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.     

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.     

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.     

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 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.     

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.     

Plasma cortisol stress response in fingerling rainbow trout, Salmo gairdneri Richardson, to various transport conditions, anaesthesia, and cold shock

Plasma cortisol levels of fingerling rainbow trout were measured as an index of the stress resulting from various procedures used for transport of the fish for stocking. When transported under 'normal' conditions, which included water at the hatchery acclimation temperature (10–11°C), O₂ saturation or supersaturation, and neutral pH, there was a marked increase in plasma cortisol levels within 0.5 h, which was maintained over the next 4 h of transport; there was a significant decrease in plasma cortisol by 8 h of transport. It was found that the plasma cortisol levels at 4 and 8 h were not appreciably altered by transport under partial O₂ desaturation, O₂ saturation, O₂ supersaturation, or 0.5% NaCl, or by anaesthesia with tricaine methanesulfonate (MS 222) prior to capture and transport in MS 222-free water or 0.5% NaCl. A 15 min exposure to an immobilizing dose of buffered or unbuffered MS 222, or 2-phenoxyethanol, caused an increase in plasma cortisol of about 2 h duration, indicating that anaesthetics are themselves stressful. Exposure to chilled water (1° C) caused a large increase in plasma cortisol levels by 4 h after initiation of exposure; plasma cortisol had decreased at 1 day, and by 2 days a constant level was reached which was above the level in fingerling trout under 'normal' hatchery conditions. Trout acclimated to chilled water for 24 h and transported in chilled water had an increase in plasma cortisol during transport. Anaesthesia prior to transport or addition of salt did not reduce the stress of transport as judged by plasma cortisol levels. The results indicate that stress from capture and transport during stocking cannot be avoided using present methods.     

Swimming performance and blood chemistry in Atlantic salmon spawners exposed to acid river water with elevated aluminium concentrations

Exposure of sexually mature pre-spawning Atlantic salmon Salmo salar to Fossbekk water (pH 5·2) for 7 days led to a significant reduction in critical swimming speed (U-crit) in females but not in males. Exposure to Fossbekk water +A1 (as AlCl₃) for 24 h led to a significant reduction in U-crit in both males and females. In contrast to fish exposed for 7 days to Fossbekk water, fish exposed to Fossbekk+A1 had accumulated much more aluminium and mucus on their gills. Losses of plasma ions were similar in both groups exposed to acid water. Blood glucose was twice as high in fish exposed to Fossbekk water for 7 days compared with fish exposed to Fossbekk+A1 for 24 h. Plasma cortisol was still elevated compared with controls after exposure to Fossbekk water for 7 days. Possible mechanisms for the observed decrease in U-crit at the different exposures are discussed.     

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.     

Plasma and ovarian thyroxine levels in relation to sexual maturation and gestation in female Sebastes inermis

Plasma T₄ (L-thyroxine) concentrations in the viviparous rockfish Sebastes inermis showed a peak (119·3±27·8 ng ml⁻¹) during the development of embryos. Ovarian T₄ concentrations increased during vitellogenesis and final maturation and decreased during embryogenesis.However, total T₄ content within the ovary increased continuously up to the larval stages. Plasma oestradiol-17β, (E₂) concentrations peaked at the final maturation stage and then recovered to the level seen during the non-reproductive periods. Plasma testosterone (T) concentrations showed a peak (5·33±1·08 ng ml⁻¹) at the embryo stage and high levels were maintained throughout the gestation period. Plasma T₄ concentrations during the embryo stage and ovarian T₄ content during vitellogenesis were much higher than the levels reported in oviparous fishes. These data suggest that in viviparous rockfish, T₄ may be required for sustaining gestation or embryo development within the maternal body. In addition, the high content of total T₄ in the ovary implies that there probably does exist a maternal-embryo relationship during gestation as well as during vitellogenesis.     

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.     

Catecholamine release and interrenal response of brown trout, Salmo trutta, exposed to aluminium in acidic water

Cannulated brown trout, Salmo trutta , were exposed for 36 h to synthetic water with a low calcium content of pH 5 and similar synthetic water dosed with aluminium to raise the filterable A1 from 5 to 290 μg 1⁻¹ over the 36-h period. There were no significant disturbances of plasma concentrations of glucose, cortisol or catecholamine (adrenaline and noradrenaline) in fish held in water of pH 5. The addition of aluminium to this acidic synthetic water resulted in a generalized endocrine stress response with a four-fold increase in plasma glucose concentration after 18 h and a significant increase in plasma cortisol concentration from 24 h onwards when filterable A1 exceeded 200 μg 1⁻¹. Plasma catecholamine concentration indicated an adrenergic stress response in aluminium-exposed brown trout. A transient doubling in noradrenaline after 6 h in A1 was followed by a larger increase in both plasma adrenaline and noradrenaline concentrations in fish surviving the 36-h exposure to A1.     

Metabolic adjustments of an air-breathing teleost, Channa maculata, to acute and prolonged exposure to hypoxic water

Plasma and tissue metabolite levels were measured in the air-breathing Channa maculata during acute and prolonged exposure to normoxic and hypoxic water. Exposure of the fish to hypoxic water (water oxygen partial pressure, PwO ₂= 50 mmHg) for 1 h caused increases in plasma glucose and lactate, liver and brain lactate, liver a-amino acid, heart and brain alanine and brain succinate levels. The metabolic changes in heart, brain and muscle could only be detected when Pw O₂ was 30 or 10 mmHg. Heart glycogen and liver lipid decreased during acute exposure. Prolonged exposure to hypoxic water ( Pw O₂= 30 mmHg) for 3 days caused an increase in plasma glycerol and liver lactate dehydrogenase activity, and a depletion of glycogen store in all tissues investigated. However, metabolite levels which had been elevated during acute hypoxic exposure were observed to return to their normoxic values after prolonged exposure. It was concluded that anaerobic metabolism was triggered by acute exposure to hypoxic water. Prolonged exposure to hypoxic water induced a metabolic readjustment involving mobilisation of lipid and glycogen stores, which is probably a reflection of the high metabolic load of aerial respiration imposed on the fish during exposure to hypoxic water.     

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 .     

Studies on the relationship between osmotic or ionic regulation and thyroid gland activity in two salmonid fishes, Salmo gairdneri Richardson and Oncorhynchus kisutch Walbaum

In freshwater-acclimated rainbow trout a single intraperitoneal injection of ovine TSH significantly elevated plasma thyroxine (T₄) levels within 1 h after the injection. In seawater adapted trout the increase in T₄ after TSH-treatment was not evident until 6 h after the injection. TSH caused a transient fall in plasma Na⁺ and Cl^- between 3 h and 9 h after the injection in seawater-adapted fish and plasma Na⁺ was lowered in freshwater-adapted trout 24 h after the injection. Although there were clear histological changes in the thyroid gland of freshwater-adapted trout after TSH-injection, no such changes were evident in seawater-adapted fish.
Plasma thyroid hormone levels and thyroid histology in freshwater-adapted rainbow trout and coho salmon transferred to sea water, and seawater-adapted trout transferred to fresh water showed no consistent changes compared with controls.
The data are interpreted to indicate that although ambient salinity may have indirect effects on thyroid activity there is no direct involvement in ionic or osmotic regulation in the two species.     

The response to capture and confinement stress of plasma cortisol, plasma sex steroids and vitellogenic oocytes in the marine teleost, red gurnard

Reproductively active female red gurnard Chelidonichthys kumu were captured on long-lines, and placed in confinement tanks for 24, 48, 72 and 96 h to examine the effect of capture and confinement on reproductive parameters (experiment I). Plasma cortisol at the time of capture was elevated to levels typical of stressed fish in other species (53–125 ng ml⁻¹). Final plasma cortisol levels in red gurnard confined for any length of time were not significantly different from one another (ranging from 17 to 43 ng ml⁻¹), indicating that fish were chronically stressed when held in captivity for up to 96 h after capture. When initial and final plasma cortisol levels were compared within confinement groups, cortisol decreased significantly after 24 and 96 h of confinement indicating that some acclimation to captivity may have occurred. In contrast, plasma 17β-estradiol (E₂) and testosterone (T) levels decreased significantly to levels comparable to those in post-spawned fish, after any period of confinement, and remained low throughout the experiment. Another group of fish was captured and confined in the same manner as experiment I but subjected to repeated blood sampling every 24 h, until 96 h post-capture. In these fish, plasma cortisol levels decreased significantly from 127 ng ml⁻¹ after 24 h confinement and thereafter showed no change (25–45 ng ml⁻¹). Plasma E₂ decreased significantly after 72 h of confinement while plasma T showed no change from levels at capture. Increased amounts of follicular atresia were found in vitellogenic oocytes of fish confined for longer periods of time in experiment I, indicating that capture and confinement stress affect reproduction negatively in captive wild fish.     

Epinephrine, norepinephrine, and cortisol concentrations in cannulated seawater-acclimated rainbow trout (Oncorhynchus mykiss) following black-box confinement and epinephrine injection

The present study investigates the effect of cannulation and chronic'black-box' confinement, as well as epinephrine administration (4–0 μg kg⁻¹), on the degree and time-course of alterations in trout ( Oncorhynchus mykiss ) catecholamine and cortisol concentrations. Plasma cortisol concentrations in seawater trout acclimated to 3–6° C reached 104 ng ml⁻¹ 1 day after cannulation/confinement and remained elevated above resting levels (8 ng ml⁻¹) until 6 days post-confinement. Although plasma epinephrine and norepinephrine generally declined over the period of confinement (day 1 approx. 12 nM; day 7 approx. 6 nM), norepinephrine titres were usually higher and more variable. Epinephrine injection caused elevations in plasma epinephrine levels but not in norepinephrine levels; epinephrine titres reaching 107 ± 26 nM (range 65–238 nM) at 2 min post-injection and returning to pre-injection levels by 30 min post-injection. Plasma cortisol increased by 20 ng ml⁻¹ following epinephrine administration. Based on the time-course for post-confinement alterations in plasma cortisol, it appears that up to a week may be required before cannulated fish are completely acclimated to 'black-box' confinement. The findings suggest that meaningful results from experiments utilizing epinephrine injection and 'black-box confinement are contingent upon: (1) knowledge of circulating epinephrine levels shortly after injection (i.e. within 2 min post-injection); and (2) an experimental design that takes into account the elevated cortisol titres that are inherent with cannulation/confinement and epinephrine injection.     

Relationship between eyed-egg percentage and levels of cortisol and thyroid hormone in masu salmon Oncorhynchus masou

A possible cause of the low eyed-egg percentage that afflicts pond-reared masu salmon Oncorhynchus masou at a fish hatchery (Kumaishi Fish Hatchery, Hokkaido, Japan) was investigated. Serum cortisol levels of masu salmon during the spawning period were compared between individuals from Kumaishi and those from another station (Mori). Cortisol, thyroxine (T₄) and triiodothyronine (T₃) hormones were also measured in eyed eggs and their levels were compared to the eyed-egg percentage. Serum cortisol levels were significantly higher at Kumaishi than at Mori in May and July of the breeding season. At the Kumaishi station, there was a linear positive relationship between serum cortisol and fertilized-egg cortisol levels of female masu salmon. As cortisol levels in both ovulating females and eyed eggs increased, the eyed-egg percentage decreased, indicating a highly significant negative relationship between cortisol levels and eyed-egg percentage. In contrast, as T₃ and T₄ levels in eyed eggs increased, the eyed-egg percentage increased. The strong positive correlation between high cortisol levels in serum and in eyed eggs at Kumaishi indicates that the frequently higher cortisol levels may have caused the lowering of the eyed-egg percentage.     

Thyroid Hormone Replacement Restores Circulating Enkephalin Concentrations in Hypophysectomized Fetal Sheep

Abstract: We have investigated the effects of fetal hypophysectomy (HX) with or without thyroxine (T₄) replacement on the plasma concentrations of free methionine-enkephalin (free Met-Enk), noradrenaline, and adrenaline in late gestation sheep fetus. Plasma adrenaline concentrations were significantly higher in intact fetal sheep (1.05 ± 0.12 pmol/L) between 125 and 140 days of gestation when compared with the HX + saline (0.64 ± 0.10 pmol/L) and HX + T₄ (0.61 ± 0.08 pmol/L) groups. During the first 15 days of the T₄ or saline infusion, the plasma concentrations of free Met-Enk were significantly higher in the HX + T₄ group (392 ± 40 pmol/L) than in the HX + saline group (299 ± 43 pmol/L). At this stage of gestation, however, circulating concentrations of free Met-Enk were significantly higher in intact fetal sheep (556 ± 51 pmol/L) than in either of the HX groups. Between 125 and 140 days of gestation, plasma free Met-Enk concentrations were similar and significantly higher in the intact and HX + T₄ groups than those measured in the HX + saline fetal sheep. We conclude that the decrease in circulating free Met-Enk concentrations after removal of the fetal pituitary is primarily a consequence of functional athyroidism.     

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.