John F. Carragher

The effects of acute and chronic stress on the levels of reproductive hormones in the plasma of mature male brown trout, Salmo trutta L

Chronic confinement for 1 month caused a significant elevation of plasma cortisol but suppressed the levels of plasma testosterone and 11-ketotestosterone in sexually mature male brown trout. An acute handling stress for 1 hr elevated blood cortisol and ACTH levels and also suppressed circulating androgens. This androgen suppression in response to acute stress was accompanied by an elevation of plasma gonadotropin levels. These findings are discussed in relation to stress-induced suppression of reproductive function in mammals and the possible biological consequences of such a suppression in fish are outlined.

The deleterious effects of cortisol implantation on reproductive function in two species of trout, Salmo trutta L. and Salmo gairdneri Richardson

Implantation of a cortisol-releasing pellet (60 mg kg-1 fish) into the peritoneal cavity of brown trout, Salmo trutta L. (sexually maturing males and females), and rainbow trout, Salmo gairdneri Richardson (maturing males and immature fish of both sexes), significantly elevated their plasma cortisol level. At 18 days postimplantation, cortisol-implanted sexually maturing male brown trout had smaller gonads, a lower plasma testosterone level, and less gonadotropin in their pituitary gland than control fish. Plasma levels of 11-ketotestosterone and gonadotropin were not significantly affected. Cortisol-implanted sexually maturing female brown trout had smaller gonads, reduced plasma levels of 17 beta-oestradiol, testosterone, and vitellogenin, and a lower pituitary gland gonadotropin content than control fish. The plasma gonadotropin level was unaffected. At 36 days post-implantation, cortisol treatment of maturing male rainbow trout significantly suppressed plasma gonadotropin levels. Plasma levels of testosterone, 11-ketotestosterone, and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one, pituitary gonadotropin content, and gonad size were not significantly affected. In sexually immature female rainbow trout, cortisol administration suppressed the level of vitellogenin in the plasma, compared to control-implanted fish. The 17 beta-oestradiol level was not affected. Cortisol implantation did not affect the plasma testosterone level in sexually immature male trout. These results suggest that prolonged elevation of plasma cortisol, to levels well within physiological range, can affect a wide range of reproductive parameters in both brown and rainbow trout. Further, some effects are manifest in immature as well as in mature fish. These findings are discussed in relation to the effects of cortisol treatment on the state of health of the treated fish.

Effects of acute and chronic stress on the levels of circulating growth hormone in the rainbow trout, Oncorhynchus mykiss.

The acute stress of handling followed by confinement for a period of 1 or 24 hr caused a typical stress response in rainbow trout (elevation of plasma ACTH and cortisol) and a significant reduction in the concentration of circulating growth hormone. The chronic stress of low oxygen levels in both crowded and uncrowded tanks of fish caused a significant elevation of circulating GH levels, an effect which was abolished by the provision of additional aeration to the rearing tanks. This chronic elevation of GH levels was closely correlated with an elevation of plasma cortisol in the same fish. These findings are discussed in relation to stress-induced growth suppression and to the links between the hypothalamic-pituitary-interrenal axis and somatotrope activity.

The effect of starvation on growth and plasma growth hormone concentrations of rainbow trout, Oncorhynchus mykiss

Two experiments, one using 0+ the other 1+ rainbow trout, were conducted to investigate the effect of prolonged starvation on plasma growth hormone levels. The results from both experiments were essentially the same. As expected, starvation resulted in cessation of growth and in a lower coefficient of condition, whereas fed fish continued to grow and remained in good condition. Starvation had relatively little effect on the plasma cortisol level; in one experiment levels were elevated temporarily in starved fish, although by the end of the experiment there was no longer any difference between starved and fed fish, and in the other experiment plasma cortisol levels remained very low throughout the course of the experiment in both starved and fed fish. In contrast, in both experiments starvation had a pronounced effect on the plasma growth hormone level, which rose steadily during both experiments, such that it was six times higher after 1 month of starvation in 0+ fish, and five times higher after 6 weeks of starvation in 1+ fish. Thus, paradoxically, fed fish had very low plasma growth hormone levels and grew rapidly, whereas starved fish had elevated plasma growth hormone levels but did not grow. In both experiments a strong negative correlation was observed between the plasma growth hormone level and the coefficient of condition of the fish. The results are discussed with regard to the well-established metabolic changes that occur during starvation, and it is suggested that a major role of growth hormone during starvation is to aid in the mobilisation of fatty acids and glycerol from adipose stores.

The effect of cortisol on the secretion of sex steroids from cultured ovarian follicles of rainbow trout

In a series of experiments, the effect of cortisol on the ability of rainbow trout (Salmo gairdneri) ovarian follicles to secrete sex steroids in vitro was investigated. The basal rate of secretion of sex steroids (oestradiol and testosterone) was suppressed in a dose-dependent manner by the addition of cortisol to the incubation medium. This suppression was evident using physiological levels of cortisol (less than 100 ng ml-1). The possibility of elevated plasma cortisol levels (due to stress) reducing the reproductive ability of sexually maturing female trout through a direct effect of cortisol on the ovary is discussed.

Development and validation of a highly sensitive radioimmunoassay for Chinook salmon (Oncorhynchus tshawytscha) growth hormone

This study describes the development of a highly specific and very sensitive radioimmunoassay for salmonid growth hormone. Antiserum raised against chinook (Oncorhynchus tshawytscha) GH2, which did not recognize 125I-sPRL and 125I-sGTH (at 1:1000 initial dilution), was able to inhibit growth when injected into rainbow trout (Oncorhynchus mykiss). 125I-sGH2, used as tracer, was not recognized by anti-sGTH or by anti-sPRL. Mammalian GH and ACTH and salmonid GTH, TSH, and PRL did not cross-react in the sGH assay. The inhibition curves for pituitary extracts and plasma from salmonids were parallel to the salmon GH standard, whereas those from carp, tilapia, and catfish showed no significant cross reactivity. The RIA ED90 and ED50 values were 0.2 and 1.5 ng/ml, respectively. Using this RIA for measuring GH release by cultured pituitary cell we observed a strong inhibiting effect of SRIF (10(-6) M) and a stimulatory effect of hGRF (10(-6) M). This RIA allowed us also to detect daily fluctuations in the plasma GH concentration in cannulated rainbow trout.

The mobilization of calcium from calcified tissues of rainbow trout (Oncorhynchus mykiss) induced to synthesize vitellogenin

1. The effect of the induction of vitellogenin synthesis on the mobilization of 45calcium (45Ca) from prelabelled scales and bones of rainbow trout was investigated. 2. A single injection of estradiol increased the hepatosomatic index, total plasma calcium and plasma vitellogenin levels. 3. These changes were accompanied by a 3-fold increase in the plasma 45Ca level and a significant decrease in the amount of 45Ca in the scales. 4. The amount of 45Ca in the vertebrae and rib bones of these fish was not significantly affected by estradiol injection.

Response of Tetraselmis suecica to nutrient and grazer manipulation

Three methods of algal quantification (direct cell counts, chlorophyll a extraction, in vivo fluorescence) were used to evaluate the response of the unicellular green flagellate Tetraselmis suecica to nutrients and grazers. Nutrient enrichment enhanced total cell counts, chlorophyll a concentration and in vivo and DCMU-fluorescence. Photosynthetic efficiency was reduced in the complete F2 medium as indicated by the high level of in vivo fluorescence, whereas photosynthetic efficiency was increased by the introduction of mussels to the F2 medium. The addition of mussels significantly increased the proportion of non-motile cells, but did not reduce the total cell count. The effect of mussel grazing on algae could be underestimated if only total cells were counted or only the chlorophyll a concentration was measured. The results indicate that these three methods measure different properties of an algal culture and are complementary to each other in assessing the quality and quantity of an algal population. Direct algal counting offers a reliable numerical assessment for cell population abundance. Chlorophyll a concentration was closely correlated to the total cell count. In the presence of mussels, in vivo fluorescence did not correlate with either algal cell counts or chlorophyll a concentration, indicating that the measurement of in vivo fluorescence may be misleading for estimating algal abundance under different culture conditions.