCID(50) mL(-1) of VHSV. While exposure to pendimethalin was maintained throughout the experiment, mortalities were recorded during the 40 days post-infection (dpi) and organs were collected from dead fish for virological examination. At the end of the experiment, anti-VHSV antibodies and the classical pathway of complement activity were assessed in trout plasma. Exposure to pendimethalin significantly affected the distribution of cumulative mortality accelerating death in fish infected by VHSV. Pendimethalin appeared to decrease the Mean Time to Death (MTD) after virus treatment from 14.9 days (C-VHSV) to 10.2 days (P500-VHSV). Nevertheless, by the end of the experiment, differences in cumulative mortality were no longer observed between the two groups, which had reached the same stage (50 percent). Furthermore, a higher concentration of the virus was recovered from the pools of organs from the P500-VHSV group than the C-VHSV group. Moreover, at 40 dpi, although no significant difference was observed in the immune response between the two groups, more fish in the P500-VHSV group had set up an immune response by secreting antibodies than in the control viral group (C-VSHV).

The relationships between growth rate and mitochondrial metabolism varies over time.

Mitochondrial metabolism varies significantly between individuals of the same species and can influence animal performance, such as growth. However, growth rate is usually determined before the mitochondrial assay. The hypothesis that natural variation in mitochondrial metabolic traits is linked to differences in both previous and upcoming growth remains untested. Using biopsies to collect tissue in a non-lethal manner, we tested this hypothesis in a fish model (Dicentrarchus labrax) by monitoring individual growth rate, measuring mitochondrial metabolic traits in the red muscle, and monitoring the growth of the same individuals after the mitochondrial assay. Individual variation in growth rate was consistent before and after the mitochondrial assay; however, the mitochondrial traits that explained growth variation differed between the growth rates determined before and after the mitochondrial assay. While past growth was correlated with the activity of the cytochrome c oxidase, a measure of mitochondrial density, future growth was linked to mitochondrial proton leak respiration. This is the first report of temporal shift in the relationship between growth rate and mitochondrial metabolic traits, suggesting an among-individual variation in temporal changes in mitochondrial traits. Our results emphasize the need to evaluate whether mitochondrial metabolic traits of individuals can change over time.

Glyphosate-based herbicide exposure: effects on gill microbiota of rainbow trout (Oncorhynchus mykiss) and the aquatic bacterial ecosystem.

The herbicide glyphosate has been widely used in the past 40 years, under the assumption that side effects were minimal. In recent years, its impact on microbial compositions and potential indirect effects on plant, animal and human health have been strongly suspected. Glyphosate and co-formulates have been detected in various water sources, but our understanding of their potential effects on aquatic animals is still in its infancy compared with mammals. In this study, we investigated the effect of chronic exposure to an environmentally relevant concentration of glyphosate on bacterial communities of rainbow trout (Oncorhynchus mykiss). Gills, gut contents and gut epithelia were then analyzed by metabarcoding targeting the 16S rRNA gene. Our results revealed that rainbow trout has its own bacterial communities that differ from their surrounding habitats and possess microbiomes specific to these three compartments. The glyphosate-based herbicide treatment significantly affected the gill microbiome, with a decrease in diversity. Glyphosate treatments disrupted microbial taxonomic composition and some bacteria seem to be sensitive to this environmental pollutant. Lastly, co-occurrence networks showed that microbial interactions in gills tended to decrease with chemical exposure. These results demonstrate that glyphosate could affect microbiota associated with aquaculture fish.

Bioconcentration and immunotoxicity of an experimental oil spill in European sea bass (Dicentrarchus labrax L.).

The effects of Polycyclic Aromatic Hydrocarbons (PAHs) resulting from a water soluble fraction (WSF) of an Arabian crude oil were tested in vivo on the bioconcentration in muscles and on immune parameters in sea bass, Dicentrarchus labrax. After 15 days of acclimation, fish were acutely exposed (48 h) to the WSF of 25 g of oil, and then returned to clean sea water for a 15 day recovery period. PAH concentration in the WSF at the beginning of the exposure was estimated to 773±187 ng L⁻¹ similar to that observed in the marine environment after an oil spill. The WSF in the experimental system was composed by lightest PAH compounds and did not remain constant during the two days of exposure. Just after exposure to the WSF, a total mean concentration of 148±46 µg kg⁻¹ of PAHs was found in contaminated fish muscle, composed of parent and alkylated naphthalene compounds (86.5%), benzo[a]pyrene (10.1%) and benzo[b+k]fluoranthene (3.4%). In addition, a decrease of leucocytes counts due to a lymphopenia and granulopenia and an increase of the haemolytic activity of the alternative pathway (ACH50) were noted. After a 15 day recovery period, haematocrit was decreased whereas effects on the blood granulocytes of fish seemed to be reversible, contrary to the specific immune system and quality of flesh. In fact, contaminated fish had still less lymphocyte cells compared to controls fish and their flesh were still contaminated by naphthalene and benzo[a]pyrene creating a risk for human consumers.

Effect of an experimental oil spill on vertebral bone tissue quality in European sea bass (Dicentrarchus labrax L.).

In order to identify biomarkers of oil pollution in fish we tested the effects of an experimental Light Cycle Oil (LCO) exposure on vertebral bone of sea bass, Dicentrarchus labrax L. A total of 60 adult fish were acclimated for fifteen days, then twenty were collected as controls (Day 0) while 40 were exposed to a soluble fraction of LCO (1136 ng L(-1) of ten Polycyclic Aromatic Hydrocarbons, PAHs) for seven days. Twenty of them were sampled at the end of the exposure period and the twenty last after a recovery period of fourteen days in clean seawater. Vertebral abnormalities were counted and bone mineralization, total bone area and bone density profiles were established for several post-cranial and caudal vertebrae. In sea bass, seven days of LCO exposure did not affect the frequency and severity of the vertebral abnormalities. No significant differences were observed in bone density and bone repartition (parameters of bone area profiles) between unexposed (Day 0), exposed (D7) and decontaminated (D21) fish. In contrast, bone mineralization of the vertebrae decreased in contaminated sea bass, but in a reversible way, which confirms a previous study in trout showing that this parameter is an early stress indicator. Our results suggest that vertebral bone mineralization could be used as a biomarker of PAH pollution in sea bass. It would be interesting to check this new biomarker in other teleost species exposed to various xenobiotics.

Impact of chronic exposure of rainbow trout, Oncorhynchus mykiss, to low doses of glyphosate or glyphosate-based herbicides.

Glyphosate is an herbicidal active substance (AS) entering in the composition of a large diversity of pesticide products (glyphosate-based herbicides; GBH) used in modern intensive agriculture. This compound has a favorable environmental safety profile but was suspected to induce deleterious effects in aquatic organisms, with a potential effect of some associated co-formulants. This study aimed to assess the impact of direct and chronic exposure to glyphosate on the health status of rainbow trout, Oncorhynchus mykiss. A total of 36 genitors were exposed daily for 10 months to a dose of glyphosate representative of environmental concentrations (around 1 µg L-1) using the AS alone or two GBHs formulations (i.e. Roundup Innovert® and Viaglif Jardin®) and findings were compared to an unexposed control group (n=12). The effects of chemical exposure on the reproductive capacities, hemato-immunologic functions, energetic metabolism, oxidative stress and specific biomarkers of exposure were analyzed over a period of 4 months covering spawning. A limited mortality between 15% and 30% specific to the spawning occurred under all conditions. No differences were observed in reproduction parameters i.e. mean weights, relative fertility and fecundity. Red blood cell count, hematocrit index, mean corpuscular volume and white blood cell counts were similar for all the sampling dates. Significant changes were observed two months before spawning with a 70% decrease of the proportion of macrophages in trout exposed to Viaglif only and a reduction of 35% of the phagocytic activity in fish exposed to the two GBHs. Trends towards lower levels of expression of tumor necrosis factor-α (between 38% and 66%) were detected one month after the spawning for all contaminated conditions but without being statistically significant. Biomarkers of exposure, i.e. acetylcholine esterase and carbonic anhydrase activities, were not impacted and none of the chemical contaminants disturbed the oxidative stress or metabolism parameters measured. These results suggest that a 10 months exposure of rainbow trout to a concentration of 1 µg L-1 of glyphosate administered using the pure active substance or two GBHs did not significantly modify their global health including during the spawning period. The immunological disturbances observed will need to be further explored because they could have a major impact in response to infectious stress.

Comparative analysis of the course of infection and the immune response in rainbow trout (Oncorhynchus mykiss) infected with the 5 genotypes of infectious hematopoietic necrosis virus.

Infectious hematopoietic necrosis virus (IHNV) is a pathogen of importance for salmonid aquaculture. In this study, we aimed to characterize virus behavior and defense mechanisms developed in rainbow trout (RT, Oncorhynchus mykiss) experimentally infected with isolates belonging to the five described genotypes of IHNV, i.e. L, U, M, E and J. Mortality was monitored for two months, and blood and target organs were sampled at different times post-infection to assess viral load and cellular and humoral immune responses. Profiles of virulence were highly linked to precocious viral replication but also to the innate and specific immunity elicited in the host. Seroneutralization test (SNT) used for specific antibodies detection exhibited reliable results, with efficient cross-neutralization observed in heterologous systems except for the Asian representative. These data bring new insights about IHNV/RT interaction and reinforce the interest of SNT as preventive and epidemiological tool.

Immunological and metabolic effects of acute sublethal exposure to glyphosate or glyphosate-based herbicides on juvenile rainbow trout, Oncorhynchus mykiss.

Glyphosate is a commonly used agrochemical active substance co-formulated in glyphosate-based herbicides (GBHs) whose environmental safety is still a subject of debate in the European Union. We evaluated the effects of acute sublethal exposure to glyphosate on rainbow trout by measuring changes in their metabolic and hemato-immunologic functions and their ability to survive a viral challenge. Juvenile fish were exposed for 96 h to 500 µg L-1 of glyphosate through the active substance alone or two GHBs, Roundup Innovert® and Viaglif Jardin®, and fish were then infected with the infectious hematopoietic necrosis virus. Red and white blood cell counts (RBCC and WBCC), as well as several enzymatic activities (citrate synthase, CS; cytochrome-c oxidase, CCO; lactate dehydrogenase, LDH; glucose-6-phosphate dehydrogenase, G6PDH; acetylcholinesterase, AChE), were measured 96 h after chemical contamination (S1), and 96 h post-viral infection (S2). Mortality rates were monitored, and virus titers at the mortality peaks and seropositivity of the survivors were analyzed at 60 days post-viral infection (S3). Cumulative mortalities, viral titers, and seropositivity induced by virus infection were similar among conditions. Hematological analysis revealed significant increases of 30% for RBCC for Roundup at S1, and of 22% for WBCC at S2. No changes were observed in metabolic enzyme activities at S1. At S2, CCO and G6PDH activities were significantly higher than controls in all the chemically contaminated groups (+61 to 62% and +65 to 138%, respectively). LDH and AChE activities were increased for the Viaglif (p = 0.07; +55%) and for glyphosate and Roundup conditions (p < 0.05, +62 to 79%), respectively. Rainbow trout acutely exposed to glyphosate or GBHs presented no major physiological changes. Viral infection revealed disruptions, potentially modulated by co-formulants, of hematological and metabolic parameters, showing that it is essential to consider the stressful natural environment of fish in the chemical assessment.

Developmental effect of parental or direct chronic exposure to environmental concentration of glyphosate on the larvae of rainbow trout, Oncorhynchus mykiss.

The environmental safety profile of glyphosate, the most commonly used herbicide worldwide, is still a subject of debate and little is known about the generational toxicity of this active substance (AS) and the associated commercial formulations called "glyphosate-based herbicides" (GBHs). This study investigated the impact of parental and direct exposure to 1µgL-1 of glyphosate using the AS alone or one of two GBH formulations (i.e. Roundup Innovert® and Viaglif Jardin®) in the early developmental stages of rainbow trout. Three different modes of exposure on the F1 generation were studied: (1) intergenerational (i.e. fish only exposed through their parents); (2) direct (i.e. fish exposed only directly) and (3) multigenerational (i.e. fish both exposed intergenerationally and directly). The impact of chemical treatments on embryo-larval development (survival, biometry and malformations), swimming behaviour, biochemical markers of oxidative stress equilibrium (TBARS and catalase), acetylcholine esterase (AChE) and energy metabolism (citrate synthase, CS; cytochrome-c oxidase, CCO; lactate dehydrogenase, LDH; glucose-6-phosphate dehydrogenase, G6PDH) was explored. Chemical exposure did not affect the survival of F1 embryos or malformation rates. Direct exposure to the AS induced some biometric changes, such as reduction in head size (with a 10% decrease in head length), independently of co-formulants. Intergenerational exposure to the AS or the Roundup GBH increased swimming activity of the larvae, with increase of between 78 and 102% in travel speeds. Viaglif co-formulants appear to have counteracted this behavioural change. The minor changes detected in the assayed biochemical markers suggested that observed effects were not due to oxidative damage, AChE inhibition or alterations to energy metabolism. Nonetheless, multi- and intergenerational exposure to Roundup increased CS:CCO and LDH:CS ratios by 46% and 9%, respectively, with a potential modification of the aerobic-to-anaerobic energy production balance. These biochemical effects were not correlated with those observed on individual level of biological organization. Therefore, further studies on generational toxicity of glyphosate and its co-formulants are needed to identify the other mechanisms of glyphosate toxicity at the cellular level.

Generational effects of a chronic exposure to a low environmentally relevant concentration of glyphosate on rainbow trout, Oncorhynchus mykiss.

In the past few decades, glyphosate became the most used herbicide substance worldwide. As a result, the substance is ubiquitous in surface waters. Concerns have been raised about its ecotoxicological impact, but little is known about its generational toxicity. In this study, we investigate the impact of an environmentally relevant concentration of glyphosate and its co-formulants on an F2 generation issued from exposed generations F0 and F1. Trans, inter and multigenerational toxicity of 1 µgL-1 of the active substance was evaluated on early stages of development and juvenile rainbow trout (Oncorhynchus mykiss) using different molecular, biochemical, immuno-hematologic, and biometric parameters, behavior analysis, and a viral challenge. Reproductive parameters of generation F1 were not affected. However, developmental toxicity in generation F2 due to glyphosate alone or co-formulated was observed with head size changes (e.g. head surface up to +10%), and metabolic disruptions (e.g. 35% reduction in cytochrome-c-oxidase). Moreover, larvae exposed transgenerationally to Viaglif and intergenerationally to glyphosate and Roundup presented a reduced response to light, potentially indicating altered escape behavior. Overall methylation was, however, not altered and further experiments using gene-specific DNA metylation analyses are required. After several months, biochemical parameters measured in juvenile fish were no longer impacted, only intergenerational exposure to glyphosate drastically increased the susceptibility of rainbow trout to hematopoietic necrosis virus. This result might be due to a lower antibody response in exposed fish. In conclusion, our results show that generational exposure to glyphosate induces developmental toxicity and increases viral susceptibility. Co-formulants present in glyphosate-based herbicides can modulate the toxicity of the active substance. Further investigations are required to study the specific mechanisms of transmission but our results suggest that both non-genetic mechanisms and exposure during germinal stage could be involved.

Tissue distribution, metabolism, and elimination of Victoria Pure Blue BO in rainbow trout: Main metabolite as an appropriate residue marker.

Victoria Pure Blue BO is a dye that bears some therapeutic activity and that can be retrieved in effluent or may be used in aquaculture as a prohibited drug. In this study, the metabolism and tissue distribution during uptake and depuration of VPBO were investigated in order to propose a residue marker of illegal treatment in fish. The dye was administered to rainbow trout (oncorhynchus mykiss) for one day by water bath at a dose of 0.1 mg.L-1. The concentrations of VPBO in all tissues increased rapidly during the treatment period, reaching a Cmax of 567 ± 301 µg.L-1 in plasma and 1846 µg kg-1 ±517 for liver after 2 h. After placing the rainbow trout in a clean water bath for a 64 day-period of depuration, the concentrations in the tissues and plasma decreased to reach comparable levels for muscle and for skin after 33 days. The concentrations measured were still above the LOQ at 2.26 ± 0.48 µg kg-1 for muscle and 2.85 ± 1.99 µg kg-1 for skin at the end of the depuration period. The results indicated the existence of 14 phase I metabolites and one glucuronide conjugated metabolite. Non-compartmental analysis was applied to assess the pharmacokinetic parameters. The half-life in edible muscle of the main metabolite detected, deethyl-leuco-VPBO, was found to be 22.5 days compared to a half-life of 19.7 days for the parent VPBO. This study provides new information to predict a VPBO drug treatment of aquacultured species via a proposed new residue marker.

Why Do Some Rainbow Trout Genotypes Grow Better With a Complete Plant-Based Diet? Transcriptomic and Physiological Analyses on Three Isogenic Lines.

Within the context of a growing aquaculture production coupled with a plateau of the production in the main components of aquafeeds (fish oil and fishmeal), recent studies have typically focused on replacing these feedstuffs with terrestrial plant ingredients for cultured carnivorous aquatic species, such as rainbow trout (Oncorhynchus mykiss). Substitution rates without adverse effects have, however, reached their limit. One potential way forward would be to take advantage of the genetic variability that exists in the salmonid population. However, to date, little is known about the underlying molecular mechanisms responsible for this genetic variability. The aim of the present research was to understand why some genotypes are better able to utilize plant-based diets devoid of marine resources. In this regard, three isogenic lines of rainbow trout (R23h, AB1h, and A22h), with similar growth when fed marine resources-based diets and which differ greatly in their responses to a plant-based diet, were fed with either a complete plant-based diet (V diet) or a marine resources-based diet (M diet) since first-feeding. Fish traits and the hepatic transcriptome of these three genotypes were compared after 5 months of feeding. First, differences in the ability to grow with the V diet observed amongst genotypes was not due to higher feed intake, but instead due to differences in feed efficiency. The comparison of transcriptome profiles revealed 575 (R23h vs. AB1h), 1,770 (R23h vs. A22h), and 2,973 (AB1h vs. A22h) probes differentially expressed amongst the three genotypes when fed the V diet. Interestingly, R23h and AB1h fish, which were the least affected by the V diet, exhibited the highest growth. These results demonstrate that these fish were able to maintain a high level of energy production and protein synthesis. Moreover, these genotypes were also able to activate pathways linked to lipid and cholesterol metabolisms, such as the biosynthesis of long-chain polyunsaturated fatty acids. Finally, as previously, immunity seems to also play an important role in the ability of fish to use the V diet, and further studies are needed to understand the mechanisms by which immunity interacts with growth.

Environmental samples of microplastics induce significant toxic effects in fish larvae.

Microplastics (MPs) are present throughout aquatic ecosystems, and can be ingested by a wide variety of organisms. At present, the physical and chemical effects of environmental MPs on aquatic organisms are poorly documented. This study aims to examine the physiological and behavioral effects caused by fish consuming environmental microplastics at different life stages. MP samples were collected from beaches on three islands (Easter Island, Guam and Hawaii) located near the North and South gyres of the Pacific Ocean. Larvae and juveniles of Japanese Medaka were fed for 30days with three doses of MPs (0.01, 0.1 and 1% w/w in fish food) approximate to the concentrations measured in moderately and heavily contaminated ocean areas. Ingestion of MPs by medaka larvae caused (variously) death, decreased head/body ratios, increased EROD activity and DNA breaks and, alterations to swimming behavior. A diet of 0.1% MPs was the most toxic. Two-month-old juveniles fed with 0.01% MPs did not exhibit any symptoms except an increase in DNA breaks. Our results demonstrate ingestion and mainly sublethal effects of environmental MPs in early life stages of fish at realistic MP concentrations. The toxicity of microplastics varies from one sample to another, depending on polymer composition, weathering and pollutant content. This study examines the ecological consequences microplastic build-up in aquatic ecosystems, more particularly in coastal marine areas, which serve as breeding and growing grounds for a number of aquatic species.