Antisomatostatin-induced growth acceleration in chinook salmon (Oncorhynchus tshawytscha)

Since somatostatin (SRIF) inhibits the release of growth hormone (GH), its immunoneutralization may provide an alternative to GH therapy as a means of enhancing somatic growth in fish. The present study examined the feasibility of accelerating growth in juvenile chinook salmon by means of antiSRIF administration. Yearling salmon of Nicola River stock (BC, Canada) were injected intraperitoneally every 5 days, for a total of 40 days, with either SRIF (1 μg g-1 body wt.), antiSRIF (SOMA-10, 1 μg g⁻¹), recombinant bovine GH (rbGH, 2.5 μg g⁻¹), recombinant porcine GH (rpGH, 2.5 μg g⁻¹) or saline (controls). No significant differences were observed in length, weight or final condition factor (k) between the SRIF-treated and control fish over the experimental period. However, the fish treated with the antiSRIF were significantly (p ≤ 0.05) longer and heavier than the control salmon after 25 and 30 days respectively. Furthermore, antiSRIF treatment caused a lowering in k when compared to the control salmon. Fish injected with rbGH or rpGH were significantly longer and heavier than all other groups (p ≤ 0.05), after only 5 days. GH treated groups also returned higher k when compared against all other treatments (p ≤ 0.05). No differences were observed in growth between the two rGH treatments over the experimental period.     

Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study

A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer (pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated. The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes.     

Genetic variation in freshwater survival and growth of chinook salmon (Oncorhynchus tshawytscha)

Five pairs of paternal half-sib families were obtained from each of six stocks of chinook salmon (Oncorhynchus tshawytscha) in British Columbia. Life history patterns and size and age at maturity varied among the parental fish used from each stock. The freshwater survival and growth of progeny were monitored under controlled hatchery conditions.Survival of uneyed eggs, alevins, fry, smolts, and cumulative survival to 125–140 days after ponding differed among stocks and among families within stocks. The overall heritabilities (sire component) of survival at all four developmental stages were less than 0.05. Alevin weight, smolt weight, and growth rate differed among stocks and among families within stocks. The heritability (sire component) of smolt weight ranged from 0.0±0.59 to 0.88±0.72 among stocks, with an overall value of 0.0. The success of selection for smolt weight might be stock-dependent.     

Oxidative stress in juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Juvenile chinook salmon, Oncorhynchus tshawytscha (Walbaum), were held in 8–11°C freshwater, starved for 3 days and subjected to a low‐water stressor to determine the relationship between the general stress response and oxidative stress. Lipid peroxidation (LPO) levels (lipid hydroperoxides) were measured in kidney, liver and brain samples taken at the beginning of the experiment (0‐h unstressed controls) and at 6, 24 and 48 h after application of a continuous low‐water stressor. Tissue samples were also taken at 48 h from fish that had not been exposed to the stressor (48‐h unstressed controls). Exposure to the low‐water stressor affected LPO in kidney and brain tissues. In kidney, LPO decreased 6 h after imposition of the stressor; similar but less pronounced decreases also occurred in the liver and brain. At 48 h, LPO increased (in comparison with 6‐h stressed tissues) in the kidney and brain. In comparison with 48‐h unstressed controls, LPO levels were higher in the kidney and brain of stressed fish. Although preliminary, results suggest that stress can cause oxidative tissue damage in juvenile chinook salmon. Measures of oxidative stress have shown similar responses to stress in mammals; however, further research is needed to determine the extent of the stress–oxidative stress relationship and the underlying physiological mechanisms in fish.     

Expression of insulin-like growth factor-I in the gastrointestinal tract of Atlantic salmon (Salmo salar L.)

The expression of insulin-like growth factor (IGF-I) in the gut of Atlantic salmon (Salmo salar L.) smolt was investigated at the cellular level using in situ hybridization. A hybridization protocol was adapted for use on formalin fixed, paraffin embedded tissues and was based on a digoxigenin-labelled oligoprobe that recognised a conserved region of IGF-I mRNA. The specificity of the anti-sense IGF-I probe and the performance of the hybridization protocol were evaluated using a sense IGF-I probe, an Ep stein-Barr virus probe and an insulin probe cocktail on serial sections of fish gut, mouse thyroid and human lymph node tissue. In the gut of Atlantic salmon smolt, IGF-I was found to be expressed in single epithelial cells or small clusters of epithelial cells in the pyloric ampulla and in the pyloric caeca. Expression was not detected in the lamina propria of the gut or in epithelial cells of the stomach, midgut or hindgut. The restriction of IGF-I expression to the pyloric ampulla and pyloric caeca is consistent with the role of these gut segments in osmoregulation and the scattered epithelial expression supports the putative autocrine/paracrine mechanism of action of IGF-I in osmoregulation.     

Isolation and in vitro characteristics of chinook salmon (Oncorhynchus tshawytscha) rosette agent

Isolation and in vitro characteristics of a significant obligate intracellular eukaryotic pathogen (rosette agent) of chinook salmon (Oncorhynchus tshawytscha) are described. The pathogen was isolated by primary culture of infected spleen tissue followed by inoculation onto the chinook salmon embryo CHSE-214 cell line. Chicken and rabbit antibody was produced to a soluble antigen derived from the rosette agent. Mortalities and the disease were experimentally reproduced by inoculating chinook salmon with the isolate. The pathogen was reisolated 27 days post-inoculation from moribund fish. Identity of the isolate and the disease-causing organism was further supported by documenting antigenic and morphologic identity. Tested in CHSE-214 cells, the organisms are sensitive to the antibiotics oxytetracycline, chloramphenicol, amphotericin-B and nystatin. A ratio of about one organism per tissue culture cell at inoculation was required to develop an easily detectable infection. Adsorption of organisms to tissue culture cells at inoculation did not enhance the pathogen's infectivity. The pathogen replicated at an increasing rate from 5°C to 20°C. It could not be stored by freezing but storage in tissue culture at 5°C for at least 5 months proved possible. Early passages of the organism showed a high degree of host specificity in tissue culture which decreased in later passages. The taxonomic affinities of the organism are discussed.     

Gastric dilation and air sacculitis in farmed chinook salmon, Oncorhynchus tshawytscha (Walbaum)

A syndrome affecting cultured chinook salmon, Oncorhynchus tshawytscha (Walbaum), characterized by distended abdomens, gastric dilation, air sacculitis (GDAS), increased feed conversion rates and increased mortality has been recognized in New Zealand. Affected fish were most obvious in sea cages but were also present in fresh water. Mortality rates associated with this condition were highest in late summer and approached 6% per month. A dilated and flaccid stomach, without visible rugal folds containing copious oil, watery fluid or undigested feed was typical. Gastric mucosal ulceration or inflammation were not present. The air sacculitis consisted of a thickened, dilated bladder with a mixed mucosal inflammatory infiltrate and a luminal exudate associated with large numbers of morphologically diverse bacteria. Gastric dilation or air sacculitis occurred alone or together in the same fish. In a group of 20 subclinically affected fish with or without gastric dilation, there were no significant differences in weight, length, serum osmolality, sodium, total protein or packed cell volume. Twenty-three severely affected fish had significantly ( P  < 0.05) higher serum osmolality but similar sodium and total protein to that of clinically normal fish.     

Evaluation of an experimental Aeromonas salmonicida epidemic in chinook salmon, Oncorhynchus tshawytscha (Walbaum)

To determine the dynamics of the transmission of Aeromonas salmonicida ssp. salmonicida infection, chinook salmon, Oncorhynchus tshawytscha, were exposed to bacteria by cohabitation. The latent period (time between exposure and infectivity) was determined by exposing a group of chinook salmonid fingerlings to A. salmonicida by bath, then, at daily intervals, by holding five exposed (donor) fish with approximately 50 naive fish for 24 h. The latent period was 3 days post-infection and the time period between the initial exposure to bacteria and the beginning of bacterial shedding was 4.5 days for the same animals. The prevalence and intensity of infection in the donor fish, to which recipient fish were exposed, i.e. the level of exposure, was highly correlated with the development of disease in recipient (susceptible) chinook salmon (r2 = 0.57). An experiment was conducted to determine the daily progress of infection and development of a furunculosis epidemic among recipient fish by cohabiting a single exposed fish with 43 unexposed salmon. At daily intervals, all fish (in seven treatment tanks and one control tank daily) were sacrificed and tested for the presence of A. salmonicida in the kidney (n = 3520). Over 10 days, mean prevalence among recipient fish reached 75% and disease related mortality exceeded 50%. Bacterial concentrations in the water continued to increase over the duration of the experiment in concert with the number of infected animals present in the population.     

Age,ration level,and exercise affect the fatty acid profile of chinook salmon (Oncorhynchus tshawytscha) muscle differently

This study was undertaken to investigate the effect of sustained physical exercise (SS, swimming speed) on the fatty acid profile of muscle in PIT-tagged all-female chinook salmon (Oncorhynchus tshawytscha) in relation to their age and ration level (RL; i.e., maximum ration, RL100 or 75% of maximum, RL75). Accordingly, body weight (BW), specific growth rate (SGR), and total muscle lipid content (TLC) were considered as covariates in data analyses. In addition, plasma levels of thyroid hormones (thyroxine, T₄ and 3,5,3′-triiodo-l-thyronine, T₃) were compared to the muscle fatty acid (FA) compositions of individual fish to determine if there were any associations between thyroidal status and FA percentages. During the 300-day study, body weight increased from 75 to 440 g. Fish age explained most of the changes found in muscle FA composition [that is, R² was 0.23 for saturated fatty acids (SAFAs), 0.65 for monounsaturated fatty acids (MUFAs), and 0.71 for polyunsaturated fatty acids (PUFAs) with p<0.0007 in all cases]. Reduction in RL had less influence on FA composition (p>0.15 for SAFA, and R² was 0.27 for MUFA and 0.34 for PUFA, with p<0.0001 in each of the latter cases). SS only affected MUFA significantly (p<0.07 with R²=0.09). The observed effects of fish age and SS on FA composition were to a large degree uninfluenced by BW, SGR, or TLC, while the effects of RL were markedly diminished when these covariates were included in the statistical model. Thus, the changes in body weight and/or lipid content were viewed as being the direct cause for the alteration in muscle FA profile seen with the main factor RL. The effect of SS was pronounced only if fish with a reduced RL were compared. In this situation, the level of PUFA, especially docosahexaenoic acid (DHA; 22:6 n-3), decreased and MUFA increased. Plasma T₄ was unrelated to muscle FA levels, but plasma T₃ was correlated positively with muscle MUFA and negatively with PUFA. We conclude that in order for exercise to affect chinook salmon fillet FA composition, it has to be combined with restricted feeding. The data also imply that in accordance with higher vertebrates, T₃ is also involved in the regulation of FA metabolism in fish.     

Plasma insulin-like growth factor-I concentrations in yearling chinook salmon (Oncorhynchus tshawytscha) migrating from the Snake River Basin, USA

During the parr-to-smolt transformation (smoltification) of juvenile salmonids, preadaptive changes in osmoregulatory and ionoregulatory ability are regulated in part by the growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis. If food intake is sufficient, plasma IGF-I increases during smoltification. On the other hand, plasma IGF-I typically decreases in fasting fish and other vertebrate animals. Because food availability is limited for juvenile salmonids undertaking an extended 6- to 12-week springmigration to and through the Snake-Columbia River hydropower system (northwestern USA), IGF-I concentrations might be expected to decrease, potentially compromising seawater tolerance. To address this possibility,yearling chinook salmon Oncorhynchus tshawytscha reared in three Snake River Basin hatcheries were sampled before release and at two downstream dams. Dry masses ofmigrating fish either did not increase during themigration (in 2000, an average-flow year), or decreased significantly (in 2001, a low-flow year). In both years, plasma IGF-I levels were significantly higher (1.6-fold in 2000, 3.7-fold in 2001) for fish sampled at the last dam on the lower Columbia River than for fish sampled prior to release. Plasma IGF-I concentrations inmigrating fish may, nonetheless, have been nutritionally down-regulated to some degree, because plasma IGF-I concentrations in juvenile chinook salmon captured at a Snake River dam and transported to the laboratory increased in fed groups, but decreased in unfed groups. The ability ofmigrating smolts to maintain relatively elevated IGF-I levels despiteRestricted food intake and loss of body mass is likely related to smoltification-associated changes in hormonal balance.     

ATP levels of chinook salmon (Oncorhynchus tshawytscha) sperm following in vitro exposure to various oxygen tensions

Adenosine triphosphate (ATP) levels in sperm from chinook salmon (Oncorhynchus tshawytscha) were found to be 12.1±1.9 pmol ATP per 10⁶ sperm cells (mean±SEM, n=18). Sperm were stored at 0–2 °C for up to 72 h under 100, 21, and 0% O₂. Changes in sperm ATP content of samples maintained under 100 and 21% O₂ were indistinguishable, decreasing to 50% of initial values after 72 h. ATP levels of sperm stored under 0% O₂ decreased to 10% of initial values within 8 h and remained unchanged throughout the following 64 h. The initial percent of living sperm (sperm viability) was 94.7±1.7% (mean±SEM, n=12) and was unaltered at 24 h under all O₂ tensions. At 72 h, sperm viability remained unchanged under 21% O₂ and had decreased by 20% under 100% O₂ and by 30% under 0% O₂. Therefore, reductions in sperm ATP levels could not be attributed to cell lysis, as viability decreased only modestly over these durations. Fertilizing ability correlated positively with sperm ATP levels. Spermatocrit values increased by 20% for the samples maintained under 100 and 21% O₂ after 72 h. However, a 20% increase in spermatocrit occurred within 24 h in samples maintained under 0% O₂, rising 70% after 72 h under 0% O₂. These data indicate that chinook sperm ATP (1) can be altered and manipulated without significant loss of viability, (2) is positively associated with fertilization, and (3) probably plays a role in sperm packing and cell flexibility.     

Utility of genetically based health indicators for selection purposes in captive-reared chinook salmon, Oncorhynchus tshawytscha, Walbaum

Three health indicators, plasma lysozyme activity, PCR‐based detection of Renibacterium salmoninarum (a causative agent of bacterial kidney disease), and a necropsy‐based Health Assessment Index (HAI), were used to examined genetically based variation in a captive population of chinook salmon (Oncorhynchus tshawytscha W.). The study group consisted of four distinct genetic cross‐types: two purebred cross‐types originating from mating wild parents (Big Qualicum River, BC, Canada) and domestic parents (Yellow Island Aquaculture, Ltd, Quadra Island, BC, Canada) and two reciprocal hybrid cross‐types from the mating of wild and domestic parents. Narrow‐sense heritability estimates for plasma lysozyme activity and the incidence of R. salmoninarum were calculated, and the genetic correlation of health indicator response with survival and growth was estimated. Significant differences among cross‐types were found for plasma lysozyme activity, HAI, survival after a natural outbreak of vibriosis (but not after a vibriosis disease challenge), relative growth rate, size‐at‐age (420 and 615 days post fertilization), and R. salmoninarum presence. Despite a significant sire component of heritability for plasma lysozyme activity, the lack of significant heritability estimates for R. salmoninarum presence, and non‐significant genetic correlations with performance variables indicates that selection to improve the health status of fish stock using the three health indicators examined here would likely not result in a measurable correlated response in survival or growth.     

Changes in plasma insulin-like growth factor-I levels in the precociously maturing amago salmon, Oncorhynchus masou ishikawai

Seasonal changes in plasma levels of insulin-like growth factor I (IGF-I) in precociously maturing amago salmon (Oncorhynchus masou ishikawai), which matured as 1-year-olds, have been investigated. Profiles of plasma IGF-I levels were compared with changes in growth and maturity, and plasma growth hormone (GH) and thyroxine (T₄) concentrations. The maturity of the fish was determined by calculating the gonadosomatic index; in November, 100% of males and 89% females matured. In both males and females, plasma IGF-I levels increased from March to August, and subsequently, plasma IGF-I levels in the early maturing males and females declined gradually and were maintained at lower levels during the spawning period in November. Plasma GH levels were high in April, and then declined gradually through September. Thereafter, in early maturing fish, a slight increase in plasma GH levels was observed in October and November. No significant changes in plasma T₄ levels were found in the precociously maturing fish. In sharp contrast, plasma IGF-I levels in immature fish remained elevated through September, reaching a peak in October, and then gradually declined in November. In immature females, plasma T₄ and GH levels were elevated in August, reached their maximum in September and then gradually declined until November.     

An immune-complex glomerulonephritis of Chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Chinook salmon from New Zealand were shown to have a generalized membranous glomerulonephritis that was most severe in large fish. Marked thickening of the glomerular basement membrane was the most consistent lesion, with the presence of an electron-dense deposit beneath the capillary endothelium.Severely affected glomeruli also had expansion of the mesangium and loss of capillaries,synechiae of the visceral and parietal epithelium and mild fibrosis of Bowmans capsule. Chinook salmon from British Columbia, Canada with bacterial kidney disease caused by Renibacterium salmoninarum had similar histological lesions. They also had thickened glomerular basement membranes that were recognized by rabbit antiserum to rainbow trout immunoglobulin. This was true only when frozen sections of kidney were used and not formalin-fixed tissue. An attempt to experimentally produce a glomerulopathy in rainbow trout by repeated immunization with killed R. salmoninarum was not successful. Case records from the Fish Pathology Laboratory at the University of Guelph over a 10-year period revealed that a range of species were diagnosed with glomerulopathies similar to those seen in Chinook salmon. The majority of these cases were determined to have chronic inflammatory disease. This report has identified the presence of immunoglobulin within thickened basement membranes of Chinook salmon with glomerulonephritis and supports the existence of type III hypersensitivity in fish.     

Juvenile chinook salmon (Oncorhynchus tshawytscha) growth on the central California coast during the 1998 El Niño and 1999 La Niña

We assessed growth in subyearling chinook salmon (Oncorhynchus tshawytscha) during the 1998 El Niño and 1999 La Niña in the Gulf of the Farallones, a region of the continental shelf off central California seaward of the Golden Gate and the southernmost ocean entry point for the species in North America. Juvenile salmon demonstrated greater growth during this strong El Niño, when water temperature anomalies of more than +3°C were recorded at local buoys, than during the similarly strong 1999 La Niña. Slopes of regressions of weight on length, length on age, and weight on age were all significantly greater for juvenile salmon during the 1998 El Niño compared with those in the 1999 La Niña. Daily otolith increment widths, an estimator of somatic growth, corroborated population data. Between June 1 and August 9, mean increment widths for juvenile chinook salmon in 1998 were 3.54 ± 0.03 μm, significantly larger than the 3.13 ± 0.03 μm found in juveniles during the same time interval in 1999. Condition factor for juvenile chinook salmon entering the ocean at the Golden Gate was the same in both years, but became significantly greater in ocean fish during the 1998 El Niño than in ocean fish during the 1999 La Niña. Energy storage was significantly greater in ocean juvenile salmon during the 1998 El Niño as well. Mean triacylglycerol/cholesterol ratios increased following ocean entry in 1998, whereas they declined in ocean juveniles during 1999. Thus, not only was growth better in the El Niño period compared with La Niña, but lipid accumulation was also better. Oceanographic data for 1998 indicated elevated temperatures, lower salinity, greater freshwater outflow from San Francisco Bay, northerly flowing coastal currents, and positive upwelling index anomalies. This combination of environmental factors resulted in greater zooplankton productivity that, in conjunction with higher temperatures, allowed metabolic processes to enhance growth. Although El Niño events have certainly produced large-scale, and often adverse, effects on ecosystems, the results of this study emphasize the importance of local oceanographic conditions to growth and other physiological and ecological processes.     

Characteristics of carotenoid distribution in various tissues from red- and white-fleshed chinook salmon, Oncorhynchus tshawytscha (Walbaum)

Abstract. Carotenoid distribution was examined in muscle, ovary, integument and serum from red- and white-fleshed chinook salmon, Oncorhynchus tshawytscha (Walbaum), along with their muscle proximate compositions. There were no significant differences in muscle compositions of moisture, protein and lipid except for the carotenoid levels between red- and white-fleshed salmon. Similar carotenoid levels of ovaries were found between red- and white-fleshed chinook salmon, although the muscle carotenoid level of white-fleshed salmon was apparently low compared to red-fleshed salmon. This suggests that white-fleshed chinook salmon have the ability to accumulate the carotenoids in their ovaries but not muscle. The white-fleshed chinook salmon may be devoid of carotenoid receptors in their muscle and this seems to be the reason for the failure to accumulate the carotenoids in their muscle.     

Adenosine triphosphate levels of chinook salmon (Oncorhynchus tshawytscha) eggs following in vitro maintenance and activation/fertilization

To further examine the concept of egg quality and the physiology of stored salmonid eggs, we investigated the effects of different oxygen tensions on the adenosine triphosphate (ATP) levels of unfertilized, activated, and fertilized chinook salmon (Oncorhynchus tshawytscha) eggs. The ATP levels of unfertilized chinook salmon eggs were 2.61±0.14 nmol ATP per egg (17.6±0.9 mol l^–1 relative to cell water) and ranged from 1.98 to 3.63 nmol ATP per egg. The ATP content of unfertilized eggs maintained at 10 °C under 100% O₂, 21% O₂, and 100% N₂ remained unaltered throughout a 120 h storage period. Storing eggs under identical conditions at 20 °C (in an effort to speed egg metabolism and ATP turnover) resulted in significant O₂-independent decreases in ATP levels. However, ATP levels of unfertilized eggs exposed to 1 mmol l^–1 potassium cyanide (a potent inhibitor of oxidative phosphorylation) at 10 °C were significantly decreased after 24 h and continued to decline throughout the 120 h maintenance period to about 30% of time=0 values. Maintenance with exogenous nutrients (5 mmol l^–1 acetate plus 5 mmol l^–1 pyruvate) over 120 h at 10 °C did not alter the ATP content of unfertilized eggs. Eggs activated by exposing them to 10 °C water for a few minutes showed a rapid decrease in ATP values, regardless of whether the eggs were fertilized or not. Following an initial 25% drop after fertilization, the ATP levels remained stable for the remainder (5 d) of the incubation period in eggs maintained in 10 °C water. Therefore, unfertilized chinook salmon egg ATP levels appear to be relatively stable and maintained by a low, cyanide-inhibitable metabolism. The stability of egg ATP levels may be one reason that salmonid eggs can be stored for several days while eggs from other fishes cannot.     

Carcass selenium loss as an indicator of stress in barge transported chinook salmon (Oncorhynchus tshawytscha Walbaum) smolts

Two salmon smolt barge transport experiments were conducted to measure tissue trace element selenium (Se) loss during the 30 h transport 500 km downriver past seven hydroelectric dams on the Columbia River. Carcass Se was measured before and after the barge trip. Liver glutathione peroxidase (GTPX) activity and total ascorbate concentrations were assayed to correlate Se loss with GTPX levels. Hatchery‐reared smolt chinook salmon, Oncorhynchus tshawytscha (Walbaum), were tested before and after barging. Liver ascorbate concentrations, measured by high‐performance liquid chromatography techniques, showed levels adequate to support GTPX activity. Salmon smolts lost up to 20% of carcass tissue Se during the 30 h barge confinement. Selenium was analysed by analytical polarography. Samples collected after two barging episodes showed Se loss and concomitant elevated liver GTPX activity after the transport. Tissue Se loss may be useful to measure stress in fish, and increasing tissue Se in hatchery salmon smolts prior to release and transport may be warranted.