A Checklist of Metazoan Parasites from Rainbow Trout (Oncorhynchus mykiss)

An extensive literature survey on metazoan parasites from rainbow trout Oncorhynchus mykiss has been conducted. The taxa Monogenea, Cestoda, Digenea, Nematoda, Acanthocephala, Crustacea and Hirudinea are covered. A total of 169 taxonomic entities are recorded in rainbow trout worldwide although few of these may prove synonyms in future analyses of the parasite specimens. These records include Monogenea (15), Cestoda (27), Digenea (37), Nematoda (39), Acanthocephala (23), Crustacea (17), Mollusca (6) and Hirudinea (5). The large number of parasites in this salmonid reflects its cosmopolitan distribution.     

Linking Personality to Larval Energy Reserves in Rainbow Trout (Oncorhynchus mykiss)

There is a surging interest in the evolution, ecology and physiology of personality differences. However, most of the studies in this research area have been performed in adult animals. Trait variations expressed early in development and how they are related to the ontogeny of an animal’s personality are far less studied. Genetic differences as well as environmental factors causing functional variability of the central serotonergic system have been related to personality differences in vertebrates, including humans. Such gene-environment interplay suggests that the central serotonergic system plays an important role in the ontogeny of personality traits. In salmonid fishes, the timing of emergence from spawning nests is related to energy reserves, aggression, and social dominance. However, it is currently unknown how the size of the yolk reserve is reflected on aggression and dominance, or if these traits are linked to differences in serotonergic transmission in newly emerged larvae. In this study we investigated the relationship between yolk reserves, social dominance, and serotonergic transmission in newly emerged rainbow trout (Oncorhynchus mykiss) larvae. This was conducted by allowing larvae with the same emergence time, but with different yolk sizes, to interact in pairs for 24 h. The results show that individuals with larger yolks performed more aggressive acts, resulting in a suppression of aggression in individuals with smaller yolks. A higher brain serotonergic activity confirmed subordination in larvae with small yolks. The relationship between social dominance and yolk size was present in siblings, demonstrating a link between interfamily variation in energy reserves and aggression, and suggests that larger yolk reserves fuel a more aggressive personality during the initial territorial establishment in salmonid fishes. Furthermore, socially naïve larvae with big yolks had lower serotonin levels, suggesting that other factors than the social environment causes variation in serotonergic transmission, underlying individual variation in aggressive behavior.     

Investigating Fear in Rainbow Trout (Oncorhynchus mykiss) Using the Conditioned-Suppression Paradigm

Trout learned the operant task of pendulum-pressing for a food-reward in a mean of 4.3 sessions lasting 1 hr. In a separate phase, fish also learned—through classical conditioning—to associate a neutral light cue with an aversive stimulus. When again allowed to pendulum-press for food, after aversive classical conditioning, there was a drop in the rate of responding. The mean rate dropped from 3.6–2.9 responses per min. Most important, when the light-stimulus was superimposed on a steady bout of pendulum-pressing, trout ceased to press the pendulum and did not resume activity until termination of the light-stimulus (mean number of responses during a 3-min interval immediately prior to light-stimulus = 14.3 vs. during 3-min light-stimulus = 0.1). Psychologists have used this decrease in operant responding, or “conditioned emotional response,” as a tool to examine the psychological nature of this type of aversive conditioning. In this study, the fish demonstrated various results under this paradigm similar to those shown by “higher” nonhuman animals, therefore challenging the view of fish as unconscious, nonsentient animals.     

Intersex Occurrence in Rainbow Trout (Oncorhynchus mykiss) Male Fry Chronically Exposed to Ethynylestradiol

This study aimed to investigate the male-to-female morphological and physiological transdifferentiation process in rainbow trout (Oncorhynchus mykiss) exposed to exogenous estrogens. The first objective was to elucidate whether trout develop intersex gonads under exposure to low levels of estrogen. To this end, the gonads of an all-male population of fry exposed chronically (from 60 to 136 days post fertilization – dpf) to several doses (from environmentally relevant 0.01 µg/L to supra-environmental levels: 0.1, 1 and 10 µg/L) of the potent synthetic estrogen ethynylestradiol (EE2) were examined histologically. The morphological evaluations were underpinned by the analysis of gonad steroid (testosterone, estradiol and 11-ketotestosterone) levels and of brain and gonad gene expression, including estrogen-responsive genes and genes involved in sex differentiation in (gonads: cyp19a1a, ER isoforms, vtg, dmrt1, sox9a2; sdY; cyp11b; brain: cyp19a1b, ER isoforms). Intersex gonads were observed from the first concentration used (0.01 µg EE2/L) and sexual inversion could be detected from 0.1 µg EE2/L. This was accompanied by a linear decrease in 11-KT levels, whereas no effect on E2 and T levels was observed. Q-PCR results from the gonads showed downregulation of testicular markers (dmrt1, sox9a2; sdY; cyp11b) with increasing EE2 exposure concentrations, and upregulation of the female vtg gene. No evidence was found for a direct involvement of aromatase in the sex conversion process. The results from this study provide evidence that gonads of male trout respond to estrogen exposure by intersex formation and, with increasing concentration, by morphological and physiological conversion to phenotypic ovaries. However, supra-environmental estrogen concentrations are needed to induce these changes.     

Morphology and Growth of Lateral Line Organs of the Rainbow Trout (Oncorhynchus mykiss)

Abstract The morphology and growth of selected lateral line organs of the rainbow trout (Oncorhynchus mykiss) are described. Canal neuromasts of the infraorbital and operculo–mandibular canal of three different–sized trout have been examined by light microscopy. The number of neuromasts and pores, as well as their distribution, is constant in all sizes of fish. However, the area and estimated number of hair cells (HC) of the examined neuromasts increase with size and with a correlation coefficient (r²) of 0.90 and 0.87, respectively. It was found that area and estimated hair cell number of neuromasts increase 6-fold in fish from 10 to 30 cm total length (TL). Based on calculation of the hair cell number in small and large fish, a net addition of 1 and 6 HC per day is suggested for IO and OM neuromasts, respectively.     

Effect of Diet Change on Rheoreaction in Young Rainbow Trout Oncorhynchus mykiss

Journal of Ichthyology - The motivational component of rheoreaction has been determined in 4-month-old juvenile rainbow trout Oncorhynchus mykiss on different diets. It is shown that the diet...     

Skeletal Anomaly Monitoring in Rainbow Trout (Oncorhynchus mykiss,Walbaum 1792) Reared under Different Conditions

The incidence of skeletal anomalies could be used as an indicator of the “quality” of rearing conditions as these anomalies are thought to result from the inability of homeostatic mechanisms to compensate for environmentally-induced stress and/or altered genetic factors. Identification of rearing conditions that lower the rate of anomalies can be an important step toward profitable aquaculture as malformed market-size fish have to be discarded, thus reducing fish farmers’ profits. In this study, the occurrence of skeletal anomalies in adult rainbow trout grown under intensive and organic conditions was monitored. As organic aquaculture animal production is in its early stages, organic broodstock is not available in sufficient quantities. Non-organic juveniles could, therefore, be used for on-growing purposes in organic aquaculture production cycle. Thus, the adult fish analysed in this study experienced intensive conditions during juvenile rearing. Significant differences in the pattern of anomalies were detected between organically and intensively-ongrown specimens, although the occurrence of severe, commercially important anomalies, affecting 2–12.5% of individuals, was comparable in the two systems. Thus, organic aquaculture needs to be improved in order to significantly reduce the incidence of severe anomalies in rainbow trout.     

Evidence for Hox Gene Duplication in Rainbow Trout (Oncorhynchus mykiss): A Tetraploid Model Species

We examined the genomic organization of Hox genes in rainbow trout (Oncorhynchus mykiss), a tetraploid teleost derivative species, in order to test models of presumptive genomic duplications during vertebrate evolution. Thirteen putative clusters were localized in the current rainbow trout genetic map; however, analysis of the sequence data suggests the presence of at least 14 Hox clusters. Many duplicated genes appear to have been retained in the genome and share a high percentage of amino acid similarity with one another. We characterized two Hox genes located within the HoxCb cluster that may have been lost independently in other teleost species studied to date. Finally, we identified conserved syntenic blocks between salmonids and human, and provide data supporting two new linkage group homeologies (i.e., RT-3/16, RT-12/29) and three previously described homeologies (RT-2/9, RT-17/22, and RT-27/31) in rainbow trout. Electronic Supplementary Material Electronic Supplementary material is available for this article at and accessible for authorised users. The sequence data for this study have been submitted to GenBank under the following accession numbers: AY567792, AY567793, AY567794, AY567795, AY567796, AY567797, AY567798, AY567799, AY567800, AY567801, AY567802, AY567803, AY567804, AY567805, AY567806, AY567807, AY567808, AY567809, AY567810, AY567812, AY567813, AY567814, AY567815, AY567816, and AY567817. [Reviewing Editor : Dr. Axel Meyer]     

Enantiomer‐Specific In Vitro Biotransformation of Select Pharmaceuticals in Rainbow Trout (Oncorhynchus mykiss)

The occurrence of pharmaceuticals in the environment represents a challenge of emerging concern. Many pharmaceuticals are chiral compounds; however, few studies have examined the relative toxicity of pharmaceutical enantiomers to wildlife. Further, our understanding of stereospecific pharmacokinetics remains largely informed by research on humans and a few well‐studied laboratory test animals, and not by studies conducted with environmentally relevant species, including fish. The objective of this study was to investigate whether rainbow trout display stereospecific in vitro metabolism of three common chiral pharmaceuticals. Metabolism by trout liver S9 fractions was evaluated using a substrate depletion approach, which provides an estimate of intrinsic hepatic clearance (CL_{IN VITRO,INT}). No biotransformation was observed for rac‐, R‐, or S‐fluoxetine. Ibuprofen, including both enantiomers and the racemic mixture, appeared to undergo slow metabolism, but the resulting substrate depletion curves did not differ significantly from those of inactive controls. Contrary to relative clearance rates in humans, S(?)‐propranolol was more rapidly cleared than the R(+)‐ enantiomer. This work demonstrates that relative clearance rates and the effects of racemic mixtures in trout could not have been predicted based on human data. Additional research describing species differences and exploring tools for species extrapolation in biomedical and environmental studies is needed. Chirality 25:763–767, 2013, © 2013 Wiley Periodicals, Inc.     

Consistency in Context-specific Measures of Shyness and Boldness in Rainbow Trout, Oncorhynchus mykiss

Shyness and boldness has been considered a fundamental axis of human behavioural variation. At the extreme ends of this behavioural continuum subjects vary from being bold and assertive to shy and timid. Analogous patterns of individual variation have been noted in a number of species including fish. There has been debate on the nature of this continuum as to whether it depends on context. That is, whether it is domain‐general (as in humans), or context‐specific. The purpose of our study was to test if shyness and boldness depends on context in rainbow trout, Oncorhynchus mykiss and to this end we estimated boldness in five different situations. Our data provide evidence of a shy–bold behavioural syndrome in rainbow trout. Bold trout tended to be bold in four situations when the context was similar (when the context concerned foraging). However, in a different context, exploring a swim flume, the ranking was entirely different. We suggest that shyness and boldness depends on context in rainbow trout.     

Effects of Desacetyl-alpha-MSH on Lipid Mobilization in the Rainbow Trout, Oncorhynchus mykiss

Effects of melanocyte-stimulating hormone (MSH) and beta-endorphin on lipid mobilization were examined in the rainbow trout (Oncorhynchus mykiss). Plasma levels of fatty acid (FA) were measured after intra-arterial administration of alpha-MSH, desacetyl-alpha-MSH, beta-MSH, or beta-endorphin through a cannula in the dorsal aorta. Desacetyl-alpha-MSH at 1 ng/g body weight resulted in an increase in plasma FA levels 1-3 hr after the injection, whereas the other three peptides showed no significant effect at the same dose. There was no significant change in plasma levels of cortisol after administration of any of the peptides. Lipolytic enzyme activity in the liver was significantly increased in a dose-related manner 1 hr after single intra-peritoneal injection of desacetyl-alpha-MSH. The direct effect of desacetyl-alpha-MSH on lipolysis was examined in liver slices incubated in vitro. Lipase activity in the liver slice was stimulated in the medium containing desacetyl-alpha-MSH in a dose-related manner. The results indicate that desacetyl-alpha-MSH is a potent stimulator of lipid mobilization in the rainbow trout.     

Rainbow trout (Oncorhynchus mykiss) neuropeptide Y.

Neuropeptide Y (NPY) has been isolated from brain extracts of the rainbow trout (Oncorhynchus mykiss) and subjected to structural analyses. Plasma desorption mass spectroscopy estimated the molecular mass of the purified peptide as 4303.9 Da. Automated Edman degradation unequivocally established the sequence of a 36 amino acid residue peptide as: Tyr-Pro-Pro-Lys-Pro-Glu-Asn-Pro-Gly-Glu-Asp-Ala-Pro-Pro-Glu-Glu-Leu-Ala-Lys-Tyr-Tyr-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-Thr-Arg-Gln-Arg-Tyr. The molecular mass calculated from this sequence (4304 Da) is consistent with that obtained by mass spectroscopy. The presence of a C-terminal amide was established by radioimmunoassay. Rainbow trout NPY is identical in primary structure to coho salmon (Oncorhynchus kisutch) pancreatic polypeptide (PP). These data may indicate that, in this group of salmonid fishes, a single member of the NPY/PP peptide family is expressed in both neurons and peripheral endocrine cells.     

Molecular Markers for Variation in Spawning Date in a Hatchery Population of Rainbow Trout (Oncorhynchus mykiss)

We examined the distribution of alleles at 63 microsatellite loci distributed across 29 linkage groups in broodstock females from a commercial population of rainbow trout spawning on different dates throughout the season (August to January). A total of 368 females, 184 and 117 females from each of the tail-ends of the spawning distribution and a subsample of 67 females spawning in the middle, were used to detect marker–trait associations. Twenty-one loci in a subset of genomic regions (RT-5, 7, 8, 10, 12, 14, 15, 22, 23, 24, 25, 29, 30, and 31) were significantly associated with variation in spawning date. Many of these markers localize to regions with known spawning date quantitative trait loci based on previous studies. An individual assignment analysis was used to test how well the molecular data could be used to assign individuals to their correct spawning group, and markers were given a ranking reflecting their contribution to the accuracy of assignment. The top 15 ranked markers were successful at assigning the majority of females to the correct spawning group based on genotype with an average accuracy of 76 %. The most likely genes that could contribute to these differences in spawning date are discussed. Together, these data indicate that the loci could be incorporated into a selection index with phenotype data to increase the accuracy of selection for spawning date.     

Heritable Targeted Inactivation of the Rainbow Trout (Oncorhynchus mykiss) Master Sex-Determining Gene Using Zinc-Finger Nucleases

Gene targeting is a powerful tool for analyzing gene function. Recently, new technology for gene targeting using engineered zinc-finger nucleases (ZFNs) has been described in fish species. However, it has not yet been widely used for cold water and slow developing species, such as Salmonidae. Here, we present the results of successful ZFN-mediated disruption of the sex-determining gene sdY (sexually dimorphic on the Y chromosome) in rainbow trout (Oncorhynchus mykiss). Three pairs of ZFN mRNA targeted to different regions of the sdY gene were injected into fertilized rainbow trout eggs. Sperm from 1-year-old male founders (parental generation one or P1) carrying a ZFN-induced mutation in their germline were then used to produce F1 non-mosaic animals. In these F1 populations, we characterized 14 different mutations in the sdY gene, including one mutation leading to the deletion of leucine 43 (L43) and 13 mutations at other target sites that had different effects on the SdY protein, i.e., amino acid insertions, deletions, and frameshift mutations producing premature stop codons in the mRNA. The gonadal phenotype analysis of the F1-mutated animals revealed that the single L43 amino acid deletion did not lead to a male-to-female sex reversal, but all other mutations induced a clear ovarian phenotype. These results show that targeted gene disruption using ZFN is efficient in rainbow trout but depends on the ZFN design. We also characterized new sdY mutations resulting in male-to-female sex reversal, and we conclude that L43 seems dispensable for SdY function.