High efficiency gene transfer in rainbow trout (Salmo gairdneri Rich.) by microinjection into egg cytoplasm

A plasmid containing human growth hormone cDNA sequence was injected into cytoplasm of fertilized rainbow trout (Salmo gairdneri Rich.) eggs before first cleavage. Survival rates at hatching (stage of analysis) were high (average: 77%; range: 50–100%). Foreign DNA was detected in pooled and individual embryos either by dot blot or by southern electrophoresis procedures using the labelled plasmid as probe; it comigrated with the high molecular weight DNA and was therefore most likely integrated in the trout genome. Transformation yields were higher with linearized than with circular plasmid (75% versus 40%). These data suggest that transgenic trout can be routinely obtained in large proportions.     

The effect of short-term starvation on ovarian development and egg viability in rainbow trout (Salmo gairdneri)

Adult female rainbow trout (Salmo gairdneri) were fed a commercial trout production diet until approximately 40 days prior to spawning at which time the fish were divided in two groups. One group continued to be fed while the other group was starved until spawning. Egg size, egg number, ovary weight, and egg proximate composition were examined periodically starting 5 months before spawning. Muscle tissues of the fish were sampled at spawning to measure somatic depletion of proximate consituents in the two groups. Egg hatchability was determined by incubating test lots from each spawning female. No significant differences (P > 0.05) were found in egg hatchability between the fed and starved groups. Changes in proximate composition were observed in the eggs during the months prior to spawning, but no differences were detected between the eggs of the starved and the fed groups. Short-term starvation did not significantly alter the proximate composition of muscle tissue of the spawned female trout.     

Genetic variance for growth in rainbow trout (Salmo gairdneri)

The possibilities of genetic improvement of growth of rainbow trout were investigated. Two half-diallel crossing experiments with three strains were performed. In the second experiment families within a strain or strain combinations could be identified. One hundred fish of each strain, strain cross or family were measured at five or four different ages. Differences between strains, crosses and between families within each of them were highly significant at all ages. In juvenile stages heterosis seemed to be present, although it was not statistically significant. Differences between strains became more pronounced as age advanced. Genetic variance within strains and strain crosses was large and appeared to increase with age. Genetic improvement by selection appears promising provided mass selection is not performed too early. The small F-values of preliminary expt. 2 may indicate strong competition within each group which tends to increase residual variability.     

Immunoglobulin heterogeneity in the rainbow trout, Salmo gairdneri Richardson

Abstract. Hetereogeneity of rainbow trout immunoglobulins was demonstrated by using monoclonal antibody 1A6 and polyacrylamide-gradient gel electrophoresis. Immunoglobulins defined by elisa using monoclonal antibody 1A6 were about 30% of the total immunoglobulins, detected by elisa using polyclonal antibodies, in healthy rainbow trout. In trout obtained from farms with a previous history of infectious viral diseases, 1A6-immunoglobulins were only about 14% of the total. Several serum pools from infected trout could be totally depleted of 1A6-immunoglobulins (about 12% of total immunoglobulins) by affinity chromatography over Sepharose immobilized monoclonal antibody 1A6. Polyacrylamide-gradient gel electrophoresis under denaturing conditions of total immunoglobulins, 1A6 immunoglobulins and no-1A6 immunoglobulins purified by affinity chromatography, showed a majority heavy chain of 70 KDa and a minority heavy chain of about 60 KDa, two light chains of 24 and 26 KDa, and a 11–14 KDa polypeptide.     

Strawberry disease in rainbow trout, Salmo gairdneri Richardson

Abstract. Strawberry disease is a subchronic, non–debilitating, and non–fatal inflammatory skin disease of rainbow trout that has been recognized for the past 25 years. Morbidity ranges up to 80% and the disease affects fishes from 15cm in length to brood stock size. Epizootiological data indicate that strawberry disease usually appears, first in the autumn and reaches peak incidence during the following winter and spring. Data also indicate that the disease is found only in certain trout farms in the western and Pacific north–western regions of the United States. Although the cause of strawberry disease is unknown, the normal clinical course of 8 weeks can be reduced by oral treatment with oxytetracycline. Clinical signs include appearance of bright red, raised, circumscribed, ulcerated and indurated lesions in the skin that usually range up to 3 cm or more in diameter. Microscopically, the disease is characterized as a subchronic, focal, non–suppurative dermatitis with ulceration and extensive infiltration of mononuclear inflammatory cells.     

Proliferative kidney disease in rainbow trout Salmo gairdneri Richardson

Abstract. Two entirely separate outbreaks of PKD are described in an attempt to properly define the condition and enable it to be differentiated from other diseases of a similar nature. Losses of up to 75% of fingerling rainbow trout were recorded in one outbreak, illustrating the potentially serious nature of the disease. Although the kidneys showed the most obvious pathological changes, other organs were also affected. The host response was granulomatous in type, the foci of each cellular reaction being 5-20 μm eosinophilic cells considered to be parasitic. The cytoplasm of these cells contained numerous electron dense bodies some with a tail, but each with an electron lucent bar always orientated at right angles to the plasmalemma whenever they were near it. The most obvious cytoplasmic inclusion of these 'parasites' were other cells numbering up to five or six, each with its own nucleus and nucleolus, but devoid of any of the small electron dense bodies. The significance of either of these inclusions is unknown.
A limited differential diagnosis is given, and the argument presented that the condition PKD is the result of infection by a parasite, possibly an amoeba, in combination with poor quality water.     

Effects of interaction between strains and environment on growth traits in rainbow trout (Salmo gairdneri)

Three strains and their three crosses were tested in two environments, floating nets and ponds. The genetic growth potential was reduced considerably in the much poorer pond environment. In addition, interactions independent of the average performance of the particular genotype were observed for weight and length but not for height and head length.     

Control of gallbladder motility in the rainbow trout,Salmo gairdneri

Immunohistochemistry revealed nerves containing VIP-like and 5-HT-like material in both gallbladder wall and bile duct of the rainbow trout, while endocrine cells containing gastrin/CCK-like and substance P-like material were present in the mucosa of the bile duct and the duodenum. Fluorescence histochemistry showed adrenergic nerves close to the muscle layer of the gallbladder.Sulphated CCK8, caerulein and non-sulphated CCK8 (in this order of potency), 5-HT and acetylcholine were excitatory on isolated strip preparations, while VIP and adrenergic agonists were inhibitory. The adrenergic drugs were probably actingvia a beta-adrenergic receptor, while the effects of 5-HT and cholinergic drugs were antagonized by methysergide and atropine respectively.Electrical stimulation of the gallbladder nervesin situ failed to show any effect or under certain conditions induced a rebound effect.It is concluded that the motility control of the rainbow trout gallbladder may involve an inhibitory innervation by adrenergic and possibly VIP-releasing nerves, while 5-HT, acetylcholine and a CCK-like substance may be involved in the excitatory control.     

Thyroidal compensation in rainbow trout (Salmo gairdneri) fed canola meal

Rainbow trout (Salmo gairdneri Richardson) were fed either a soybean mealbased (SM) or canola meal-based (CM) diet for up to 20 weeks. Plasma thyroxine (T₄) and triiodothryonine (T₃) levels were significantly lower in the CM-fed fish sampled after 12 weeks. However, there appeared to be some compensation after 12 and 20 weeks in that the thyroid hormone levels in trout fed the CM were not significantly different from those of the SM-fed fish. Nevertheless, there was marked thyroid hyperplasia and hypertrophy in the CM-fed fish sampled at 12, 16 and 20 weeks after commencement of the experiment. Moreover, the growth rate was significantly lower in the CM-fed fish in comparison to the SM-fed fish throughout the 20 week study period.Plasma T₄ levels were similar in SM-fed fish sampled 12, 16 and 20 weeks after commencement of the experiment, but plasma T₃ levels progressively increased over this period, as did the apparent activity of the thyroid tissue based on histological criteria.Fasting for up to 8 weeks resulted in the arrested growth of the SM-fed fish, and a loss in body weight of the CM-fed animals over the 8 week period of the fast. In addition, the plasma thyroid hormone levels in the fasted fish tended to be lower than in fish fed both the SM and CM diets prior to fasting, and there was histological evidence indicating a reduced activity of the pituitary-thyroid axis. However, thyroid hyperplasia and hypertrophy were still evident in the fasted fish previously fed the CM diet indicating that the adverse affects of CM diets are not completely reversible after 8 weeks.In fish fed the CM diet for 12 weeks and then the SM diet for up to a further 8 weeks (diet C-S) there was a compensatory increase in plasma thyroid hormone levels evident within 4 weeks after the change in diet, but no apparent decrease in thyroid hyperplasia or hypertrophy. In addition, in the fish fed the C-S diet there was a marked compensatory growth rate, and an increased feed: gain ratio; body weights of this group of fish were not significantly different from those of the SM-fed animals after 20 weeks of study, indicating a considerably higher growth rate over the last 8 week period.     

Effects of adrenaline on ionic equilibria in red blood cells of rainbow trout (Salmo gairdneri)

Effects of adrenaline on the equilibrium distributions of Na⁺ , K⁺ , H⁺ , Cl^– , and H₂O across the cell membrane of rainbow trout (Salmo gairdneri) erythrocytes were determinedin vitro, as a function of P CO₂ (1.76–7.77 torr). CO₂-carrying capacity of the blood was also examined. Plasma catecholamine concentrations inunanaesthetized, unrestrained trout were 3.1 nM adrenaline and 1.2 nM noradrenaline. Elevation of the plasma adrenaline concentrationin vitro to 4.6 × 10³ nM resulted in net gains of Na⁺ , Cl^– and H₂O by red cells, a net loss of H⁺ from red cells, and a pronounced red cell swelling. Adrenaline also reduced the CO₂-carrying capacity of trout bloodin vitro. The magnitudes of these effects increased with P_CO2 and, thus, were sensitive to blood HCO₃ ^– concentrations. The distribution of K⁺ between red cells and plasma was unaffected by adrenaline. Adrenergic-mediated ion movements and red cell swelling were sensitive to both propranolol and SITS. These results are consistent with the symport NaCl uptake model for adrenergic-mediated swelling of Baroinet al. (1984). The adrenergic response of fish erythrocytes may function to ameliorate the effects of blood acidoses on O₂-carrying capacity by maintaining red cell pH in the face of a decrease in plasma pH.     

Protein sparing effects in large rainbow trout,Salmo gairdneri

Large rainbow trout (250–550 g) were fed to satiation or 0.5% live weight/day, one of six balanced diets varying in protein (30%, 43%, 52%) and lipid levels (12%, 24%). Growth and changes in body composition were measured over a 60-day feeding period. Mean appetite varied inversely with dietary digestible energy. Total weight gain over the 60-day period was not significantly different (P = 0.05) for fish fed to satiation on diets containing 43 or 52% protein at the 24% lipid level. Weight gain was significantly lower among fish fed to satiation on the 30% protein, 12% lipid diet than those fed diets containing 43 and 52% protein, at either lipid level. When feeding level was limited to 0.5% of live body weight/day, weight gain was significantly higher for those fish fed diets containing 43 or 52% protein and 24% lipid, than fish fed all other diets. An increase in dietary lipid from 12 to 24% resulted in a significant increase in gross conversion efficiency of both dietary energy and protein, at all three dietary protein levels when fish were fed to satiation. A similar trend resulted when feeding level was reduced to 0.5% live weight/day. Dietary protein concentration did not significantly affect gross conversion efficiency of protein within either feeding rate. Increasing dietary protein tended to result in an increase in gross conversion efficiency of lipid at both dietary lipid levels within each feeding rate.     

Phagocytic cells in the epidermis of rainbow trout, Salmo gairdneri Richardson

Abstract. Electron microscopical observations of skin of rainbow trout revealed the presence of several types of phagocytic cells in the epidermis, most notably macrophages, some of which were seen crossing the basal lamina. Occasionally, filament-containing cells showed either melanin granules or phagosomes, suggesting these cells were also capable of phagocytosis. No cells with the characteristics of the Langerhans cells were observed, which does not necessarily preclude the possibility of macrophages being able to play an identical role as antigen-trapping cells.     

Behaviour of rainbow trout, Salmo gairdneri Richardson, in marine cages

Abstract. Underwater television was used to observe the distribution and swimming behaviour of groups of farmed rainbow trout, Salmo gairdneri Richardson, in marine net cages. Undisturbed rainbow trout (mean total length 240 mm) aggregated near the surface, remaining motionless in slack water and formed polarized shoals at higher current velocities. Larger fish (mean total length 317 mm) occupied deeper parts of the cage and were more active during periods of slack water. The results are compared with previous work on farmed Atlantic salmon, Salmo salar L., and discussed in relation to the biology of wild rainbow trout and recent work on exercise, growth and condition in salmonids.     

Relationship between transmembrane potential and activation of motility in rainbow trout (Salmo gairdneri)

A hypothesis is developed that activation of motility in rainbow trout spermatozoa is a result of membrane hyperpolarization. This hypothesis was developed to explain experimental observations of a relationship between membrane potential and motility as revealed by the use of voltage sensitive fluorescent dyes. The results lead to the following conclusions: a) Transmembrane potential hyperpolarizes with decreasing KCl concentration in 100 mM NaCl. b) Transmembrane potential hyperpolarizes with decreasing NaCl concentration. c) NaCl is three time less effective in changing transmembrane potential and two orders of magnitude less effective in inhibiting activation of motility than KCl. d) Chloride ions have little effect on transmembrane potential or motility. e) Increases in osmotic pressure with the non-ionic molecule sucrose increased the amount of KCl required to inhibit activation. f) The major effect of Na⁺ on K⁺ inhibition may be osmotic.It is suggested that while sperm cells are in the seminal plasma in the reproductive tract of the male rainbow trout their transmembrane potential is maintained above an activation threshold, probably through Na/K pumps which are found in almost all animal cells. Since K⁺ is the most important ion in determining the transmembrane potential, hyperpolarization of the plasma membrane below an activation threshold occurs when the sperm cells are diluted, during spawning, into the low K⁺ environment of freshwater.