Effect of rearing density on thyroid and interrenal gland activity and plasma and hepatic metabolite levels in rainbow trout, Salmo gairdneri Richardson

There were significant inverse correlations between rearing density of rainbow trout, Salmo gairdneri Richardson, and final body weight, plasma L-thyroxine (T₄), trüodo-L-tryronine (T₃), cortisol and protein concentrations, plasma T₄/T₃ ratios and thyroid epithelial cell height. In addition, hepatosomatic indices and plasma free fatty acid concentrations were higher in fish reared at low (134 g 1⁻¹) density compared with groups reared at medium (210g1⁻¹) or high density (299g 1⁻¹), and the post-feeding (3.5-4h) elevation in plasma glucose and triglyceride levels evident in trout maintained at low rearing density was not found in those fish reared at higher densities. There were no significant effects of rearing density on hematocrit, carcass composition, hepatic glycogen and lipid levels and interregnal nucleus size.     

Influences of photoperiod and alternate days of feeding on plasma growth hormone and thyroid hormone levels in juvenile rainbow trout

The diurnal patterns of plasma growth hormone (GH), thyroid hormone and cortisol concentrations in rainbow trout Oncorhynchus mykiss held under three photoperiod (L : D) regimes (6 : 18, 12 : 12 and 18 : 6), and fed either daily (DF) or on alternate days (ADF) with 2·0% body mass per day of a commercial trout diet were determined. The ADF groups had reduced total mass gain and specific growth rates compared with DF fish, but photoperiod had no affect on growth for either of the feeding regime groups. In the ADF groups, the mean 24 h plasma thyroxine (T₄) and triiodothyronine (T₃) concentrations were significantly lower, both on days of feeding and days of fasting, than in DF fish held under all three photoperiod regimes, but for GH, only the 18L : 6D DF group was higher than the comparable ADF groups. There were no significant differences in mean 24 h plasma cortisol concentrations of DF and ADF groups. Diurnal patterns of plasma GH, cortisol, T₄ and T₃ were found in DF fish held under all three photoperiod regimes. Increases in plasma cortisol changes were associated with the onset of the light phase; elevations in plasma GH and T₄ concentrations were more closely associated with clock time, regardless of photoperiod; increases in plasma T₃ concentrations were strongly associated with time of feeding. In ADF groups, these diurnal changes in plasma GH, T₄ and T₃ concentrations were suppressed for both the fed and fast days, and plasma cortisol concentrations were suppressed on the fasting day. The observations are discussed in terms of the proposed anabolic, catabolic and growth regulating roles of these hormones in different growth and metabolic modifying situations in teleosts.     

Dose-related effects of 17βoestradiol (E2) on liver weight, plasma E2, protein, calcium and thyroid hormone levels, and measurement of the binding of thyroid hormones to vitellogenin in rainbow trout, Salmo gairdneri Richardson

Administration of 17β-oestradiol (E₂) to rainbow trout, in the form of hydrogenated coconut oil implants produced a stable, long-term elevation in plasma E₂ levels. The elevation was doserelated (over the range 1–10mg kg-¹ body weight) both 4 and 8 weeks after implantation. Dose-related increases were also observed with respect to liver weight-body weight ratios and plasma protein levels. Plasma T₃ and total calcium levels were depressed and elevated, respectively, by E₂ treatment but the responses were not linearly related to the dose of E₂ administered; there was no significant effect of E₂ on plasma T₄ levels.
E₂ induced a shift in the binding of T₃ to plasma proteins, with T₃ binding to smaller molecular weight proteins; neither T₄ nor T₃ bound to vitellogenin which was present at high levels in the plasma of E₂-treated fish.     

Migration tendency in juvenile steelhead trout, Salmo gairdneri Richardson, injected with thyroxine and thiourea

Juvenile steelhead trout, Salmo gairdneri injected with thyroxine (T₄) near the time of seaward migration exhibited a significantly lower migration tendency than untreated controls of fish injected with saline, thiourea, or a combination of T₄ and thiourea. Fish injected with thiourea alone or in combination with T₄ prior to the time of maximum migration tendency showed enhanced migration over untreated and saline injected controls and those injected with T₄ alone. Injections of T₄ or a combination of T₄ and thiourea elevated plasma T₄ and tri-iodothyronine (T₃) concentrations, while injection of thiourea alone depressed thyroid hormone levels relative to saline-injected controls. Plasma concentrations of T₃ and T₄ returned to control levels within 10 days in all groups. We suggest that thyroid hormones are antagonistic to mechanisms underlying seaward migration of steelhead trout and that these antagonistic effects must be overcome before migration tendency is expressed.     

Seasonal changes of thyroid hormones in field-collected Atlantic cod in relation to condition indices, water temperature and photoperiod

Serum T₄ and T₃ in wild Atlantic cod Gadus morhua ranged from 1 to 12 ng ml⁻¹ and from 2 to 27 ng ml⁻¹ respectively over a 3-year period. In general, the concentrations increased from summer (T₃) or early autumn (T₄) to maxima in mid-winter and declined abruptly during spring. The T₄/T₃ monthly means were lowest in summer and highest in winter. The seasonal patterns of thyroid hormones were weakly correlated with changes in water temperature. However, both T₄ and T₃ co-varied simultaneously with photoperiod. In addition, T₃ was correlated with the hepatosomatic index and condition factor during summer and autumn. It is suggested that the seasonal changes in the release of T₄ from the thyroid were photoperioddriven, and that the course of T3 was regulated by the metabolic state of the fish during the somatic growth period.     

Cortisol and thyroid hormone responses to acid stress in the brown trout, Salmo trutta L.

Individual cannulated brown trout monitored during exposure to acidic water showed increased plasma cortisol after 3 h at pH 4.0 with low (0.05 mm) or high (2.8 mm) calcium (Ca) content, and after 2 days in acidic water with a high Ca content. Most fish did not survive for 2 days in acidic water with a low Ca content. Non-cannulated fish showed a similar increase in mean plasma cortisol after 2 days in high-Ca acidic water (pH 4.0–4.6), but not in acidic water of a low Ca content. After 7 days of exposure to acidic water, plasma cortisol appeared to recover when there was a high Ca content but increased 20-fold when Ca content was low. In cannulated fish severe acid stress resulted in a marked and rapid thyroid response. Plasma thyroxine (T₄) was elevated after 3 h exposure to acidic water of both low and high Ca content and remained elevated for 2 days of acid exposure with high Ca. In non-cannulated fish an increase in mean T₄ was apparent only after 7 days in low-Ca acidic water. Plasma triiodothyronine (T₃) levels were not significantly altered by any of the acid regimes. Plasma glucose of cannulated fish was elevated within 3 h of acid-exposure and remained elevated after 2 days in high-Ca acidic water.     

Influence of plasma lipid changes in response to 17β-oestradiol stimulation on plasma growth hormone, somatostatin, and thyroid hormone levels in immature rainbow trout

Plasma total lipids were significantly higher in 17β-oestradiol(E₂)-treated immature rainbow trout Oncorhynchus mykiss at week 4 after implantation, due to increases in polar and neutral lipids. The lipid classes responding were phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, sterols and sterol esters, in a proportion that approximately reflected the increase in plasma vitellogenin (VtG) levels as measured by a non-competitive enzyme-linked immunosorbent assay (ELISA). Plasma non-esterified fatty acids and triacylglycerol were not affected by E₂ treatment. Plasma growth hormone GH levels were increased, and plasma somatostatin-14 (SRIF) levels decreased in E₂-treated fish, responses which could be secondary to elevated plasma lipid (VtG) content, although a direct E₂ action on somatotroph function is possible. Plasma T₄ concentrations were not affected by E₂ treatment, but plasma T₃ concentrations were significantly lower than in controls 1 week after implantation when plasma E₂ concentrations were the highest; this is in support of the hypothesis that E₂ has a suppressive action on T₃ production.     

Seasonal and diurnal rhythms of thyroidal status in the rainbow trout, Salmo gairdneri Richardson

The concentrations of thyroxine (T₄) and triiodothyronine (T₃) in the blood plasma of rainbow trout ( Salmo gairdneri Richardson) at intervals throughout the year have been measured by a chemically specific gas-liquid chromatographic (GLC) method. Mean hormone levels showed a seasonal variation, maximal levels of both hormones occurring in winter and minimal concentrations in mid summer. An apparent secondary maximum in mean T₄ and T₃ concentrations was observed in spring. (T₄)/(T₃) ratios have been found to be highest in winter and lowest in summer. A radioimmunoassay procedure, validated by GLC analyses, revealed the presence of a diurnal rhythm in serum T₄ levels of trout sampled in September and in April.     

Poor water quality suppresses the cortisol response of salmonid fish to handling and confinement

Confinement of brown trout in small troughs of static water for 1 h at a density of six fish 251⁻¹ stimulated the hypothalamic-pituitary-interrenal axis and resulted in an elevation of plasma cortisol from basal levels (less than 2 ng m1⁻¹) to about 100 ng m1⁻¹, the degree of stimulation being dependent upon water temperature. Confinement at a density of 30 fish 251⁻¹ resulted in a 50% suppression of this response. It is demonstrated that this effect is mediated by changes in water chemistry and not by crowding per se . Experimental manipulation of the water chemistry showed that reduced pH (7.1 → 6.3), elevated free CO₂ (63 → 520 μmoll⁻¹) or elevated ammonia (8 → 1300 μg 1⁻¹ as total ammonia nitrogen) had no individual effects on the interrenal response to acute confinement. Elevated ammonia in combination with reduced pH significantly increased the plasma cortisol levels in response to acute confinement, whereas a combination of reduced oxygen (100 → 20% saturation), elevated free CO₂ and elevated ammonia markedly suppressed (∼ 50%) the cortisol response of both brown trout and rainbow trout to acute confinement in a manner similar to that observed with trout at high densities. A compensatory increase in plasma cortisol levels was observed during the subsequent recovery of fish which had been confined for 1 h in water of poor quality. These findings are discussed in relation to the exposure of fish to multiple stresses and to the role of corticosteroids in the stress response.     

Plasma Ca2+ concentration limits melatonin night production in two fish species

In freshwater (FW) rainbow trout Oncorhynchus mykiss of spontaneously low plasma calcium concentrations ([Ca²⁺]_pl), plasma melatonin at night was significantly lower than that measured in FW fish with the highest [Ca²⁺]_pl. In brackish water adapted rainbow trout with originally high [Ca²⁺]_pl, plasma melatonin concentration at night was elevated. In cannulated flounder Platichthys flesus , night plasma melatonin increases (ΔMel) corresponded to [Ca²⁺]_pl. It is postulated that in physiological steady-state conditions, melatonin synthesis capacity is coupled to free calcium concentration in plasma of O. mykiss and P. flesus .     

Changes in thyroid function and gonadotrophin activity during sexual development in ducks Anas sp. (Aves: Anatidae)

Age-related changes in plasma iodohormone concentrations in male and female Aylesbury and Khaki Campbell ducklings were determined between the second week after hatching and sexual maturity.
In both sexes and strains the concentration of total thyroxine (T₄) and the free thyroxine index (FT₄1) were low during the first 6–8 weeks after hatching. A marked increase in the T₄ and FT₄1 occurred when the ducklings were nine weeks old; T₄ progressively increased over the following four weeks and remained high thereafter. The T₄ concentration in both sexes was directly related to age.
Concentrations of total tri-iodothyronine (T₃) were more variable than T₄ but were highest before sexual maturation and inversely age-related. An age-related decline in free tri-iodothyronine index (FT₃1) was observed during the first nine weeks.
The possibility that these maturational changes in thyroid function may reflect changes in gonadal activity was assessed by measuring the concentration of luteinizing hormone (LH) in the same plasma samples. Although plasma LH increased with age, in all cases the increase occurred after changes in the thyroid hormones were observed. Moreover, the maturational pattern of LH was related to sex and strain whereas thyroid function was unaffected by sex or strain.
These results demonstrate reciprocal changes in plasma T₃ and T₄ levels in ducks during sexual development, although the stimuli responsible and physiological significance are uncertain.     

Cohort sampling, anaesthesia and stocking-density effects on plasma cortisol, thyroid hormone, metabolite and ion levels in rainbow trout, Salmo gairdneri Richardson

The effect of serial removal of fish from aquaria, anaesthesia and stocking density on plasma cortisol, thyroid hormone, metabolite and ion levels was examined in rainbow trout, Salmo gairdneri , to determine the consequences of normal handling and maintenance procedures on the activity of the pituitary-adrenal and pituitary-thyroid axes in the species.
The capture and handling associated with serial removal of fish from an aquarium resulted in a rapid elevation of plasma cortisol concentrations and a slower rise in plasma glucose, Mg²⁺ and Na⁺ concentrations; plasma K.+ levels showed a transient fall shortly after commencing the sampling, followed by a significant increase.
A lethal concentration of 3-aminobenzoic acid ethyl ester (MS 222) of 125 mgl⁻¹ (but not a sublethal concentration of 62.5 ng 1⁻¹) prevented the capture stressor-associated changes in most of the measured parameters, including plasma cortisol levels, although plasma protein and ion concentrations were elevated in fish sampled after MS 222-anaesthesia.
Stocking densities from 1 to 64 fish 80 1⁻¹ aquarium did not appear to effect changes in the measured parameters, except for plasma T3/T4 ratios which decreased with increasing stocking density. The exception was found in fish which were stocked in pairs: one member of each pair had a markedly elevated plasma cortisol level and there was a high incidence of mortality, probably related to aggressive social interaction.     

Effect of excess vitamin D3 on calcium metabolism in rainbow trout Salmo gairdneri Richardson

The effect of excess vitamin D₃ on calcium metabolism in rainbow trout was investigated. Trout were reared for 24 weeks on diets containing 4000, 104 000 or 1 004 000 iu of supplemental vitamin D₃ kg^-1 dry diet. No effect of excess vitamin D₃ was detected in the weight gain, feed efficiency or total mortalities of trout in the different groups. None of the fish showed any signs of hypercalcaemia and no significant difference was detected in the calcium, magnesium, phosphorus and sodium content of the trout vertebrae or total carcass. However, there was a significant increase in the calcium, magnesium and phosphorus content of the skin in trout reared on the highest supplemental level of vitamin D₃ (1 004 000 iu kg^-1 diet). No overt signs of renal calcinosis were detected in any of the trout. This study indicates that trout are resistant to excess vitamin D₃, and that excess vitamin D₃ does not appear to be related to the increased incidence of renal calcinosis in rainbow trout.     

Glomerular ultrastructure of the trout,Salmo gairdneri: Effects of angiotensin II and adaptation to seawater

Summary The effect of angiotensin infusion on the glomerular ultrastructure of freshwater- and seawater-adapted rainbow trout, Salmo gairdneri, has been examined by scanning and transmission electron microscopy. Adaptation of trout to seawater resulted in epithelial podocyte flattening, primary process broadening and apparent loss of foot processes in almost all glomeruli, features which were uncommon in freshwater-adapted trout. Similar changes were induced by infusion of freshwater-adapted animals with angiotensin, suggesting that the renin-angiotensin system plays a role in the modification of glomerular epithelial ultrastructure. Adaptation of trout to seawater also reduced glomerular diameter, but infusion of freshwater-adapted animals with angiotensin did not mirror this effect. Infusion of angiotensin into seawater-adapted animals increased the overall thickness of glomerular basement membrane by increasing the lamina rara interna and lamina densa. This did not occur when freshwater-adapted fish were either infused with angiotensin or adapted to seawater. These findings suggest that other humoral systems are involved in the control of glomerular diameter and basement membrane thickness as part of an integrated response to increased environmental salinity.     

Changes in plasma thyroid hormone levels in pink salmon, Oncorhynchus gorbuscha, during their spawning migration in the Fraser River (Canada)

Plasma thyroid hormone levels were measured in pink salmon, Oncorhynchus gorbuscha , during their spawning migration in the Fraser River, British Columbia. The plasma levels of both l-thyroxine (T₄) and triiodo-l-thyronine (T₃) were significantly higher in males than in females. In both sexes the hormone levels were maintained, or increased somewhat, during the early stages of migration, but fell thereafter. In females the plasma T₄ and T₃ levels of salmon collected on the spawning grounds were at or below detectable levels of the assays. The changes in thyroid hormone levels are correlated with changes in plasma insulin, gonadotropin, gonadal steroid hormones, cortisol and vitellogenin levels measured in the same specimens.     

Constraints of body size and swimming velocity on the ability of juvenile rainbow trout to endure periods without food

The hypothesis that body size and swimming velocity affect proximate body composition, wet mass and size‐selective mortality of fasted fish was evaluated using small (107 mm mean total length, L _T) and medium (168 mm mean L _T) juvenile rainbow trout Oncorhynchus mykiss that were sedentary or swimming ( c . 1 or 2 body length s⁻¹) and fasted for 147 days. The initial amount of energy reserves in the bodies of fish varied with L _T. Initially having less lipid mass and relatively higher mass‐specific metabolic rates caused small rainbow trout that were sedentary to die of starvation sooner and more frequently than medium‐length fish that were sedentary. Swimming at 2 body length s⁻¹ slightly increased the rate of lipid catabolism relative to 1 body length s⁻¹, but did not increase the occurrence of mortality among medium fish. Death from starvation occurred when fish had <3·2% lipid remaining in their bodies. Juvenile rainbow trout endured long periods without food, but their ability to resist death from starvation was limited by their length and initial lipid reserves.     

Plasma ionic regulation and gill Na+/K+-ATPase changes during rapid transfer to sea water of yearling rainbow trout, Salmo gairdneri: time course and seasonal variation

Danish rainbow trout, Salmo gairdneri Richardson, (40–65 g) were transferred to 28%o sea water at intervals throughout the early spring and summer. Gill Na⁺/K⁺-ATPase of fish kept in fresh water surged distinctly during May. Simultaneously, a body silvering occurred and plasma concentrations of Cl⁻, Na⁺ and total thyroxine (T4) decreased. The seawater transfer-induced adaptive response in plasma electrolytes comprised a biphasic change, i.e., an adjustive peak phase and a regulatory phase lasting for 2 days and 1 week after transfer, respectively. Further, gill Na⁺/K⁺-ATPase activity increased to a new level after an initial lag phase of 2–3 days, but electrolyte regulation was mostly initiated prior to the adaptive change in ATPase activity. In spite of increasing pre-transfer freshwater Na⁺/K⁺-ATPase activity during the spring, the electrolyte peak level, the degree of muscle dehydration and the mortality of fish transferred to sea water increased from April to July. The apparent uncoupling of freshwater Na⁺/K⁺-ATPase activity and plasma electrolyte regulation in sea water is discussed in relation to smelting and prediction of hypo-osmoregulatory performance.     

Nitrogen excretion and determination of nitrogen and energy budgets in rainbow trout (Oncorhynchus mykiss R.) under different feeding regimes

The postprandial excretion pattern of ammonia in dependence of feeding regime (fasting, 1 ×/day, 2 ×/day, 4 ×/day and continuous feeding), was determined in rainbow trout ( Oncorhynchus mykiss ), using continuous flow analysis. In fed fish ammonia peaked c. 7 h after the first meal with no differences in pattern between treatments. Fasting fish did not show a pattern. Overall production rates (NH₄ and NO₂+ NO₃) ranged from 0.29–0.31 g kg⁻¹ BW/d in fed fish and were around 0.07 g kg⁻¹ BW/d in fasting fish. Additionally determined total N (Kjeldahl) showed much higher values in fed fish (0.78–1.05 g kg⁻¹ BW/d) but only slightly higher values in fasting fish (0.11 g kg⁻¹ BW/d). Budgets of nitrogen (N) and energy (E) showed low recoveries ( c. 50% and between 50% and 70%, respectively). When correcting ammonia excretion (NH₄ and NO₂+ NO₃) using literature data on urea excretion of O. mykiss and assuming that total N partly stemmed from uneaten but undetected feed, both N and E budgets reached a recovery of around 100% in all four fed groups. Implications of this approach are discussed in the light of incomplete budgets as determined in earlier studies.     

Endocrine, osmoregulatory, respiratory and haematological parameters in flounder exposed to the water soluble fraction of crude oil

Flounders Pleuronectes flesus with an implanted vascular catheter were exposed to a 50% dilution of the water soluble fraction (WSF) of Omani crude oil (c. 6ppm total aromatic hydrocarbons) and serial blood samples taken to determine their endocrine status (cortisol, catecholamines and thyroid hormones) and the resultant and/or causal physiological (haematological, ionoregulatory and respiratory) disturbances. This resulted in a progressive increase in plasma cortisol concentrations from 3 h onwards (rising from 18 to 51 ng ml⁻¹ after 48-h exposure), and increased plasma glucose concentrations indicating a generalized stress response. Plasma T3 and T4 concentrations of both control and WSF-exposed groups declined progressively over the experimental period; exposure to the WSF of crude oil further depressed plasma T4 concentrations but not plasma T₃ concentrations relative to those of control fish. Plasma osmolality and sodium and chloride concentrations were stable in WSF-exposed fish, however, plasma potassium concentrations were increased significantly at the 24-and 48-h sampling points. The most profound physiological disturbance in WSF-exposed fish was a dramatic decline in blood oxygen content (CvO₂) (from 2–8 to 0–8 ml 100 ml⁻¹ after 48-h exposure), which is likely to be the cause of the increased plasma noradrenaline concentrations from 3 h onwards. Increased noradrenaline is likely in turn to have been responsible for the significant increase in blood haematocrit and blood haemoglobin at the 3-h sampling point, although the dominant effect in the longer-term was a significant decline in both of these haematological parameters.     

Effect of feeding time on postprandial nitrogen excretion and energy expenditure in rainbow trout

The effect of feeding time (dawn or midnight) on nitrogen excretion and energy expenditure was studied in immature rainbow trout using measurements of respiratory gas exchange. Fish (mean individual weight 70 g) were maintained indoors under natural photoperiod and fed by hand (commercial food pellets) at a rate of 1% weight/day⁻¹. Rates of ammonia and CO₂ excretion and O₂ uptake were measured every hour. Ammonia excretion increased immediately after feeding in fish fed at midnight, and 2h after feeding in fish fed at dawn. Ammonia excretion and energy supply from protein catabolism, were higher in trout fed at midnight than in those fed at dawn, while total energy expenditure was the same in both groups. The results suggested that trout fed in phase with their natural feeding rhythm use dietary protein more efficiently for growth than do trout fed out of phase with the natural rhythm.