Insulin-like growth factor I (IGF-I) mRNA expression in coho salmon,Oncorhynchus kisutch: Tissue distribution and effects of growth hormone/prolactin family proteins

To examine the hormonal and nutritional regulation of insulin-like growth factor I (IGF-I) mRNA expression, a sequence-specific solution hybridization/RNase protection assay for coho salmon IGF-I mRNA was developed. This assay is both rapid and sensitive and has low inter- (less than 15%) and intra-assay variations (less than 5%). Using this assay, the tissue distribution of IGF-I mRNA and effects of growth hormone (GH), prolactin (PRL) and somatolactin (SL) on hepatic IGF-I mRNA expression in coho salmon were examined in vivo. Liver had the highest IGF-I mRNA level of 16 pg/μg DNA. Significant amounts of IGF-I mRNA were also found in all other tissues examined (intestine 4.1, kidney 3.8, gill arch 2.4, brain 2.4, ovary 2.3, muscle 2.1, spleen 1.7 and fat 1.1 pg/μg DNA). Injection of coho salmon GH at doses of 0.1 and 1 μg/g body weight significantly increased the hepatic IGF-I mRNA levels in a dose-dependent manner. Injection of coho salmon SL, a recently discovered member of the GH/PRL family, stimulated the IGF-I mRNA expression at the higher dose (1 μg/g), whereas coho salmon PRL had no effect at either dose. Concentration-dependent stimulation by coho salmon GH was also obtained in vitro in primary culture of salmon hepatocytes in concentrations ranging from 0.01 to 1 μg/ml. These results indicate that IGF-I mRNA expression occurs in a variety of tissues in coho salmon, and that at least the hepatic expression is under the regulation of GH and possibly other hormones. The sequence-specific assay established in the present study can be used for accurate quantitation of IGF-I mRNA in salmonid species, and can contribute to a better understanding of the physiology of IGF-I in salmonids.

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Résumé Afin d'étudier les régulations homronales et nutritionnelles de l'expression des ARNm de l'IGF-I (insulin-like growth factor I), un dosage spécifique par hybridation en solution des ARNm d'IGF-I de saumon coho et protégé des RNases, a été développé. Ce dosage, à la fois rapide et sensible, présente un faible coefficient de variation inter- (< 15%) et intra- (< 5%) dosage. L'étude de la distribution tissulaire des ARNm de l'IGF-I et des effets de l'hormone de croissance (GH), de la prolactine (Prl) et de la somatolactine (SI) sur l'expression hépatique des ARNm de l'IGF-I, a été entreprise in vivo chez le saumon coho en utilisant ce dosage. Le foie présente les plus grandes quantités d'ARNm d'IGF-I (16 pg/μg d'ADN). Des quantités significatives d'ARNm d'IGF-I ont été également détectées dans tous les autres tissus étudiés (intestin 4,1; rein 3,8; branchie 2,4; ovaire 2,3; muscle 2,1; rate 1,7 et graisse 1,1 pg/μg d'ADN). L'injection à des saumons coho, de GH à des doses de 0,1 et 1 μg/g de poids vif, augmente significativement et de manière dose dépendante les niveaux hépatiques d'ARNm d'IGF-I. L'injection de SI de saumon coho, un membre récemment découvert de la famille GH/Prl, stimule avec la plus haute dose utilisée, l'expression des ARNm d'IGF-I alors que la Prl n'a aucun effet. La GH augmente de manière dose dépendante (0,01–1 μg/ml) l'expression in vitro des ARNm d'IGF-I par des ARNm d'IGF-I par des hépatocytes de saumon coho en culture. Ces résultats indiquent que, chez le saumon coho, l'expression des ARNm d'IGF-I est présente dans le nombreaux tissus et que, l'expression hépatique est, au moins en partie, régulée par la GH et peut-être par d'autres hormones. Le dosage par séquence spécifique mise au point dans le présent travail, peut-être utilisé pour la quantification précise des ARNm, d'IGF-I de salmonidés et devrait permettre une meilleure connaissance de la physiologie de L'IGF-I chez les salmonidés.

     

Identification and expression analysis of two growth hormone receptors in zanzibar tilapia (Oreochromis hornorum)

Growth hormone plays important roles in various physiological processes such as growth, metabolism, and reproduction. In this study, two cDNAs encoding growth hormone receptor (GHR) were isolated from the liver of zanzibar tilapia (Oreochromis hornornum). The two cDNAs were 2,831 and 2,044 bp in length and named GHR1 and GHR2, respectively. GHR1 and GHR2 shared 57.4% similarity in nucleotide sequences and 33.5% similarity in deduced amino acid sequences. Consequently, it was presumed that they were two different genes. Conserved regions of GHR1 and GHR2 in zanzibar tilapia were different from those of other vertebrates. For example, conserved box2 regions of GHR1 and GHR2 in zanzibar tilapia were, respectively, WVELM and WVEFT, while it was WVEFI for GHRs in other vertebrates. Similar to other fish species, GHR1 and GHR2 were expressed in brain, gill, liver, muscle, spleen, gonad, stomach, kidney, and pituitary in zanzibar tilapia. The expression levels were the highest in liver. Unlike fathead minnow (Pimephales promelas) and mossambique tilapia (O. mossambicus), the expression levels of GHR1 in most female fish tissues were higher than those in male fish. No significant difference in GHR2 expression was found in all the tissues in male and female of zanzibar tilapia. Under fasting condition, the expressions of GHRs and IGF-II were significantly up-regulated (P < 0.05) in liver, while the expression of IGF-I remained stable. This observation would contribute to understanding the evolution of the GHR family in further investigation of growth regulation of zanzibar tilapia.     

Disrupting actions of bisphenol A and malachite green on growth hormone receptor gene expression and signal transduction in seabream

Environmental estrogen could mimic natural estrogens thereby disrupting the endocrine systems of human and animals. The actions of such endocrine disruptors have been studied mainly on reproduction and development. However, estrogen could also affect the somatotropic axis via multiple targets such as growth hormone (GH). In the present study, two endocrine disruptors were chosen to investigate their effects on the expression level and signal transduction of growth hormone receptor (GHR) in fish. Using real-time PCR, it was found that exposure to both the estrogenic (bisphenol A) and anti-estrogenic (malachite green) compounds could attenuate the expression levels of GHR1 and GHR2 in black seabream (Acanthopagrus schlegeli) hepatocytes. The expression level of IGF-I, the downstream effector of GHR activation in the liver, was decreased by bisphenol A but not by malachite green. Luciferase reporter assay of the β-casein promoter was used to monitor GHR signaling in transfected cells. In the fish liver cell line Hepa-T1, both GHR1 and GHR2 signaling were attenuated by bisphenol A and malachite green. This attenuation could only occur in the presence of estrogen receptor, indicating that these agents probably produce their actions via the estrogen receptor. Results of the present study demonstrated that estrogenic or anti-estrogenic compounds could down-regulate the somatotropic axis in fish by affecting both the gene expression and signaling of GHR. In view of the increasing prevalence of these compounds in the environment, the impact on fish growth and development both in the wild and in aquaculture would be considerable.     

Increased ability to compete for food by growth hormone-transgenic coho salmon Oncorhynchus kisutch (Walbaum)

In salmonids, growth hormone (GH) stimulates growth, appetite and the ability to compete for food. This study tested the hypothesis that increased GH levels in GH-transgenic coho salmon Oncorhynchus kisutch (Walbaum) increase competitive ability through higher feeding motivation. The transgenic strain of salmon used contained a gene construct consisting of the sockeye metallothionein-B promoter fused to the type 1 growth gene coding region. The transgenic animals (mean size = 250 g) were F₁ individuals. In six consecutive feeding trials, the intake of contested food pellets by size-matched pairs consisting of one control (1 year older non-transgenic coho salmon) and one GH-transgenic coho salmon was compared. Pellets were provided sequentially until neither fish took three consecutive pellets; the identity of the fish taking each pellet was noted. Calculated on the three first pellets offered at each feeding trial, the transgenic coho salmon consumed 2.5 times more contested pellets than the controls, supporting the hypothesis that GH transgenesis increases the ability to compete for food. Overall, the transgenic fish consumed 2.9 times more pellets that the non-transgenic controls, indicating a high feeding motivation of the transgenic fish throughout the feeding trials. It appears that GH transgenesis and GH treatments can induce similar changes in the feeding behaviour of salmonids. Depending on how transgenic and wild individuals differ in other fitness-related characters, escaped GH transgenic fish may compete successfully with native fish in the wild.     

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.     

Juvenile coho salmon movement,growth and survival in a coastal basin of southern Oregon

Juvenile salmonids display highly variable spatial and temporal patterns of early dispersal that are influenced by density‐dependent and density‐independent factors. Although juvenile coho salmon (Oncorhynchus kisutch) movement patterns in streams and their relationship with body mass and growth have been examined in previous studies, most observations were limited to one season or one stream section. In this study, we monitored the movement of juvenile coho salmon throughout their period of residence in a coastal basin to identify prevalent dispersal strategies and their relationships with body mass, growth rates and survival. Our results revealed seasonally and spatially variable movement patterns. Juvenile coho salmon that dispersed to tidally affected reaches soon after emergence remained more mobile and expressed lower site fidelity than those individuals that remained in upper riverine reaches. We did not detect significantly different growth rates between sedentary and mobile individuals. Although a greater proportion of sedentary than mobile fish survived winter to emigrate from the creek in the spring, reach of residence at the onset of winter influenced these survival estimates. Hence, apparent summer‐to‐smolt survival for mobile individuals was greater than for sedentary fish in tidally influenced reaches, whereas in riverine reaches the sedentary strategy seemed to be favoured. Our research identified complex movement patterns that reflect phenotypic and life history variation, and underscores the importance of maintaining diverse freshwater and estuarine habitats that support juvenile coho salmon before marine migration.     

Effects of habitat and internal prey subsidies on juvenile coho salmon growth: implications for stream productive capacity

Abstract –  To evaluate the effects of habitat, foraging strategy (drift vs. limnetic feeding) and internal prey subsidies (downstream transport of invertebrate drift between habitats) on fish production, we measured the growth of juvenile coho salmon confined to enclosures in flowing (pond inlets and outlets) or standing water (centre of pond) habitats in a constructed river side-channel. The effects of habitat and foraging strategy on fish growth were mediated primarily through habitat effects on prey abundance. Invertebrate drift biomass was nearly an order of magnitude higher at pond inlets relative to outlets. Drift-feeding coho in inlet enclosures grew 50% faster than drift-feeding coho at pond outlets or limnetic feeding coho in the centre of ponds, suggesting that elevated drift at inlets was sufficient to account for higher inlet growth rates. Forty per cent of prey biomass in stomachs was terrestrial in origin. These results indicate that, in addition to dependence on external terrestrial subsidies, streams with alternating slow and fast water (i.e., pool-riffle) sequences are also characterised by internal prey subsidies based on transport of drifting invertebrates from refuge habitats (high velocity riffles) to habitats more suitable for drift-feeding predators (e.g., pools), which may result in higher maximum fish growth in systems where internal subsidises are large. Restoration of small streams to maximise productive capacity for pool-rearing salmonids will require a better understanding of the length and interspersion of habitats that maximises both internal prey subsidies and available rearing habitat for juvenile salmon.     

An autumn profile of growth regulatory hormones in chinook salmon (Oncorhynchus tshawytscha)

We examined trends in the growth regulatory hormones growth hormone (GH) and insulin-like growth factor-I (IGF-I) from August to December in chinook salmon. Fish on 100% (ad libitum) and 64% rations of a low fat high protein diet, and a 64% ration of commercial feed (BioOregon-grower) were sampled twice a month. Fish were kept on simulated natural photoperiod at constant temperature. GH declined in late August and early September, consistent with photoperiodic regulation. No effects of ration or diet composition on GH were found. IGF-I increased to a peak on 4 October 1998 and declined thereafter. High dietary ration and the higher fat commercial diet increased IGF-I. Fish length and IGF-I level were positively correlated. This study shows that a peak in IGF-I may occur in the fall in chinook salmon. This revised version was published online in August 2006 with corrections to the Cover Date.     

Growth Hormone Endocrinology of Salmonids: Regulatory Mechanisms and Mode of Action

The focus of this review is on the regulatory mechanisms and the mode of action of GH in salmonids. To stimulate further research, it aims at highlighting areas where numerous important breakthroughs have recently been made, as well as where data are currently lacking. The regulation of GH secretion is under complex hypothalamic control, as well as under negative feedback control by GH and IGF-I. Further, the recently characterized ghrelin is a potent GH secretagogue, and may prove to be a link between feed intake and growth regulation. GH plasma profiles show indications of diurnal changes, but whether salmonids have true pulsatile GH secretion remains to be elucidated. The recent cloning and characterization of the salmon GH receptor (GHR) is a major research break-through which will give new insights into the mechanisms of GH action. It should also stimulate research into circulating GH-binding proteins (GHBPs), as they appear to be a soluble form of the GHR. The salmonid GHR sequences show evolutionary divergence from other fish species, but with a high degree of identity within the salmonid group. Radioreceptorassay studies have found GHR present in all tissues examined, which is in line with the highly pleiotropic action of GH. Data are currently scarce on the plasma dynamics of GH in salmonids, and further studies on GHR and GHBPs dynamics coupled to assessments of GH clearance rates and pathways are needed. The direct versus indirect nature of GH action remains to be clarified, but GH appears to act both locally at the target tissue level to stimulate the autocrine/paracrine action of IGF-I, as well as on the liver to increase plasma IGF-I levels. In addition, GH interacts with other hormones such as cortisol, thyroid hormones, insulin, and reproductive hormones, generating a wide range of physiological effects. GH may act both peripherally and directly at the level of the central nervous system to modify behavior, probably by altering the dopaminergic activity in the brain.     

Modelling the reestablishment of coho salmon (Oncorhynchus kisutch) in Klamath River tributaries after dam removal

The planned removal of four dams on the Klamath River (anticipated 2024) will be the largest river restoration effort ever undertaken on the planet. Dam removal will restore access to >50 km of the Klamath River mainstem for coho salmon, but mainstem habitat may not be suitable for rearing juvenile coho salmon. Instead, small tributaries may provide most rearing habitat for reestablishing coho salmon. We used four approaches to evaluate six Klamath River tributaries above existing dams to assess their potential to support juvenile coho salmon: (1) We measured summer temperature regimes and evaluated thermal suitability. (2) We applied an Intrinsic Potential (IP) model to evaluate large-scale geomorphological constraints on coho salmon habitat. (3) We used the Habitat Limiting Factors Model (HLFM) to estimate rearing capacity for juveniles given current habitat conditions. (4) We developed an occupancy model using data from reference tributaries to predict coho salmon rearing distribution. All six streams had summer temperatures cooler than the mainstem Klamath River. However, five of the streams have barriers that will restrict coho salmon to within 5 km of the confluence with the Klamath River and two were disconnected mid-summer. Despite these constraints, the tributaries will likely produce coho salmon. Most streams had high IP in their lower reaches, the HLFM model estimated a total capacity of 105,000 juvenile coho salmon, and the occupancy model predicted juvenile coho salmon will rear throughout the accessible reaches. Protection and habitat enhancement for these tributaries will be important for coho salmon reestablishment post-dam removal.     

Dynamics of insulin and insulin-like growth factor-I (IGF-I) ovarian receptors during maturation in the brown trout (Salmo trutta)

In salmonid fishes, the role of insulin and insulin-like growth factor-I (IGF-I) in the regulation of ovarian function is not well understood. Recently, we reported that isolated follicular layers of the preovulatory ovarian follicle of coho salmon (Oncorhynchus kisutch) have specific receptors for insulin and IGF-I and that IGF-I modulates steroid production in the follicular layers. In the present study we have investigated the structural and functional characteristics of insulin and IGF-I receptors in the ovary of brown trout (Salmo trutta) and the changes in insulin and IGF-I binding throughout the reproductive cycle of this species. The specific binding for IGF-I was 8- to 15-fold higher than the specific binding for insulin. IGF-I receptors were also more specific than insulin receptors because unlabeled insulin displaced bound radiolabeled insulin at concentrations 40- to 80-fold lower than unlabeled IGF-I; whereas, unlabeled IGF-I displaced bound radiolabeled IGF-I at concentrations 4000- to 8000-fold lower than unlabeled insulin. Insulin and IGF-I receptors from the brown trout ovary were composed of 120 kDa -subunits and 90 kDa -subunits, which underwent autophosphorylation in a concentration-dependent manner. Receptor tyrosine kinase activity was also stimulated in a concentration-dependent manner by insulin and IGF-I. When ovarian insulin and IGF-I binding was determined from mid-vitellogenesis (March) until ovulation (November), maximal binding for both peptides was detected in mid-vitellogenesis and gradually decreased until the end of vitellogenesis (August). In the preovulatory period (October), a small increase of insulin and IGF-I binding was observed. After ovulation, insulin binding was no longer detectable and IGF-I binding was very low. These results suggest that insulin and IGF-I receptors in the salmonid ovary follow the structural pattern described in other vertebrate species and that insulin and IGF-I could be involved in the regulation of ovarian function during reproductive stages other than the preovulatory period.     

Evaluation of an alkane yeast (Candida sp.) as a substitute for fish meal in Oregon Moist Pellet: Feeding trials with coho salmon (Oncorhynchus kisutch) and rainbow trout (Salmo gairdneri)

Up to 40% of the fish meal portion of the Oregon Moist Pellet (OMP) was replaced with up to 40% alkane yeast single cell protein (22% of dietary protein) in diets fed to rainbow trout in fresh water, and up to 100% of the fish meal portion was replaced with alkane yeast (54% of dietary protein) in OMP diets fed to coho salmon in salt water. Salmon diets were supplemented with DL-methionine to correct for a suspected deficiency in this essential amino acid. Alkane yeast was an acceptable substitute for fish meal at all levels tested in trout. Only 25% yeast substitution was judged to be acceptable in the OMP diet for marine-cultured coho salmon, causing less than 4% reduction in growth. In marine-cultured coho salmon, growth was depressed further at fish meal substitution levels greater than 25%, while supplementation with DL-methionine enhanced growth and feed conversion only slightly.     

Survival of smaller sport caught chinook, Oncorhynchus tshawytscha (Walbaum), and coho, Oncorhynchus kisutch (Walbaum), salmon from Lake Michigan and its management implications

Abstract The survival of small-sized (<50.8 cm) chinook salmon, Oncorhynchus tshawytscha (Walbaum), and coho salmon, Oncorhynchus kitsutch (Walbaum), caught by sport fishing was determined to assess the potential for increasing the size limit for these fish. Fishermen were recently catching smaller salmon than in the 1970s, but salmon growth rates had not changed. To be an effective management option, the survival rate of hooked and returned fish must be high. The overall survival rates were high: 76% for chinook salmon and 70% for coho salmon. There was no significant difference in survival of the coho salmon with size of hook used ( P = 0.31). Any mortality among fish was generally acute; fish hooked deep in the mouth or gills generally bled and died shortly after capture. Fish hooked in the gills had a significantly greater mortality ( P = 0.0002). The overall high survival rate for these species was the result of a small proportion of fish being hooked in the gills or deep in the mouth. Since the survival rate of the salmon was high, the size limit could be increased to allow smaller fish to grow to sizes preferred by sport fishermen.     

Relationship between fish growth rate and progression of artificially induced erythrocytic inclusion body syndrome in coho salmon, Oncorhynchus kisutch (Walbaum)

Abstract. Studies were conducted to clarify the relationship between growth rate and disease progression of erythrocytic inclusion body syndrome (EIBS) in artificially infected coho salmon, Oncorhynchus kisutch (Walbaum). In the diseased state, the haematocrit values decreased and the number of erythrocytes with inclusions was higher in faster growing fish. Rapid growth was accompanied by an abundance of immature erythrocytes, which had the greatest incidence of inclusion bodies.     

Infectious haemolytic anaemia causes jaundice outbreaks in seawater-cultured coho salmon, Oncorhynchus kisutch (Walbaum), in Chile

In the last 9 years, epizootics of an icterus condition has affected coho salmon, Oncorhynchus kisutch (Walbaum), reared in seawater cages in southern regions of Chile. At necropsy, fish from field cases exhibited signs of jaundice accompanied by pale light-brown livers and dark spleens. Histopathological and haematological results indicated that these fish presented haemolytic anaemia. After microbiological examination no bacterial or viral agents could be identified as aetiological agents of this disease. In an infectivity trial, coho salmon, Atlantic salmon, Salmo salar L., and rainbow trout, Oncorhynchus mykiss (Walbaum), were inoculated intraperitoneally with a filtrate of an organ homogenate (0.45 microm) from a diseased coho salmon and held for 60 days in tanks supplied with fresh water. The disease was only reproduced in coho salmon in which mortalities, beginning at day 23 post-inoculation (p.i.), reached a cumulative value of 24% at day 27 p.i. This condition was transmitted to non-inoculated cohabiting coho salmon suggesting that it is a waterborne disease. Thus, this icteric condition is caused by an infectious form of haemolytic anaemia, probably of viral aetiology, and coho salmon are more susceptible than either Atlantic salmon or rainbow trout.     

Cloning of two Atlantic salmon growth hormone receptor isoforms and <Emphasis Type="Italic">in vitro</Emphasis> ligand-binding response

Two isoforms of the full-length cDNA of the growth hormone receptor (GHR) of the Atlantic salmon (Salmo salar; ss) were cloned by a PCR approach using RACE. Respectively, the cDNA sequences of ssGHR isoforms 1 and 2 are 2654 and 2608 nucleotides long, with 1782 and 1773 nucleotide ORFs. The resulting coded proteins are 594 and 590 aa long, with 19 and 20 aa signal peptides. The two isoforms share 86% protein and 87% cDNA sequence similarity. Isoform 1 is most similar to other salmonid GHR isoforms 1 while isoform 2 is most similar to salmonid GHR isoforms 2 (93–95%). Similarity with other teleost species was lower (37–44%). The bioactivity of the cloned ssGHR was tested by transfecting the ssGHR isoform 1 cDNA into CHO-K1 hamster cells, incubating with recombinant salmon GH (sGH) or native ovine prolactin (oPRL), and measuring cell proliferation by the MTT assay. The ssGHR-transfected cells significantly increased proliferation when stimulated by sGH at all concentrations. oPRL stimulated ssGHR-transfected cells at higher concentrations due to receptor cross reaction. ssGHR isoforms 1 and 2 contain a single transmembrane domain and the typical conserved motifs found in other teleost GHRs, including four paired cysteine residues and five potential N-glycosylation sites in the extracellular domain, Box I and Box II, as well as seven potential tyrosine phosphorylation sites in the intracellular domain. However, in salmonids, these motifs differ from those of other teleosts, and could be responsible for differentiated hormone binding, signal transduction and response.     

Immunogenicity of vaccines from a virulent and an a virulent strain of Aeromonas salmonicida

Abstract. Injected vaccines consisting of formalin-killed cells and extracellular antigens prepared from a virulent and an avirulent strain of Aeromonas salmonicida were tested for their efficacy in protecting juvenile coho salmon. Oncorhychus kisutch (Walbaum). Against experimental furunculosis following active and passive immunization. Sera used in the passive immunization experiments were raised in subadult coho salmon and in rabbits. Results indicated that the avirulent strain was inferior to the virulent strain in its immunogenicity for coho salmon. Thus, even though avirulent cells possessed at least one immunogen–an immunogen that was show in passive immunization experiments to be well recognized by rabbits–the immunogen was only inefficiently protection when vaccinated with avirulent cells. Furher, extracellular antigens of the avirulent strain were not protective for coho salmon even though they elicited the production of anti- A. salmonicida agglutinins. In contrast, the killed cells and extracellular antigens of the virulent strain were both immunogenic in coho salmon. By passively immunizing coho salmon with rabbit sera raised against the virulent and the virulent strain produced at least two immunogens, only one of which was shared in common with the avirulent strain. The extra immunogen possessed by the virulent strain is presumably responsible for its superior immunogenicity in coho salmon; the immunogen had the properties of a protein and is believed to be the A-protein.     

Food habits and marine survival of juvenile Chinook and coho salmon from marine waters of Southeast Alaska

Little is known about the food habits of juvenile Chinook (Oncorhynchus tshawytscha) and coho (Oncorhynchus kisutch) salmon in marine environments of Alaska, or whether their diets may have contributed to extremely high marine survival rates for coho salmon from Southeast Alaska and much more modest survival rates for Southeast Alaskan Chinook salmon. To address these issues, we documented the spatial and temporal variability of diets of both species collected from marine waters of Southeast Alaska during summers of 1997–2000. Food habits were similar: major prey items of both species included fishes, crab larvae, hyperiid amphipods, insects, and euphausiids. Multivariate analyses of diet composition indicated that the most distinct groups were formed at the smallest spatial and temporal scales (the haul), although groups also formed at larger scales, such as by month or habitat type. Our expectations for how food habits would influence survival were only partially supported. As predicted, Southeast Alaskan coho salmon had more prey in their stomachs overall [1.8% of body weight (BW)] and proportionally far fewer empty stomachs (0.7%) than either Alaskan Chinook (1.4% BW, 5.1% empty) or coho salmon from other regions. However, contrary to our expectations, coho salmon diets contained surprisingly few fish (49% by weight). Apparently, Alaskan coho salmon achieved extremely high marine survival rates despite a diet consisting largely of small, less energetically‐efficient crustacean prey. Our results suggest that diet quantity (how much is eaten) rather than diet quality (what is eaten) is important to marine survival.     

Propylthiouracil-induced hypothyroidism in coho salmon, Oncorhynchus kisutch: effects on plasma total thyroxine, total triiodothyronine, free thyroxine, and growth hormone

Thyroid hormones transiently increase during parr-smolt transformation in coho salmon, Oncorhynchus kisutch, and are believed to trigger morphological, physiological, behavioural, and neural changes. The effectiveness of propylthiouracil (PTU) to induce hypothyroidism in smolting coho salmon was determined by immersing coho salmon, Oncorhynchus kisutch, in 30 mg l–1 PTU from May 1, two weeks prior to the consistent annual total thyroxine (TT4) peak in mid-May, until the last sampling date. Plasma was obtained at two sampling dates from control and PTU -treated coho salmon: May 15, during the plasma TT4 peak; and May 26, after the TT4 peak. Radioimmunoassays were used to measure plasma TT4, total triiodothyronine (TT3), free thyroxine (FT4), and salmon growth hormone (sGH). The PTU -treatment inhibited the natural smoltification-related increases in plasma TT4, TT3 and GH levels compared with controls, but PTU-treatment did not affect these hormone levels when they were low. PTU -treatment increased FT4 and decreased TT3 and sGH levels in the May 26 sample. In the May 15 sample, FT4 levels were unaffected by PTU-treatment, whereas TT4 levels were decreased. These data demonstrate the ability of PTU to induce hypothyroidism in salmonids as shown by the decrease in TT4 and TT3. These data demonstrate that PTU treatment by immersion can induce hypothyroidism in salmonids as shown by: (1) the inhibition of the natural increases of TT4 and TT3; (2) the increase in FT4 levels corresponding to the lowered TT3 levels, suggesting an inhibition of thyroxine 5-monodeiodinase activity. We also show for the first time that PTU treatment can lower plasma GH levels in salmonids. This lowering of plasma GH level is associated with the decrease in TT3 levels and the increase in FT4 levels. The PTU induced lowering in GH levels may contribute to the observed changes in FT4 and TT3, since GH is known to increase thyroxine 5-monodeiodinase activity.