Growth Hormone Transgenic Coho Salmon Research Articles

Growth and endocrine effects of recombinant bovine growth hormone treatment in non-transgenic and growth hormone transgenic coho salmon

To examine the relative growth, endocrine, and gene expression effects of growth hormone (GH) transgenesis vs. GH protein treatment, wild-type non-transgenic and GH transgenic coho salmon were treated with a sustained-release formulation of recombinant bovine GH (bGH; Posilac™). Fish size, specific growth rate (SGR), and condition factor (CF) were monitored for 14weeks, after which endocrine parameters were measured. Transgenic fish had much higher growth, SGR and CF than non-transgenic fish, and bGH injection significantly increased weight and SGR in non-transgenic but not transgenic fish. Plasma salmon GH concentrations decreased with bGH treatment in non-transgenic but not in transgenic fish where levels were similar to controls. Higher GH mRNA levels were detected in transgenic muscle and liver but no differences were observed in GH receptor (GHR) mRNA levels. In non-transgenic pituitary, GH and GHR mRNA levels per mg pituitary decreased with bGH dose to levels seen in transgenic salmon. Plasma IGF-I was elevated with bGH dose only in non-transgenic fish, while transgenic fish maintained an elevated level of IGF-I with or without bGH treatment. A similar trend was seen for liver IGF-I mRNA levels. Thus, bGH treatment increased fish growth and influenced feedback on endocrine parameters in non-transgenic but not in transgenic fish. A lack of further growth stimulation of GH transgenic fish suggests that these fish are experiencing maximal growth stimulation via GH pathways.

Genetically modified growth affects allometry of eye and brain in salmonids

Effects of growth acceleration on eye development have been examined in genetically modified salmonids. Growth hormone (GH) transgenic coho salmon ( Oncorhynchus kisutch (Walbaum, 1792)) show dramatically elevated overall body growth and an absolute increase in eye size, but relative eye growth is shifted from negatively allometric to more isometric. Thus, transgenic fish possess significantly smaller eyes relative to nontransgenic fish of the same size. Ration-restricted limitation of growth in transgenic salmon to that of wild type restores relative eye growth rate, suggesting that effects on eyes are an indirect consequence of modification of growth rate rather than a direct effect of GH overexpression. Heart, spleen, and liver did not show changes in proportion among groups, whereas total brain size showed the same response as eye. Relative eye and brain size were also reduced in a fast-growing domesticated strain of rainbow trout ( Oncorhynchus mykiss (Walbaum, 1792)), suggesting modifications of allometry are a more general effect of growth acceleration. GH mRNA levels from the transgene were elevated in eyes, whereas IGF-I mRNA was not, suggesting this organ may be regulated in a different fashion than other organs. Neural tissues with critical structural requirements for optimal function may be subject to less modification of growth rate than are other organ systems.

Cultured growth hormone transgenic salmon are reproductively out-competed by wild-reared salmon in semi-natural mating arenas

The production of growth hormone (GH) transgenic animals has raised a host of social and scientific concerns regarding their potential impacts on ecosystems should they escape into nature. Indeed, theoretical models suggest that GH transgenic animals could, under specific conditions, decimate local populations. However, while there are numerous laboratory examinations of factors affecting survival of transgenic and wild animals, we know little about the competitive reproductive capacity of transgenic animals. Here, we examined the reproductive capabilities of cultured GH transgenic coho salmon ( Oncorhynchus kisutch) when in competition with wild coho derived from nature using semi-natural mating arenas (within a contained facility). To account for the well-known reproductive impairments associated with culturing salmon in laboratory facilities we contrasted the competitive reproductive success of GH transgenic coho against that of cultured non-transgenic coho. We also performed in vitro sperm analyses to assess the postcopulatory competitive ability of GH transgenic coho. In competitive mating arenas, transgenic coho performed fewer courtship and aggressive behaviours than coho from nature and sired less than 6% of offspring. Non-transgenic cultured coho, despite their smaller body size, sired more than twice as many offspring than transgenic coho when competing against wild coho in mating arenas. Transgenic males also face a postcopulatory reproductive disadvantage as their ejaculates contained fewer sperm that swam slower and for shorter durations than sperm from wild males. Together, these findings suggest limited potential for the transmission of transgenes from cultured GH transgenic coho salmon through natural matings should they escape from a contained culture facility into nature and reproductively interact with a local wild coho salmon strain. However, as responses of wild-reared fish can differ greatly from those of cultured fish, we stress the importance of understanding genotype-by-environment interactions for reproductive phenotypes when developing risk assessment information.

Increased intrinsic growth rate is advantageous even under ecologically stressful conditions in coho salmon (Oncorhynchus kisutch)

Growth rate is an ecologically important trait, affecting the energy acquisition from, and provisioning to, the surrounding community. One of many costs suggested to counteract the evolution of increased intrinsic growth rate is an associated reduction in tolerance to conditions of nutrient stress. Here we test this concept with individuals possessing experimentally increased intrinsic growth rates (growth hormone transgenic coho salmon, Oncorhynchus kisutch) relative to wild genotypes. Using a series of three experiments, survival and growth of both genotypes were assessed on a physiological and behavioral level while varying food abundance, social interactions, and predation risk. Only in complete absence of exogenous food in newly emerged fry did the high intrinsic growth rate appear costly with a shorter average survival time compared to wild-type (Exp. 1). In experiment 2, genotypes with elevated intrinsic growth showed equal or higher survival and growth than wild-type genotypes In a third experiment, adding very limited amounts of food and allowing for social interactions in a simulated natural environment benefited transgenic individuals relative to wild-types, but at similar magnitudes in both the absence and presence of predators. Populations with transgenic individuals present did not crash under these competitive conditions as previously reported when studied in simple environments where hiding and attack escape were not possible. Our data suggest that transgenic fish have a greater scope for growth under most conditions, but are not obligated to use this capability. Physiological (e.g. appetite and conversion efficiency) and behavioral traits (e.g. competitive ability and risk-taking) found previously to correlate positively with intrinsic growth rate in the transgenic strain likely aided in their survival and growth, even under food limited conditions. Hence, at least in coho salmon, intrinsic growth rate does not appear to strongly affect survival under nutrient stress.

Migration and growth potential of coho salmon smolts: implications for ecological impacts from growth-enhanced fish

Wild-genotype and growth hormone (GH) transgenic coho salmon (with dramatically enhanced growth potential) were used to examine the influence of genotype, age, body size, growth, and rearing conditions on the onset of seaward migration and to assess the potential consequences of the introduction of such transgenic fish on natural ecosystems and wild populations. When reared from the first feeding stage under naturalized stream conditions, there was no difference in survival or migratory timing between the two genotypes. However, larger fish migrated earlier in the season than smaller fish of both genotypes, and transgenic fish with higher specific spring growth rates migrated earlier in the season than slower-growing transgenic fish. Stream-reared fish of both genotypes also displayed increased migratory activity at dawn and dusk. Fish reared in the hatchery for 3 and 15 months before being released into the stream in August differed in onset of seaward migration due mainly to age (older fish migrated earlier in the season) and genotype (transgenic fish migrated before wild-type in younger fish). Further, hatchery-reared fish showed no diel pattern in activity during migration. In older fish, larger individuals migrated later in the season than small individuals, whereas there was no clear size effect in younger individuals. Thus, although small differences in spring migration timing were observed among groups, seaward migration in coho salmon was largely independent of major shifts in size and growth rate induced by GH transgenesis (i.e., transgenic fish migrated at approximately the normal time in the spring, rather than at the typical size). Further, early rearing conditions had a stronger effect on migratory behavior than did the growth-promoting transgene. Taking into account effects of migratory timing, growth, survival, and differential food conversion efficiencies, these data suggest that transgenic fish escaped from hatcheries would have a greater impact on stream ecosystems during early life compared to escaped wild-type fish. However, this difference may be reduced if rearing occurred in subsequent generations under wild conditions where growth rates of transgenic fish are reduced compared with hatchery conditions.

Expression profile for metabolic and growth-related genes in domesticated and transgenic coho salmon ( Oncorhynchus kisutch) modified for increased growth hormone production

To gain a better understanding of the aspects underlying growth in salmonids, quantitative expression analysis was performed for a number of genes related to muscle growth, metabolism, immunology and energy regulation in the liver and muscle of wild-type coho salmon ( Oncorhynchus kisutch), domesticated coho salmon selected for growth, growth hormone-transgenic coho salmon fed to satiation, and growth hormone-transgenic coho salmon reared on restricted rations. In comparisons among these four experimental groups, our findings show a significant and correlative up-regulation in the expression of a number of muscle determination and development factors in full-fed (FF) transgenic and domestic coho salmon. Expression of several metabolic genes involved with amino acid turnover and utilization also was correlated between these two faster-growing groups. However, distinct differences between domesticated and FF-transgenic fish were found for some genes involved with muscle cell differentiation, innate immune system, metabolism, and amino acid regulation. Wild-type and ration-restricted growth hormone-transgenic fish correlated very highly in the expression of most genes tested, suggesting that limiting available energy reduces the effects of elevated growth hormone and down-regulates muscle and metabolic regulatory pathways similarly to the reduced potential for growth hormone production seen in wild-type fish.

Genotype-Temperature Interaction in the Regulation of Development, Growth, and Morphometrics in Wild-Type, and Growth-Hormone Transgenic Coho Salmon

BackgroundThe neuroendocrine system is an important modulator of phenotype, directing cellular genetic responses to external cues such as temperature. Behavioural and physiological processes in poikilothermic organisms (e.g. most fishes), are particularly influenced by surrounding temperatures.Methodology/Principal FindingsBy comparing the development and growth of two genotypes of coho salmon (wild-type and transgenic with greatly enhanced growth hormone production) at six different temperatures, ranging between 8° and 18°C, we observed a genotype-temperature interaction and possible trend in directed neuroendocrine selection. Differences in growth patterns of the two genotypes were compared by using mathematical models, and morphometric analyses of juvenile salmon were performed to detect differences in body shape. The maximum hatching and alevin survival rates of both genotypes occurred at 12°C. At lower temperatures, eggs containing embryos with enhanced GH production hatched after a shorter incubation period than wild-type eggs, but this difference was not apparent at and above 16°C. GH transgenesis led to lower body weights at the time when the yolk sack was completely absorbed compared to the wild genotype. The growth of juvenile GH-enhanced salmon was to a greater extent stimulated by higher temperatures than the growth of the wild-type. Increased GH production significantly influenced the shape of the salmon growth curves.ConclusionsGrowth hormone overexpression by transgenesis is able to stimulate the growth of coho salmon over a wide range of temperatures. Temperature was found to affect growth rate, survival, and body morphology between GH transgenic and wild genotype coho salmon, and differential responses to temperature observed between the genotypes suggests they would experience different selective forces should they ever enter natural ecosystems. Thus, GH transgenic fish would be expected to differentially respond and adapt to shifts in environmental conditions compared with wild type, influencing their ability to survive and interact in ecosystems. Understanding these relationships would assist environmental risk assessments evaluating potential ecological effects.

Sustained predation effects of hatchery‐reared transgenic coho salmon Oncorhynchus kisutch in semi‐natural environments

Summary 1. The potential risks of accidental or intentional introduction of transgenic organisms to nature are unknown. We have previously shown that, after being reared in the hatchery, growth hormone transgenic coho salmon Oncorhynchus kisutch can exert a stronger predation effect on natural prey under simulated natural conditions compared with hatchery‐reared genetically wild conspecifics. However, when reared in a simulated natural environment, the difference between the two genotypes was greatly reduced. 2. Here, we tested if the difference in predation rate between the two genotypes after being reared in the hatchery diminishes with time spent in a simulated natural environment. Genetically wild coho salmon reared in the hatchery were size matched to younger satiation‐fed (fast‐growing) and same age restricted‐fed (growth rates matched to wild‐type) transgenic fish. These three types of predators were released into simulated natural habitats and their effects on prey survival and growth were monitored over a 2‐month period. 3. Restricted‐fed, but not satiation‐fed, transgenic predators consumed significantly more prey (×2·7) relative to wild predators during the first month, with an increase to 3.8 times during the second period. Prey biomass decreased more in the presence of restricted‐fed predators during the first period, but a faster growth of these prey during the second period compensated for the higher predation rate resulting in no significant difference in prey biomass among predators after 2 months. Transgenic predators grew more in length than wild type, but all three types converged in weight and condition over time. 4. Synthesis and applications. Behavioural phenotypes developed in the hatchery showed little plasticity after release. Hence, development of policy on the use of transgenic organisms must incorporate knowledge of the effect of environmental conditions, experienced by the organism prior to escape or release, on phenotype. The risk posed by culture‐reared organisms may depend on the frequency of escapes, number of individuals escaping and age of escapees. Important to note is that wild‐reared organisms may be biologically different from culture‐reared and thus require separate evaluation. Our results will be important for science policy makers and regulators to consider when deciding whether to allow commercial application of transgenic species in aquaculture.

Influence of dietary concentrations of protein, lipid and carbohydrate on growth, protein and energy utilization, body composition, and plasma titres of growth hormone and insulin-like growth factor-1 in non-transgenic and growth hormone transgenic coho salmon, Oncorhynchus kisutch (Walbaum)

This study compared the ability of juvenile non-transgenic (NT) and transgenic growth hormone (T; gene construct OnMTGH1) coho salmon ( Oncorhynchus kisutch) to metabolically utilize energy from lipid and carbohydrate for growth at two dietary protein concentrations. Triplicate groups of size-matched (initial weight, 28.2–29.1 g) NT and T salmon held in 10.5–10.8 °C well water on a natural phototocycle (12.25 h → 8.25 h) were each fed one of four isoenergetic (~ 17.5 MJ of digestible energy (DE)/kg) dry diets twice daily to satiation for 83 days. These diets contained 340 g (LP) or 430 g (HP) of estimated digestible protein (DP)/kg and either 123 g (LL) or 164 g (HL) of estimated digestible lipid (DL)/kg at each DP level (dry weight basis). Estimated digestible carbohydrate (DCHO) concentrations ranged from 45 to 269 g/kg to equalize dietary DE. Under the preceding conditions 46–66% of NT and 18.7–27.5% of T coho, depending upon diet treatment, exhibited no growth or lost weight during the study. NT coho are known to grow slowly or lose weight between the fall equinox and winter solstice but this was unexpected for T coho. Considering all fish T coho, regardless of diet treatment, exhibited significantly higher specific growth rates (SGR) than NT fish due to enhanced feed intake, feed and protein (gross and available deposited) utilization and generally improved available energy utilization. Diet treatment did not affect the growth performance of T fish. Within NT fish, the HP diets supported best gross and available protein and energy utilization. SGR values for growing fish (F G) only followed identical trends to those for all fish in relation to diet treatment. Regardless of fish genotype, terminal hepatosomatic indices for F G were directly related to dietary DCHO content and maximum values were noted in NT fish fed LP–LL (significantly higher than observed in NT or T fish fed HP–HL). Final whole body protein concentrations adjusted for dissimilar fish size were higher in T fish fed HP–HL than in T fish fed LP diets and NT fish fed LP–LL. This was also true for T fish fed HP–LL versus T fish fed the LP diets. Within HP groups, whole body lipid and energy contents were generally higher in NT versus T fish whereas within the LP groups energy content was significantly higher in T versus NT fish and was highest in T fish fed LP–LL. Terminal plasma titres for GH and IGF–1 (all fish) were significantly higher in T than in NT fish and were generally uninfluenced by diet treatment. Non-growing NT and T fish had elevated GH relative to NT and T fish that grew. Also, GH was higher in growing T fish relative to growing NT fish whereas the opposite was true in non-growing fish. Plasma IGF-1 levels were higher in growing NT and T fish than in their respective non-growing counterparts. Within F G only, IGF-1 levels were higher in T versus NT fish. It is concluded that T coho have enhanced ability to effectively utilize DE from DCHO relative to NT fish especially when DP is near the bottom of the optimal range and DL is concurrently suboptimal for NT fish. This is likely due to the elevated titres of GH, IGF-1 and 3,5,3′-triiodo- l-thyronine (measured in another study) in T fish. Also, the enhanced ability of T fish to store energy under these dietary conditions suggests enzymatic improvements in their anaerobic and aerobic metabolism of glucose.

Disruption of seasonality in growth hormone-transgenic coho salmon ( Oncorhynchus kisutch) and the role of cholecystokinin in seasonal feeding behavior

Seasonal variation in daily food intake is a well-documented phenomenon in many organisms including wild-type coho salmon where the appetite is noticeably reduced during periods of decreased day length and low water temperature. This reduction may in part be explained by altered production of cholecystokinin (CCK) and growth hormone (GH). CCK is a hormone produced in the brain and gut that mediates a feeling of satiety and thus has an inhibitory effect on food intake and foraging behaviour. Growth hormone (GH) enhances feeding behaviour and consequently growth, but its production is reduced during winter. The objectives of this study were: first, to compare the seasonal feeding behaviour of wild and GH-transgenic coho salmon; second, to determine the behavioural effect of blocking the action of CCK (by using devazepide) on the seasonal food intake; and third, to measure CCK expression in brain and gut tissues between the two genotypes across seasons. We found that, in contrast to wild salmon, food intake in transgenic salmon was not reduced during winter indicating that seasonal control of appetite regulation has been disrupted by constitutive production of GH in transgenic animals. Blocking of CCK increased food intake in both genotypes in all seasons. The increase was stronger in wild genotypes than transgenic fish; however blocking CCK in wild-type fish in winter did not elevate appetites to levels observed in the summer. The response to devazepide was generally faster in transgenic than in wild salmon with more rapid effects observed during summer than during winter, possibly due to a higher temperature in summer. Overall, a seasonal effect on CCK mRNA levels was observed in telencephalon with levels during winter being higher compared to the summer in wild fish, but with no seasonal effect in transgenic fish. No differences in seasonal CCK expression were found in hypothalamus. Higher levels of CCK were detected in the gut of both genotypes in winter compared to summer. Thus, CCK appears to mediate food intake among seasons in both wild-type and GH-transgenic salmon, and an altered CCK regulation may be responsible at least in part for the seasonal regulation of food intake.

Gene–environment interactions influence ecological consequences of transgenic animals

Production of transgenic animals has raised concern regarding their potential ecological impact should they escape or be released to the natural environment. This concern has arisen mainly from research on laboratory-reared animals and theoretical modeling exercises. In this study, we used biocontained naturalized stream environments and conventional hatchery environments to show that differences in phenotype between transgenic and wild genotypes depend on rearing conditions and, critically, that such genotype-by-environment interactions may influence subsequent ecological effects in nature. Genetically wild and growth hormone transgenic coho salmon (Oncorhynchus kisutch) were reared from the fry stage under either standard hatchery conditions or under naturalized stream conditions. When reared under standard hatchery conditions, the transgenic fish grew almost three times longer than wild conspecifics and had (under simulated natural conditions) stronger predation effects on prey than wild genotypes (even after compensation for size differences). In contrast, when fish were reared under naturalized stream conditions, transgenic fish were only 20% longer than the wild fish, and the magnitude of difference in relative predation effects was much reduced. These data show that genotype-by-environment interactions can influence the relative phenotype of transgenic and wild-type organisms and that extrapolations of ecological consequences from phenotypes developed in the unnatural laboratory environment may lead to an overestimation or underestimation of ecological risk. Thus, for transgenic organisms that may not be released to nature, the establishment of a range of highly naturalized environments will be critical for acquiring reliable experimental data to be used in risk assessments.

The glutathione antioxidant system is enhanced in growth hormone transgenic coho salmon (Oncorhynchus kisutch)

Insertion of a growth hormone (GH) transgene in coho salmon results in accelerated growth, and increased feeding and metabolic rates. Whether other physiological systems within the fish are adjusted to this accelerated growth has not been well explored. We examined the effects of a GH transgene and feeding level on the antioxidant glutathione and its associated enzymes in various tissues of coho salmon. When transgenic and control salmon were fed to satiation, transgenic fish had increased tissue glutathione, increased hepatic glutathione reductase activity, decreased hepatic activity of the glutathione synthesis enzyme gamma-glutamylcysteine synthetase, and increased intestinal activity of the glutathione catabolic enzyme gamma-glutamyltranspeptidase. However, these differences were mostly abolished by ration restriction and fasting, indicating that upregulation of the glutathione antioxidant system was due to accelerated growth, and not to intrinsic effects of the transgene. Increased food intake and ability to digest potential dietary glutathione, and not increased activity of glutathione synthesis enzymes, likely contributed to the higher levels of glutathione in transgenic fish. Components of the glutathione antioxidant system are likely upregulated to combat potentially higher reactive oxygen species production from increased metabolic rates in GH transgenic salmon.

SELECTION ON INCREASED INTRINSIC GROWTH RATES IN COHO SALMON, ONCORHYNCHUS KISUTCH

Substantial evidence from the animal kingdom shows that there is a trade-off between benefits and costs associated with rapid somatic growth. One would therefore expect growth rates under natural conditions to be close to an evolutionary optimum. Nevertheless, natural selection in many salmonid species appears to be toward larger size and earlier emergence from spawning redds, indicating a potential for increased growth rate to evolve. We tested how selection for genetic variants (growth hormone transgenic coho salmon, Oncorhynchus kisutch, with more than doubled daily growth rate potential relative to wild genotypes) depended on predator timing and food abundance during the early period of life (fry stage). In artificial redds, fry of the fast-growing genotypes showed a highly significant developmental shift, emerging from gravel nests approximately two weeks sooner, but with an 18.6% reduced survival, relative to wild-genotype fry. In seminatural streams, fry of the fast-growing genotypes suffered higher predation than those of wild genotypes when predators were present at the time of fry emergence, but this difference was less pronounced when food was scarce. In streams where predators were introduced after emergence, fry survived equally well regardless of food availability. Surviving fry grew faster in habitats provided with more food, and fast-growing genotypes also grew faster than wild genotypes when predators arrived late and food was abundant. Fewer fish migrated downstream past a waterfall when food availability was high and in the presence of predators, and wild-genotype fry were more likely to migrate than fry of the fast-growing genotypes. After being returned to the experimental streams after migration, fast-growing genotypes survived equally well as those of the same genotypes that did not migrate, whereas migrating wild genotypes experienced higher mortality relative to those of the same genotypes that did not migrate. Comparisons of growth rates between siblings retained under hatchery conditions and those from habitats with the fastest growth in the experimental stream revealed that growth rates were similar for wild genotypes in both environments, whereas the fast-growing genotypes in the streams only realized 90% of their growth potential. The present study has shown that a major shift in developmental timing can alter critical early stages affecting survival and can have a significant effect on fitness. Furthermore, ecological conditions such as food abundance and predation pressure can strongly influence the potential for fast-growing variants to survive under natural conditions. The large-scale removal of many predatory species around the world may augment the evolution of increased intrinsic growth rates in some taxa.

Gut size in GH‐transgenic coho salmon is enhanced by both the GHtransgene and increased food intake

Growth hormone transgenic coho salmon Oncorhynchus kisutch fed at the same ration level as non‐transgenic controls (Tc) had the same growth rate as non‐transgenic controls (Nt). In contrast, growth hormone transgenic coho salmon (Tf) fed ad libitum ate about twice as much and had much higher growth rates than the other two groups. The most obvious result was the significantly larger caeca in the Tf group relative to both Nt or pair‐fed Tc. The Tf fish had more caeca that were longer. The results suggested that the effect was indirect and the enlarged caeca required both the GHtransgene and hyperphagia to cause enlarged caecal capacity. A small part of the results, however, also suggested that there was a direct effect of the GHtransgene on some gut tissues, particularly the intestine.

SELECTION ON INCREASED INTRINSIC GROWTH RATES IN COHO SALMON, ONCORHYNCHUS KISUTCH

Substantial evidence from the animal kingdom shows that there is a trade-off between benefits and costs associated with rapid somatic growth. One would therefore expect growth rates under natural conditions to be close to an evolutionary optimum. Nevertheless, natural selection in many salmonid species appears to be toward larger size and earlier emergence from spawning redds, indicating a potential for increased growth rate to evolve. We tested how selection for genetic variants (growth hormone transgenic coho salmon, Oncorhynchus kisutch, with more than doubled daily growth rate potential relative to wild genotypes) depended on predator timing and food abundance during the early period of life (fry stage). In artificial redds, fry of the fast-growing genotypes showed a highly significant developmental shift, emerging from gravel nests approximately two weeks sooner, but with an 18.6% reduced survival, relative to wild-genotype fry. In seminatural streams, fry of the fast-growing genotypes suffered higher predation than those of wild genotypes when predators were present at the time of fry emergence, but this difference was less pronounced when food was scarce. In streams where predators were introduced after emergence, fry survived equally well regardless of food availability. Surviving fry grew faster in habitats provided with more food, and fast-growing genotypes also grew faster than wild genotypes when predators arrived late and food was abundant. Fewer fish migrated downstream past a waterfall when food availability was high and in the presence of predators, and wild-genotype fry were more likely to migrate than fry of the fast-growing genotypes. After being returned to the experimental streams after migration, fast-growing genotypes survived equally well as those of the same genotypes that did not migrate, whereas migrating wild genotypes experienced higher mortality relative to those of the same genotypes that did not migrate. Comparisons of growth rates between siblings retained under hatchery conditions and those from habitats with the fastest growth in the experimental stream revealed that growth rates were similar for wild genotypes in both environments, whereas the fast-growing genotypes in the streams only realized 90% of their growth potential. The present study has shown that a major shift in developmental timing can alter critical early stages affecting survival and can have a significant effect on fitness. Furthermore, ecological conditions such as food abundance and predation pressure can strongly influence the potential for fast-growing variants to survive under natural conditions. The large-scale removal of many predatory species around the world may augment the evolution of increased intrinsic growth rates in some taxa.

Growth, viability and genetic characteristics of GH transgenic coho salmon strains

Transgenic coho salmon strains containing an all-salmon growth hormone (GH) gene construct (OnMTGH1) have been examined. The transgene utilized is comprised of the metallothionein-B promoter driving the expression of the type-I growth hormone gene from sockeye salmon. Transgene DNA is integrated into the salmon genome, but is transmitted at low frequency from founder transgenic animals consistent with late integration following microinjection resulting in mosaic animals, whereas subsequent generations transmit transgene DNA as a stable Mendelian trait. Different families established from separate founder animals yield lines with unique growth characteristics suggesting important site-of-integration effects on transgene expression. Growth enhancement of transgenic salmon is initiated early, with advanced hatch timing but occurring also throughout the life history, particularly during the early phase in fresh water. GH transgenic fish showed precocious smoltification and onset of sexual maturation, but approximately normal adult body size, indicating that compression of the normal coho salmon life history is occurring. The viability of diploid GH transgenic salmon ranges from reductions to greater than that of controls among strains, and triploid transgenic animals had normal viability relative to diploid transgenic salmon. Triploid transgenic salmon display a reduction in growth rate relative to transgenic diploids, but are still significantly growth enhanced compared with nontransgenic controls. The distinct phenotypic characteristics of GH transgenic families suggest that evaluation for aquaculture and for risk assessments requires examination of strains on a case-by-case basis. Furthermore, strong effects of size at maturity in culture conditions were observed for nontransgenic wild strain coho salmon which were not apparent in GH transgenic salmon, indicating that strong genotype by environment effects strongly influence phenotype and complicate risk assessments.

Disease resistance, stress response and effects of triploidy in growth hormone transgenic coho salmon

Diploid and triploid coho salmon Oncorhynchus kisutch transgenic for growth hormone (GH) and control coho salmon were compared for differences in disease resistance and stress response. Resistance to the bacterial pathogen Vibrio anguillarum was not affected in transgenic fish relative to their non‐transgenic counterparts when they were infected at the fry stage, but was lower in transgenic fish when infected near smolting. Vaccination against vibriosis provided equal protection to both transgenic and non‐transgenic fish. Triploid fish showed a lower resistance to vibriosis than their diploid counterparts. Diploid transgenic fish and non‐transgenic fish appeared to show similar physiological and cellular stress responses to a heat shock. These studies provide information useful for both performance and ecological risk assessments of growth‐accelerated coho salmon.

Oxygen uptake of growth hormone transgenic coho salmon during starvation and feeding

Oxygen uptake of growth hormone transgenic coho salmon Oncorhynchus kisutch was measured in individual fish with a closed‐system respirometer and was compared with that of similar‐sized non‐transgenic control coho salmon during starvation and when fed a fixed ration or to satiation. Transgenic and control fish did not differ in their standard oxygen uptake after 4 days of starvation, although control fish had a higher routine oxygen uptake, scope for spontaneous activity and initial acclimation oxygen uptake. During feeding, transgenic fish ate significantly more than control fish, and had an overall oxygen uptake that was 1·7 times greater than control fish. When fish that had eaten the same per cent body mass were compared, transgenic fish had an oxygen uptake that was 1·4 times greater than control fish. Differences in oxygen uptake in growth hormone transgenic coho salmon and non‐transgenic fish appear to be due to the effects of feeding, acclimation and activity level, and not to a difference in basal metabolism.

Intestinal morphology in growth hormone transgenic coho salmon

Intestinal morphology in growth hormone transgenic coho salmon

Intestinal morphology in growth hormone transgenic coho salmon

In two GH transgenic coho salmon Oncorhynchus kisutch, the surface area of the intestine was 2·2 times that of control salmon and the growth rate was about twice that of controls. It seems likely that the enhanced intestinal surface area is a compensatory feature that is manifested in fast growing salmonids.