Direct and indirect evidence for polypeptide absorption by the teleost gastrointestinal tract

The ability of the gastrointestinal tract of chinook salmon, Oncorhynchus tshawytscha , to absorb polypeptides in vivo was investigated by reference to the appearance of orally administered adrenocorticotropic hormone (ACTH) within the blood of fish previously treated with dexamethasone (3μg g^−l body weight) in order to suppress endogenous ACTH secretion. Further, since cortisol presence within plasma is dependent upon the availability of ACTH, dexamethasone blockade of endogenous ACTH secretion, in conjunction with subsequent measurements of plasma cortisol levels, provides a means by which biological patency of absorbed exogenous ACTH may be demonstrated. Levels of ACTH and cortisol in plasma of dexamethasone-treated salmon were therefore measured for a period of 360 min immediately following oral intubation of ACTH (15μg g⁻¹ body weight). Peak plasma presence of ACTH-like immunoreactivity (676.6 ± 33.6 pg ml⁻¹ plasma) and cortisol (227.1 ± 29.0 ng ml⁻¹ plasma) were recorded 120 min after ACTH administration. Results from the experimental groups were compared to those of 15 control treatments. Since administration of ACTH to chinook salmon elicited a consistent and significant elevation in not only plasma ACTH but also cortisol presence, it is contended that the salmonid gut expresses an ability to absorb polypeptides of dietary origin. The significance of these findings with respect to the oral administration of biologically active peptides and proteins to fish of importance to aquaculture is discussed.     

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.     

Primary and secondary stress responses to line capture in the blue mao mao

Primary and secondary stress responses were measured in wild Scorpis violaceus subjected to burst swimming from angling. Fish were blood sampled from 20 s to 30 min after hooking. Consequent rises in plasma adrenaline (14–316 nmol l⁻¹), noradrenaline (25–345 nmol l⁻¹), and cortisol (0.4–197 ng ml⁻¹) correlated with time since capture, and plasma lactate (0.1–12.2 mmol l⁻¹) reflected work done during intense exercise. Haemoglobin concentration and haematocrit also increased with exercise, and erythrocyte swelling occurred. Wild S. violaceus demonstrated a spontaneity and intensity of exercise not seen in fish acclimatized to aquarium conditions. By contrast, the stress responses of fish in captivity, despite careful husbandry, differed qualitatively and quantitatively from those in the wild. Cannulated fish had higher resting plasma cortisol concentrations (61.9±9.5 ng ml⁻¹) than did rapidly caught wild fish (<5 ng ml⁻¹) and these values were not significantly changed with burst swimming. Catecholamine secretion, possibly suppressed by cortisol, was insufficient to cause erythrocyte swelling. Erythrocyte nucleotides do not play a role in exercise, but are elevated in captive fish. It is hypothesized that primary endocrine responses are triggered by higher cortical processing of sensory information which is fundamentally different in the natural environment.     

Cortisol and glucose responses in juvenile brown trout subjected to a fluctuating flow regime in an artificial stream

In an artificial stream channel, wild 1 year old brown trout Salmo trutta were exposed to fluctuations in flow and water level to simulate hydro-peaking conditions downstream of a hydropower installation. Blood plasma cortisol concentrations reached a maximum of 59·4 ± 35·3 ng ml⁻¹ (mean ± 95% CL) 2h after the end of down-ramping. Return to the pre-exposure cortisol level was achieved within 6 h. When subjected to daily cyclical fluctuations over 7 days, plasma cortisol levels were significantly elevated (61·3 ± 26·8 ng ml⁻¹) on the first day compared to undisturbed fish (4·9 ± 3·7 ng ml⁻¹). On the fourth and seventh day, no elevation in plasma cortisol above control levels was observed. No changes in blood glucose that could be attributed to the stressor were found. There was no correlation between plasma cortisol and blood glucose levels. The short-lived cortisol response to daily fluctuations indicates a rapid habituation to this Stressor.     

Differences in plasma cortisol and cortisone dynamics during stress in two strains of rainbow trout (Oncorhynchus mykiss)

Plasma levels of cortisone, a steroid hormone of potential physiological significance in fish, have rarely been measured. This study examines the interrelationship between circulating levels of cortisone and the major teleost corticosteroid, cortisol, in the blood of two strains of rainbow trout subject to confinement stress, a condition know to stimulate corticosteroidogenic activity. In unstressed fish from both strains, mean plasma cortisol levels were within the range 0.4–7.5 ng ml⁻¹. Mean plasma cortisone levels were within the range 7.1–15.9 ng ml⁻¹. Plasma cortisol levels were elevated within 5 min of the onset of strees and reached peak values within 45 min, although there was a marked difference betweed the maxima observed in the two strains (strain 1:70 ng ml⁻¹; strain 2:150 ng ml⁻¹). The rate of increase of plasma cortisone levels during strees was more rapid than that of cortisol, maximum values (strain 1:100ng ml⁻¹; strain 2:160 ng ml⁻¹) being reached within 10 to 20 min of the onset of stress. This rapid stress-induced elevation of plasme cortisone has not previously been reported in fish. We suggest that rapid conversion of cortisol to cortisone during the initial response to stress accounts for the appearance of large amounts of cortisone in the blood, indicating that circulating for the appearance of large amounts of cortisone in the blood, indicating that circulating levels of cortisol alone do not fully reflect the secretory activity of the interregnal during the initial of cortisol alone do not fully the secretory activity of the interregnal during the initial phase of the stress response. The results also indicate that the rate of clearance of cortisone from the circulation may be a major factor in determining stress-stimulated levels of plasma cortisol.     

Physiological stress effects of continuous- and pulsed-DC electroshock on juvenile bull trout

Juvenile bull trout Salvelinus confluentus exposed to continuous- or pulsed-DC electroshock exhibited rapid elevations in plasma cortisol and glucose, but plasma chloride did not change. In a 1-h experiment using 240 V at 1·4 A of 60-Hz pulsed DC (voltage gradient 0·81 V cm⁻¹), which proved lethal, plasma cortisol and glucose rose significantly within 15 min of a 10-s electroshock. Plasma cortisol reached a peak level of 156 ± 18 ng ml⁻¹ at 45 min and then decreased, whereas plasma glucose reached its highest level of 179 ± 7·5mg dl⁻¹ at 1 h. In a 24-h experiment using lower dosages, plasma cortisol increased from 6·1-16 ng ml⁻¹ to peak levels of 155–161 ng ml⁻¹ in 1 h in response to a 10-s electroshock of continuous (130 V, 0·5 A, 1·45 V cm⁻¹) or pulsed (120 V, 0·5 A, 60 Hz, 0·55 V cm⁻¹) DC. Although plasma concentrations declined thereafter, levels remained above control values at 24 h. Plasma glucose was elevated from 60–65 to 120–134 mg dl⁻¹ after 1h by both electroshock treatments and remained near or above those levels for the 24-h duration. Plasma cortisol and glucose levels were much higher in electroshocked bull trout at 1 h compared with those in fish 1 h after receiving a 30-s handling stressor (cortisol, 90 ± 12 ng ml⁻¹; glucose, 82 ± 6·1 mg dl⁻¹). The results indicate that both continuous and pulsed DC were more stressful to juvenile bull trout than handling and that recovery, at least for pulsed DC, may take longer than 24 h.     

The effect of exercise on plasma cortisol and blood sugar levels in the rainbow trout, Salmo gairdnerii Richardson

Preliminary experiments were performed to determine the diurnal variation in cortisol, using trout which had been cannulated three days previously. These results indicated that cortisol levels were reasonably stable between 10.00 and 18.00 hours, thus permitting experimentation during this period without diurnal fluctuations masking the cortisol response. Uncannulated fish were exercised in a flume for 2 h at 1, 2.6 and 5 bl s^-1 and plasma samples taken from groups of five animals at 15, 30, 60 and 120 min after the start of exercise and at 1½, 12 and 24 h after the exercise ceased. The cortisol levels in all cases were elevated after 15 min, but the magnitude of the elevation increased with swimming speed. At 1 bl s^-1 the cortisol levels increased from 76.4 (± 20.4) to 129.2 (± 20.4) ng ml^-1 [mean (± s.d. )]. At 2.6 bl s^-1 the increase was from 72.4 (± 17.1) to 254.4 (± 34.4) ng ml^-1 and at 5 bl s^-1 the increase was from 69.5 (± 27.5) to 326.4 (± 39.0) ng ml^-1. The cortisol levels were stable over the exercise period and all groups recovered to baseline levels after 24 h, though the sample taken 12 h after the termination of exercise was elevated due to regular nocturnal increases in cortisol levels. There were no dramatic changes in blood sugar levels during and after exercise at 1 and 3.2 bl s^-1.     

Response of the interrenal to adrenocorticotropic hormone after short-term thyroxine treatment of coho salmon (Oncorhynchus kisutch)

The effects of experimentally elevated plasma thyroxine levels on the subsequent response of interrenals of coho salmon (Oncorhynchus kisutch) to adrenocorticotropic hormone (ACTH) in vitro were examined. Animals were treated with thyroxine by immersion (200 micrograms/L) for 3 days, which resulted in physiological elevations in circulating thyroxine. In animals treated before the parr-smolt transformation was completed (early smolts), thyroxine had no effect on plasma cortisol levels but significantly enhanced the sensitivity of the interrenal to ACTH in vitro. In animals treated after the period of smoltification (postsmolts), plasma cortisol levels were significantly higher than those of controls; both experimental and control animals had plasma cortisol levels higher than normally observed at this stage of development. The response of the interrenals of thyroxine-treated postsmolts to ACTH in vitro was significantly lower than that of controls. Results from the experiments using early smolts are in agreement with studies in other vertebrates showing that thyroid hormones are involved in maintaining the normal functioning of corticosteroidogenic tissue and suggest that thyroid hormones may be involved in the activation of the interrenal that occurs during smoltification. The results obtained using postsmolts are more difficult to interpret because of the possibility that these animals were physiologically stressed by the treatment. Increased ACTH release in vivo resulting from stress may have led to a depression of interrenal sensitivity to ACTH in vitro and may have masked a refractoriness of the pituitary-interrenal axis to thyroxine.     

Acclimation of the interrenal tissue of the brown trout, Salmo trutta L., to chronic crowding stress

Brown trout kept under chronically crowded conditions grew more slowly over a 110 day period than did uncrowded fish given similar rations. Crowding resulted in a significant elevation of plasma cortisol for at least 25 days but by day 39 the cortisol levels in both crowded and uncrowded fish were similar. After 110 days, plasma cortisol levels in both groups were low (<2ng ml⁻¹) and no significant differences in interrenal histology were detected. It is concluded that prolonged growth rate suppression as a result of crowding is not mediated by a chronic elevation of plasma corticosteroids. Under the conditions of the present study, water quality deterioration due to crowding was not sufficient to account for any of the changes in interrenal activity of the fish. These findings are discussed in relation to social interactions between saimonid fish maintained at unnaturally high stocking densities.     

Responsiveness of the interrenal tissue of Jundiá (Rhamdia quelen) to an in vivo ACTH test following acute exposure to sublethal concentrations of agrichemicals

As in many aquatic environments, pollution is a widespread problem in Southern Brazil. In our previous work, we demonstrated that sublethal contamination with some agrichemicals impairs the capacity of fishes to elevate cortisol levels in response to an additional acute stressor. In earlier experiments, the experimental design did not allow us to conclude where this effect occurs. In the present work, we used the adrenocorticotropic hormone (ACTH) challenge test to help us identify if the impairment occur in the interrenal tissue. For this purpose, five experiments were conducted, each with one specific agrichemical (methyl-parathion, atrazine+simazine, atrazine, tebuconazole, and glyphosate) in sublethal concentrations of 16.6% of the LC(50-96h), as previously determined. Fish were subjected to the ACTH challenge test protocol as follows: group 1, were non-injected and maintained as the specific control group; group 2 received an injection of the vehicle alone (the saline group); and group 3 receive an injection of ACTH. One hour later, blood samples were taken from the caudal plexus, using sterile syringes. In all specific control groups, the injection of ACTH induced a strong rise in plasma cortisol, compared with the fish injected only with the vehicle and the non-injected group. Fish exposed to methyl-parathion and tebuconazole did not elevate cortisol in response to the ACTH injection, with values significantly lower than the control fish. Fish exposed to sublethal concentrations of atrazine+simazine, atrazine, and glyphosate showed a rise in plasma cortisol very similar to the control fish. We conclude that the ACTH challenge test revealed that R. quelen exposed to sublethal concentrations of tebuconazole and methyl-parathion had a reduced ability to elevate plasma cortisol in response to an intraperitoneal (i.p.) injection of exogenous ACTH, indicating that the interrenal tissue is the site of the impairment within the HPI axis. These ACTH challenge tests also revealed that the impairment of the cortisol response verified in fish exposed to atrazine+simazine and glyphosate, as shown in our previous work, seems to be related to steps of cortisol secretion in higher levels within the HPI axis.     

The endocrinology of 1α-hydroxycorticosterone in elasmobranch fish: a review

The endocrine underpinnings of the stress response in fish have been the subject of intense research for well over 50years. Much of the research has focussed on teleost fish and so the endocrine mechanisms for cortisol production, transport and action at the target site have received significant attention. However, corticosteroidogenesis in elasmobranchs is exceptional on a number of levels. Unlike teleost fish the interrenal tissue is anatomically distinct from both renal and chromaffin (catecholamine producing) tissue; further the final product, 1α-hydroxycorticosterone (1α-OH-B), is unique to chondrichthyans where the carbon atom at position 1 of corticosterone has a hydroxyl group attached in the α orientation. The homologous nature of interrenal tissue in elasmobranchs presents an obvious advantage in the study of corticosteroidogenesis, however, the unique chemical nature of 1α-OH-B has presented distinct disadvantages as it has proven to be difficult to synthesise, and therefore studies examining the mineralocorticoid and glucocorticoid actions of this steroid are limited. Over the last decade molecular techniques have provided significant insight in the involvement of corticosteroiogenic enzymes in the elasmobranch interrenal in addition to the evolution of corticosteroid receptors. Given the number of excellent reviews focussing on the role of cortisol in the stress response of teleost fish, this short review aims to synthesise the endocrine basis for the synthesis, release, and action, of the enigmatic 1α-OH-B in elasmobranch fish.     

Sequence of gonadal events and oestradiol levels in Sparus aurata (L.) under two photoperiod regimes

Individually tagged Sparus aurata were kept in tanks with running sea water (21°± 2°C) during their second and third years of life. Gonads were biopsied and blood was sampled at monthly intervals. Thirteen of 50 fish in the second year and 24 of 35 fish in the third year were phenotypically females. Fish kept under 16L/8D photoperiod from July 1981 to August 1982 did not reach maturation; waves of initial ovarian growth alternated with waves of atresia. When photoperiod was shortened as from August 1982, gonadal development commenced within a month and proceeded at a rate higher than that of the control fish reared under natural photoperiod. Under natural photoperiod oestradiol serum levels (E₂) were relatively low during the resting phase, May-October (108 ± 11 pg ml⁻¹), and during the late vitellogenic phase (502 ± 76 pg ml⁻¹). High levels (1669 ± 312 and 1240 ± 172pg ml⁻¹) occurred during the early vitellogenic and the maturational phases. The high level of E₂ during the spawning season of S. aurata is explained by earlier reports indicating a prolonged breeding season with daily release of eggs, and alternating daily surges of E₂ and 17α, 20β, dihydroxy-4-pregnen-3-one, possibly a 'maturational progestin' in this fish. In phenotypic males, E₂ was highest during early spermatogenic phases (745 ± 142pg ml⁻¹) but was low (155 ± 11 pg ml⁻¹) in males with running sperm.     

Response to confinement in sea bass (Dicentrarchus labrax) is characterised by an increased biosynthetic capacity of interrenal tissue with no effect on ACTH sensitivity

Two experiments were carried out to investigate the influence of confinement stress on plasma cortisol levels and on the sensitivity of the interrenal cells to adrenocorticotropic hormone (ACTH) stimulation in sea bass Dicentrarchus labrax. Confining sea bass at 70 kg m(-3) for 24 h resulted in elevated plasma cortisol levels at all times (0.1, 1, 4 and 24 h) and corresponded to a reduced cortisol content in head-kidney homogenates after 0.1 and 1 h of confinement. An increased activity of the interrenal cells was also indicated by the enlarged nuclear diameters measured after 1 and 4 h of confinement. In vitro superfusion experiments showed that 4 h of confinement resulted in an increased basal unstimulated release of cortisol from head-kidney tissues compared with that in unstressed control fish. Although the stimulation factor (cortisol release as percent increase above basal) of the stressed fish was significantly lower than in controls, no difference in the maximal stimulated release (in absolute amounts) was evident between stressed and control fish. Care must be taken when interpreting superfusion data, as to whether the stressor actually leads to a reduction in interrenal sensitivity, or is due to an alteration in the basal release of cortisol.     

Plasma catecholamines and accumulation of adrenaline in the atrial cardiac tissue of aquacultured Atlantic salmon (Salmo salar) during stress

The effects of stress on plasma catecholamines (CA) and capacity for tissue accumulation of CA were studied in cardiac and skeletal muscle of cultured Atlantic salmon ( Salmo salar L.). Adrenaline (A) and noradrenaline (NA) were quantified by high performance liquid chromatography.
Plasma A and NA levels were 56±10 nmoll⁻¹ and 77±17 nmoll⁻¹ (± s.e.m. ), respectively, in a control group living under normal rearing conditions in a fish farm. Following a ±3 h period of pre-slaughter crowding and handling in the fish farm, plasma A reached 221 ± 72 nmol1⁻¹ with no increase in plasma NA. An 0.5 h period of struggling out of water led to even higher level of plasma A (480 ± 89 nmol1⁻¹), without change in NA.
Skeletal muscle was low in CA (A, 0.07 ± 0.02 and NA, 0.06 ± 0.01 nmol g⁻¹ wet wt). Tissue CA was higher in the atrium (A, 0.47 ± 0.04 and NA 0.94 ± 0.10 nmol g⁻¹) than in the ventricle (A, 0.25 ± 0.03 and NA, 0.30 ± 0.02 nmol g⁻¹). The 0.5 h period out of water and the 0.5-3 h period of pre-slaughter crowding led to accumulation of A, but not NA, in the atrium.
These data show that A, released during stress, accumulates in an undegraded form in the atrial tissue of the Atlantic salmon. This suggests a potent uptake mechanism for A in the atria presumably in the sympathetic nerve terminals. The acumulation of A in the atrium appears to reflect the period of high plasma A during stress.     

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.     

Characterization of the cortisol response following an acute challenge with lipopolysaccharide in yellow perch and the influence of rearing density

Two experiments were performed to characterize the corticosteroid response of yellow perch Perca flavescens following an intraperitoneal injection of lipopolysaccharide (LPS) and determine if sustained differences in rearing density alter this response. In the first experiment, yellow perch were injected with LPS (3 mg kg⁻¹), saline, or handled without receiving any injection. Concentrations of cortisol in plasma were elevated in all groups relative to non‐disturbed fish at 1·5 and 3 h after handling but by 6 h after injection the mean concentration of cortisol in plasma from LPS‐injected yellow perch were three to five times higher than fish before injection and significantly larger than groups of fish not treated with LPS. In the second test, yellow perch were held at different rearing densities (9 v . 18-19 kg m⁻³) for 3, 7 and 14 days before injection with LPS (3 mg kg⁻¹). The cortisol response of yellow perch following LPS injection of fish held for 14 days at high density was significantly lower than that of fish held at the low density for the same duration. Additionally a trend of a decreased cortisol response to LPS injection as duration of holding increased was observed among fish held at high density relative to fish held at low density. These data illustrate that the corticosteroid response of yellow perch following LPS injection is distinct from handling alone and that the magnitude of the response is impacted by rearing density.     

Physiological impact of sea lice on swimming performance of Atlantic salmon

Atlantic salmon Salmo salar were infected with two levels of sea lice Lepeophtheirus salmonis (0·13 ± 0·02 and 0·02 ± 0·00 sea lice g⁻¹). Once sea lice became adults, the ventral aorta of each fish was fitted with a Doppler cuff to measure cardiac output ( ̇ ), heart rate ( f _H) and stroke volume ( V _S) during swimming. Critical swimming speeds ( U _crit) of fish with higher sea lice numbers [2·1 ± 0·1 BL (body lengths) s⁻¹] were significantly lower ( P  < 0·05) than fish with lower numbers (2·4 ± 0·1 BL s⁻¹) and controls (sham infected, 2·6 ± 0·1 BL s⁻¹). After swimming, chloride levels in fish with higher sea lice numbers (184·4 ± 11·3 mmol l⁻¹) increased significantly (54%) from levels at rest and were significantly higher than fish with fewer lice (142·0 ± 3·7 mmol l⁻¹) or control fish (159·5 ± 3·5 mmol l⁻¹). The f _H of fish with more lice was 9% slower than the other two groups at U _crit. This decrease resulted in ̇ not increasing from resting levels. Sublethal infection by sea lice compromised the overall fitness of Atlantic salmon. The level of sea lice infection used in the present study was lower than has previously been reported to be detrimental to wild Atlantic salmon.     

Physiological responses to handling in the turbot

Turbot Scophthalmus maximus were cannulated via the afferent branchial artery and were either net-confined in sea water or in air for 9 min to monitor the stress response of a hatchery-reared marine flatfish. No mortality was observed. Aerial exposure appeared to mobilize plasma free fatty acids and stimulate the interrenal tissue but had no effect on circulating glucose or lactate levels. This pattern was qualitatively similar to that induced by net-confinement in sea water, although the magnitude and duration of the changes were more marked in the turbot handled in sea water. Aerial exposure had no effect on plasma osmolality, protein, or Cl⁻ concentrations and only a minor effect on plasma Na⁺ concentrations, which was in sharp contrast to the ionoregulatory disturbance noted in turbot which were net-confined in sea water. However, plasma K⁺ and cortisol concentrations were similarly elevated by both handling procedures. The results from these experiments suggest that whilst similarities with the salmonid physiological stress response are apparent, the lack of a plasma glucose response may represent a fundamental difference in turbot stress physiology.     

The effect of sublethal endrin exposure on rainbow trout, Salmo gairdneri Richardson. II. The effect of altering serum cortisol concentrations on the immune response

The effect of altering serum cortisol concentrations on the immune response was elucidated in endrin- and non-endrin-exposed rainbow trout, Salmo gairdneri. Fish were immunized with 10 μg of Yersinia ruckeri O-antigen following 30 days of treatment. The migration inhibition factor assay (MIF), plaque-forming cell assay (PFC) and serum agglutination titres (SAG) were performed 2, 14 and 30 days post-antigen inoculation. Endrin exposure was continued subsequent to antigen inoculation. Control fish were fed 20 and 35 mg kg⁻¹ body weight day⁻¹ of cortisol and metyrapone, respectively. Endrin-exposed fish received 35 mg kg⁻¹ body weight day⁻¹ of metyrapone in their diet. Control fish receiving cortisol had significantly reduced MIF, PFC and SAG responses. The MIF response was completely restored in endrin-exposed fish receiving dietary metyrapone. The PFC response and SAG titres were partially restored, 61 and 69% respectively, in endrin-exposed fish receiving metyrapone. The results indicate that elevated serum cortisol concentration obtained in endrin-exposed fish has a central role in repression of the immune response.     

PCB disruption of the hypothalamus-pituitary-interrenal axis involves brain glucocorticoid receptor downregulation in anadromous Arctic charr

We examined whether brain glucocorticoid receptor (GR) modulation by polychlorinated biphenyls (PCBs) was involved in the abnormal cortisol response to stress seen in anadromous Arctic charr (Salvelinus alpinus). Fish treated with Aroclor 1254 (0, 1, 10, and 100 mg/kg body mass) were maintained for 5 mo without feeding in the winter to mimic their seasonal fasting cycle, whereas a fed group with 0 and 100 mg/kg Aroclor was maintained for comparison. Fasting elevated plasma cortisol levels and brain GR content but depressed heat shock protein 90 (hsp90) and interrenal cortisol production capacity. Exposure of fasted fish to Aroclor 1254 resulted in a dose-dependent increase in brain total PCB content. This accumulation in fish with high PCB dose was threefold higher in fasted fish compared with fed fish. PCBs depressed plasma cortisol levels but did not affect in vitro interrenal cortisol production capacity in fasted charr. At high PCB dose, the brain GR content was significantly lower in the fasted fish and this corresponded with a lower brain hsp70 and hsp90 content. The elevation of plasma cortisol levels and upregulation of brain GR content may be an important adaptation to extended fasting in anadromous Arctic charr, and this response was disrupted by PCBs. Taken together, the hypothalamus-pituitary-interrenal axis is a target for PCB impact during winter emaciation in anadromous Arctic charr.