Effects of Rhizopus extract administration on somatic growth and sexual maturation in lacustrine sockeye salmon Oncorhynchus nerka

ABSTRACT: The filamentous fungi Rhizopus , like many fungal species, possesses physiologically active substances. Rhizopus extract (RU) is reported to be effective for various aspects of growth and reproduction in many vertebrates. The effects of RU administration on body growth and plasma levels of steroid hormones were investigated in lacustrine sockeye salmon Oncorhynchus nerka . One-year-old fish were fed daily with RU (20 mg/kg feed) from July 1999 to October 2000 for 15 months. Fish were sampled every month and plasma levels of testosterone (T), 11-ketotestosterone (11-KT), estradiol-17β (E₂) and 17α,20β-dihydroxy-4-pregnen-3-one (DHP) were measured by time-resolved fluoroimmunoassay. Body growth of RU-fed fish of both sexes increased significantly in 1⁺ and 2⁺ October, and 2⁺ January–March and July. All RU-fed males and one female matured in 2⁺ October. RU-fed 1⁺ precociously mature males showed increased plasma levels of T, 11-KT and DHP in 1⁺ October. In 2⁺ males, RU significantly elevated plasma levels of T from May to June, 11-KT from June to July, and DHP in October. In sockeye salmon, administration of RU accelerated body growth of both sexes and sexual maturation in males, suggesting physiologically active substances present in RU enhance somatic growth and sexual maturation by sex-specific mechanisms.     

Changes in GnRH levels in the brain and pituitary gland during migrations of sockeye salmon and chum salmon

Levels of two moleculer types of gonadotropin-releasing hormone (GnRH), salmon GnRH (sGnRH) and chicken GnRH–II (cGnRH–II) in the various brain regions and pituitary gland of sockeye salmon (Oncorhynchus nerka) and chum salmon (O. keta) during smoltification and spawning migration, respectively, were measured using specific time-resolved fluoroimmunoassay (TR-FIA) systems. Changes in sGnRH levels in different brain regions tended to be specifically synchronized with serum thyroid hormone or pituitary gonadotropin (GTH) levels during smoltification and spawning migration, respectively. In contrast, cGnRH–II levels did not show such synchronized changes. SGnRH and cGnRH–II in various brain regions might have different roles during smoltification and spawning migration of salmonid fishes.     

Protein metabolism during sexual maturation in female Atlantic salmon (Salmo salar L)

Body composition and fractional rates of protein synthesis (percentage of the protein mass synthesized per day) were determined in female Atlantic salmon returning to the River Tay, Scotland in July and in October after a 95 day period without food, during which time the animals became sexually mature. During the 95 day period of starvation/sexual maturation the ventricle and red muscle remained as a constant proportion of fresh weight whereas the liver, gill and ovary increased and the stomach and white muscle decreased. Fractional rates of protein synthesis increased markedly in the liver, stomach and ovary during the period of starvation/sexual maturation. In the gill, ventricle and white muscle fractional protein synthesis rates increased slightly or remained constant. From the estimated rates of protein loss or gain in the various tissues it is concluded that there is considerable protein turnover and repartitioning of amino acids during the period of starvation and sexual maturation. The absolute rate of protein synthesis rates in the ovary indicates that this tissue made the largest contribution to the energy and amino acid demands of the fish, whilst most of the amino acids required for maturation of the ovary were derived from white muscle, principally as the result of increased muscle protein degradation.     

Plasma somatolactin levels in coho salmon (Oncorhynchus kisutch) during smoltification and sexual maturation

Somatolactin (SL) is a novel pituitary hormone recently characterized in several fish species. Structural analyses have shown that SL belongs to the growth hormone/prolactin family, and that it is a highly conserved protein. SL is synthesized by the periodic acid/Schiff-positive cells in the pars intermedia, but has an as yet unidentified function(s). We have recently developed a homologous radioimmunoassay for coho salmon SL and measured plasma levels of SL during two stages of the coho salmon life cycle, smoltification and sexual maturation. During smoltification, plasma levels of SL changed almost in parallel with plasma levels of thyroxine; levels increased as morphological indices of smoltification appeared and decreased as smoltification was completed. Following this period, SL levels remained low until the spring prior to spawning. In a separate study, plasma levels of SL were measured in sexually maturing coho salmon that remained in fresh water throughout their life cycle. During the year of sexual maturation, plasma levels of SL gradually increased from the spring onward, reaching peak levels at the time of spawning in November and December. These data are similar to those previously reported for sexually maturing coho salmon that were maintained in seawater prior to spawning (Rand-Weaver et al. 1992). Therefore, increases in plasma SL levels occurred in sexually maturing fish irrespective of whether they were maintained in fresh water or seawater. Peak levels at spawning were higher than those observed during smoltification. Possible roles for SL in metabolism and reproduction are discussed.     

Photoperiod-induced precocious male sexual maturation in Atlantic salmon (Salmo salar)

Precocious sexual maturation of male Atlantic salmon (Salmo salar) was induced by photoperiod manipulation to shorten the time required for optimisation of sex inversion treatments. Exposure to a short-day photoperiod (8L:16D) prior to the summer solstice reduced the time to completion of sexual maturation from 3 years to 8–9 months for the majority of fish.     

Leptin in fish: possible role in sexual maturation in male Atlantic salmon

Leptin plays an important role in energy homeostasis and reproductive function in mammals, while its function in fish is still poorly understood, especially its role in reproduction. In the present study, leptin gene expression and circulating leptin plasma levels were measured during sexual maturation in male Atlantic salmon parr. In maturing male salmon, higher hepatic leptin (lepa1) gene expression levels were observed during mid-spermatogenesis compared to immature fish. An upregulation of leptin during sexual maturation has also been observed in a few other teleost species. The physiological significance of elevated leptin levels during the maturational process in teleosts remains to be explored.     

Seasonal changes of hormones and muscle enzymes in adult lacustrine masu (Oncorhynchus masou) and sockeye salmon (O. nerka)

The upstream migration of adult anadromous fishes is characterized by physiological changes in responses to reproductive and energetic challenges. This study analyzed the physiological responses of lake-resident anadromous masu salmon (Oncorhynchus masou) and sockeye salmon (O. nerka) to migration in order to determine if these fish might serve as a suitable model for ocean-running populations and to differentiate between physiological responses to reproduction and to exercise-linked aspects of migration. Reproductive (estradiol, testosterone, 11-ketotestosterone, 17α,20β-dihydroxy-4-pregnen-3-one) and metabolically-linked (thyroxine, triiodothyronine) hormones showed similar patterns to ocean-running anadromous populations. White muscle pyruvate kinase, lactate dehydrogenase and malate dehydrogenase decreased with the onset of spawning season while white muscle citrate synthase, β-hydroxyacetyl CoA dehydrogenase, phosphofructokinase and glutamate oxaloacetate transaminase did not, suggesting that the former group of enzymes are responding to reproductive or food intake signals while the second group, which typically change during anadromous migration, may be responding to exercise-linked aspects of migration. This revised version was published online in August 2006 with corrections to the Cover Date.     

Approaches for delaying sexual maturation in salmon and their possible ecological and ethical implications

ABSTRACT

The aquaculture industry is under pressure to satisfy global demand for marine foods. Atlantic salmon has been bred for more than 40 years, and substantial progress has been made within the culturing and breeding programs. The improved growth rate of Atlantic salmon has been accompanied by an earlier onset of maturation. Among the factors controlling maturation in salmon are photoperiod, temperature, and body composition. Early sexual maturation is detrimental to fish health and quality when viewed from an aquacultural viewpoint. There are several approaches for alleviating this problem: (1) traditional selection, (2) manipulation of external factors affecting puberty (e.g., light), (3) novel biotechnological methods for improving breeding methods, (4) induction of polyploidy, and (5) genetic modification controlling maturation. This article presents the ecological and ethical issues connected to these approaches and argues the importance of acknowledging and discussing such issues in order to ensure that all stakeholder concerns are considered.     

Rearing conditions to develop seawater tolerance in underyearling sockeye salmon smolt

ABSTRACT:   In order to confirm the conditions required to produce underyearling smolts, juvenile sockeye salmon Oncorhynchus nerka were reared under different photoperiod and temperature conditions. The level of smoltification was assessed by plasma sodium concentration 24 h after transfer to sea water (plasma-Na), gill Na⁺,K⁺-ATPase activity (gill-ATPase), and plasma thyroxine (plasma-T4) and growth hormone (plasma-GH) concentrations. Mean fork length of all groups reached more than approximately 8.5 cm at the end of the experiment. The fish that experienced an alteration in photoperiod from short day (8:16 h light–dark cycle) to long day (16:8 h light–dark cycle) showed significant elevation in gill-ATPase from 9 to 16 µmol inorganic phosphate per milligram protein per hour (µmol Pi/mg pro/h) and reduction in plasma-Na from 194 to 154 mM. This group displayed distinct pigmentation on their fin margins. However, in groups reared under constant short day and constant long day, changes in gill-ATPase and plasma-Na occurred vaguely at each different time and pigmentation on fin margins were not observed. The observed photoperiod-induced changes were greater at 7°C than at 11°C. Further, the alteration of the photoperiod induced two–threefold increases in plasma-GH and plasma-T4. These results suggest that lengthening of photoperiod from short to long daylight at 7°C causes smoltification accompanied by active secretion of growth hormone and thyroxine in underyearling sockeye salmon when the fish are larger than 8.5 cm in fork length.     

Thyroid hormone concentrations in the gonads of wild chum salmon during maturation

Changes in gonadal and plasma concentrations of thyroid hormones were examined at various stages of maturation in chum salmon (Oncorhynchus keta) caught in the Bering Sea and the Bay of Alaska. Plasma concentrations of thyroxine (T₄) were less than 5 ng ml⁻¹, and those of 3,5,3′-triiodo-L-thyroxine (T₃) were less than 2 ng ml⁻¹ I in both males and females, regardless of the degree of sexual maturity or the gonadosomatic index (GSI). There was no clear relationships between circulating thyroid hormone levels and tissue levels. The ovarian T₄ concentrations were undetectable (less than 0.2 ng g⁻¹) or less than 2 ng g⁻¹ when GSI was lower than 1%, but increased thereafter and reached a plateau of 8–10 ng g⁻¹ when GSI became 2%. The ovarian T₃ concentrations were about 5 ng g⁻¹ when GSI was 1%, increased to a maximum level (20 ng g⁻¹) when GSI was about 2%, and decreased to a constant level of 10 ng g⁻¹ thereafter. The T₄ and T₃ content in single oocyte increased proportionally to the oocyte volume, indicating a constant incorporation of the hormones into the oocyte. The T₄ concentrations in the testis were 1 ng g⁻¹ or less regardless of the GS1. On the other hand, the T₃ concentrations were highest (15 ng g⁻¹) when the GSI was less than 1%, decreased thereafter when spermatocytes appeared in the testis, and became about 5 ng g⁻¹ I in testes containing spermatozoa, raising the possibility of a role for T₃ during early gamete and/or gonad maturation of testes.     

Effects of aromatase inhibitors on sexual maturation in Atlantic salmon,Salmo salar,male parr

Atlantic salmon, Salmo salar, male parr were implanted with Silastic capsules filled with different aromatase inhibitors: 1,4,6-androstatriene-3,17-dione (ATD), 4-hydroxy-4-androstene-3,17-dione (4OH), and the non-steroidal CGS16949 A, 4-benzonitrile monohydrochloride (CGS). Aromatization in brain homogenates were lower in salmon implanted with CGS and ATD than in controls. This was not the case for 4OH, but administration of 4OH to brain homogenates reduced the aromatase activity. All three aromatase inhibitors had effected gonadal weights in fish sampled in the summer, but the effects were markedly different among inhibitors. Plasma levels of the androgen 11-ketotestosterone (11KT) and the progestin 17, 20-dihydroxy-4-pregnen-3-one (17,20P) were measured by means of radioimmunoassay. CGS and ATD, but not 4OH, significantly decreased the plasma 17,20P levels in the autumn. Plasma levels of 11 KT were not influenced by ATD or CGS treatment, but 4OH had a lowering effect in one autumn sampling. ATD and 4OH (CGS not tested) increased the proportion of maturing males.These findings suggest that aromatization is of physiological importance in different mechanisms controlling reproduction in salmon.     

Epizootiology of Parvicapsulaminibicornis in Fraser River sockeye salmon, Oncorhynchusnerka (Walbaum)

Late-spawning Fraser River sockeye salmon, Oncorhynchus nerka , stocks have suffered significant prespawn mortality associated with an unusually early freshwater migration pattern and the myxosporean parasite Parvicapsula minibicornis . Surveys of migrating adult salmon from several spawning populations were conducted in 1999 and 2000 to determine the extent of infection with P. minibicornis , when and where the parasite first becomes detectable during migration, and whether early migrating stocks might be used as sentinels to assess risk of infection in late-spawning stocks. Posterior kidney, preserved in 95% ethanol, was examined for P. minibicornis in stained histological sections and using a polymerase chain reaction (PCR) test. The prevalence of this parasite in all Fraser River sockeye salmon stocks examined was high (range 47–100% infected). In contrast, P. minibicornis was not detected in the fish tested from the two sockeye salmon stocks outside the Fraser River drainage in either 1999 or 2000. The parasite was also not detected histologically or by PCR in the kidney tissue of the fish from the Fraser River that were sampled in salt water or early during their freshwater migration up the river. These findings and the progression in the prevalence and intensity of infection as the fish from three stocks (early Stuart, Weaver Creek and Cultus Lake) were monitored over time, suggest salmon acquired the parasite either in the lower Strait of Georgia or in the lower Fraser River before the confluence of the Harrison River. In both 1999 and 2000 the parasite was present in all Fraser River sockeye salmon stocks sampled, which suggests that early Stuart salmon may be valuable as a sentinel stock for the presence of the parasite in later-spawning stocks.     

Environmental conditions and physiological state influence estuarine movements of homing sockeye salmon

The reproductive migration of anadromous salmonids through estuarine waters is one of the most challenging stages of their life cycle, yet little is known about the environmental and physiological conditions that influence migratory behaviour. We captured, sampled tissues, tagged and released 365 sockeye salmon (Oncorhynchus nerka) homing through inner coastal waters towards the Fraser River, British Columbia, Canada. Biotelemetry was used to assess the behaviour of individual sockeye salmon approaching estuarine waters and at river entry, which were related to both fish physiological condition at release and to prevailing environmental conditions. Sockeye salmon tended to stay close to the shore, migrated during the day, and movements were related to tide. Sockeye salmon migration rate was linked to wind‐induced currents, salinity and an individual's physiological state, but these factors were specific to location and stock. We propose that wind‐induced currents exposed sockeye salmon entering the estuary to stronger olfactory cues associated with Fraser River water, which in turn resulted in faster migration rates presumably due to either an increased ability for olfactory navigation and/or advanced reproductive schedule through a neuroendocrine response to olfactory cues. However, once the migration had progressed further into more concentrated freshwater of the river plume, sockeye salmon presumably used wind‐induced currents to aid in movements towards the river, which may be associated with energy conservation. Results from this study improve our biological understanding of the movements of Fraser River sockeye salmon and are also broadly relevant to other anadromous salmonids homing in marine environments.     

GnRH affects activity and jumping frequency in adult sockeye salmon, Oncorhynchus nerka

We tested whether waterflow, gender, maturity or treatment with a GnRH analogue affected jumping frequency, an acclimatization response and swimming activity in three-year-old sockeye salmon when confronted with an artificial waterfall. In our first behavioural experiment, increased waterflow and the stage of sexual maturity led to a significant increase in the number of fish that overcame an artificial waterfall, while sex had no significant effect. In a second experiment, we demonstrated that GnRH treatment affected behavioural responses in a waterfall channel as follows: (i) GnRH injected fish did not display an increase in their time of acclimatization while the acclimatization period in control fish increased significantly. (ii) GnRH significantly enhanced swimming activity and jumping motivation. We suggest that waterflow, degree of sexual maturity and injections with a GnRH analogue, lead to behavioural patterns that are correlated with migratory behaviour of adult sockeye salmon.     

Effects of rearing density on sexual maturation and growth in sea-cage reared Atlantic salmon, Salmo salar L.

Abstract. Paired subgroups of fish were derived in January from each of two parent sea-cages of Atlantic salmon, Salmo salar L. The proportion of fish which later became sexually mature in each parent group, after one winter (as grilse) and under commercial rearing density, was determined. Maturity rates in the subgroups, reared in sea-cages at lower density, were significantly greater than in the parent groups. Rearing at reduced density was associated with increased growth in some, hut not all, comparisons. Periodic anaesthesia, handling and sampling of blood for steroid hormone determinations did not consistently affect maturation rate or growth among fish in one of each pair of subgroups. Sexual development was assessed by determining levels of the steroid hormones 11-oxotestosterone or 17β-oestradiol in samples of blood serum taken monthly from individually marked fish in one of each pair of subgroups. Between February and April specific growth rates in maturing male grilse were significantly higher than in fish subsequently shown to have remained immature. Growth rates in maturing female grilse differed similarly between March and April. Between July and August, however, growth rates of non-maturing salmon exceeded those of grilse.     

Changes in alkaline phosphatase isoenzyme activity in tissues and plasma of Atlantic salmon (Salmo salar) before and during smoltification and gonadal maturation

Changes in tissue and plasma isoenzymes of alkaline phosphatase (ALP) were qualitatively and quantitatively determined for male and female Atlantic salmon parr, silvery parr, smolt, immature grilse, prespawning grilse and postspawning grilse using cellulose acetate electrophoresis, densitometry and spectrophotometry. Tissue ALP isoenzymes were isolated from intestine, kidney, bone, liver, and gonad and compared to plasma isoenzymes. Parr plasma displayed three isoenzymes from bone and liver (slow and fast). During smoltification, ALP activity increased in tissue extracts from liver, gonad, and kidney of males and females. Total plasma ALP activity also increased and was due to slow and fast liver isoenzymes. During ovarian development, total ALP plasma activity increased in females and was due mostly to liver isoenzymes and an incompletely identified isoenzyme or isoenzyme mixture (band 2). However, in males total ALP plasma activity did not increase during maturation and no band 2 was evident. In male and female maturing adult grilse, bone ALP activity declined and the isoenzyme band evident in parr plasma could not be detected. ALP activity declined in the plasma of postspawning males and females. In females this was due partly to the total clearance of band 2 from the plasma, together with lowered levels of liver isoenzymes. Treatment of postspawned grilse in February and March with triiodothyronine and thyroxine elevated plasma thyroid hormone levels and increased plasma ALP levels. In conclusion, plasma ALP isoenzyme activities change with physiological state, and knowledge of the conditions governing these changes is important when using these enzymes as a diagnostic tool.     

Chronic low pH exposure affects the seawater readiness of juvenile Pacific sockeye salmon

Chronic exposure to water of low pH during the freshwater life stage of Pacific salmonids is presently the cause for concern due to its potential to reduce subsequent performance in the marine environment. Sockeye fry (0+) were raised under sublethal long-term, low pH conditions (pH 4.8-6.8) in soft water and assessed for effects on freshwater growth, stress physiology, and seawater tolerance following smoltification. Fish gained significantly lower mass (average 46% of control [pH 6.8] values) and had lower condition factor and liver somatic index values than control fish following a 126-days exposure to water at pH 5.0. Liver glycogen concentrations (49% of control values) and whole-body lipid content (65% of control values) were also significantly lower. Low pH exposure also resulted in a sustained organismal stress response that included significant and substantial increases in plasma cortisol concentrations. Fish exposed to pH 5.0 in freshwater for 30 days exhibited an average of 14% mortality in a seawater challenge, as well as a significant osmoregulatory stress measured by increases in plasma Na? and Cl? concentrations as well as osmolality compared to controls. Significantly lower critical swimming speed values (U(crit)) were also seen (22% reductions compared to controls). The data generated indicate that sockeye salmon are sensitive and do not acclimate to low pH under long-term exposure conditions, potentially decreasing the probability of survival in the marine environment.     

Synergistic osmoregulatory dysfunction during salmon lice (Lepeophtheirus salmonis) and infectious hematopoietic necrosis virus co‐infection in sockeye salmon (Oncorhynchus nerka) smolts

While co‐infections are common in both wild and cultured fish, knowledge of the interactive effects of multiple pathogens on host physiology, gene expression and immune response is limited. To evaluate the impact of co‐infection on host survival, physiology and gene expression, sockeye salmon Oncorhynchus nerka smolts were infected with the salmon louse Lepeophtheirus salmonis (V?/SL+), infectious hematopoietic necrosis virus (IHNV; V+/SL?), both (V+/SL+), or neither (V?/SL?). Survival in the V+/SL+ group was significantly lower than the V?/SL? and V?/SL+ groups (p = 0.024). Co‐infected salmon had elevated osmoregulatory indicators and lowered haematocrit values as compared to the uninfected control. Expression of 12 genes associated with the host immune response was analysed in anterior kidney and skin. The only evidence of L. salmonis‐induced modulation of the host antiviral response was down‐regulation of mhc I although the possibility of modulation cannot be ruled out for mx‐1 and rsad2. Co‐infection did not influence the expression of genes associated with the host response to L. salmonis. Therefore, we conclude that the reduced survival in co‐infected sockeye salmon resulted from the osmoregulatory consequences of the sea lice infections which were amplified due to infection with IHNV.