Ce travail de thèse a eu pour objectif d’identifier les mécanismes majeurs intervenant dans la régulation de l’adiposité musculaire chez la truite arc-en-ciel. Pour cela, nous avons analysé les effets combinés de la sélection génétique et de l’alimentation, facteurs prépondérants de variation de l'adiposité. Deux lignées de truites arc-en-ciel sélectionnées sur la teneur en lipides du muscle dorsal ("muscle gras" et "muscle maigre"), ont été nourries pendant 6 mois avec un régime contenant 10 ou 23% de lipides (% de la matière sèche). Nous avons mesuré l'activité et/ou l'expression d’enzymes clé des principales voies métaboliques intervenant dans l'utilisation de l'énergie, puis développé une analyse différentielle globale à l’échelle du transcriptome (microarray nylon) et du protéome (électrophorèse bidimensionnelle). Ces analyses portent sur le muscle blanc, tissu cible de la sélection, et le foie, carrefour métabolique et site majeur de la lipogenèse chez les poissons. Les résultats obtenus confirment l’effet inhibiteur d’un apport alimentaire riche en lipides sur la lipogénèse et la désaturation des acides gras dans le foie, déjà observé chez des individus de plus grande taille, et fournissent de nouvelles connaissances sur l’effet exercé sur les autres voies, en particulier la protéolyse. Ces analyses ont également permis de mettre en évidence des différences métaboliques existant entre lignées, qui concernent non seulement le métabolisme des lipides mais aussi celui des autres substrats énergétiques. Il apparaît que les deux moyens utilisé pour augmenter la teneur en lipides du muscle mettent en jeu des mécanismes moléculaires différents. Nos travaux ont permis d’identifier deux gènes dont l’expression est augmentée dans le muscle en réponse à un apport alimentaire riche en lipides et par la sélection génétique en faveur d’un indice d’adiposité musculaire élevé, et qui pourraient être des marqueurs moléculaires de l’adiposité musculaire. / The objective of the study was to identify genes and proteins that are involved in the control of muscle fat deposition in rainbow trout. We analyzed the combined effects exerted by genetic selection and dietary treatment, which are the two main factors that can be used to manage body fat content. Two lines of rainbow trout, obtained after 3 generations of divergent selection for high or low muscle fat content, were fed diets containing either 10% or 23% lipids (% dry matter), for six months. We analyzed the activity and gene expression of key enzymes involved in energy utilization, and performed a more global approach through transcriptome (nylon microarray) and proteome (two- dimensional electrophoresis) analysis. We analyzed the liver, which is the centre of intermediary metabolism and the main site of lipogenesis in fish, and the muscle, the target tissue of the selection provedure. The results confirmed the depressing effect exerted by a lipid rich diet on lipogenesis and fatty acid desaturation, already described in larger size fish, and provided new insight about the effect exerted on the other metabolic pathways, in particular the proteolysis. These analyses pointed out metabolic differences existing between lines. They involved not only lipid metabolism, but also the other pathways of nutrient utilization. With regard to their muscle-fattening effect, the dietary treatment and the genetic selection appear to act through different molecular mechanisms. These analyses allowed the identification of two genes that are over-expressed in muscle upon both high dietary lipid supply and upward selection for muscle fat content, suggesting that these two genes could be relevant molecular markers of muscle fattening.

L'acide rétinoïque (AR) régule la myogénèse embryonnaire. Dans le cadre de ce projet de thèse, nous avons d'une part utilisé l'AR pour moduler la myogénèse embryonnaire, dans la perspective d'étudier les conséquences de cette modulation sur le potentiel ultérieur de croissance et identifier les mécanismes moléculaires mobilisés.D'autre part, nous avons étudié la fonction de deux gènes régulés par l'AR et susceptibles de participer au contrôle de la myogénèse embryonnaire.La première partie du travail a été réalisée sur les modèles truite et poisson-zèbre. Nous avons montré que chez la truite comme chez le poisson zèbre, une incubation dans l'AR entrainait une activation de l'expression de Fgf8et de la différenciation des fibres musculaires rapides. Toutefois, chez la truite, nous n'avons pas pu mettre en évidence de régulation des MRF, indiquant qu'une autre voie est utilisée pour activer la myogénèse chez cette espèce.Dans la seconde partie de ce travail, la fonction de deux gènes régulés par l'AR et exprimés dans le mésoderme a été étudiée chez le poisson-zèbre. Le gène vertnin est exprimé essentiellement dans le tailbud. Quand il est inactivé par injection d'un oligo nucléotide morpholino antisens, on observe une altération de la formation des somites (mais pas de modification apparente du processus de segmentation) et une altération de l'intégrité des fibres lentes. Les fibres lentes sont en effet irrégulièrement espacées et les espaces au niveau des myoseptes verticaux peuvent être anormalement larges et les jonctions myotendineuses mal formées. Le gène arrestine β2aest exprimé dans les somites néo-formés puis également dans le mésoderme présomitique et le tailbud. Son inactivation par injection d'OM antisens entraine l'apparition du phénotype U-type et une altération de la morphologie des fibres lentes avec des fibres qui se détachent des jonctions myotendineuses. / Retinoic acid (RA) regulates embryonic myogenesis. During this thesis project, we first used RA to modulate embryonic myogenesis in order to study consequences of this modulation on the future potential for growth and to identify the underlying molecular mechanisms. Second part deals with the characterisation of the function of two genes regulated by the RA which may be involved in the control of embryonic myogenesis.The first part of the work was performed on the trout and zebrafish models. We have shown that in trout as in zebrafish, incubation in RA produced an activation of Fgf8 expression and differentiation of fast muscle fibers.However in trout, we did not observed regulation of MRF expression indicating that an alternative pathway isused to activate myogenesis in this species.In the second part of this work, the function of two genes regulated by the RA and expressed in the mesodermwas studied in zebrafish. The vertnin gene is expressed primarily in the tailbud. When it is inactivated by injection of antisense morpholino oligonucleotide, there is an alteration in the somites morphogenesis (but no apparent change in the process of segmentation) and impairment of the integrity of the slow muscle fibers. Slowfibers are indeed irregularly spaced and the vertical myosepta can be abnormally large. In addition myotendinous junctions display some abnormal branches. The arrestin β 2a gene is expressed in last formed somites and then also in the presomitic mesoderm and the tailbud. Its inactivation by injection of antisense MO leads to the appearance of the U-type phenotype and alteration of the slow muscle fibers morphology which detach frommyotendinous junctions

Over recent decades recreational fisheries have grown substantially throughout the world. Despite this increase, catches from recreational fisheries have often been ignored in fisheries management, although this is now being remedied. Monitoring recreational fisheries can be expensive, and the primary means used for monitoring is angler (creel) surveys, typically funded from sales of fishing licences. The studies presented in this thesis examine different approaches to monitoring recreational trolling fisheries’ catch and effort, where fishing licenses are not required and there are no reporting requirements. I present results from a complemented roving/mail-in survey undertaken during 2013-2014 to estimate recreational effort and catch of Atlantic salmon (Salmo salar) and brown trout (S. trutta) in the largest lake in the European Union, Lake Vänern, Sweden. I also evaluate different angler catch reporting methods (mail-in, tournament reports and face-to-face interviews) and compare catch rates within and among spring and fall fishing periods. In addition, mail-in survey data are examined for recall bias.   I estimate that 28.7 tonnes of salmon and trout combined were harvested by the recreational trolling fishery in 2014, more than the commercial and subsistence fisheries combined. Seasonal differences in both recreational effort and catch were observed. Effort, in boat hours, was significantly higher in spring than in fall. Catch rates of trout were higher in fall than in spring, but there were no seasonal differences in catches of salmon. Harvest per boat day did not differ significantly among catch reporting methods, indicating that all three methods could be useful for managers interested in harvest rates. In contrast, total and released catch per boat day differed among reporting methods, with tournament anglers catching more fish in total. Finally, there was little evidence for recall bias in mail-in surveys, indicating that mail-in surveys are useful for collecting unbiased catch data. My study is the most comprehensive angler survey to date for Lake Vänern, and my results should be of immediate use to local fisheries managers and should also be of interest to researchers and managers interested in estimating catch and effort for fisheries at large spatial scales. / Over the past several decades recreational fisheries have grown substantially throughout the world. Until recently, however, recreational catch has been ignored in the management of many important fisheries. The studies presented in this thesis examine different approaches to monitoring recreational trolling fisheries’ catch and effort in Lake Vänern, Sweden, the largest lake in the European Union. Paper I presents results from a complemented roving/mail-in survey, designed to estimate recreational effort and catch of Atlantic salmon (Salmo salar) and brown trout (S. trutta). The results show that the recreational trolling fishery today harvests more salmon and trout annually than the commercial and subsistence fisheries combined, and that there are seasonal patterns in effort and catch. Paper II evaluates different angler catch reporting methods (mail-in, tournament reports, and face-to-face interviews), compares catch rates within and among spring and fall fishing periods and examines mail-in data for recall bias. Harvest per boat day did not differ significantly among catch reporting methods, indicating that all three could be useful for estimating harvest rates. However, tournament anglers had higher rates for released catch. Finally, there was little evidence for recall bias in mail-in surveys. In summary, this thesis has developed the framework for a recreational angler survey program for Lake Vänern, and should be of interest to researchers and managers interested in estimating catch and effort for fisheries at large spatial scales.

L'impact de la nutrition sur l'immunité intestinale de la truite arc-en-ciel est encore mal connu. C'est pourquoi cette thèse avait pour objectifs de mieux caractériser son système immunitaire intestinal et d'évaluer les possibilités de modulation de la réponse immune intestinale par l'ajout de nucléotides libres dans son alimentation. Nos résultats indiquent que les phagocytes intestinaux présentent une activité de phagocytose plus faible que ceux du rein antérieur. La cytotoxicité naturelle mesurée au niveau intestinal est deux fois plus élevée que celle du rein antérieur et cette observation est corrélée à une augmentation du transcrit codant NKEF (Natural Killer Enhancement Factor). Nous avons également montré que les lymphocytes intestinaux ne répondent pas à une stimulation mitogénique in vitro et que ceci n'est pas due à l'apoptose des cellules. Une forte expression des transcrits codant CD8a et CD3 a été détectée dans les leucocytes intestinaux, ce qui suggère une importante proportion de lymphocytes T exprimant la forme homodimérique aa de CD8 dans ce tissu. Enfin, nous avons montré que l'ajout de nucléotides libres à l'alimentation de truites saines stimule la prolifération spontanée ainsi que la phagocytose des leucocytes intestinaux in vitro. Par contre, aucune modulation de la cytotoxicité naturelle ou de l'expression des transcrits codant les marqueurs spécifiques des lymphocytes T et B et les cytokines inflammatoires n'a été observée. Il serait à présent intéressant de renouveler ces essais en utilisant des poissons infectés afin de pouvoir observer l'effet des nucléotides sur la réponse inflammatoire et sur la réponse spécifique / The impact of nutrition on rainbow trout intestinal immunity, a farmed fish with high economic value, remains unclear. Consequently, the objectives of this thesis were to better characterize the intestinal immune system of that fish and to determine if it is possible to modulate its intestinal immune response by dietary free nucleotides. Our results show that intestinal phagocytes are less activated by yeast cells but when they are activated they can ingest as many yeast cells as their head kidney (HK) counterparts. We noted that the natural cytotoxic activity of intestinal leukocytes is twice higher than the one of HK leukocytes. This natural cytotoxic activity is correlated with an increase of transcripts encoding the natural killer enhancement factor (NKEF). Intestinal leukocytes did not respond to an in vitro mitogenic stimulation. This lack of response is not due to apoptosis. We also observed a high expression of CD8a and CD3 transcripts in gut leukocytes, suggesting that the intestine could contain a high proportion of T cells expressing the aa homodimeric form of CD8. Finally, we observed that dietary free nucleotides stimulate the spontaneous proliferation and the phagocytic activity of intestinal leucocytes in vitro. However, they did not modulate natural cytotoxicity activity nor did they affect the amounts of transcripts encoding specific markers of T and B lymphocytes and inflammatory cytokines. In the future, it will be interesting to repeat these experiments using infected fish in order to study the effect of nucleotides on the inflammatory and specific immune responses

Dentre as espécies exóticas de peixes produzidas no Brasil, a truta arco-íris, Oncorhynchus mykiss, tem se destacado. Diante da expansão desse setor, intensifica-se a necessidade de estudos visando à diminuição dos problemas produtivos e sanitários apresentados na piscicultura, assim como a busca por tratamentos alternativos aos controversos antimicrobianos. Nesse contexto, o uso de probióticos, constituídos por microrganismos vivos, que quando consumidos adequadamente beneficiam a saúde do hospedeiro, demonstra ser uma abordagem promissora no aprimoramento na criação de trutas. Deste modo, o presente trabalho buscou contribuir para o desenvolvimento de uma tecnologia alternativa na criação de truta arco-íris, avaliando o efeito de preparações probióticas sobre o desempenho dos animais submetidos às condições convencionais de manejo ou de estresse crônico. O estresse consistiu em aumentar diariamente o adensamento dos tanques de 15k g/m3 para 45 kg/m3 por 90 minutos. O tratamento consistiu na avaliação de um pool de células composto por 4 cepas de Lactobacillus com propriedades probióticas, que foi incorporado à alimentação por meio de veículo oleoso, totalizando 4 x 108 UFC/g de ração. O delineamento experimental consistiu na utilização de 140 trutas com peso médio de 360,8 ± 22,67 g e comprimento de 30,2 ± 0,8 cm, mantidas em tanques de 1500L e distribuídas em 4 grupos, a saber: Grupo NR - ração controle; Grupo ER - estresse de manejo e ração controle; Grupo NP - ração contendo a preparação probiótica; Grupo EP - estresse e ração contendo a preparação probiótica. Foram coletados 8 indivíduos de cada grupo nos dias inicial, 30 e 60 para avaliação das variáveis hematológicas, bioquímicas e de desempenho zootécnico. A possibilidade de incorporação de cepas probióticas, não liofilizadas, na ração extrusada de peixes foi demonstrada. Os resultados mostraram que a suplementação da dieta de truta com probiótico aumentou significativamente (P<0,05) a contagem de eritrócitos e monócitos. Não foram observadas alterações estatisticamente discerníveis nos parâmetros bioquímicos e no desempenho, bem como não ficou comprovado que o tratamento avaliado mitigou as respostas de estresse. / Among the exotic species of fish produced in Brazil, the rainbow trout, Oncorhynchus mykiss, has excelled. Given the expansion of this sector, intensifies the need for studies to decrease production and health problems presented in aquaculture, as well as the search for alternatives to controversial antimicrobial treatments. In this context, the use of probiotics and consist of live microorganisms, that when properly consumed benefit the health of the host, proves to be a promising approach to the improvement in the creation of trout. Thus, the present study sought to contribute to the development of an alternative technology in creating rainbow trout, evaluating the effect of probiotic preparations on the performance of the animals subjected to conventional management conditions or chronic stress. The stress consisted of daily increasing densification of 15k g/m3 tanks to 45 kg/m3 for 90 minutes. The treatment consisted in evaluating a pool of cells composed of four Lactobacillus strains with probiotic properties, was incorporated by feeding oil vehicle, totaling 4 x 108 CFU/g of feed. The experiment consisted in using 140 trout with an average weight of 360.8 ± 22.67 g and length of 30.2 ± 0.8 cm, kept in tanks of 1500L and divided into 4 groups, namely: NR Group - feed control; CWT - stress management and diet control; NP Group - feed containing probiotic preparation; EP group - stress and diet containing the probiotic preparation. Were collected eight subjects in each group in the initial, 30 and 60 for assessment of hematological, biochemical and growth performance variables. The possibility of incorporating not freeze-dried probiotic strains in extruded fish feed was demonstrated. The results show that dietary supplementation with probiotic trout increased significantly (P <0.05) the erythrocyte count and monocytes. No statistically discernible changes in biochemical parameters and performance were observed, and it was not proven that evaluated treatment mitigated the stress response. Keywords:

Cette thèse a pour but de déterminer les espèces séléniées (spéciation) issues du métabolisme du poisson (truite) suite à l’enrichissement en sélénium, organique ou inorganique, des aliments piscicoles. Ces informations, complémentaires à celles utilisées en nutrition, sont nécessaires pour juger de la nécessité et de l’adéquation d’une supplémentation en sélénium des aliments végétaux afin de garantir les fonctions biologiques dépendantes de Se, ainsi que la qualité nutritionnelle des poissons. δa capacité équivalente d’un apport en sélénium inorganique ou organique à relever les niveaux de sélénoprotéines en cas de carence a ainsi été mise en évidence, ce qui diffère des conclusions habituellement obtenues sur la base de mesures du sélénium total. δ’utilisation de traceurs mono-spécifiques et mono-isotopiqueslors de la préparation d’échantillons (sang/plasma de truite) a montré l’existence de réactions co-précipitation et/ou d’interactions entre analytes séléniés et protéines démontrant l’attention particulière qui doit être portée à l’étape de préparation d’échantillon. / This PhD aims at the determination of selenium species (speciation) of fish metabolism after inorganic or organic selenium enrichment of aquaculture feeds. This information, complementary to the one obtained in the nutrition field, is necessary to assess the requirement and the suitability of the selenium supplementation of plant based feed in order toensure selenium dependent biological functions, as well as the nutritional quality of fish. The equivalent ability of inorganic and organic selenium to raise selenoproteins levels in case of deficiency was revealed, which differs with conclusion usually obtained based on total selenium measurements. The use of monospecific and monoisotopic tracers during sample preparation (trout plasma/blood) showed the existence of co-precipitation reactions and interactions between selenized analytes and proteins demonstrating thus that attention has to be paid to the sample preparation step.

Très peu de données sont aujourd’hui disponibles sur le rôle des nutriments et de leurs interactions in vivo dans la régulation de l’autophagie. L'objectif principal de cette thèse était donc d’étudier l’effet de la qualité nutritionnelle de l’aliment sur le contrôle de cette fonction cellulaire. Une première étude a permis de montrer que les différents ratios en protéines et en glucides influencent significativement les mécanismes de contrôle de l’autophagie dans le muscle de truite arc-en-ciel (Oncorhynchus mykiss). Ces résultats ont été renforcés par une étude sur culture primaire de myoblaste de truite montrant que l’addition d’acides aminés dans le milieu de culture inhibe l’autophagie alors que le glucose à un effet inverse (article 1). Une seconde étude a porté sur la fraction protéique de l’aliment et notamment sur la teneur en méthionine, dont le niveau est trop faible dans les aliments pour poissons à base de végétaux. Les résultats obtenus indiquent que la machinerie autophagosomale mais également les principaux facteurs du renouvellement des protéines musculaires sont sensible aux variations de la teneur en méthionine de l’aliment et que la réponse qui en résulte peut fortement affecter la croissance (article 2). L’ensemble des données obtenues dans les deux premières études reposaient sur la mesure du taux d’un marqueur de l’autophagie (LC3-II) qui est à la fois produite et dégradée au cours du processus (flux) autophagique. Ainsi, dans l’optique de préciser les résultats obtenus dans les deux premiers articles, une troisième étude a été effectuée afin de déterminer s’il est possible de bloquer le flux autophagique dans le muscle de truite par l’emploi de différents agents pharmacologiques (inhibiteurs du flux autophagique). Il s’agissait également de déterminer les limites de l’utilisation de tels inhibiteurs chez cette espèce. Les essais effectués n’ont pas permis de mesurer le flux autophagique dans le muscle. En revanche, l’injection intrapéritonéale de colchicine a bien bloqué le flux autophagique dans le foie, ouvrant ainsi un nouveau champ d’investigations sur le rôle de l’autophagie dans le métabolisme intermédiaire. En conclusion, l’ensemble de ces travaux montre que l’autophagie n’est pas uniquement sensible à l’état nutritionnel (jeûne/nourris) mais également à la nature des aliments consommés. Outre leurs intérêts agronomique et thérapeutique, ces résultats ouvrent de nouvelles perspectives pour une meilleure compréhension des mécanismes d’action de l’autophagie au niveau cellulaire et métabolique mais également de son rôle dans l’adaptation des espèces au cours l’évolution. / Few data has been published on the role of nutrients and their interactions in vivo in the regulation of autophagy. The main objective of this thesis was therefore to characterize the response of the autophagic/lysosomal pathway to the macronutrients composition of the diets. The first study showed that different ratio of proteins and carbohydrates in the diet significantly affect the controls of autophagy in the muscle of rainbow trout (Oncorhynchus mykiss). These results were strengthened by study on primary culture of trout myoblasts showing that the addition of amino acids in cell culture medium inhibited the formation of autophagosomes, whereas the addition of glucose had an opposite effect (paper 1). A second study focused on the protein fraction of the diet and specifically on the content of methionine, whose the levels are low in fish feed plant-based diets. The obtained results showed that both autophagy machinery and the main factors of muscle protein turnover are significantly sensitive to change in dietary methionine levels and the resulting response may strongly affect growth and feed utilization (paper 2). The data obtained in these two first studies were based on measuring the level of autophagy marker (LC3-II), which is both produced and degraded during autophagic (flux) process. Thus, in view to clarify the results obtained in the two first studies, we conducted a third study to determine whether it is possible to block the autophagic flux in trout muscle by using different lysosomotropic agents (autophagic flux inhibitors). The objective was also to determine the limits of autophagic flux inhibitors utilisation in vivo. Different tests failed to measure autophagic flux in the muscle of rainbow trout. In contrast, intraperitoneal injection of colchicine blocked the autophagic flux in the liver. This study allowed us to investigate the function of autophagy in the intermediary metabolism. In conclusion, these studies show that autophagy is not only sensitive to the nutritional status (fasting/fed) but also to the nature of the consumed diets. In addition to their therapeutic and agricultural interests, these results open new perspectives to better understand the mechanisms of autophagy at metabolic and cellular level but also its role in the adaptation of species during evolution.

Dentre as espécies exóticas de peixes produzidas no Brasil, a truta arco-íris, Oncorhynchus mykiss, tem se destacado. Diante da expansão desse setor, intensifica-se a necessidade de estudos visando à diminuição dos problemas produtivos e sanitários apresentados na piscicultura, assim como a busca por tratamentos alternativos aos controversos antimicrobianos. Nesse contexto, o uso de probióticos, constituídos por microrganismos vivos, que quando consumidos adequadamente beneficiam a saúde do hospedeiro, demonstra ser uma abordagem promissora no aprimoramento na criação de trutas. Deste modo, o presente trabalho buscou contribuir para o desenvolvimento de uma tecnologia alternativa na criação de truta arco-íris, avaliando o efeito de preparações probióticas sobre o desempenho dos animais submetidos às condições convencionais de manejo ou de estresse crônico. O estresse consistiu em aumentar diariamente o adensamento dos tanques de 15k g/m3 para 45 kg/m3 por 90 minutos. O tratamento consistiu na avaliação de um pool de células composto por 4 cepas de Lactobacillus com propriedades probióticas, que foi incorporado à alimentação por meio de veículo oleoso, totalizando 4 x 108 UFC/g de ração. O delineamento experimental consistiu na utilização de 140 trutas com peso médio de 360,8 ± 22,67 g e comprimento de 30,2 ± 0,8 cm, mantidas em tanques de 1500L e distribuídas em 4 grupos, a saber: Grupo NR - ração controle; Grupo ER - estresse de manejo e ração controle; Grupo NP - ração contendo a preparação probiótica; Grupo EP - estresse e ração contendo a preparação probiótica. Foram coletados 8 indivíduos de cada grupo nos dias inicial, 30 e 60 para avaliação das variáveis hematológicas, bioquímicas e de desempenho zootécnico. A possibilidade de incorporação de cepas probióticas, não liofilizadas, na ração extrusada de peixes foi demonstrada. Os resultados mostraram que a suplementação da dieta de truta com probiótico aumentou significativamente (P<0,05) a contagem de eritrócitos e monócitos. Não foram observadas alterações estatisticamente discerníveis nos parâmetros bioquímicos e no desempenho, bem como não ficou comprovado que o tratamento avaliado mitigou as respostas de estresse. / Among the exotic species of fish produced in Brazil, the rainbow trout, Oncorhynchus mykiss, has excelled. Given the expansion of this sector, intensifies the need for studies to decrease production and health problems presented in aquaculture, as well as the search for alternatives to controversial antimicrobial treatments. In this context, the use of probiotics and consist of live microorganisms, that when properly consumed benefit the health of the host, proves to be a promising approach to the improvement in the creation of trout. Thus, the present study sought to contribute to the development of an alternative technology in creating rainbow trout, evaluating the effect of probiotic preparations on the performance of the animals subjected to conventional management conditions or chronic stress. The stress consisted of daily increasing densification of 15k g/m3 tanks to 45 kg/m3 for 90 minutes. The treatment consisted in evaluating a pool of cells composed of four Lactobacillus strains with probiotic properties, was incorporated by feeding oil vehicle, totaling 4 x 108 CFU/g of feed. The experiment consisted in using 140 trout with an average weight of 360.8 ± 22.67 g and length of 30.2 ± 0.8 cm, kept in tanks of 1500L and divided into 4 groups, namely: NR Group - feed control; CWT - stress management and diet control; NP Group - feed containing probiotic preparation; EP group - stress and diet containing the probiotic preparation. Were collected eight subjects in each group in the initial, 30 and 60 for assessment of hematological, biochemical and growth performance variables. The possibility of incorporating not freeze-dried probiotic strains in extruded fish feed was demonstrated. The results show that dietary supplementation with probiotic trout increased significantly (P <0.05) the erythrocyte count and monocytes. No statistically discernible changes in biochemical parameters and performance were observed, and it was not proven that evaluated treatment mitigated the stress response. Keywords:

The Pacific Northwest is known for its once-abundant wild salmonid populations that have been in decline for more than 50 years due to habitat destruction and commercial overexploitation. To compensate, federal and state agencies annually release hundreds of thousands of hatchery-reared fish into the wild. However, accumulating data indicate that hatchery fish have lower fitness in natural environments, and that hatchery rearing negatively influences return rates of anadromous salmonids. Recently, mounting evidence revealed that the richness and diversity of intestinal microbial species influence host health. We examined the gut microbiota of pre-migratory hatchery-reared steelhead (Oncorhynchus mykiss) to assess microbial community diversity. The Cascade Mountains serve as an allopatric border between two distinct clades of steelhead that show significant differences in genomic and mitochondrial diversity. We identified differences in core microbiota of hatchery-reared fish that correlate with this divergent phylogeographic distribution. Steelhead sampled from hatcheries east of the Cascades had overall greater core gut microbiota diversity. These differences were found despite similarities in diet and rearing conditions. In addition to taxonomic variation across the geographic divide, we identified significant differences in metabolic pathways using PICRUSt gene prediction software. Our analysis revealed significant enrichment of genes associated with lipid metabolism in the gut microbiome of western fish. 8 of 19 individual lipid metabolism pathways were more prominent in western populations. Lipids are a vital nutritional component for teleost species involved in migration and subsequent return for spawning in natal environments. We hypothesize that the observed differences in lipid metabolism across this phylogenetic divide results from an increased ability of eastern Cascade (O. m. gairdneri) fish to utilize lipids taken in via the diet. This increased absorption and utilization would make lipids less available for the intestinal microbiota of the eastern fish, as evidenced by the lower abundance of lipid metabolism genes in the east. Our research utilizes information from the microbiome to understand the phenotypic implications occurring in segregated populations of hatchery-reared steelhead, further confirming elements of coevolution between an organism and its internal environment.

Effects of exercise, social facilitation, and delayed conditioning after vi exercise on the learning behavior of 5 to 6-inch rainbow trout, Salmo gairdneri, was measured in a conditioned avoidance response apparatus. The conditioning schedule contained an intertrial interval between alternate presentations of conditioned (light) and unconditioned (shock) stimuli. This randomization eliminated learning losses found in a previous study. Mean percentage avoidance, the measurement of learning, did not decrease significantly during conditioning trials. Exercised fish learned avoidance better than did non-exercised fish. Mean percentage avoidance for fish exercised at 0.5 ft/sec was 66.3; at 1.0 ft/ sec, 1.5 ft/sec, and no-exercise, mean percentage avoidance was 68.2, 68.9 and 65.0, respectively. Social facilitation affected learning in the one, two, and three fish per cell tests, where mean percentage avoidance was 55.5, 68.9, and 81.0 percent, respectively. A delay of 24 hours between exercise and conditioning resulted in decreased learning levels. Mean percentage avoidance was 60.3, 63.5, 67.7, and 53.7 for the 1-, 2-, 2-, and 24-hour delay tests, respectively; however, mean percentage avoidance for the last 60 trials of each test indicated the 1-, 2-, and 4-hour delay tests were all similar, over 70 percent, while mean percentage avoidance for the 24-hour delay test was only 57.8 percent.