Adult Triploids in a Rainbow Trout Family

Six triploid individuals were found in a full-sib family of 11 adult rainbow trout (Salmo gairdneri) from a domesticated hatchery stock. The triploid individuals were normal in size and external appearance, had underdeveloped gonads, and showed no evidence of 3n/2n chimerism or mosaicism. XXY triploids were males, suggesting that the Y chromosome is male determining in trout. Because they may avoid production losses associated with sexual maturation in normal fish, triploid trout and salmon could potentially be useful in fish culture.     

Occurrence and Phenotypic Characterization of Yersinia ruckeri Strains with Biofilm-Forming Capacity in a Rainbow Trout Farm

The presence of Yersinia ruckeri in a French fish farm was investigated. Y. ruckeri was isolated mainly from algae and sediment samples rather than from water. Twenty-two Y. ruckeri isolates were obtained, and three strains were distinguished by enterobacterial repetitive intergenic consensus PCR amplification. These strains were able to adhere to solid supports. This characteristic was correlated with flagellum-mediated motility. Killing experiments showed that sessile cells were more resistant to oxolinic acid than their planktonic counterparts. Our results demonstrate that surface colonization of fish farm tanks by Y. ruckeri biofilms is a potential source of recurrent infection for extended periods of time.     

Histidine Absorption across Apical Surfaces of Freshwater Rainbow Trout Intestine: Mechanistic Characterization and the Influence of Copper

The essential amino acid histidine performs critical roles in health and disease. These functions are generally attributed to the amino acid itself, but could also be mediated by a positive effect on trace element bioavailability. Mechanistic information regarding the absorption of histidine across the gastrointestinal tract is essential for understanding the interplay between amino acid and mineral nutrients and the implications of these interactions for nutrition and toxicology. Using intestinal brush-border membrane vesicles obtained from freshwater rainbow trout, absorption of histidine over the range 0.78–780 μm was found to be saturable, with a maximal transport rate (J _max) of 9.1 ± 0.8 nmol mg protein⁻¹ min⁻¹ and a K _m (histidine concentration required to reach 50% of this level) of 339 ± 68 μm. Histidine uptake was highly specific as 10-fold elevated levels of a variety of amino acids with putative shared transporters failed to significantly inhibit uptake. Elevated levels of d-histidine, however, impaired uptake of the natural l-isomer. The presence of “luminal” copper (8.3 μm) significantly increased both the J _max and K _m of histidine transport. This suggests that chelated copper–histidine species cross the brush-border epithelium through transport pathways distinct from those used by histidine alone.     

Purification and Characterization of Glycoprotein from the Skin Mucus of the Rainbow Trout,Salmo gaivdneri

Mucus glycoprotein (RGP) was purified and characterized from the skin mucus of rainbow trout, Salmo gairdneri. RGP was found to contain 30.1% NeuAc, 26.0% GalNAc, 5.0% Gal, and 26.0% amino acids. The protein moiety of RGP is very rich in Thr (32.4 mol%). Neither NeuGc nor KDN (2-keto-3-deoxy-d-glycero-d-galacto-nononic acid) was found in RGP. Alkaline borohydride treatment of RGP yielded a major disaccharide alditol, NeuAcα2→6GalNAc-ol and more than 4 minor oligosaccharide alditols including NeuAc→(GalNAcα1→)GalNAc-ol. It was evident that an average RGP molecule has approximately 500 NeuAc-containing oligosaccharide chains, which are attached to the Thr and Ser residues of the protein moiety and spaced at an average of 3 amino acids apart.     

Genetics of Growth Reaction Norms in Farmed Rainbow Trout

Rainbow trout is farmed globally under diverse uncontrollable environments. Fish with low macroenvironmental sensitivity (ES) of growth is important to thrive and grow under these uncontrollable environments. The ES may evolve as a correlated response to selection for growth in one environment when the genetic correlation between ES and growth is nonzero. The aims of this study were to quantify additive genetic variance for ES of body weight (BW), defined as the slope of reaction norm across breeding environment (BE) and production environment (PE), and to estimate the genetic correlation (r _{g(int, sl)}) between BW and ES. To estimate heritable variance of ES, the coheritability of ES was derived using selection index theory. The BW records from 43,040 rainbow trout performing either in freshwater or seawater were analysed using a reaction norm model. High additive genetic variance for ES (9584) was observed, inferring that genetic changes in ES can be expected. The coheritability for ES was either -0.06 (intercept at PE) or -0.08 (intercept at BE), suggesting that BW observation in either PE or BE results in low accuracy of selection for ES. Yet, the r _{g(int, sl)} was negative (-0.41 to -0.33) indicating that selection for BW in one environment is expected to result in more sensitive fish. To avoid an increase of ES while selecting for BW, it is possible to have equal genetic gain in BW in both environments so that ES is maintained stable.     

Elucidation of the Molecular Basis of a Null Allele in a Rainbow Trout Microsatellite

The use of microsatellites for studies of population structure, as markers in genome mapping, and for parentage control has become increasingly popular in recent years. However, the presence of null alleles can lead to confounding results when using microsatellites. In the Omy3DIAS microsatellite, the presence of a null allele was discovered by analysis of family material. The null allele was sequenced after amplification with new primers located farther away from the repeat sequence. The null allele was shown to be caused by a deletion of a 4-bp sequence, which was part of a repetitive sequence within one of the primer recognition sites. As this phenomenon has been seen in other cases of null alleles, this observation leads to the recommendation to avoid repetitive sequences of any kind within primer sequences. Allele-specific amplification of the null allele revealed the presence of a single variant of this allele. Received January 31, 2000; accepted May 5, 2000.     

Gene-Centromere Mapping in Rainbow Trout: High Interference over Long Map Distances

Ten enzyme loci were mapped in relation to their centromeres in gynogenetic diploid rainbow trout. Gene-centromere map distances, calculated under the assumption of complete interference, range from 1.1 cM for Ldh4 to 50 cM for Sod1. The Idh2 and Est1 loci are linked on the same chromosome arm.—The observation of close to 100% heterozygous gynogenetic diploids for the Sod1 and Mdh3,4 loci suggests that near-complete interference occurs on the chromosome arms carrying these loci. The high interference observed in this study and in several other species of fish may be related to the small size of fish chromosome arms.—Comparisons of map locations for the Ldh3 and Ldh4 and the Mdh3 and Mdh4 loci, which were duplicated by a tetraploid event in the evolution of salmonid fish, reveal that they are located at similar distances from their centromeres. Comparative mapping of loci duplicated longer ago shows more variation in map location.—The high proportion of heterozygotes for some loci after gynogenesis involving second polar body retention demonstrates that this is not a practical method for producing homozygous inbred lines in rainbow trout; treatments suppressing the first cell division are more promising for this purpose.     

Development of Rainbow Trout Microsatellite Markers from Repeat Enriched Libraries

The efficiency of developing polymorphic microsatellite markers from 2 repeat enriched libraries was evaluated. Thirty-six polymorphic microsatellite markers were developed for rainbow trout, 27 of which were informative in a mapping family. The ability of each marker to amplify genomic DNA from other salmonids was also observed. Received March 1, 2001; accepted May 1, 2001.     

Identification of a Gene Regulating the Tissue Expression of a Phosphoglucomutase Locus in Rainbow Trout

Nine percent of the rainbow trout (Salmo gairdneri) from a hatchery source have a greater than 100-fold increase in expression of a phosphoglucomutase (PGM) locus, Pgm1, in the liver but have normal expression of this locus in other tissues. The results of genetic crosses are consistent with a single regulatory gene with additive inheritance being responsible for the differences in the amount of PGM activity in the liver.—The allele responsible for the expression of Pgm1 in the liver is apparently a recent mutation. This is supported by its restricted distribution in rainbow trout and the absence of liver Pgm1 expression in closely related species. This genetic system is valuable for future analysis of the control of gene expression and in determining the relative evolutionary importance of genetic variation at structural and regulatory genes.     

The Influence of Infective Dose on the Virulence of a Generalist Pathogen in Rainbow Trout (Oncorhynchus mykiss) and Zebra Fish (Danio rerio)

Pathogen density and genetic diversity fluctuate in the outside-host environment during and between epidemics, affecting disease emergence and the severity and probability of infections. Although the importance of these factors for pathogen virulence and infection probability has been acknowledged, their interactive effects are not well understood. We studied how an infective dose in an environmentally transmitted opportunistic fish pathogen, Flavobacterium columnare, affects its virulence both in rainbow trout, which are frequently infected at fish farms, and in zebra fish, a host that is not naturally infected by F. columnare. We used previously isolated strains of confirmed high and low virulence in a single infection and in a co-infection. Infection success (measured as host morbidity) correlated positively with dose when the hosts were exposed to the high-virulence strain, but no response for the dose increase was found when the hosts were exposed to the low-virulence strain. Interestingly, the co-infection resulted in poorer infection success than the single infection with the high-virulence strain. The rainbow trout were more susceptible to the infection than the zebra fish but, in both species, the effects of the doses and the strains were qualitatively similar. We suggest that as an increase in dose can lead to increased host morbidity, both the interstrain interactions and differences in infectivity in different hosts may influence the severity and consequently the evolution of disease. Our results also confirm that the zebra fish is a good laboratory model to study F. columnare infection.     

Visual and Chemical Release of Feeding Behavior in Adult Rainbow Trout

Feeding behavior of adult rainbow trout (Oncorhynchus mykiss)is released by visual and/or chemical stimuli. Detection ofeither a conditioned visual or a conditioned chemical stimuluscreates an excitatory feeding state within the central nervoussystem which turns on feeding behavior composed of swimming,turning and biting/snapping actions. Particular amino acidsthat are highly effective physiological taste stimuli that arealso detected through olfaction (e.g. L-proline, L-alanine,L-leucine) release the initial sequence of food searching andbiting/snapping behaviors; however, an effective olfactory,but poor gustatory, stimulus (e.g. L-arginine) is rarely effectivebehaviorally. After bilateral removal of the paired olfactoryorgans, visual stimuli alone release the entire set of feedingbehavior patterns. Since amino acids that are highly potentphysiological taste stimuli do not release either feeding behavioror reflex biting/snapping actions in adult anosmic rainbow trout,it is postulated that the olfactory system detects potent tastestimuli and provides the afferent input for arousal and therelease of all feeding activity patterns. Chem. Senses 22: 375–382,1997.     

Computational Study of Evolutionary Selection Pressure on Rainbow Trout Estrogen Receptors

Molecular dynamics simulations were used to determine the binding affinities between the hormone 17-estradiol (E2) and different estrogen receptor (ER) isoforms in the rainbow trout, Oncorhynchus mykiss. Previous phylogenetic analysis indicates that a whole genome duplication prior to the divergence of ray-finned fish led to two distinct ER isoforms, ER and ER, and the recent whole genome duplication in the ancestral salmonid created two ER isoforms, ER and ER. The objective of our computational studies is to provide insight into the underlying evolutionary pressures on these isoforms. For the ER subtype our results show that E2 binds preferentially to ER over ER. Tests of lineage specific N/S ratios indicate that the ligand binding domain of the ER gene is evolving under relaxed selection relative to all other ER genes. Comparison with the highly conserved DNA binding domain suggests that ER may be undergoing neofunctionalization possibly by binding to another ligand. By contrast, both ER and ER bind similarly to E2 and the best fitting model of selection indicates that the ligand binding domain of all ER genes are evolving under the same level of purifying selection, comparable to ER.     

New species of Rhabdochona Railliet, 1916 (Nemata: Rhabdochonidae) from Rainbow Trout in California Streams

Three new species of Rhabdochona Railliet, 1916 are described and illustrated from Salmo gairdneri Richardson (rainbow trout) in freshwater streams in California: Rhabdochona californiemis n. sp., R. paxmani n. sp., and R. satmonis n. sp. Rhabdochona californiensis n. sp. is characterized by 14 anteriorly directed teeth in the prostome, egg devoid of filaments or floats, male and female tail terminus with a single mucro, left (long) spicule slender with a moderate distended podoid terminal end, spicular ratio 1:3.8. Rhabdochona paxmani n. sp. is characterized by 10 teeth in the prostome, eggs with polar floats, left (long) spicule slender with podoid terminus distended and having a minute subterminal spine; right spicule with prominent gorgeret (barb), spicular ratio 1:4.3, male and female tail terminus with a cuticular conical rounded short projection. Rhabdochona salmoni, n. sp. is characterized by 10 teeth anteriorly directed in the prostome, eggs with polar floats, left spicule slender with a distended podoid terminus; right spicule with a sharply indented gorgeret, spicular ratio 1:4.3, male and female tail terminus with a conical or rounded tip.     

Effects of Some Metals on Carbonic Anhydrase from Brains of Rainbow Trout

Carbonic anhydrase (CA) enzyme was purified from rainbow trout brain by Sepharose-4B-L: -tyrosine-sulfanilamide affinity chromatography. The enzyme was obtained with a specific activity of 2,275 EU mg(-1) and a yield of 22.5%. The sample obtained from the affinity column was used for kinetic properties and inhibition studies. Both optimum and stable pH were found as 9.0 in 1 M Tris-SO(4) at 4 degrees C, respectively. To check the purity and subunit molecular weight of enzyme, sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis was performed, and MW was found as approximately 29.0 kDa. The molecular weight of native enzyme was estimated to be approximately 27.3 kDa by gel filtration chromatography. The purified enzyme had apparent K (m),V (max), and k (cat) as follows: 0.92 mM, 0.207 micromol.min(-1) and 43.6 s(-1) for p-nitrophenylacetate. The inhibitory effects of Co(II), Cu(II), Zn(II), Ag(I), and Cd(II) on CA enzyme activity were determined using the esterase method under in vitro conditions at low concentrations of the corresponding metals. The obtained IC(50) values, which cause 50% inhibition on in vitro enzyme activity, were 0.05, 30, 0.31, 159, and 82.5 mM for cobalt, copper, zinc, silver, and cadmium, respectively. K ( i ) values were also calculated from Linewaever-Burk plots for these substances as 0.014, 27.68, 2.15, 193.86, and 94.18 for cobalt, copper, zinc, silver, and cadmium, respectively; it was determined that cobalt, silver and cadmium inhibited the enzyme competitively, copper inhibited noncompetitively while zinc inhibited the enzyme uncompetitively.