Disruption of gonadal maturation in cultured Senegalese sole Solea senegalensis Kaup by continuous light and/or constant temperature regimes

A major problem in the development of Senegalese sole Solea senegalensis intensive culture is the poor control on reproduction, in part due to the lack of knowledge on the precise role of photoperiod and temperature. Thus, gonadal maturation was evaluated by assessing the sequential changes in plasma levels of 17β-estradiol (E₂), 11-ketotestosterone (11-KT), and testosterone (T) in both female and male cultured Senegalese sole (F1 generation) exposed to various combinations of constant or naturally-fluctuating daylength and water temperature. Under natural photoperiod (NP; 36° N), exposure to constant temperature (t0; 18-20 °C) disrupted gonadal development, as indicated by a lower incidence (in comparison with naturally-fluctuating water temperature; 14-24 °C) of females at advanced maturation (from February to April: 12 vs. 33%) and running males (from February to May: 46% vs. 57%), and the reduced mean (± S.E.M.) sex steroid plasma levels (female peak E₂ levels: 2.9 ± 0.28 vs. 1.8 ± 0.3 ng ml^− 1; male peak T levels: 1.5 ± 0.14 vs. 0.9 ± 0.06 ng ml^− 1). Therefore, the onset and progression of gonadal development in this species seem to be strongly (“proximally”) influenced by fluctuating water temperature. When compared to NP and t0, exposure to continuous light (LL) under t0 significantly reduced steroid production (female peak E₂ levels: 1.8 ± 0.28 vs. 0.5 ± 0.05 ng ml^− 1; male peak 11-KT levels: 9.4 ± 1.06 vs. 5.4 ± 1.33 ng ml^− 1) and subsequently gonadal development (lower proportions of females at intermediate [46 vs. 6%] and advanced maturation [12 vs. 0%] from February to April and of RM [46 vs. 33%] from February to May). Thus, the seasonal changes of daylength would be crucial for normal gonadal development, being its cueing effects of higher magnitude than those of water temperature. The present report constitutes the first systematic study focused on the environmental control of reproductive events in Senegalese sole.     

Intra‐annual changes in reproductive indices of male and female Himalayan snow trout,Schizothorax richardsonii (Gray, 1832)

Periodic changes in reproductive hormone levels, gonadal histology and gonadosomatic index (GSI) of snow trout, Schizothorax richardsonii, were examined to ascertain annual cycle of gonadal development and reproductive status in their natural habitat. In females, there were coherent changes in plasma 17β‐oestradiol and vitellogenin along with GSI, oocyte maturation and vitellogenic progression, collectively indicating two distinct maturation peaks during the months of September and February. Coinciding with this, in males, plasma 11‐keto testosterone was also noticeably higher during September and February, with highest GSI values in September. However, plasma 17α, 20β‐dihydroxyprogesterone levels in males were found to be persistently high from September to February. This observation suggests the potential presence of matured oozing males over a longer period, unlike in females. Overall, the close association between reproductive hormone levels, GSI and gonadal maturation stages in males and females (particularly, the presence of postovulatory follicle complexes) with apparent natural synchronization clearly indicates that S. richardsonii breeds twice in a year, possibly during late September to early November and late February to early April in the coldwater riverine habitats of the Indian Himalayan region.     

The influence of reproductive status on the stimulatory action of N-methyl-D,L-aspartate on growth hormone secretion, in vitro in rainbow trout, Oncorhynchus mykiss

The glutamate agonist, N-methyl-D,L-aspartate (NMA) stimulates the secretion of growth hormone (GH) from pituitary fragments in vitro and increases plasma GH levels in vivo in rainbow trout, Oncorhynchus mykiss (Flett et al. 1994; Holloway and Leatherland 1997a,b); however gonadal steroid hormones appear to modulate this response in experimental situations. This study examines whether steroid hormones also modulate the GH-regulatory actions of NMA during the normal reproductive cycle of rainbow trout by examining the relationship between the stage of sexual maturation and the pituitary release of GH in vitro in response to an NMA (10^-8 M) challenge. NMA had no effect on mean GH release from the pituitary glands of fish that were immature (GSI <1.0), from males during early development (GSI 1.0-3.0), or from sexually mature males (with free running milt) and females (ovulated). However, NMA significantly increased GH release from pituitary glands taken from females during the early stages of gonadal growth (GSI 1.0-9.0) and from males and females sampled during the later stages of gonadal growth (males GSI 3.01-6.0; females GSI 9.01-15.0). The GH-stimulatory action of NMA in males and females progressed to a maximum effect during the late stages of gonadal growth, and disappeared in ovulated females and free running males. Moreover, in female fish, the maximal GH release in response to the NMA challenge is positively correlated with plasma 17β-estradiol levels; no such correlation was evident for plasma testosterone levels in males. Changes in the GH response to NMA during maturation while gonadal steroid levels fluctuate provides further evidence to suggest that the effects of NMA on GH secretion are intimately linked to endogenous gonadal steroid hormone levels. This revised version was published online in July 2006 with corrections to the Cover Date.     

Sex differences in circulating steroid hormone levels in the red drum, Sciaenops ocellatus L.

Steroid profiles of cultured and captive red drum (Sciaenops ocellatus L.) were investigated to evaluate the potential use of circulating sex steroid levels as a tool for gender identification in this species. Cultured 18‐month‐old fish were maintained on a 120‐day shortened photothermal cycle to induce precocious maturation. Additionally, wild‐caught fish were maintained in captivity under simulated natural photothermal conditions from late spring to early fall. Circulating 11‐ketotestosterone (11‐KT) levels were significantly higher in males compared with females during the early stages of gonadal growth in both cultured and captive fish. Plasma testosterone (T) levels showed a similar trend; however, the differences were significant only when males were already producing sperm. 17β‐estradiol (E₂) concentrations were low in males and females before gonadal recrudescence but increased significantly with the progression of vitellogenesis in females. These results show that a test using a minimum concentration of circulating 11‐KT could be developed to differentiate between sexes in the early stages of gonadal maturation in red drum. Moreover, plasma E₂ concentrations could be used to identify vitellogenic females. The two steroids considered together could help avoid possible error in gender identification due to unusually high levels of certain steroids encountered in some individuals.     

Reproduction of striped bass, Morone saxatilis (Walbaum), broodstock: monitoring maturation and hormonal induction of spawning

Abstract Levels of gonadal steroid hormones were quantified in an adult striped bass, Morone saxatilis (Walbaum), broodstock during their gametogenic cycle. Blood plasma concentrations of Estradiol (E₂) and testosterone in females, or 11-ketotestosterone(11-KT) and testosterone (T) in males, were used as indicators of maturation. In both sexes, hormone levels were low in summer but increased significantly by late October to intermediate levels which were then maintained until late January. They then increased again rapidly to maximum pre-spawning values attained in late February or March, and subsequently decreased during the spawning period (April and May) with an increased incidence of spent fish with low hormone levels. The changes in blood hormone concentrations coincided with annual changes in photoperiod and water temperature that may be useful landmarks for maturation in captive broodstock. Mature females were implanted with pellets containing a dose of approximately 20 μg/kg body weight of [D-Ala⁶-Pro⁹-Net]-LHRH (GnRHa) in a matrix of cholesterol (CH) and cellulose. In April, they had not yet begun final oocyte maturation (FOM) and were too immature for conventional induction of spawning by injection with human chorionic gonadotropin (hCG). In early April, females given two 95% CH (slow hormone-release) GnRHa pellets (95/95) or females given one 80% CH (fast hormone-release) GnRHa pellet and one 95% CH GnRHa pellet (80/95) spawned within 13 days treatment (n= 4) with good egg fertility (76 ± 7% of total) and hatch rates (62 ± 15% of fertile). Females given dual fast-release GnRHa pellets (80/80) or control (Sham) pellets did not spawn or show evidence of increased oocyte diameter or development. In late April, four of six females given the 80/95 GnRHa pellet combination spawned within 9 days. Three fish produced fertile eggs (54 ± 18%). one spawned overripe eggs, and the remaining two increased oocyte diameter and maturation. Three corresponding controls did not spawn, and two of these showed clear signs of atresia within 11 days. In early May, some females were undergoing early FOM and were mature enough to be spawned by hCG injection. Three were given a single 80% CH GnRHa pellet and spawned within 6 days of treatment to produce fertile eggs (44 ± 6%). Of two other females given dual 80% CH GnRHa pellets, one spawned infertile eggs and the other failed to spawn within 9 days. GnRHa implants show promise as a technique for inducing spawning of captive striped bass broodstock although the optimum hormone delivery systems, dosages and release rates should be verified for fish at specific maturational stages.     

Environmental Manipulation of Growth and Sexual Maturation in Yellow Perch, Perca flavescens

Yellow perch, Perca flavescens, exhibit sexually dimorphic growth detected as the fish enter puberty. The purpose of this study was to evaluate the differential influences of photoperiod and temperature on the sexual maturation and dimorphic growth pattern of yellow perch. Four combinations of constant (CP) or natural (NP) photoperiod with constant (CT) or natural (NT) temperature were evaluated (CPCT, CPNT, NPCT, NPNT). Following 6 months of growth, results indicate that (1) CT is the strongest promoter of overall growth regardless of photoperiod. Perch reared under CT were 26.4 ± 8.1 g heavier than perch reared under NT (P < 0.0001). (2) CP also promotes overall growth regardless of temperature. CP perch outgrew NP perch by 7.0 ± 8.1 g (P < 0.0872). (3) NP (decreasing) initiates maturation in yellow perch regardless of temperature. (4) NT is required for late maturational processes. While both NP groups initiated maturation, only NPNT females were able to recruit oocytes to a vitellogenic stage. Only NPNT males were able to sustain spermiation to the end of the study. (5) CPCT confer the best overall growth performance, fully inhibit maturation in both genders, and suppress a sexually dimorphic growth pattern up to 53 g average weight. These results indicate that using a CPCT regime during intensive fingerling production will produce a larger, more uniform fish population that could alleviate losses as a result of cannibalism. Perch that remained immature did not exhibit a dimorphic growth pattern.     

Changes in levels of GnRH in the brain and pituitary and GTH in the pituitary in male masu salmon,Oncorhynchus masou,from hatching to maturation

Levels of two types of gonadotropin-releasing hormone (salmon GnRH and chicken GnRH-II) in the brain and pituitary, and content of gonadotropin (GTHIβ and IIβ) in the pituitary were measured in male masu salmon from hatching to gonadal maturation for three years in order to clarify the involvement of GnRHs in precocious maturation. Underyearling precocious males were distinguishable in summer of year 1 and were marked by an increased GSI. Spermiation was observed among these individuals thereafter every autumn. Pituitary GTHIβ content in both precocious and immature males, and GTHIIβ content in precocious males showed seasonal fluctuations — high in autumn and low in winter. Pituitary GTHIIβ content was low in immature males. Pituitary sGnRH content in precocious males increased from spring to autumn during the three-year period. sGnRH concentrations in discrete brain areas showed seasonal changes — high during autumn to winter and low in summer. Concentrations in the olfactory bulbs and hypothalamus increased significantly in association with testicular maturation during year 3. sGnRH concentrations in the hypothalamus were significantly higher in precocious males than in immature males; this was possibly due to positive feedback of steroid hormones. cGnRH-II was undetectable in the pituitary and no distinct changes were observed in its concentration in the brain in relation to maturation. The phenomenon of underyearling precocious maturation is considered to be triggered before the onset of early summer. It is suggested that males which mature precociously are larger in size and contain much sGnRH in the pituitary before the outward signs of precocity appear; sGnRH may stimulate GTH II synthesis and induce precocious maturation.     

Growth,sexual maturation,and spawning season of Atlantic halibut,Hippoglossus hippoglossus,in Faroese waters

Biological data on Atlantic halibut, Hippoglossus hippoglossus, caught in an exploratory bottom trawl fishery in November–February 1983–1986 in a recently discovered deep water spawning area and in other both coastal and bank areas in Faroese waters, are presented.Studies of increases in length and weight confirmed the sex-dependent growth patterns found in earlier works, i.e. a higher growth rate in females than males especially after sexual maturity. The data show that Faroese halibut grow much more rapidly and mature at lower average ages (c. 4.5 years in males and slightly above 7 years in females) than in any other area investigated in the North Atlantic. Average length and round weight at sexual maturity were slightly above 55 cm and c. 1.7 kg in males, while the corresponding values in females were 110–115 cm and c. 18 kg.Spawning in the area appeared to commence in January and intensify in February. The peak in spawning activity was apparently not attained during the study period (November–February), and most probably occurred later in winter or early in spring.     

Hematological parameters and phagocytic activity in fat snook (Centropomus parallelus): Seasonal variation, sex and gonadal maturation

The objective of this work was to determine hematological parameters and the phagocytic capacity of peritoneal macrophages of the fat snook related to sex, stage of gonadal maturation and seasonal cycle. The hematological results did not show significant differences between males and females. With respect to stage of gonadal maturation, only erythrocytes numbers (RBC), in females, was found to be significantly elevated in stage III (maturing) and decreased in V (spent). An analysis of the results of the erythrocyte and leukocyte series, thrombocytes and phagocytic activity related to sex and seasonal cycle showed statistically significant differences: a) hematocrit (Ht) and hemoglobin concentration (Hb) were elevated in the spring and low in the winter in males; b) mean corpuscular volume (MCV) was high in the spring and summer and low in the fall in both sexes; c) thrombocytes numbers were elevated in the fall and low in the other seasons in females; d) phagocytic capacity (PC) and phagocytic index (PI) were higher in the summer and lower in the fall in females. The results showed that spring and summer correspond to seasons of the year for better hematological and phagocytic responses for survival of the fat snook in its natural habitat. The parameters studied could be utilized for evaluation of the health status fat snook in its own habitat or in captivity.     

Effect of stocking density,oxygen level,light regime and swimming velocity on the incidence of sexual maturation in adult Atlantic salmon (Salmo salar)

The effects of various environmental parameters on sexual maturation of two sea-winter Atlantic salmon (Salmo salar) were tested in two separate experiments. In the first experiment Atlantic salmon with initial mean individual weight 1.5 kg (smolt 13 months before) were reared for 8 months from June to February at different oxygen levels and stocking densities using continuous light. Oxygen levels of 5–7, 7.5–9.5 and 10–12 mg O₂ l⁻¹ and stocking densities starting at about 20, 30 and 40 kg m⁻³ and increasing as the fish grew to 80–90 kg m⁻³ for the highest densities were tested in a factorial design. Only male fish matured, and incidence of maturation among males varied from 4.1% to 25% between tanks. The highest percentage of mature males was found in the tanks with low stocking density. No clear effect on oxygen level was found.The second experiment lasted 20 months from seawater transfer in May until the fish weighed 3.3–3.5 kg. Two water current speeds (14–16 and 20–24 cm s⁻¹) and two photoperiod regimes (LD 20:4) and continuous light (LL) were tested in a factorial design. Neither swimming velocity nor photoperiod affected growth rate. Continuous light reduced the incidence of sexual maturation. The average proportion of maturation among males was 8% and 25% under the LL and LD 20:4 regimes respectively. The fish reared under the LD 20:4 light regime had a significant lower condition factor and significant larger hearts than the fish reared under continuous light. Swimming velocity had no significant effect on the incidence of maturation. The results indicate that the swimming velocity must be higher than 0.5 BL s⁻¹ in order to influence the energy stores. An important finding in this study is that light cues are not required for gonadal growth. The results also indicate that environmental factors can affect maturation even after the first sea-winter.     

Evidence that elevated water temperature affects the reproductive physiology of the European bullhead Cottus gobio

Climate change is predicted to increase the average water temperature and alter the ecology and physiology of several organisms including fish species. To examine the effects of increased water temperature on freshwater fish reproduction, adult European bullhead Cottus gobio of both genders were maintained under three temperature regimes (T1: 6–10, T2: 10–14 and T3: 14–18°C) and assessed for gonad development (gonadosomatic index—GSI and gonad histology), sex steroids (testosterone—T, 17β-estradiol—E2 and 11-ketotestosterone—11-KT) and vitellogenin (alkali-labile phosphoprotein phosphorus—ALP) dynamics in December, January, February and March. The results indicate that a 8°C rise in water temperature (T3) deeply disrupted the gonadal maturation in both genders. This observation was associated with the absence of GSI peak from January to March, and low levels of plasma sex steroids compared with T1-exposed fish. Nevertheless, exposure to an increasing temperature of 4°C (T2) appeared to accelerate oogenesis with an early peak value in GSI and level of plasma T recorded in January relative to T1-exposed females. In males, the low GSI, reduced level of plasma 11-KT and the absence of GSI increase from January to March support the deleterious effects of increasing water temperature on spermatogenesis. The findings of the present study suggest that exposure to elevated temperatures within the context of climate warming might affect the reproductive success of C. gobio. Specifically, a 4°C rise in water temperature affects gametogenesis by advancing the spawning, and a complete reproductive failure is observed at an elevated temperature of 8°C.     

Development and validation of an enzyme immunoassay for testosterone: Effects of photoperiod on plasma testosterone levels and gonadal development in male sea bass (Dicentrarchus labrax, L.) at puberty

A specific immunoassay was developed for the quantification of testosterone (T) in sea bass plasma. Specific primary antibody against T was produced using an antigen BSA conjugated with T. The enzyme immunoassay (EIA) had a sensitivity of 5–0.009 ng ml^–1 and 6.2% intra-assay variation; inter-assay variation was 9.5% for sea bass plasma. The effects of two different accelerating photoperiod regimes, compressed photoperiod (CO; 6 months), and constant short photoperiod (9L:15D) with a long photoperiod (15L:9D) in March (SLmar), on T plasma levels and sexual maturation were examined during the onset of puberty in male sea bass. Natural photoperiod (NP) and SLmar groups exhibited the highest T values in December (8.69±1.03 and 10.85±1.04 ng ml^–1, respectively). However, SLmar group showed the first significant decrease in T plasma levels in January, two months earlier than the NP group, which presented elevated T levels until February. The CO group displayed two significant T peaks, one in October (8.90±1.60 ng ml^–1) and the other in January (9.60±1.10 ng ml^–1). Gonadosomatic index (GSI) in the NP and SLmar groups displayed the highest values from December to February (>2.5%). However, the SLmar group showed the first significant increase in GSI in November, one month before the controls, indicating a clear advancement of gonadal development with respect to the NP group. In the CO group, a bimodal pattern was observed with two peaks, one in October–November (1.30±0.25%) and the second in March–April (0.97±0.33%) (P<0.05). In NP group, the percentage of running males was about 80% from December to March while the percentage of running males in the SLmar group (70%) lasted only three months (December to February) decreasing (P<0.05) in March. In the CO group, spermiation began in October (60%), decreased during the next months, and increased again in March–April (30%) (P<0.05). These results indicate the advancement of puberty by either one or two months with respect to the control group in the SLmar and CO groups, respectively, and the presence of a second reproductive surge in the CO group. Collectively, these results suggest that exposure of fish to these photoperiod regimes may affect both the time of the onset of puberty and the pattern of gonadal development in prepuberal male sea bass.     

Effects of altered photoperiod regimes during winter on growth and gonadosomatic index in Arctic charr (Salvelinus alpinus) reared in freshwater

The short‐ and long‐term effects of altered photoperiods during winter on growth and final gonadosomatic index (GSI) were investigated in 178 individually tagged 2‐year‐old smolt Arctic charr from an anadromous strain. The fish were reared at ambient temperature (2.3–12.5°C) for 18 months and reared at five different photoperiods. One group was reared on constant LD16:8 (light–dark, N = 40) photoperiod and a second group on continuous light (LD24:0, N = 32) throughout the experimental period. Three groups of fish were moved from LD16:8 to LD24:0 for 44 days and subsequently back to LD16:8, that is early winter light group (Early WL: 17 November–5 January; N = 35), Mid WL group (5 January–23 February; N = 38) and Late WL group (23 February–6 April; N = 33). No differences in growth were found for females, whereas males reared at constant LD24:0 were larger (mean ± SEM, 1,780 g ± 180) compared with the Late (1,264 g ± 101) and Mid WL (1,413 g ± 120) groups towards the end the study. Exposure to continuous light during early winter significantly influenced the GSI in female Arctic charr, whereas no differences were found in the males. Female GSI (%) was lowest in the Mid WL group (1.7) and highest in the LD24:0 group (7.0). In conclusion, the present study demonstrated that application of brief continuous light treatments during January and February can possibly be used as a tool to lower subsequent female maturation in Arctic charr farming.     

Oogenesis and lipids in gonad and liver of daubed shanny (Leptoclinus maculatus) females from Svalbard waters

Oocyte and liver histomorphology of the daubed shanny (Leptoclinus maculatus) from Isfjorden and Kongsfjorden in Svalbard were investigated during three Arctic seasons: summer (July), autumn (October) and winter (April). Three oocyte developmental phases were observed: primary growth phase, secondary growth phase and maturation phase. We observed four different developmental stages: (1) perinucleolus stage with cortical alveolus formation, (2) lipid droplets formation, (3) vitellogenesis stage and (4) maturation. Late maturation stage of oocytes in the ovaries was from the autumn season. Females accumulated lipids in liver (up to 35.2?% dw) and deposited large amounts of lipids into gonads (up to 52.2?% dw) during maturation. Lipid classes in female gonads changed seasonally, with relative increase in cholesterol during summer and depletion of storage lipids (triacylglycerols and wax esters/cholesterol esters) during the winter. Lipid composition in liver changed during oocyte development and spawning, as neutral lipids were transferred to developing oocytes during summer to autumn. During winter, storage lipids were depleted during starvation. Based on the increase in gonadosomatic index (GSI) with length and observed maturation stages, females seem to mature at a length of about 125-130?mm. The GSI and hepatosomatic index of large females sampled in autumn (September-October) were significantly higher than for females in late winter (April) and spring (May). These results indicate that spawning takes place during winter in Isfjorden and that energy reserves stored in the liver are utilized by females during gonadal development and reproduction.     

Effects of light and temperature conditions on the expression of GnRH and GtH genes and levels of plasma steroids in <Emphasis Type="Italic">Odontesthes bonariensis</Emphasis> females

In this study we examined the endocrine mediation between environmental factors (temperature and photoperiod) and the brain–pituitary–gonadal axis in females of pejerrey Odontesthes bonariensis. Changes in the expression of brain gonadotropin-releasing hormones (GnRHs) and gonadotropin (GtH) subunit [follicle stimulating-β (FSH-β), luteinizing hormone-β (LH-β), glycoprotein hormone-α (GPH-α)] genes, plasma gonadal steroids [estradiol (E₂) and testosterone (T)], gonadal histology, and gonadosomatic index (GSI) in adult females exposed to combinations of short-day (8 h) or long-day (16 h) photoperiods and low (12°C) or high (20°C) temperatures after winter conditions (8 h light, 12°C) were analyzed. Pejerrey females kept under the short photoperiod had low GSIs, and their ovaries contained only previtellogenic oocytes regardless of the experimental temperature. In contrast, females exposed to the long photoperiod had high GSIs and ovaries with vitellogenic oocytes at both temperatures. These fish also showed a significantly higher expression of sGnRH, pjGnRH, cGnRH-II (the three different GnRH variants found to date in the pejerrey brain), FSH-β, LH-β and GPH-α genes and plasma E₂ levels than those at the shorter photoperiod. No significant changes were observed in plasma T levels. Based on these results, we concluded that the increase in day length but not that of temperature triggers the maturation of pejerrey females after the winter period of gonadal rest and that this occurs by an integrated stimulation of the various components of the brain–pituitary–gonad axis.     

The effect of whole body lipid on early sexual maturation of 1+ age male chinook salmon (Oncorhynchus tshawytscha)

Early sexual maturation of male chinook salmon (maturation 1 to 4 years prior to females in the same age class) results in reduced effectiveness of stock enhancement programs and a financial loss to the salmon farming industry. Previous studies in Atlantic salmon have shown that the age of maturity in males is affected by growth and/or body energy stores, but the relative roles of these two factors are not well understood. Therefore, an experiment was designed to determine when spermatogenesis was initiated, to characterize the endocrine changes during the onset of puberty in male salmon, and to determine if the level of whole-body lipid affects the incidence of early male maturation in a wild stock (Yakima River) of 1+ spring chinook salmon. Fry were fed a commercial diet from February until August and were then divided into groups of 320 fish (mean weight, 5.6 g) and fed one of five experimental diets (two replicate groups/diet) containing 4%, 9%, 14%, 18% or 22% lipid and 82%, 77%, 73%, 69%, or 65% protein for 13 months. Fish were reared on natural photoperiod and ambient temperature (6°C to 16°C), and pair-fed to a level based on the tank with the lowest feed consumption. Fish were weighed monthly and sampled to determine body composition, pituitary follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels, plasma insulin-like growth factor I (IGF-I) levels, and stage of gonadal development.

Throughout the experimental period the mean fish weight was similar among treatment groups. However, from December through the end of the experiment in the following September, maturing males were significantly larger than nonmaturing fish. Initial lipid levels in 0-age experimental fish were near 6%, which is similar to wild fish of the same stock and age captured in the Yakima River during August. Fish fed diets containing more than 4% lipid increased in whole-body lipid content during the first 2 months of feeding and then maintained at relatively constant levels during the course of the experiment. Whole-body lipid levels for the dietary treatment groups averaged 5.6%, 7.1%, 8.2%, 9.4%, and 9.6% from October through the following September.

Based on histological examination of the testes of experimental fish, type B spermatogonia and primary spermatocytes were first observed in some of the yearling males during November. These were designated maturing males. Pituitary FSH levels were significantly higher in maturing than nonmaturing males at this time and for the remainder of the study. Pituitary FSH levels increased as spermatogenesis proceeded in maturing fish, whereas pituitary LH levels increased in maturing 1+ males only during July and August, when testes were in late stages of spermatogenesis and in September during spermiation. Plasma IGF-I levels were significantly higher in maturing males than nonmaturing fish from December through the end of experiment. Since maturing males were significantly larger than nonmaturing fish of both sexes from December through September, the difference in IGF-I levels could be due to differences in growth or due to maturation.

The percentage of maturing males was significantly influenced by whole-body lipid, increasing from 34% in fish fed the 4% lipid diet to 45% in fish fed the 22% lipid diet. These data suggest that whole-body lipid levels influenced the incidence of maturation of male spring chinook salmon. In addition, both endocrine and histological indicators suggest that maturation was initiated in males approximately a full year prior to the time the fish will spawn.     

Growth, osmoregulation and sexual maturation of underyearling Atlantic salmon smolt Salmo salar L. exposed to different intensities of continuous light in sea cages

Atlantic salmon smolts are usually transferred to sea water during early spring, but photoperiod treatment can be used to produce underyearling (0+) smolts for transfer to sea water in late autumn, 7–8 months earlier than usual. This study investigated the effect of exposure to additional continuous light (LL) of different intensities on 0+ smolts after transfer to sea water. 0+ smolts transferred to sea cages in mid-October were exposed to natural light (NL) or LL of one of three intensities (low, medium or high) until January, whereupon they experienced NL until harvest in December. Fish in all groups displayed a normal ability to hypoosmoregulate, assessed by monitoring plasma chloride concentrations after transfer to sea water. No consistent differences in length, weight or condition were found between LL groups. The LL groups had greater growth in length than the NL group during the time of LL exposure, followed by an increase in weight when exposed to NL. This resulted in fish of larger size than the NL group. By August, the fish in the NL group had caught up with the LL groups in respect of length, weight and condition factors. This suggests that the LL treatment led to increased winter growth and phase-shifted a seasonal pattern of growth. The incidence of sexual maturation was low (< 1%), with no differences between groups.     

Conditioning of the European flat oyster (Ostrea edulis,Linnaeus 1758): effect of food ration

To establish a reliable protocol for the broodstock conditioning of the oyster Ostrea edulis (Linnaeus 1758) in hatcheries is important to obtain the best number of viable larvae all year even outside the natural season for maturation and spawning. Hatchery production of bivalves depends on their initial gonadal maturation state, which is linked to season and several other factors, such as temperature, daylight, salinity and food supply. Ration and diet composition are important in broodstock conditioning because food availability affects fecundity and gametogenic development. In this work, the effect of food ration on O. edulis conditioning was studied in autumn and winter. The results of this study showed that the broodstock conditioned with a food ration of 6% and 9% produces the highest number of larvae in autumn and winter. These food rations produced rapid gonadal development and larval release in a short period with a similar larval production. The oysters conditioned with a food ration of 3% delayed larval release and produced a significantly lower number of larvae in autumn. In the 9% ration treatment, the percentages of mortality were significantly higher in both autumn and winter. Thus, a 6% food ration is likely to be the most suitable for flat oyster conditioning combined with a gradient of temperature (14–18°C) and a gradient of daylight (8–16 h).     

Long photoperiod delayed spawning and increased somatic growth in gilthead seabream (Sparus aurata)

The effects of extended photoperiods, mimicking the longest day of the year, were studied in 1- and 2-year seabream. The photoperiod regimes started in late July, 36 and 39 days after the summer solstice and continued for 11 months, well beyond the natural reproductive season of December–March. Regime 1 (long day, 15.5L:8.5D), which used natural and fluorescent light, reduced the incidence of maturity in both year classes and females did not spawn although some gonadal development was observed. Among all 1-year sampled fish of regime 1, a maximum of 5% became spermiating males (March) and 5% reached the yolk granule stage of vitellogenesis (VO3; 250–400 μm diameter) by May. Among 2-year sampled fish of regime 1, 45% became spermiating males and 25% were females, which reached the advanced vitellogenesis stage (VO4; 400–600 μm) by April. Regime 2 (skeleton photoperiod), consisting of natural light and a 1.5-h pulse of fluorescent light during the period 14–15.5 h after sunrise, postponed gonadal development and spawning for up to 3 months. In this regime, a maximum of 80% of 1-year sampled fish were spermiating males in February and a maximum of 10% were VO3 stage females in March. In the sampled 2-year fish, the maximum levels were 50% spermiating males in February and 25% VO3 stage females in March. Control fish, which were exposed to the natural photoperiod (29°34′N), spawned during their natural season. The maximum levels for 1-year sampled control fish were 95% spermiating males and no females in December, while among 2-year sampled fish, maxima of 75% males in February and 45% VO4 stage females in November. Final average weights of photoperiod treated fish (1-year=430 g—regime 1, 400 g—regime 2; 2-year=582 g—regime 1, 518 g—regime 2) were significantly greater (p<0.05) than control fish (1-year=341 g; 2-year=476 g). Daily feed consumption (g/100 g fish) dropped from an average of 1.83 to 0.93 g for 1-year fish during August–December and from 0.88 to 0.54 g for 2-year fish during the same period. This was correlated with reduced autumn and winter water temperatures (26–20°C summer to winter change) and increased fish weight in all treatments. Daily feed consumption was also affected by the onset of spawning in the control and regime 2 (skeleton photoperiod) treatments of both 1- and 2-year fish. The efficiency of feed utilization (FCR) and protein and energy retention were all affected by the photoperiod regimes and followed the same pattern of decrease as weight gain, regime 1 (long day)>regime 2 (skeleton photoperiod)>control.     

Seasonal changes in plasma gonadal steroid concentrations and gonadal morphology of male and female tench (Tinca tinca, L.)

In order to gain a better understanding of the reproductive cycles of male and female tench (Tinca tinca), gonadosomatic index, gonad histology and plasma concentrations of estradiol‐17β (E₂), testosterone, an drostenedione, 11‐ketotestosterone (11‐KT), 17,20β, 21‐trihydroxy‐4‐pregnen‐3‐one (17,20β,21‐P), 17,20β‐dihydroxy‐4‐pregnen‐3‐one (17,20β‐P) and 17,20α‐dihydroxy‐4‐pregnen‐3‐one (17,20α‐P) were measured at the four seasons of the year, plus a further sampling coincident with the peak of spawning in early July. As expected, in both males and females, the plasma concentrations of androgens (excluding 11‐KT in females – undetectable) and C₂₁ steroids were significantly more elevated in the spring and summer (when most gonadal development took place) than in the autumn and winter. The only unexpected finding was that 17,20β‐P and 17,20β,21‐P, the steroids that are normally associated with oocyte final maturation in females and spermiation in males, were found in substantial amounts in both pre‐vitellogenic, pre‐spermatogenic and post‐spawning fish. This suggests that these steroids may have other as yet unidentified roles in this species.