Incorporation of 1-14C-acetate into fatty acids and sterols by isolated hepatocytes of thermally acclimated rainbow trout (Salmo gairdneri)

Summary Rates of 1-¹⁴C-acetate incorporation into specific fatty acids and sterol fractions were determined at assay temperatures of 5°C and 20°C in hepatocytes isolated from warm (20°C)- and cold (5°C)- acclimated rainbow trout (Salmo gairdneri). Rates of sterol lipogenesis were 2.5- to 3-fold higher in hepatocytes from cold-acclimated trout. Rates of acetate oxidation and of total fatty acid lipogenesis did not differ significantly between acclimation groups. Fatty acid compositions did not change significantly during the experiment (9–12 h), but hepatocytes from cold-acclimated trout possessed significantly higher levels of polyunsaturates and unsaturates of the linolenic acid (n-3) family, and significantly lower levels of monounsaturates than did hepatocytes from warm-acclimated animals. Hepatocytes from cold-acclimated trout channeled a larger percentage of their total acetate incorporation into unsaturated fatty acids at 5°C than at 20°C due primarily to increased recovery of acetate in polyunsaturates and monoenes at 5°C. In contrast, hepatocytes from warm-acclimated trout channeled a slightly smaller percentage of their total acetate incorporation into unsaturates at 5°C than at 20°C. Hepatocytes from warm-acclimated trout incorporated significantly more 1-¹⁴C-acetate into the unsaturated fatty acid fraction (due primarily to incorporation into the diene fraction and less importantly all other classes of unsaturates) and significantly less into the saturated fatty acid fraction than hepatocytes from cold-acclimated trout when assayed at 20°C; similar but less dramatic differences were observed at 5°C. Consequently, unsaturated/saturated ratios for acetate incorporation ranked: warm-acclimated at 20°Cwarm-acclimated at 5°Ccold-acclimated at 5°C>cold-acclimated at 20°C. These results suggest that regulation of the relative rates of unsaturated and saturated fatty acid synthesis is involved in lipid restructuringduring adaptation from one temperature regime to another, but that other mechanisms must be invoked to explain the maintenance of observed steady state differences between the fatty acid compositions of warm- and cold-acclimated trout.This work was supported by grant PCM-76-04313-AO1 from the National Science Foundation     

Haemodynamic effects of adenosine on gills of the trout (Salmo gairdneri)

Summary The haemodynamic effects of adenosine on gills of the trout (Salmo gairdneri) were studied with in vitro and in vivo preparations.On the isolated head preparation, adenosine induced a decrease of the ventral aortic inflow and of the dorsal aortic outflow. Simultaneously the venous outflow increased. These effects were antagonized by theophylline. Adenosine induced a vasoconstriction in gill arches without filaments perfused by the afferent or the efferent branchial arteries. The efferent vessels were more sensitive to adenosine than afferent vessels. The whole systemic circulation of the isolated trunk did not show any response to adenosine. When adenosine was infused into the ventral aorta of living trout, the gill resistance to blood flow was greatly increased.These results suggest that adenosine is able to control the arterious and venous blood pathways in the trout gills by modulating their vascular resistance.     

Identification of glycoproteins in goblet cells of epidermis and gill of plaice (Pleuronectes platessa L.), flounder (Platichthys flesus (L.)) and rainbow trout (Salmo gairdneri Richardson)

Synopsis A quantitative analysis has been made of the glycoproteins present in the goblet cells of the epidermis, gill filaments and gill lamellae of three species of teleost fish. The glycoproteins have been identified by a combination of techniques, including the use of the enzyme sialidase followed by Alcian Blue staining, at pH 2.6 or I. o, in combination with periodic acid-Schiff. The selected fish were representative of species living in marine, freshwater and estuarine environments.The range of glycoproteins identified in these fish was similar to that found in mammalian tissue in that both neutral and acid glycoproteins were present, the latter included both sialomucins sensitive and resistant to sialidase, and sulphomucin. A single goblet cell contained either neutral or acid glycoproteins alone or in combination. Only the epidermis of the plaice and rainbow trout contained uniform cell populations producing acid glycoproteins, the former sulphomucin and the latter mainly sialomucin. At each site in the flounder and in the gill epithelia of the plaice and rainbow trout, the goblet cell population was mixed, with cells producing each type of glycoprotein. The number of goblet cells producing each type of glycoprotein varied at each tissue site.     

The entrainment and gating of the endogenous circannual rhythm of reproduction in the female rainbow trout (Salmo gairdneri)

Summary The endogenous circannual rhythm of maturation in the female rainbow trout and associated changes in serum oestradiol-17 and vitellogenin (as calcium) can be entrained by abrupt changes in the photoperiod. Groups of virgin fish (which naturally spawn in December) maintained under LD 186 from mid-January (Year 1), and then subjected to a reduction to LD 618 either on 1st March (Group A), 1st April (Group B), 1st May (Group C) or 1st June (Group D), commenced spawning on 31st July, 13th and 30th August, and the 16th September, respectively. Fish maintained on LD 186 throughout the experiment (Group E) commenced spawning on 10th October. These advances in the timing of spawning can be described in the form of a phase-response curve analogous to the entrainment behaviour of circadian oscillators. It is concluded that under natural conditions the annual change in photoperiod serves to continuously entrain the circannual clock thus ensuring that maturation and spawning occur at the optimal season for the survival of the species.In Groups A, B, C and D, respectively, 74%, 48%, 23% and 8% of the fish failed to mature in Year 1. Maintaining a sample of both maturing and non-maturing trout under constant LD 618 for a further year resulted in all the fish spawning in August of Year 2, approximately one year after the spawning members of this group matured in Year 1. It is proposed that the reduction from LD 186 to LD 618 phase-advanced a circannual timing mechanism in all the fish which then free-ran; this advance was only overtly expressed by the earlier spawning of a certain percentage of the fish in Year 1, but in all the fish in Year 2. These results indicate that the internal timing mechanism can be dissociated from the neuroendocrine mechanisms controlling maturation and thus it can be considered as an autonomous circannual clock. The relationship between the timing of the reduction in photoperiod and the percentage of virgin fish attaining maturation can be explained by a gating model. It is hypothesized that the energetically demanding process of gonadal maturation is only allowed in virgin fish which have reached a certain threshold stage of development and when the clock is at a specific (gate open) phase of the circannual cycle.     

Effects of elevated water temperature on the critical swim speeds of yearling rainbow trout Salmo gairdneri

Effects of elevated water temperature on the critical swim speeds of rainbow trout, Salmo gairdneri, were investigated. Trout acclimated to 10°C were exposed to 10, 15, 20 and 20°C while swimming and at rest. Initial swim speed of 20 cms⁻¹ was increased in 10 cms⁻¹ increments every 20 min until the fish fatigued. Critical swim speeds were calculated in absolute values (cms⁻¹) and relative performance values (body lengths s⁻¹). Critical swim speeds were similar at 10, 15 and 25°C. Swimming performance was significantly decreased at 25°C. Performance measured as critical swim speed was unaffected by temperature elevations up to 10°C above acclimation temperature of 10°C.     

Plasma cortisol measurements in precocious male and nonprecocious male juvenile steelhead trout (Salmo gairdneri)

Summary The mean biweekly plasma cortisol levels of juvenile nonmigratory precocious male steelhead trout,Salmo gairdneri, maintained from June through November in our laboratory under natural photoperiodic conditions ranged from 2.4–27.2 g/dl. Greatest mean cortisol occurred during the month of greatest gonadal mass, suggesting a direct interrenal-testicular relationship in this population of trout. The monthly mean plasma cortisol level of nonprecocious juvenile steelhead males captured prior to their downstream migration to the ocean from an Idaho hatchery and maintained in the laboratory simultaneously with the precocious males was low. For some months plasma cortisol was less than 0.1 g/dl. In June, however, plasma cortisol in nonprecocious males was 11.0 g/dl.     

Blood respiratory properties of rainbow trout,Salmo gairdneri: Responses to hypoxia acclimation and anoxic incubation of blood in vitro

Summary Blood respiratory properties of rainbow trout were determined following acclimation to normoxia and two levels of hypoxia.The most prominent response appeared to be an increase in blood O₂ affinity graded to the level of hypoxia. TheP ₅₀ values (at pH 7.8 and 20°C) were 24.1 21.7 and 16.8 mm Hg when specimens were acclimated to water O₂ tensions of 150, 80 and 50 mm Hg, respectively. The blood O₂ affinity was closely correlated with the erythrocytic ATP concentration. The stepwise correlation of ATP andP ₅₀, when trout were exposed to graded oxygen lack in the water, indicates that the blood O₂ affinity is precisely regulated.Anoxic incubation of trout blood in vitro induced a rapid reduction in erythrocytic ATP concentration (t _1/2=75 min), which was closely correlated to theP ₅₀ value. The drop inP ₅₀ value during anoxic exposure can be explained partly by the direct allosteric effect of a decreased erythrocytic ATP concentration and partly by the modified Donnan distribution of protons across the red cell membrane. Reoxygenation of the incubated blood, however, only partly re-established the erythrocytic ATP concentration, with a concurrent rise inP ₅₀ value.The results invite discussion about the mechanism, by which fish regulate their blood O₂ affinity. It is concluded, that it is regulated at the organismal rather than at the red cell level.Abbreviation (E) erythrocytes, erythrocytic     

The palatability of amino acids and related compounds to rainbow trout, Salmo gairdneri Richardson

Using a behavioural assay, the palatability of urea, taurine, betaine and the 20 common amino acids (L-isomers) to rainbow trout was determined. The trout responded positively only to aqueous solutions of leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glutamate, arginine, taurine and urea. Proline, the most effective compound, was active at 10 ^-4 m but not 10^-5m. Palatibility was not directly related to test solution pH; concentration and molecular structure apparently played a greater role. There is good, though not perfect, agreement between the behavioural results obtained here and published electrophysiological data on the palatal sensitivity of rainbow trout to these substances.     

Intestinal helminths in rainbow trout, Salmo gairdneri (Richardson): Absence of effect on nutrient absorption and fish growth

Rainbow trout. Salmo gairdneri (Richardson), from a fish farm, were found to be infected with two cestodes, Eubothrium crassum and Proteocephalus sp. Observations are described which suggest that these parasites have no effect on fish growth or on the intestinal absorption of L-leucine and D-glucose.     

Time course of osmotic adaptation and gill energetics of rainbow trout (Salmo gairdneri R.) following abrupt changes in external salinity

Summary The physiological and biochemical responses of rainbow trout (mean weight 250 g) to abrupt increases in salinity have been investigated. An initial crisis period lasting about 30 h was characterized by an increase in plasma and muscle ions, a rapid gill dehydration and a pronounced acidosis following a transient alkalosis. Mortality was low during this period. During the following days, gradual changes resulted in new steady state levels for most parameters examined.Analysis of adenylate pool (ATP, ATP/ADP ratio and energy charge) in the gills demonstrated an increased energy demand exhibiting two phases (4 h and 3 days), and a return to freshwater values. The gill respiration rate was constant during 3 days in sea water and decreased slightly later on. It was not influenced during the reverse transfer of seawater adapted fish into fresh water at the level of either isolated gills or perfused heads.     

Effect of ovine TSH, thiourea, ovine prolactin and bovine growth hormone on plasma thyroxine and tri-iodothyronine levels in rainbow trout,Salmo gairdneri

Summary A single injection of ovine TSH (1.1 g or 2.2 g/g body weight) effected a significant (P<0.01) increase in plasma T₄ levels in rainbow trout within 9 h and 1 h respectively. In fish given the higher dose the T₄ levels were still significantly higher (P<0.01) than in the controls 24 h after the injection. Ovine TSH had no significant effect on plasma T₃ levels. Plasma T₃/T₄ ratios in fish given the higher TSH dose were significantly lower than the controls at 3 h (P<0.01), 6 h (P=0.01), 9 h and 24 h (P<0.05) while in fish given the lower dose they were significantly lower than in controls at 3 h (P<0.01), 6 h (P=0.01) and 9 h (P<0.05).T₄ levels in TSH-injected fish were significantly higher (P<0.05) than in controls after 3 and 11 injections but were not significantly different after 16 injections. Plasma T₃ and T₃/T₄ ratios did not differ significantly in fish given multiple injections of either TSH or saline.Plasma T₄ levels in fish treated with thiourea were significantly lower (Pº0.01) than in comparable controls after 6, 9 and 15 days and plasma T₃ levels were lower (P<0.05) after 15 days treatment. T₃/T₄ ratios did not differ significantly at any time period.A single injection of bovine STH (10 g/g body weight) effect a significant rise in plasma T₄ levels 9 h (P=0.05) and 24 h (P<0.01) after the injection while ovine prolactin in a similar dose was without effect. Plasma T₃ levels in saline injected fish showed a steady decline over the 24 h following the injection whereas levels in prolactin and STH injected animals tended to rise. T₃ levels in the saline-injected fish were significantly higher (P<0.01) than in prolactin and STH injected animals 1 h after the injection but were significantly lower (P<0.01) than in the hormone-treated fish 24 h after the injection. There were no significant differences in T₃/T₄ ratios in any of the groups. Multiple injections of STH effected a significant increase (P<0.05) in plasma T₄ compared with the controls; prolactin had no significant effect. There was no significant differences in plasma T₃ concentration or T₃/T₄ ratios in any of the groups.     

Turnover of the fatty acyl and glycerol moieties of microsomal membrane lipids from liver, gill and muscle tissue of thermally acclimated rainbow trout,Salmo gairdneri

Summary Rainbow trout (Salmo gairdneri) acclimated to 5°C or 20°C were administered 2-³H-glycerol and 1-¹⁴C-acetate (63 Ci of each isotope/100 g body weight) via intraperitoneal injection, and subsequently maintained at their respective acclimation temperatures. Total lipid extracts (>80% phospholipid) were prepared from isolated microsomes of liver, gill and muscle tissue at various times over a three week period. Half-lives were determined independently for the fatty acyl and glycerol moieties from slopes of regression lines relating dpm/nmole phospholipidP _i vs time. In liver tissue, rates of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) turnover were also determined. Membrane turnover was most rapid in liver followed by gill and muscle. In liver, membrane fatty acids turned over more rapidly in warm-(t _1/2=3.4 days) than in cold-(t _1/2=6.8 days) acclimated fish, whereas in gill, rates of fatty acid turnover, did not differ significantly between acclimation groups. In contrast, rates of glycerol turnover were independent of acclimation temperature in liver, but faster (t _1/2=6.7 days) in warm- than cold- (t _1/2=15.1 days) acclimated fish in gill. In total lipid extracts, rates of fatty acid and glycerol turnover were equivalent in warm-acclimated fish, however, in cold-acclimated trout, there was a tendency for fatty acids (t _1/2=9.1 days) to turnover more rapidly than glycerol (t _1/2=15.1 days) in gill tissue, but more slowly (t _1/2=6.82 days) than glycerol (t _1/2=4.1 days) in liver. Although rates of glycerol turnover were equivalent in PC and PE of liver microsomes, the fatty acyl component turned over significantly more rapidly in PC at both acclimation temperatures. In cold-acclimated trout, rates of fatty acid and glycerol turnover were equivalent in PE, but the fatty acyl moiety of PC (t _1/2=4.7 days) turned over significantly more rapidly than glycerol (t _1/2=7.5 days). These results were interpreted as indicating that: (1) acclimation temperature independently influenced rates of fatty acid and glycerol turnover in a tissue specific manner, (2) a deacylation-reacylation pathway was activated in both liver and gill as a consequence of cold acclimation, but that liver tissue was more effective than gill in reutilizing the fatty acids released by phospholipase activity, and (3), in liver microsomes, patterns of turnover were phospholipid specific, with PC and PE differing either in the susceptibility of their acyl groups to degradation, or in their ability to reutilize fatty acids cleaved during membrane turnover at cold temperatures.     

Ventilation in rainbow trout (Salmo gairdneri, Richardson) with damaged gills

An account is given of damage to the gills of trout which involved reduction in total filament length and consequently reduced respiratory surface area. It was noted that fish with the greatest gill damage tended to ventilate the gills by active swimming (ram ventilation) to a greater extent than those with more normal gills.     

Studies on the uptake of (14C) amino acids derived from both dietary (14C) protein and dietary (14C) amino acids by rainbow trout, Salmo gairdneri Richardson

Previous studies have shown that rainbow trout fed on diets containing whole protein have superior growth rates compared to fish fed on diets of similar amino acid composition but containing a high proportion of free amino acids. The influence of several nutritional factors on the uptake of radioactivity from food pellets containing either [U-^I4C] protein or [U-¹⁴C] amino acids into the systemic blood of trout has been investigated. The time taken for radioactivity in the free amino acid fraction of blood to reach a peak after a meal containing [U-¹⁴C] protein had been given was much shorter, and the level of radioactivity in the blood higher, in trout with almost empty stomachs than in fish with almost full stomachs; uptake of radioactivity into blood amino acids was also more rapid and reached much higher concentrations when pellets containing [U-¹⁴C] amino acids were fed than when [U-¹⁴C] protein was fed. Incorporation of radioactivity into blood protein continued for a much longer period and reached higher levels when a pellet containing [U-¹⁴C] protein was fed than when a pellet containing [U-¹⁴C] amino acids was fed. Previous dietary history (low or high protein intake) did not appear to affect the rate of absorption of amino acids from either protein or free amino acid pellets. The uptake rates from pellets containing free amino acids could be slowed by mixing the dietary amino acids with albumin. The distribution, postabsorption, of radioactivity in the different fractions of blood and liver suggested that incorporation of carbon residues into glycogen and lipid from an amino acid diet was greater than from a protein diet. The converse was true of incorporation of radioactivity into tissue protein.     

Chromosome polymorphism in the rainbow trout (Salmo gairdneri Richardson)

Chromosome preparations from lymphocyte cultures of 50 rainbow trout were studied. Diploid chromosome numbers of 59, 60, 61, 62 and 63 were found in different individuals in which the arm number (NF) was 104. Intraindividual polymorphism was found at a low level in 25 of the fish. The results suggest that numerous chromosome polymorphisms exist in rainbow trout.     

Surface specializations of the olfactory epithelium of rainbow trout, Salmo gairdneri

Receptors for olfactory stimulus molecules appear to be located at the surface of olfactory receptor cells. The ultrastructure of the distal region of rainbow trout (Salmo gairdneri) olfactory epithelium was examined by transmission electron microscopy. On the sensory olfactory epithelium, which occurs in the depressions of secondary folds of the lamellae of the rosettes, five cell types were present. Type I cells have a knob-like apical projection which is unique in this species because it frequently contains cilia axonemes within its cytoplasm in addition to being surrounded by cilia. Type II cells bear many cilia oriented unidirectionally on a wide, flat surface. Type III cells have microvilli on a constricted apical surface and centrioles in the subapical cytoplasm. Type IV cells contain a rod-like apical projection filled with a bundle of filaments, and type V cells are supporting cells. Cilia on the sensory epithelium contain the 9 + 2 microtubule fiber pattern. Dynein arms are clearly present on the outer doublet fibers, which suggests that the cilia in the olfactory region are motile. Their presence in olfactory cilia of vertebrates has been controversial. The cilia membrane in this species is unusual in often showing outfoldings, within which are included small, irregular vesicles or channels. In addition, cilia on type II cells frequently contain dense-staining bodies closely apposed to the membranes, along with a densely stained crown at the cilia tip. Previous biochemical evidence indicates that odorant receptors are associated with the cilia.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Some of the blood constituents of male, female and immature rainbow trout of the Shasta variety, maintained in known environmental and dietetic conditions, were examined and the results statistically treated. The parameters for male and female trout were similar, only the erythrocyte count exhibiting a significant difference. Large but expected differences were evident between the parameters for mature and immature fish.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Fifty rainbow trout of the Kamloops strain were examined for 12 haematological parameters: erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, leukocyte count, differential leukocyte count, plasma total protein and plasma glucose concentration. The fish had been held under known environmental and dietetic conditions, and at the time of sampling were 14 months old. The majority of results for erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, total protein and differential leukocyte count fell within narrow ranges. The total leukocyte counts and glucose levels were more widely spread. The results are discussed and compared with those already published for Idaho and Shasta strains. It is impossible to say whether the differences that were observed between Kamloops and these other varieties were due to strain alone, since other variables were present. Some problems associated with establishing normal ranges for these parameters in fish are discussed.