A comparative study of glycolysis in red and white muscles of the trout (Salmo gairdneri) and mirror carp (Cyprinus carpio)

The activities of some glycolytic and associated enzymes have been determined in the muscles of trout and carp to investigate the possibility that the discrepancies previously reported between lactate accumulation and anoxic tolerance in these two fish result from underlying differences in glycolytic potential. Steady state concentrations of certain glycolytic intermediates were also determined in freeze-clamped muscles from tankrested fish. The activities of hexokinase, phosphorylase and phosphofructokinase were approximately 2–3 times lower in carp than trout white muscles. Pyruvate kinase and lactate dehydrogenase activities were 5 times lower in carp white muscle. The lower, broader pH optima of lactate dehydrogenase and pyruvate kinase from carp compared to trout muscles is thought to be correlated with the greater anoxic tolerance of the carp. Glycolytic enzyme profiles were markedly different between the red and white muscles of the rainbow trout but broadly similar, with the exception of hexokinase activity, for the corresponding muscles of the carp. The results are discussed in relation to what is known about anaerobiosis in these two species and the comparative physiology of red and white muscles in fish.     

Red muscle recruitment during steady swimming correlates with rostral-caudal patterns of power production in trout

Rainbow trout (Oncorhynchus mykiss) and brook trout (or charr, Salvelinus fontinalis) display different rostral-caudal patterns of power production by the red or aerobic muscle during steady swimming. The anterior muscle of rainbow trout produces much less power for swimming than the posterior, while in brook trout there is no variation in power output. To determine if red muscle recruitment is associated with anterior-posterior patterns of power production, electromyography (EMG) was used to record red muscle activity at three body positions across a range of swimming speeds in fish of each species. The initial recruitment of the anterior red muscle in swimming rainbow trout was predicted to lag behind, i.e. occur at higher speeds, that of the posterior due to the variation in power production, but no variation in recruitment was expected for brook trout. Burst of red muscle EMG activity occurring with each tailbeat was analyzed for frequency (tailbeat frequency), duty cycle (DC) (duration of burst relative to the period of the tailbeat) and burst intensity (BI) (magnitude of the measured EMG activity). Brook trout swam with higher tailbeat frequencies and longer values of DC than rainbow trout. Both species showed a pattern of longitudinal variation in DC, with longer DC values in the anterior red muscle. BI also differed significantly along the length of rainbow trout but not brook trout. In the former, BI of anterior muscle was significantly less than the posterior at lower steady swimming speeds. The EMG data suggest that power production and muscle recruitment are related. In rainbow trout, where there is longitudinal variation in muscle power output, there are also significant rostral-caudal differences in red muscle recruitment.     

Hybridization dynamics between Colorado's native cutthroat trout and introduced rainbow trout

Newly formed hybrid populations provide an opportunity to examine the initial consequences of secondary contact between species and identify genetic patterns that may be important early in the evolution of hybrid inviability. Widespread introductions of rainbow trout (Oncorhynchus mykiss) into watersheds with native cutthroat trout (Oncorhynchus clarkii) have resulted in hybridization. These introductions have contributed to the decline of native cutthroat trout populations. Here, we examine the pattern of hybridization between introduced rainbow trout and 2 populations of cutthroat trout native to Colorado. For this study, we utilized 7 diagnostic, codominant nuclear markers and a diagnostic mitochondrial marker to investigate hybridization in a population of greenback cutthroat trout (Oncorhynchus clarkii stomias) and a population of Colorado River cutthroat trout (Oncorhynchus clarkii pleuriticus). We infer that cutthroat-rainbow trout hybrid swarms have formed in both populations. Although a mixture of hybrid genotypes was present, not all genotype combinations were detected at expected frequencies. We found evidence that mitochondrial DNA introgression in hybrids is asymmetric and more likely from rainbow trout than from cutthroat trout. A difference in spawning time of the 2 species or differences in the fitness between the reciprocal crosses may explain the asymmetry. Additionally, the presence of intraspecific cytonuclear associations found in both populations is concordant with current hypotheses regarding coevolution of mitochondrial and nuclear genomes.     

Mitochondrial cytochrome b sequence variation in Korean salmonids

The complete 1141 bp mitochondrial cytochrome b sequences were determined for lenok Brachymystax lenok , cherry salmon Oncorhynchus masou masou , Ishikawa's cherry salmon O. m. ishikawai , chum salmon O. keta , rainbow trout O. mykiss , and an albino mutant of rainbow trout. Common substitutions detected in these species were transitional mutations. There were no significant differences in the intraspecific variation of the cytochrome b genes. Interspecific divergences were greater than intraspecific variation. The level of variation ranged from 8·026–15·686%. The cherry salmon was closer to chum salmon than to rainbow trout, and the lenok was the most distantly related species.     

Intraspecific sequence variation in the mitochondrial genome of rainbow trout (Oncorhynchus mykiss)

We examined the sequence of a 2214 base pair HindIII fragment from the mitochondrial genome of six rainbow trout. The fragment encodes four proteins and two tRNAs. Sequences for two fish from a single locality were identical. Those from separate localities differed by from 1 to 7 nucleotide substitutions. Of 13 variable sites, 12 were synonymous and 1 led to a conservative amino acid substitution. Transitions accounted for 12 of the 13 variants. In contrast to interspecific comparisons (Thomas and Beckenbach. 1989. J. Mol. Evol. 29: 233-245), the intraspecific divergence estimates based on sequence are less than those estimated from restriction fragment analysis, suggesting a complex, dynamic process for the accumulation of variation in the mitochondrial genome.     

The relationships among sprint performance, voluntary swimming activity, and social dominance in juvenile rainbow trout

The specific objectives of this study were to determine whether sprint performance in juvenile rainbow trout is correlated with either voluntary swimming activity or aggressive behaviors and to determine the reciprocal: the effect of swimming activity and aggression on sprint performance. Sprint performance was assessed by rapidly accelerating trout (5-7-cm fork length) to a fixed velocity (40, 42, or 45 cm s(-1)) and then holding them at that velocity until fatigue. There was considerable interindividual variation in sprint performance not explained by variations in body size, but intraindividual performance was highly repeatable over at least 2 mo. Voluntary swimming was measured as the frequency of transits (voluntary transit activity, VTA) between two identical tanks via a connecting channel with two different flow regimes: zero or minimum velocity (0 or 2.5 cm s(-1)) and high velocity (84 cm s(-1)). There was a strong correlation between sprint performance and VTA in minimal current but no correlation in high current. Furthermore, sprint performance did not predict the outcome of dominance encounters. Experience with rapid acceleration, especially when voluntary, led to a pronounced improvement in sprint performance in proportion to the number of acceleration events. Social dominance encounters had a more complex effect: a significant reduction in sprint performance in previously high-performance sprinters and the reverse for low performers. We propose that there are four independent axes of interindividual variation in juvenile rainbow trout: spontaneous and rheotaxis-stimulated locomotor activity, aggressive activity, and the trainability of sprint performance. The independence of these axes has the potential to produce a much larger diversity in behavioral and ultimately physiological phenotypes than would be produced if the axes were linked.     

AN EXPERIMENTAL STUDY OF COMPETITIVE PERFORMANCE IN BRASSICA RAPA (CRUCIFERAE)

Growth chamber experiments with rapid-cycling Brassica rapa were designed to estimate the signs and magnitudes of the genetic correlations for plant performance in each of three conditions: no-competition (isolated plants), intraspecific competition, and interspecific competition with Raphanus sativa. Biomass and flower number were highest in the no-competition treatment, intermediate under intraspecific competition, and lowest under interspecific competition. Significant among-family variation in biomass and flower number was found under each regime. The mean family performance (biomass or flower number) in the no-competition treatment was significantly positively correlated with the performance in only one of the competitive treatments (for biomass in the intraspecific treatment). For both biomass and flower number there was a significant positive correlation between family means in the intra- and interspecific regimes. These correlations were greater in magnitude than those for the comparison between no-competition and competition (intra- or interspecific) treatments. Our results suggest that the importance of traits affecting plant performance is environment-dependent; the performance of a family grown without competition was a poor predictor of performance with competition, while the performance of families grown under intra- and interspecific competition was positively correlated.     

Regional variation in the microhardness and mineralization of vertebrae from brown and rainbow trout

Regional variation in properties of vertebral bone from brown Salmo trutta and rainbow trout Oncorhychus mykiss were explored by using microhardness tests. Statistically‐significant positive correlations were identified between the microhardness of bone and its mineral content. In both brown and rainbow trout, the vertebrae from the caudal region were harder than those of the trunk region. There was a significant difference between the species; microhardness of bone from vertebrae of rainbow trout was greater than those from brown trout.     

Erosion of interspecific reproductive barriers resulting from hatchery supplementation of rainbow trout sympatric with cutthroat trout

The frequency of hybridization between cutthroat (Onchorhynchus clarki clarki) and rainbow (O. mykiss irideus) trout from coastal habitats in British Columbia, Canada, was examined in seven populations where the two species are sympatric with no history of rainbow trout stocking and compared with areas where native rainbow trout populations have been supplemented with hatchery fish (three populations). Four nuclear markers were used to identify each species and interspecific hybrids and one mitochondrial marker showed the direction of gene exchange between species. The frequency of hybrids was significantly higher (Fisher exact test, P < 0.001) in river systems where hatchery rainbow trout have been introduced (50.6% hybrids) than in populations where the two species naturally co-occur without supplementation (9.9% hybrids).     

Advancing provision of multi-species fish passage: Behaviour of adult European eel (Anguilla anguilla) and brown trout (Salmo trutta) in response to accelerating flow

The behavioural response of downstream moving European eel (Anguilla anguilla) and brown trout (Salmo trutta) to velocity gradients created by orifice weirs placed in a flume were studied. The aim was to identify interspecific variation and test two hypotheses: (1) a more abrupt velocity gradient would induce a higher avoidance response and over a greater distance, and (2) fish would acclimate to the velocity gradient by incrementally sampling conditions closer to the orifice until successful passage occurred. European eel moved along the channel floor and walls, responding after physical contact with the weir. Brown trout moved downstream head first and switched orientation at the velocity gradient without contacting the structure. Brown trout spent longer than European eel immediately upstream of a channel floor orifice, although time to pass was similar; but less time at a mid-column orifice despite taking longer to pass. The mid-column orifice delayed both species longer. European eel passed orifices head first on initial encounter with no rejections, whereas brown trout passed tail first. Positions where brown trout first switched orientation and closest point to the orifice reached during initial approach occurred closer to a channel floor than mid-column treatment. Brown trout did not appear to acclimate to the velocity gradient by sampling successively closer to the orifice. Interspecific variation in behaviour remains a key challenge in developing multi-species fish passes.     

Effects of stress on growth, cortisol and glucose levels in non-domesticated Eurasian perch (Perca fluviatilis) and domesticated rainbow trout (Oncorhynchus mykiss)

Eurasian perch (Perca fluviatilis) is a promising aquaculture candidate, but the growth performance of this non-domesticated species may be negatively affected by its stress responsiveness to intensive culture conditions. To evaluate this potential problem, juvenile Eurasian perch were exposed to a standardized handling stressor twice a week for an 8-week period. A similar study was conducted on domesticated rainbow trout (Oncorhynchus mykiss) for comparison of intra- and inter-specific differences. The stressed fish of both species showed lower body growth than the non-stressed control fish, however, the final mean body mass was 35.4% lower in the stressed Eurasian perch than in the non-stressed controls, compared to 22.8% difference between the two groups in rainbow trout. The stress responsiveness was examined by comparing the post-stress cortisol and glucose levels in repeatedly stressed fish and fish exposed to the stressor only once. The cortisol stress response in both species strongly indicated a habituation to the repeated stressor. Thus, repeatedly stressed Eurasian perch reached maximum cortisol levels of 130 ng/mL after 0.5 h compared to 200 ng/mL in the fish stressed once, while considerably smaller differences in cortisol levels were shown between the repeatedly and single stressed rainbow trout. Rainbow trout also showed lower post-stress glucose levels in the repeatedly stressed fish compared to the single stressed fish. In contrast, the glucose levels in both groups of Eurasian perch increased abruptly after stress treatment and remained elevated at approximately 19 mM for 6 h; levels were three times as high as the peak levels 3 h post-stress in rainbow trout. Together, the habituation of the stress response shown in both species did not eliminate the growth difference found in the repeatedly stressed fish versus the control fish. Further, the lower growth performance of Eurasian perch compared to rainbow trout could partly be due to the increased energy consumption in the more stress responsive Eurasian perch.     

Biochemical and physiological characteristics of semen of sex-reversed female rainbow trout (Oncorhynchus mykiss, Walbaum)

This works studies the biochemical (protein concentration, osmolality, antitrypsin activity, lactate dehydrogenase activity) and physiological characteristics (sperm motility characteristics) of semen of sex-reversed female rainbow trout (n = 42) obtained with the application of 11β-hydroksyandrostendione for sex reversal. All data were arbitrarily divided into three classes depending on the percentage of sperm motility: I XX < 25%; II XX 25-50% and III XX > 50%. The average percentage of sperm motility was 18 ± 7% n = 12 (group I XX); 42 ± 6% n = 15 (group II XX) and 65 ± 12% n = 15 for group III XX, respectively) to link the values of semen parameters to the maturation stage of semen. Semen from 12 normal males of the same age was used as a reference group. Sperm concentration as well as protein concentration, osmolality, antitrypsin activity, and lactate dehydrogenase activity in seminal plasma of sex-reversed females were higher compared with the values obtained for normal male rainbow trout. The values of these parameters declined with the increasing percentage of sperm motility toward values established for normal males. The fertilization success of semen (3 × 10⁶ spermatozoa/egg) of sex-reversed females was very high (above 90%) for both the percentage of eyed embryos and hatched larvae and was related to sperm motility classes. Correlations between the quality parameters of sex-reversed females semen corresponded to those established previously for the semen of normal male rainbow trout. Antitrypsin activity, lactate dehydrogenase, protein concentration, and osmolality were found to be characteristic of seminal plasma of sex-reversed females. The maturity of sex-reversed female spermatozoa seems to be associated with the decline in the values of those parameters toward the values characteristic for seminal plasma of normal males.     

Characterization of sex chromosomes in rainbow trout and coho salmon using fluorescence in situ hybridization (FISH)

With the aim of characterizing the sex chromosomes of rainbow trout (Oncorhynchus mykiss) and to identify the sex chromosomes of coho salmon (O. kisutch), we used molecular markers OmyP9, 5S rDNA, and a growth hormone gene fragment (GH2), as FISH probes. Metaphase chromosomes were obtained from lymphocyte cultures from farm specimens of rainbow trout and coho salmon. Rainbow trout sex marker OmyP9 hybridizes on the sex chromosomes of rainbow trout, while in coho salmon, fluorescent signals were localized in the medial region of the long arm of one subtelocentric chromosome pair. This hybridization pattern together with the hybridization of a GH2 intron probe on a chromosome pair having the same morphology, suggests that a subtelocentric pair could be the sex chromosomes in this species. We confirm that in rainbow trout, one of the two loci for 5S rDNA genes is on the X chromosome. In males of this species that lack a heteromorphic sex pair (XX males), the 5S rDNA probe hybridized to both subtelocentrics This finding is discussed in relation to the hypothesis of intraspecific polymorphism of sex chromosomes in rainbow trout.     

Exhaustive exercise does not affect the preferred temperature for recovery in juvenile rainbow trout (Oncorhynchus mykiss)

We tested the hypothesis that juvenile rainbow trout (Oncorhynchus mykiss) would select a temperature colder than their acclimation temperature (16 deg +/-1 deg C) to minimize postexhaustive exercise metabolic demands and enhance oxygen availability. After an initial 3-h exploratory period in a thermal gradient (6 degrees -25 degrees C), fish selected a temperature of approximately 14 degrees C and had a baseline exploratory swimming activity of approximately 60 cm min(-1). Subsequently, experimental (chased) fish were individually removed, exhaustively exercised for 1.5 min, and replaced. Both control (unchased) and experimental fish were allowed to explore the thermal gradient for another 2 h. Immediately after being chased, trout had a metabolic profile that was consistent with being exhausted; levels of plasma and muscle lactate were 4.38+/-0.25 mmol L(-1) and 28.0+/-2.0 mmol kg(-1), respectively, and levels of muscle glycogen, adenosine triphosphate, and phosphocreatine were 3.89+/-0.95, 4.23+/-0.62, and 3.07+/-0.73 mmol kg(-1), respectively. Although exploratory swimming activity of the chased fish was significantly lower (by 81%) as compared with control fish during the first 5 min postchase, differences in the mean, median, and mode values for selected temperatures during the next 2 h were neither large (<1 degrees C) nor significant (P>0.05). Contrary to our initial hypothesis, these findings suggest that juvenile rainbow trout do not select a colder temperature to decrease metabolic rate following exhaustive exercise. Instead, rainbow trout selected a temperature marginally cooler than their acclimation temperature (16 degrees C) regardless of whether they had been previously exhausted.     

Habitat shifts in rainbow trout: competitive influences of brown trout

Summary We compared habitat use by rainbow trout sympatric (three streams) and allopatric (two streams) with brown trout to determine whether competition occurred between these two species in the southern Appalachian Mountains. We measured water depth, water velocity, substrate, distance to overhead vegetation, sunlight, and surface turbulence both where we collected trout and for the streams in general. This enabled us to separate the effects of habitat availability from possible competitive effects. The results provided strong evidence for asymmetrical interspecific competition. Habitat use varied significantly between allopatric and sympatric rainbow trout in 68% of the comparisons made. Portions of some differences refelected differences in habitats available in the several streams. However, for all habitat variables measured except sunlight, rainbow trout used their preferred habitats less in sympatry with brown trout than in allopatry if brown trout also preferred the same habitats. Multivariate analysis indicated that water velocity and its correlates (substrate particle size and surface turbulence) were the most critical habitat variables in the interaction between the species, cover in the form of shade and close overhead vegetation was second most important, and water depth was least important.     

Use of trout serum to prepare primary attached monolayer cultures of hepatocytes from rainbow trout (Salmo gairdneri)

Summary The influence of trout serum on the attachment and spreading of isolated trout hepatocytes maintained in primary culture at different temperatures was evaluated. Hepatocytes were obtained from young rainbow trout (Salmo gairdneri) by collagenase dissociation and maintained in modified Leibowitz L15 medium at 10° or 27° C for 24 h in plastic dishes previously coated with type I bovine collagen. In the absence of serum, fewer than 10% of hepatocytes attached and none of them spread on the collagen substrate. Trout serum at concentrations as low as 1.25% in the medium resulted in a pronounced concentration-dependent increase in hepatocyte attachment, as determined by direct counts by phase contrast microscopy, or by percentage of lactate dehydrogenase activity attached to the dishes after washing away unattached cells. Attachment rates were greater at the lower temperature (10° C). Trout serum also substantially increased the proportion of attached hepatocytes that spread as monolayers on the collagen substrate, especially at 10° C. By comparison, fetal bovine serum had little influence on the attachment or spreading of trout hepatocytes. These studies demonstrate a simple inexpensive method for preparing attached monolayer trout hepatocyte cultures. This procedure may be useful in toxicologic or functional studies in which fish hepatocyte attachment is an operational requirement.     

A molecular analysis of hybridization between native westslope cutthroat trout and introduced rainbow trout in southeastern British Columbia, Canada

Restriction site variation in the Ikaros gene intron was used to assess the incidence of westslope cutthroat trout ( Oncorhynchus clarki lewisi ), rainbow trout ( O. mykiss ) and interspecific hybrids at 11 localities among eight streams tributary to the upper Kootenay River system in south-eastern British Columbia, Canada. Out of 356 fish assayed by this technique, hybrids ( n =16) were found at seven of the 11 sites across five different streams. Rainbow trout ( n =6) were found at two of the 11 sites. Analysis of hybrids with a second genetic marker (heat shock 71 intron) indicated that most represented either backcrosses to both westslope cutthroat and rainbow trout, or post F₁ hybrids. Mitochondrial DNA analysis indicated that hybrid matings occur between male rainbow trout and female westslope cutthroat trout and vice versa. Comparison of present hybridization in five tributaries relative to an allozyme-based analysis in the mid-1980s, that documented hybrids in only a single tributary of seven that were common to the two studies, suggests that hybridization and introgression has increased in upper Kootenay River tributaries. The present analysis is a conservative estimate of genetic interaction between the species because introgression was not tested in the majority of samples. Identification of genetically pure westslope cutthroat trout populations, and why they might be resistant to introgression from rainbow trout, are crucial conservation priorities for this unique subspecies of cutthroat trout.     

Susceptibility of native perca (Percichthys trucha) and exotic rainbow trout (Oncorhynchus mykiss) to high temperature in Patagonia: different physiological traits and distinctive responses

The aim of this work was to explore for the existence of a physiological basis for the apparent exclusion of salmonid fish from the littoral zone of lakes in Patagonia in response to increasing water temperature, to the benefit of the native perca (Percichthys trucha). We conducted experiments on the thermal preferences of juvenile perca and rainbow trout collected in three sites of Northern Patagonia. We characterized the relationship between preferred and acclimation temperature (PT, AT) for perca and trout and analyzed between-sites variations in PTs. Both species responded with increasing PT to higher AT, but with differences in overall PT for perca from different sites (lower PTs for fish from cooler lake). In general, rainbow trout displayed lower PT and a narrower range of PT than perca. Our results provide the first comparative view of PTs of one important native fish and the main sport fish species of Patagonia. Native perca may be better equipped to cope with warmer waters than exotic rainbow trout, in agreement with their geographic distribution. Also, perca may have an advantage at the local level, particularly in littoral areas of lakes, through the alleviation of predation by and trophic competition with trout.     

Heterochrony and the development of the escape response: prehatching movements in the rainbow trout Oncorhynchus mykiss

Teleost fishes produce coordinated escape responses (C-starts) at hatching. This implies that essential swimming morphologies and motor behaviors develop during the incubation interval while the embryo is in the chorion. We examined prehatching motor behaviors in rainbow trout Oncorhycus mykiss (considered morphologically mature at hatching) and compared this species with zebrafish Danio rerio (considered morphologically immature) and assessed two hypotheses concerning the development of escape behavior. (1) Escape behavior is associated with the formation of key elements of the musculoskeletal and nervous systems; thus, the escape response appears early in ontogeny, when these elements form. (2) Escape behavior is not directly associated with the formation of underlying morphological elements; instead, it appears at hatching (i.e. when needed). We find that rainbow trout, like zebrafish, respond to touch early in the incubation interval, but do not demonstrate a complete C-start (including the second, propulsive stage) until shortly before hatching. At hatching, rainbow trout and zebrafish are similar in the degree of development of the chondocranium, paired fins and visceral arches (which comprise the larval jaw and gill support); however, rainbow trout have incipient rays in their unpaired fins (dorsal, anal and caudal), whereas zebrafish retain the embryonic fin fold. Although rainbow trout are more mature in axial swimming morphology at hatching, the essential neural and musculoskeletal systems that produce a coordinated escape response are functional at hatching in both species. This finding supports the evolutionary hypothesis that an effective escape response is critical for the survival of newly hatched teleost fishes.     

Effects of sustained swimming on rainbow trout muscle structure, blood oxygen transport, and lactate dehydrogenase isozymes: evidence for increased aerobic capacity of white muscle

Groups of rainbow trout (Salmo gairdneri, Richardson) were continuously swum at 20 cm s-1 (1.0 body lengths s-1) for 0, 3, 30, and 200 days. No significant changes in fish condition factor, combined red and white muscle mass, muscle fibre size or fibre size distribution were observed. After 200 days of swimming there was a significant 2.2 fold increase in red muscle mass. Number of capillaries per red muscle fibre increased significantly in each group by a maximum of 27% after 200 days exercise. Number of capillaries per white muscle fibre increased significantly by 95% after 200 days exercise. Blood lactate, haemoglobin (Hb) concentration haematocrit, erythrocyte adenosine triphosphate, and whole blood oxygen affinity P50 were unchanged by swimming. After 30 and 200 days swimming there was a shift in expression of white muscle lactate dehydrogenase (LDH) isozymes from LDH-A to LDH-B. Within the duplicated LDH-B isozyme complex, there was a shift in expression from LDH-B to LDH-B' subunits. These results suggest that sustained swimming at 1(-1) bl s-1 increased the aerobic capacity of red and particularly white (fast) muscle of rainbow trout but did not alter the gas transport characteristics of the blood.