Lack of species-specific primer effects of odours from female Atlantic salmon, Salmo salar, and brown trout, Salmo trutta

We exposed, in two successive spawning seasons, individually placed precocious male Atlantic salmon ( Salmo salar ) and brown trout ( Salmo trutta ) parr to odour stimuli (ovarian fluid and urine mix) from ovulated conspecific or heterospecific anadromous females. Atlantic salmon parr had significantly higher plasma concentrations of the hormones 17α,20β-dihydroxy-4-pregnen-3-one (17,20β-P), 11-ketotestosterone (11-KT) and testosterone (T) after exposure to odours from conspecific females or from brown trout females compared to parr exposed to a control solution (0.9% NaCl). We did not observe any significant differences between the hormone levels in salmon parr exposed to the two female odours. The salmon parr exposed to conspecific odours had significantly higher volumes of strippable milt compared to the controls, but we did not find any significant differences when comparing the effect of the two female odours. Brown trout parr had significantly higher plasma 17,20β-P levels following exposure to heterospecific female odours compared to control males, but there was no significant difference between males exposed to the different female odours. We did not observe any significant differences in plasma levels of T and 11-KT and in milt volumes between exposed and control trout. Taken together, the results from both tested species indicate that the potency of heterospecific stimuli in stimulating increased plasma sex steroid hormone levels in male parr was as strong as stimuli from conspecific females. The results are discussed in connection to observed hybridisation between the two sympatric species.     

Stimulatory and inhibitory effects of testosterone on testes in Atlantic salmon male parr

Immature 1-year-old Atlantic salmon Salmo salar parr were implanted with Silastic capsules of different sizes filled with testosterone (T). Testosterone had both positive and negative effects on testicular weights, spermatogenesis and steroidogenesis. The positive effects: higher incidence of males with enlarged gonads, spermiation, and high plasma levels of 11-ketotestosterone (11-KT) and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P), were most pronounced in males treated with small T capsules. The negative effects: suppression of gonadal development and depressed plasma levels of 11-KT and 17,20β-P compared with mature controls, were most evident in fish treated with large T capsules.     

The role of F-series prostaglandins as reproductive priming pheromones in the brown trout

Electrophysiological studies demonstrated that the olfactory epithelium of mature male brown trout Salmo trutta parr was acutely sensitive to F-series prostaglandins (PGFs) PGF_1α and PGF_2α, with detection threshold concentrations of 10⁻¹¹ M. The olfactory epithelium was also sensitive to the PGF metabolite 15-ketoPGF_2α (threshold 10⁻⁸ m), but did not detect a further metabolite, 13,14,-dihydro-15-ketoPGF_2α Immature brown trout did not detect any of the prostaglandins tested. Exposure of mature male brown trout parr to waterborne PGF_1α and PGF_2α (concentration 10⁻⁸ m), resulted in significant increases in levels of expressible milt and the plasma concentrations of 17,20β-dihydroxy-4-pregnen-3-one, testosterone and 11-ketotestosterone. The olfactory epithelium of both immature and mature male brown trout parr was sensitive to the urine and ovarian fluid from ovulated female brown trout. Exposure of mature male brown trout parr to ovarian fluid resulted in an increase in the levels of plasma 17,20β-dihydroxy-4-pregnen-3-one whilst exposure to urine increased the levels of expressible milt. In addition, PGF_2α was found to be present within both the urine and ovarian fluid of mature female brown trout. It is suggested that the F-series prostaglandins have a role as priming pheromones in male brown trout.     

Sublethal effects of the pesticide Diazinon on olfactory function in mature male Atlantic salmon parr

Diazinon, an organophosphate pesticide, had a sublethal effect on the olfactory system of mature male Atlantic salmon parr. The olfactory responses of the parr to prostaglandin F_2a (PGF_2a) were studied after exposure of the epithelium to different concentrations of Diazinon in water. Electrophysiological recordings from the epithelium indicated that the responses to this prostaglandin were significantly reduced at nominal concentrations as low as 1.0μg l⁻¹ and the threshold of detection was reduced 10-fold at 2.0 μg 1⁻¹ . Mature male salmon parr exposed for a period of 120 h to Diazinon (nominal concentrations 0.3, 0.8, 1.7, 2.7, 5.6, 13, 28 and 45 μg 1 ⁻¹) also had significantly reduced levels of the reproductive steroids, 17,20β-dihydroxy-4-pregnen-3-one, testosterone and gonadotrophin II in the blood plasma after priming with ovulated female salmon urine. Both prostaglandin F_2a and ovulated female urine are known to have important roles in synchronizing reproductive physiology and behaviour in salmonids as well as other fish species. The results are therefore discussed in relation to the possible sublethal effects of Diazinon on reproduction in the Atlantic salmon and possible effects on populations of salmonids.     

17 alpha,20 beta-dihydroxy-4-pregnen-3-one 20-sulphate is a potent odorant in precocious male Atlantic salmon (Salmo salar L.) parr which have been pre-exposed to the urine of ovulated females.

Electrophysiological recordings from the olfactory epithelium have shown that 17 alpha,20 beta-dihydroxy-4-pregnen-3-one 20-sulphate (17,20 beta-P-sulphate; a conjugate of the oocyte-maturation-inducing steroid in teleosts) is a potent odorant in precocious male Atlantic salmon (Salmo salar L.) parr. However, the olfactory epithelium of these fish only appeared to be responsive to the steroid after stimulation with the urine of ovulated female Atlantic salmon. Immature fish did not respond at any time. Stimulation with urine from immature and precocious male Atlantic salmon parr did not make the olfactory epithelium of precocious male salmon parr responsive to the steroid. 17,20 beta-P-sulphate was found in the urines of ovulated females, precocious male parr and mature male Atlantic salmon. The findings are discussed in relation to the possible role of 17,20 beta-P-sulphate in the physiology of Atlantic salmon.     

The impact of two pesticides on olfactory-mediated endocrine function in mature male Atlantic salmon (Salmo salar L.) parr

Short-term exposure of the olfactory epithelium of mature male Atlantic salmon parr to either the pesticide simazine (concentrations 1.0 and 2.0 microg l(-1)) or the pesticide atrazine (concentration 1.0 microg l(-1)) significantly reduced the olfactory response to the female priming pheromone, prostaglandin F(2alpha). In addition, the reproductive priming effect of the pheromone on the levels of expressible milt was also reduced after exposure to the individual pesticides (simazine 0.1, 0.5, 1.0 and 2.0 microg l(-1) and atrazine 0.5 and 2.0 microg l(-1)). When the olfactory epithelium was exposed to a mixture of simazine and atrazine, (concentrations of 0.5:0.5 and 1.0:1.0 microg l(-1)), there was no significant reduction in the olfactory response when compared to the single pesticides at equivalent concentrations. In addition, exposure to a mixture of simazine and atrazine had no synergistic effect on the priming response, and plasma levels of testosterone, 11-ketotestosterone and 17,20beta-dihydroxy-4-pregnen-3-one were similar in the groups of male parr exposed to the individual pesticides. Although the levels of expressible milt were reduced in all groups, there were no significant differences between the different pesticide treatments. The results of the study suggest that the two s-triazine pesticides have an additive and not a synergistic impact on olfactory-mediated endocrine function in mature male salmon parr.     

Differential expression of Atlantic salmon thyrotropin β subunit mRNA and its cDNA sequence

To study the regulation of the thyroid system, an Atlantic salmon Salmo salar cDNA clone was isolated for thyroid stimulating hormone (TSH) β subunit gene. A cDNA (866 bp) was isolated from an adult Atlantic salmon pituitary cDNA library, this clone was sequenced and shown to be highly conserved when compared to other teleost β TSH subunit sequences. The cDNA was used as a probe for Northern blot analysis of total pituitary RNA from the different life cycle stages of Atlantic salmon. Northern blot analysis demonstrated that β TSH mRNA is expressed at all life cycle stages studied, including parr, smolt, immature fish at sea and sexually mature male fish. Densitometry of Northern blots showed that sexually mature male salmon had low levels of salmon β TSH mRNA compared to non-mature fish. Stunts, fish performing poorly in salt water, were shown to have elevated levels of β TSH mRNA when compared to healthy fish.     

Female rainbow trout urine contains a pheromone which causes a rapid rise in plasma 17,20β-dihydroxy-4-pregnen-3-one levels and milt amounts in males

Previous studies have shown that spermiating male rainbow trout respond to the presence of female urine in the water with significant increases in plasma levels of gonadotrophin II, 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) and testosterone. The present results show that males only need a single brief exposure to female urine in order to respond; levels of 17,20β-P rise significantly within 1 h of exposure, and peak between 3 and 4 h. Also, milt amounts increase significantly following exposure of males to female urine. Levels of 17,20β-P are also related positively to the amount of female urine to which the males are exposed. Furthermore, when live females are placed, out of physical and visual contact, in the same tank as males, levels of 17,20β-P rise in the same way as in males which are exposed to female urine. However, if females are fitted with urinary catheters (which drain the urine outside the tank), males respond more slowly. These results indicate that urine is the main source of the male 'priming' pheromone.     

Effect of stripping on the length, condition and gonadal state of mature male Atlantic salmon, Salmo salar L., parr during autumn and winter

Hatchery-reared mature male Atlantic salmon parr were shorter than immature parr of the same age through the winter period and this difference increased prior to smolting in spring. Stripping had no effect on growth. In autumn mature male parr had a higher condition factor (CF) than immature parr, but from early January no difference was observed between unstripped mature males and immature parr. Stripping reduced the CF of male parr but the difference had disappeared by the time of smolting when the CFs of both mature groups were lower than that of immature fish. The quantity of expressible milt in stripped males decreased from c . 3% body weight in November and December to <0.05% by 2 March, after which no further milt expression was recorded. Milt expression in previously unstripped males was about 1% of body weight on 2 March, similar to that of stripped males 1 month earlier.     

Effects of testosterone on gonadotropins, testes, and plasma 17alpha,20beta-dihydroxy-4-pregnene-3-one levels in postbreeding mature Atlantic salmon, Salmo salar, male parr.

Atlantic salmon, Salmo salar, mature male parr were implanted with testosterone (T) in small (T3) or large (T10) Silastic capsules in the breeding season or at its end in November or December, in order to find out whether the postbreeding decline in 17alpha, 20beta-dihydroxy-4-pregnene-3-one (17,20beta-P) and milt production is a consequence of declining T levels. In the first of three experiments, fish were sampled the following January and March, whereas in the second and in the third they were sampled in April. Pituitary and plasma concentrations of gonadotropic hormone (GTHs) I and II and plasma levels of T, 11-ketotestosterone (11-KT) and 17, 20beta-P were measured by radioimmunoassays, and the testes were examined histologically. Administration of T prolonged the period in which running milt was present, suppressed Sertoli cells, and prevented the postbreeding decline in testes weight in experiment two. The postbreeding decline in plasma 17,20beta-P levels was diminished by T10 in experiment one, and by both T3 and T10 in experiment two. The similar decline in 11-KT levels was not influenced by T treatment (only studied in experiment one). T treatment also prevented a decline in pituitary GTH II content (in experiments two and three) and in plasma GTH II levels (only studied in experiment three). However, pituitary GTH I content was not influenced (experiment two and three), whereas plasma GTH I levels (only studied in experiment three) were suppressed by T. To summarize, T treatment prevents postbreeding decline in 17,20beta-P levels, probably via a stimulation of GTH II secretion. J. Exp. Zool. 284:425-436, 1999.     

Metabolic recovery in Atlantic salmon fry and parr following forced activity

Atlantic salmon Salmo salar fry and parr were subjected to 5 min of forced activity and the subsequent changes in oxygen consumption and ammonia excretion rates were evaluated over a 24 h period. In a second experiment, individual Atlantic salmon fry and parr were freeze‐clamped in liquid nitrogen, before, immediately following a 5 min activity period, or after periods of recovery up to 2 h. Samples were analysed for whole body phosphocreatine (PCr), ATP and lactate. Five minutes of forced activity resulted in significant increases in both oxygen consumption and ammonia excretion rates. Changes in the oxygen consumption rates were greater in the parr compared with the fry. In contrast, the post‐exercise ammonia excretion rates were nearly twice as high for the fry compared with the parr. Exercise also caused a marked decrease in PCr levels (c. 47 and 65% in fry and parr, respectively), no change in ATP levels and a significant increase in lactate levels in Atlantic salmon fry and parr. Recovery of PCr occurred quickly (between 15 and 30 min) in fry and parr. Although the post‐activity levels of lactate were lower in fry (c. 3 μmol g^?1) compared with parr (c. 14 μmol g^?1), lactate levels returned to control levels within 60 min in fry, but it took >2 h for this metabolite to recover in parr. Compared with parr, these findings show that Atlantic salmon fry possess a reduced anaerobic capacity, and these results are consistent with the theoretical and experimental evidence that smaller fish support burst swimming through aerobic processes.     

Size variability of juvenile Atlantic salmon: links to environmental conditions

Climate change models predict a 2 to 6° C increase in air temperature within the next 100 years in the Maritime Provinces of eastern Canada. Higher air temperatures are expected to contribute to increased water temperatures, alterations in stream flow conditions, and ultimately reductions in fish growth. Mean annual size-at-age of juvenile Atlantic salmon Salmo salar decreased in the Northwest Miramichi and Southwest Miramichi Rivers between 1971–1999. Lengths-at-age of juveniles were significantly correlated between the two rivers. For Atlantic salmon parr, stronger associations between inter-cohort fork length ( L _F) than intra-cohort L _F were observed, suggesting that environmental conditions in the current year of growth have the more significant effects on size of age 2 year parr than conditions encountered the previous year by age 1 year parr of the same cohort. Fork lengths of parr were significantly and negatively associated with spring air and water temperatures. In the Miramichi River, increases in air and water temperature as predicted from climate change models may adversely affect growth of juvenile Atlantic salmon parr, reducing the overall productivity of the Atlantic salmon populations in this region.     

Effects of copper on olfactory‐mediated endocrine responses and reproductive behaviour in mature male brown trout Salmo trutta parr to conspecific females

In the present study, the effects of copper (CuSO₄) on the ability of mature male brown trout Salmo trutta parr to detect and react both physiologically and behaviourally to female pheromones were studied. The study was composed of two parts. In the first experiment, priming effects of the female pheromone prostaglandin F_2α (PGF_2α) were evaluated by determining the amount of milt produced and the blood plasma levels of 11‐ketotestosterone (11‐KT) and 17α,20β‐dihydroxy‐4‐pregnen‐3‐one (17,20β‐P) after the PGF_2α exposure. In the second experiment, male parr were placed in a large stream tank together with a group of adult males and ovulated females and their individual behaviours were recorded. In the priming experiment, the amount of expressible milt was significantly lower, less than half, in groups exposed during 4 days to 10 or 100 µg l^?1 copper compared with control parr only exposed to water. No significant differences were observed in plasma levels of 11‐KT and 17, 20β‐P. During the behavioural experiment, exposed parr spent less time with the female and had a lower number of courting events. Blood plasma levels of 11‐KT were, however, significantly higher in the group exposed to 100 µg l^?1 copper compared with the control group. Furthermore, the exposed group spent significantly less time swimming upstream than did the control group. The present study demonstrates that exposure to copper affects reproductive behaviours and endocrinology of S. trutta male parr.     

The effect of pineal removal on circulating melatonin levels in Atlantic salmon parr

Nocturnal melatonin levels showed a significant decrease in pinealectomized Atlantic salmon parr in comparison with intact control and sham-operated fish.     

Moon phase and nocturnal density of Atlantic salmon parr in the Sainte‐Marguerite River, Québec

Nocturnal underwater counts of Atlantic salmon Salmo salar parr were made on four consecutive occasions (two lunar cycles, samplings at both the full and new moon) at four 40 m long sites in the Sainte‐Marguerite River, Québec, Canada, between 30 June and 14 August 2003. Atlantic salmon parr counts did not differ significantly between moon phases. Cloud cover ranged from 0 to 100% during the study, and had no significant effect on parr counts. There were significantly more Atlantic salmon parr in the near shore than in the midstream areas. The findings of this study suggest that the sampling strategy of summer studies aimed at assessing population abundance or developing habitat quality models can be designed without taking moon phase or cloud cover into consideration, but it should account for the higher relative abundance of Atlantic salmon parr in the near shore areas as compared to areas closer to the middle of streams.     

Luteinizing hormone and sexual steroid plasma levels after treatment of European sea bass with sustained-release delivery systems for gonadotropin-releasing hormone analogue

Spermiating male European sea bass Dicentrarchus labrax were treated with gonadotropin-releasing hormone agonists (GnRHa), either a GnRHa injection (IN; 25 μg kg⁻¹ body mass) or one of three types of controlled-release GnRHa-delivery systems: fast release implants (EVAc; 1OO μg kg⁻¹), slow release implants (EVSL; lOO μg kg⁻¹) and slow release microspheres (MC; 50 μg kg⁻¹). Luteinizing hormone (LH) release was highly stimulated by all GnRHa treatments, with elevated plasma levels lasting for 2 days in injected fish (IN) and 2, 4 and 6 weeks in controlled-release-treated fish (EVAc, MC and EVSL, respectively), correlating with a 1, 3, 5 and 5 week period of stimulation of milt production, respectively. Plasma levels of the androgens testosterone (T) and 11-ketotestosterone (11-KT), were not significantly affected by the GnRHa treatments. Plasma T was high at early spermiation and declined sharply near the end of this period. Plasma 11-KT levels declined continuously throughout the experiment. Levels of 17,20 β -dihydroxy-4-pregnen-3-one (17,20 β -P), a proposed maturation-inducing steroid (MIS) in European sea bass, fluctuated around 0.2–1 ng ml⁻¹ and were not greatly affected by the treatments. These results indicated a close correlation between sustained stimulation of LH release, achieved by GnRHa-delivery systems, and long-term enhancement of milt production. They also show an absence of changes in the common sex steroids, associated with elevated LH and enhanced spermiation.     

Relative ventricular weight of wild Atlantic salmon parr in relation to sex, gonad maturation and migratory activity

Relative ventricular weight (RVW, ventricular weight/body weight · 100) was measured for five groups of Atlantic salmon parr from the Girnock Burn in Scotland. The mean RVW was: mature males, 0.087; immature resident males, 0.054; immature resident females, 0.056; immature migrant males, 0.056; immature migrant females, 0.053. For a group of mature male parr from the R. Nith, Galloway, Scotland, the mean RVW was 0.108. There were no significant differences in the RVW between groups of immature fish. The RVW of each group of mature male parr was significantly greater than those of each, group of immature parr. Condition indices (weight/length³) were closely similar between all groups of parr. The functional significance of plasticity in ventricular weight is discussed in relation to life histories of Atlantic salmon.     

Determination of body shape variation in Irish hatchery‐reared and wild Atlantic salmon

Discriminant function analysis was used to distinguish morphologically between samples of parr, smolts and adult Atlantic salmon Salmo salar from several hatchery and river systems in Ireland. The effect of habitat shift was investigated in Atlantic salmon parr. Parr grown from the eyed‐egg stage with a non‐sibling group in a hatchery environment, came to resemble the mean body shape of their host hatchery Atlantic salmon stock more closely than that of a full sibling group grown at their natal hatchery. Wild Atlantic salmon smolts differed in shape from hatchery‐reared smolts. This difference was less pronounced, but still statistically significant when wild adults were compared with hatchery‐reared adults captured in the coastal drift‐net fishery after a year spent at sea. Rearing conditions had a significant impact on the production and growth of fish body shape. This in turn may have affected adaptability and survivorship of ranched Atlantic salmon in the marine environment.     

The role of the maturation-inducing steroid, 17,20β-dihydroxypregn-4-en-3-one, in male fishes: a review

The major progestin in teleosts is not progesterone, as in tetrapods, but 17,20β-dihydroxypregn-4-en-3-one (17,20β-P) or, in certain species, 17,20β,21-trihydroxy-pregn-4-en-3-one (17,20β,21-P). Several functions for 17,20β-P and 17,20β,21-P have been proposed (and in some cases proved). These include induction of oocyte final maturation and spermiation (milt production), enhancement of sperm motility (by alteration of the pH and fluidity of the seminal fluid) and acting as a pheromone in male cyprinids. Another important function, initiation of meiosis (the first step in both spermatogenesis and oogenesis), has only very recently been proposed. This is a process that takes place at puberty in all fishes and once a year in repeat spawners. The present review critically examines the evidence to support the proposed functions of 17,20β-P in males, including listing of the evidence for the presence of 17,20β-P in the blood plasma of male fishes and discussion of why, in many species, it appears to be absent (or present at low and, in some cases, unvarying concentrations); consideration of the evidence, obtained mainly from in vitro studies, for this steroid being predominantly produced by the testis, for its production being under the control of luteinizing hormone (gonadotrophin II) and, at least in salmonids, for two cell types (Leydig cells and sperm cells) being involved in its synthesis; discussion of the factors involved in the regulation of the switch from androgen to 17,20β-P production that seems to occur in many species just at the time of spermiation; discussion of the effects of in vivo injection and application of 17,20β-P (and closely related compounds) in males; a listing of previously published evidence that supports the proposed new function of 17,20β-P as an initiator of meiosis; finally, discussion of the evidence for environmental endocrine disruption by progestins in fishes.     

Is Atlantic salmon production limited by number of territories?

Thirty Atlantic salmon Salmo salar parr (mean 135 mm total length, L _T) in the River Alta, Norway, were radio‐tagged and tracked during a 11–13 day period. The parr stayed within defined home ranges with a 95% probability of localization within an average area of 1286 m²(241–3484 m²). On average, each parr had overlapping home ranges with 7·7 other parr, and the overlap between pairs of fish covered on average 24% of their home range areas. Mean length of the river stretch used was 90 m (22–383 m). On average, during 38%(16–77%) of the tracking surveys, the fish had moved >10 m since the previous survey. Mean total distance moved during the whole study was 402 m (208–862 m). The Atlantic salmon were most often recorded in riffles, but 27% of the parr alternated between riffles and pools. The extensive movements, flexible habitat utilization and relatively large home ranges, together with the fact that all the parr had home ranges partly overlapping with other radio‐tagged parr, indicate a flexible and variable behaviour and, thereby, a more complex social structure than a rigid defence of a fixed territory. Thus, Atlantic salmon production seems not to be limited by the highest potential number of territories for large parr in a given area.