Photoperiodic manipulations of the reproductive cycle of sea bass (Dicentrarchus labrax) and their effects on gonadal development, and plasma 17ß-estradiol and vitellogenin levels

The annual profile of plasma vitellogenin (VTG) and 17ß-estradiol (E₂) levels, as well as gonadal development and spawning characteristics were investigated in captive female sea bass (Dicentrarchus labrax). Endocrine and gonadal changes were studied in fish reared under natural conditions or exposed to manipulated photothermal cycles. In natural conditions of photoperiod and temperature, sea bass spawned from February through March (East coast of Spain, 40°N 0°E). One or two months of constant long-days (15L/9D) in a constant short-day photoperiod regime (9L/15D) all-year-round, given early in the year (March and March–April), advanced spawning by 3 months. The same treatment applied later in the year (September–October) delayed spawning by 1 month, compared to controls.In all groups, changes in plasma VTG levels were correlated with E₂ levels, oocyte growth and spawning time. In control females, VTG was low (<100 ng ml^-1) during the summer, until its first surge in plasma 4 months before the beginning of spawning. The VTG (3.1 ± 0.3 mg ml^-1) and E₂ (4.1 ± 0.5 ng ml^-1) levels showed a single annual peak during late vitellogenesis, the time of the highest proportion of vitellogenic oocytes in the ovary. Constant high levels of VTG (1–1.4 mg ml^-1) and E₂ (1.6–1.9ng ml^-1) were maintained during the entire spawning time, together with the presence of vitellogenic oocytes, suggesting the existence of several waves of oocyte growth in the ovary and thus, several spawns per female. Endocrine profiles and oocyte development in fish exposed to constant photoperiods were similar to controls, but were shifted in time in relation to the displacement of the spawning time. In the fish showing advanced spawns, the duration of the gametogenic proces was compressed when compared to controls. The differences observed in the evolution of the reproductive-related factors in the advanced groups, which were exposed to a reduction in temperature to 15°C, suggest an influence of the temperature in the early stages of the reproductive cycle in sea bass.     

Effect of triiodothyronine on the growth and survival of larval striped bass (Morone saxatilis)

This study was carried out to test the effect of triiodothyronine (T₃) on the growth and survival of larval striped bass (Morone saxatilis). Growth and survival of striped bass held in 5 ppt seawater and treated with various doses of T₃ were measured beginning at 5 and 16 days after hatching. Body content of T₃ was measured by radioimmunoassay. T₃ dissolved in the 5 ppt seawater was taken up by larval striped bass in a dose-dependent manner, and affected the growth and survival of the fish. At 5 days after hatching, T₃ at 100 ng ml^–1 and 50 ng ml^–1 retarded the growth of larval striped bass and caused a lower survival rate than T₃ at 25 ng ml^–1 or the control treatment. At 16 days after hatching, T₃ at 100 ng ml^–1 retarded the growth of larval fish and caused a higher mortality. T₃ at 10 ng ml^–1 and 1 ng ml^–1 did not show any effect on either survival or growth. Body content of T₃ returns to control levels within days following end of treatment. The results indicate that exogenous T₃ can be detrimental to the growth and survival of larval striped bass.     

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.     

Fructose 1,6 bisphosphatase activity in liver and gonads of sea bass (Dicentrarchus labrax). Influence of diet composition and stage of the reproductive cycle

The fructose 1,6 biphosphatase (FBPase) activity was measured in the liver and gonads of broodstock sea bass that were fed three experimental diets over the reproductive cycle. Enzyme activity was measured at the pre-spawning (November), spawning (February) and post-spawning (May) periods. Additionally, some biometric indices were calculated at these three times. The different diets, one with standard composition (51% protein, 13% lipid, 10% digestible carbohydrates), one with a high-carbohydrate content (32% protein, 13% lipid, 32% digestible carbohydrates) and one deficient in ω3 essential fatty acids (51% protein, 13% lipid, 10% digestible carbohydrates), had similar affects on growth and gonad development, as deduced from the similar weight, condition factor, hepatosomatic index, gonadosomatic index and liposomatic index over the reproductive cycle. FBPase activity was detected at all times in the liver of both sexes, and in the ovaries but not in the testes. In general, females showed greater liver activity than males, especially at spawning time. During post-spawning, FBPase activity fell in tissues of both sexes. Enzyme activity was also affected, mostly in the liver, by the different types of feed; hepatic activity in fish fed the high-carbohydrate diet fell during the spawning and post-spawning periods, though at a different level for each sex. The diet deficient in ω3 EFA presented the lowest FBPase activity, compared with the other diets, in females during the spawning period; there was very little change in males or females over the reproductive cycle. The results showed that the highest gluconeogenic activity in sea bass broodstock occurred at the spawning time and the females had larger glucose requirements than males over the reproductive cycle, as demonstrated by the absence of FBPase activity in testes. This work also confirmed the adaptative capacity of this enzyme to different nutritional and physiological conditions.     

The Effect of Photoperiod on Sexual Maturation,Appetite and Growth in Wild Atlantic Cod (Gadus morhua L.)

An experiment was conducted to examine (a) the effects of photoperiod on timing of sexual maturation (b) the relationship between plasma steroid levels, appetite and growth in male and female Atlantic cod (Gadus morhua L.). Wild caught Norwegian coastal cod were subjected to either a 6L/18D photoperiod typical of January at 60° N-(Short day group) or a simulated natural photoperiod (Normal day group) from June 2000 until spawning started. Appetite of individual fish were measured twice weekly, while weight, length and plasma levels of the sex steroids testosterone (T), 11 keto-testosterone (11-KT) and estradiol-17β (E2) were monitored bimonthly. Cod in the Short day group matured 3 months ahead of the cod in the Normal day group and started spawning in early November. Appetite decreased in both sexes 2–3 months prior to spawning in both groups, but this reduction was stronger among males. In both sexes, length growth was reduced concurrently with the appetite loss. Overall, females had significantly higher somatic growth, put relatively less energy into length growth and had developed larger livers compared to males at the time of spawning in the Short day group. Plasma steroid levels increased in both groups throughout the experiment, reaching peak levels of ca 10 ng ml⁻¹ (T) and 15–20 ng ml⁻¹ (11-KT) in males, and 1.5–2 ng ml⁻¹ (T) and 12–18 ng ml⁻¹ (E2) in females at the onset of spawning. Steroid levels increased more rapidly among Short day cod verifying the earlier onset of maturation. These results confirm that photoperiod is a major cue to maturation in cod and imply that the high cost of spawning for females incur differences in appetite between the sexes.     

Purification,characterization and immunoassay of striped bass (Morone saxatilis) vitellogenin

The egg yolk precursor, vitellogenin (VTG), was purified from blood plasma of striped bass by chromatography on hydroxylapatite or DEAE-agarose. The fish were first implanted with estradiol-17β (E₂), which induced vitellogenesis. A rabbit antiserum (a-FSPP) raised against plasma from mature female striped bass, and then adsorbed with mature male plasma, was used to detect female-specific plasma protein (FSPP) in the chromatography fractions. Striped bass VTG (s-VTG) was collected from the peak fraction that was induced by E₂, reacted with a-FSPP, and contained all detectable phosphoprotein. It appeared as a single band (M_r ≂ 170,000) in SDS-PAGE or Western blots using a-FSPP, and as a pair of closely-spaced phospholipoprotein bands in native gradient-PAGE, suggesting that there is more than one circulating form of s-VTG. The relationship of s-VTG to the yolk proteins was verified using a-FSPP. The antiserum reacted with the main peak from gel filtration of saline ovary extracts, and it specifically immunostained the two main bands in Western blots of the extracts and the yolk granules of mature oocytes. The amino acid composition of s-VTG was similar to that of VTG from other fish and Xenopus. A radial immunodiffusion assay for s-VTG was developed using a-FSPP and purified s-VTG as standard. The s-VTG was not detected in blood plasma of males, immature females, or regressed adult females, but plasma s-VTG levels were highly correlated with plasma E₂ and testosterone levels, and oocyte growth, in maturing females. The results indicate that the maturational status of female striped bass can be identified by s-VTG immunoassay.     

Effects of dietary carnitine and lipid on growth and body composition of hybrid striped bass (Morone chrysops ♀× M. saxatilis ♂)

A study was undertaken to investigate the effects of graded dietary levels and different types of carnitine on hybrid striped bass (Morone chrysops × M. saxatilis %) fed different levels of lipid. An incomplete factorial design was utilized in which diets containing lipid at either 5 or 10% were supplemented with l-carnitine at 0, 500, or 1000 mg kg^–1 diet, dl-carnitine at 1000 mg kg^–1 diet, or carnitine chloride to provide 1000 mg carnitine kg^–1 diet. Juvenile hybrid striped bass (3.3 g fish^–1) were stocked into individual 38-l aquaria connected as a brackish water (6), recirculating system and fed each diet in triplicate for 9 weeks.Supplementation of the diet with 1000 mg carnitine kg^–1 increased muscle carnitine from 35.5 to 47.7 g g^–1 tissue. Carnitine supplementation did not result in increased weight gain regardless of carnitine level or type; however, weight gain showed a significant (p<0.05) response to dietary lipid with fish fed diets containing 10% lipid growing 34% more than fish fed diets with 5% lipid. The hepatosomatic index also was unaffected by diet, but the intraperitoneal fat (IPF) ratio was significantly elevated (5.1 vs 3.2%) in fish fed diets with 10% lipid compared to those fed diets with 5% lipid. Fish fed diets containing 1000 mg carnitine kg^–1 had increased IPF ratio values at 4.7% compared to 3.9% for fish fed the basal diet. Liver lipid also was responsive to dietary treatment, increasing from 6.7 to 8.8% of wet weight as dietary lipid increased from 5 to 10%. The relative quantities of triglycerides, free fatty acids and phospholipids in muscle and liver were not influenced by carnitine level, carnitine type or dietary lipid level. Supplementation of carnitine does not appear to be beneficial to hybrid striped bass based on either growth performance or body composition.     

Endocrine and gonadal changes during the annual reproductive cycle of the freshwater teleost,Stizostedion vitreum

The annual reproductive cycle of walleye (Stizostedion vitreum) was characterized by documenting changes in gonadal development and serum levels of estradiol-17β (E₂), testosterone (T), 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P), and 11-ketotestosterone (11-KT) in wild fish captured from upper midwestern lakes and rivers throughout the year. Fish from the populations used in this study spawn annually in early- to mid-April. Walleye showed group synchronous ovarian development with exogenous vitellogenesis beginning in autumn. Oocyte diameters increased rapidly from ∼ 200 μm in October to ∼ 1,000 μm in November, and reached a maximum of 1,500 μm just prior to spawning. Changes in gonadosomatic indices (GSIs) paralleled changes in oocyte diameters. Serum E₂ levels in females increased rapidly from low values in October (< 0.1 ng ml⁻¹) to peak levels of 3.7 ng ml⁻¹ in November, coinciding with the period of the most rapid ovarian growth. Subsequently, E₂ levels decreased from December through spawning. Serum T levels exhibited a bimodal pattern, increasing to 1.6 ng ml⁻¹ in November, and peaking again at 3.3 ng ml⁻¹ just prior to spawning. We detected 11-KT in the serum of some females at concentrations up to 5.6 ng ml⁻¹, but no seasonal pattern was apparent. In this study (unlike our results in a related study) 17,20-P was not detected. In males, differentiation of spermatogonia began in late August, and by January the testes were filled (> 95% of germ cells) with spermatozoa. Mature spermatozoa could be expressed from males from January through April. GSIs ranged from 0.2% (post-spawn) to 3.2% (pre-spawn). Serum T levels rose from undetectable levels in post-spawn males to 1.6 ng ml⁻¹ by November, remained elevated throughout the winter, and peaked at 2.8 ng ml⁻¹ I prior to spawning. Levels of 11-KT in males remained low (< 10 ng ml⁻¹, from post-spawning through January, then increased significantly by March and peaked just prior to spawning at 39.7 ng ml⁻¹. Our results indicate that vitellogenesis and spermatogenesis are complete or nearly so, in walleye by early winter, and suggest that it may be possible to induce spawning in this species several months prior to the normal spawning season by subjecting fish to relatively simple environmental and hormonal treatments.     

Changes in gonadal hormones during oocyte development in the striped bass,Morone saxatilis

Wild striped bass,Morone saxatilis, were collected from coastal waters and spawning areas to describe the endocrine correlates of oocyte development in non-captive, migratory fish. The fish were classified according to their most advanced oocytes. Serum levels of estradiol (E₂), testosterone (T) and 17-20-dihydroxyprogesterone (DHP) were measured by radioimmunoassay (RIA). Females in the primary growth phase and early secondary growth phase (pre-vitellogenic) had low levels of plasma steroids, ovarian lipid content and gonadosomatic indices (GSIs). Significant increases in E₂, T, ovarian lipid content and GSIs occurred during the vitellogenic phase. Maximum levels of all reproductive parameters were found in prespawning fish sampled in the Hudson River. Mean levels of E₂, T, ovarian lipids and GSIs for these fish were 2.0±0.5 ng/ml, 3.0±0.3 ng/ml, 24±1 mg/g, and 5.6±0.3% (mean±SEM), respectively. In fish induced to spawn with human chorionic gonadotropin (HCG), DHP levels (1.9±0.4 ng/ml) were significantly elevated. Similar levels were found in two fish captured during the spawning season, suggesting that DHP may serve as the maturation-inducing steroid in this species.     

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.     

Off-Season Spawning of Eurasian Perch Perca Fluviatilis

This study was designed to determine the effects of the selection for body weight and of two final warming durations during an out-of-season reproductive cycle on gonad recrudescence, plasma steroid levels and spawning in Eurasian perch Perca fluviatilis males and females. Two different bodyweight classes (200–250 and 350–400 g) and two different warming periods (1 or 2 months) were studied.At the end of the chilling period, a significant effect of the selection for body weight was observed on gonad development whereas fish were the same age. According to higher levels in testosterone (12 ng mL^–1), a better development was recorded for smaller breeders (P < 0.05). A short water warming period up to 14°C (1 month v.s. 2 months) allowed higher rates of females with completed gonadogenesis (33–38% v.s. 7–8%) and spawning (31–33% v.s. 0%) (P < 0.05). Spontaneous out-of-season spawning (17 ribbons) and larvae were obtained. Relative fecundity was about 100 eggs g^–1 of female body weight. Five ribbons were partly fertilized with a rate ranging from 5 to 90%. These studies have shown that delayed spawning in Eurasian perch can be obtained by a temperature manipulation, but results are not sufficient because of low stimulation rate of females, low spawning rates and high heteroge-neity of fertilization rates. The effects of other environmental factors like photoperiod should be tested in further experiment to improve the protocol inducing out-of-season spawning production.     

Potassium flux in juvenile striped bass (Morone saxatilis): influence of external concentrations of sodium chloride and calcium

Whole-body influx and efflux of K⁺ were determined for 25-day-old striped bass,Morone saxatilis, in conditions that simulated harvesting fish from ponds. During the first 5h in fresh water with combined high NaCl (80 mM) and low Ca²⁺ (0.12 mM) concentrations, a combination that is acutely lethal to this age of striped bass, K⁺ influx for fish in 0.07 mM K⁺ was 21±1.7 (SEM) compared to 3.4±0.33 nmol g^–1 h^–1 for fish in water with low Na⁺ (0.25 mM) or high Ca²⁺ (2.5 mM) concentrations. Influx of K⁺ was inhibited during the first few hours after fish were placed in flux chambers. Potassium efflux as percentage of⁴²K lost per hour was two-fold higher from fish in the high Na-low Ca treatment compared to fish in low concentrations of Na⁺ or high concentrations of Ca²⁺. Potassium efflux was probably much greater than influx, but exact values for efflux could not be calculated from the data available. Survival of fish in water with high Na-low Ca was not increased by addition of KCl to the water, indicating that the net loss of K⁺ was probably not the cause of death.     

Lipids in female northern bluefin tuna (Thunnus thynnus thynnus L.) during sexual maturation

Total lipids, lipid classes, fatty acids and vitamin E contents were measured in ovary, liver, dark muscle, white muscle and mesenteric perigonadal fat in four maturational stages of Atlantic female northern buefin tuna (Thunnus thynnus thynnus L.) during migration to Mediterranean spawning grounds through the Strait of Gibraltar. Ovary lipid content increased significantly by 37.6-fold from inmature to spawning stage, accumulating large quantities of steryl/wax esters, whereas the ratio of mesenteric perigonadal fat (primarily triacylglycerols) weight to ovary weight decreased linearly throughout maturation. In contrast, liver, dark red and white muscles lipid content did not show a major depletion during maturation compared to that of perigonadal fat. The fatty acid composition of ovary from bluefin tuna showed that docosahexaenoic acid (DHA; 22:6n-3) was the most abundant fatty acid and DHA: EPA: AA and DHA: EPA ratios were 19.3/4.3/1 and 4.5/1, respectively. Neutral lipids predominated over polar lipids (2-3:1) in ovary and triacylglycerol and steryl/wax ester were major lipid classes. In general, fatty acid composition of most tissues examined were similar to each other but, in contrast, the fatty acid profile of perigonadal fat was rich in monoenes such as 18:1n-9, 20:1n-9 and 22:1n-11, and DHA decreased significantly at spawning stage. The total lipid balance suggested for the disappearance of lipid from perigonadal fat depots and, to a lesser degree, from muscle and liver depots, reflected lipid utilization for gonadal development on the one hand and non-gonadal metabolism in the other. The former included lipid transferred to the gonad and lipid catabolized to provide metabolic energy for the biosynthesis of gonadal constituents and the later was likely to be lipid catabolism to provide energy for swimming during spawning migration. This revised version was published online in August 2006 with corrections to the Cover Date.     

Endocrine changes during the annual reproductive cycle of the red porgy, Pagrus pagrus (Teleostei, Sparidae)

Changes in 17-estradiol (E2), estrone (E1), testosterone (T), 11-ketotestosterone (11KT), and 17,20-dihydroxy-4-pregnen-3-one (17,20-P) levels were correlated to changes in gonadosomatic index (GSI), vitellogenin concentration (Vg), ovarian and testicular histology during the annual reproductive cycle of the red porgy, Pagrus pagrus. The production of E2, E1, T and 17,20-P was confirmed by analysis of the steroidogenic activity of ovaries. In females, the average concentration of E2 was lower than 2 ng ml^–1. E2 values first increased significantly at the stage of endogenous vitellogenesis and remained high during exogenous vitellogenesis. E1 levels were lower values than E2 (less than 300 pg ml^–1), but they increased at the beginning of exogenous vitellogenesis. Estrogens concentrations followed similar pattern to Vg and were significantly correlated. Mean levels of T were mostly lower than 1 ng ml^–1. They followed a pattern similar to that of E2 except for a further increase observed at the stage of final maturation. T and E2 levels were significantly correlated. The concentration of 11KT did not change significantly. The levels of 17,20-P ranged between 0.22 and 1.22 ng ml^–1 but changes were not related to gametogenesis. In males, the concentrations of T and 11KT fluctuated significantly during the sexual maturity stages, showing a similar pattern and were significantly correlated to GSI changes. T levels increased during spermiogenesis and spermiation stages to reach about 3 ng ml^–1. 11KT levels stayed about half those of T. The levels of estrogens showed no significant changes. Level of 17,20-P showed no significant variation related to male maturity. Results are discussed in relation to changes in plasma steroid levels during gametogenesis of other multiple spawner species.     

Marine wax ester digestion in salmonid fish: a review

Alternative marine resources from lower trophic levels could partly cover the rapidly increasing needs for marine proteins and oils in the future. The North Atlantic calanoid copepod, Calanus finmarchicus, has a high level of lipids rich in n‐3 fatty acids. However, these animals have wax esters as the main lipid storage component rather than triacylglycerol (TAG). Although these esters are considered difficult to digest by many fish, is it well known that juvenile Atlantic salmon (Salmo salar) feed on zooplankton species. It is therefore possible that the capacity to utilize these lipids should be well developed in salmonids. Nonetheless, salmon hydrolyse wax esters slower than TAG and absorb fatty alcohols slower than fatty acids. However, salmon have several adaptations to digest diets rich in wax esters. These includes increased feed conversion, higher production of bile and higher activity of lipolytic enzymes in the midgut. Atlantic salmon has been shown to feed and grow on diets with a medium amount of wax esters (30% of the lipid) with results comparable to fish maintained on fish oil diets. Ingestion of higher level of wax esters (50% of the lipid) cause, however, poorer lipid digestibility and growth, so that optimal utilization of wax esters in Atlantic salmon is closer to 30% than 50% of the dietary lipid.     

Plasma levels of Gonadotropin-II and gonadal sex steroids in triploid catfish, Heteropneustes fossilis (Bloch)

Triploid fish have under-developed gonads due to altered reproductive endocrinology. Triploids of Indian catfish (H. fossilis) showed significantly reduced plasma levels of gonadotropin (GtH-II), testosterone (T) and estradiol-17 (E₂) than that of diploids throughout the year, except for the resting phase, irrespective of sex. Plasma levels of GtH-II were significantly different (p<0.001) between diploid and triploid fish during preparatory, prespawning and spawning phase. The plasma testosterone contents in triploids were significantly less (p<0.001) than that of diploids, except during the resting phase. Triploid females showed very low titres of estradiol-17 (<1 ng ml^–1) throughout the annual reproductive cycle in contrast to highly fluctuating levels in diploid females. Thus, this study for the first time provides information on reduced levels of GtH-II and sex steroids in plasma of male triploid fish and additional information on species-specific alteration of sex hormones in female triploids.     

Utilization of lipids by Dentex dentex L. (Osteichthyes, Sparidae) larvae during lecitotrophia and subsequent starvation

Total lipids, lipid classes and their associated fatty acids were measured in developing eggs, yolksac larvae and starving larvae (from fertilized egg to day 9 after hatch) of the common dentex Dentex dentex (L., 1758). The larvae of common dentex during lecitotrophia and subsequent starvation consumed 1.6 g of total lipid per larvae per day. The overall decrease was mainly due to utilization of the major neutral lipids, TAG and SE (0.5 and 0.6 g larvae^–1 day^–1, respectively) which was 3.4-fold greater than that of the the major phosphoglycerides (primarily PC by 0.2 g larvae^–1 day^–1). There was net synthesis/conservation of PE during the first half of the study period before it decreased rapidly (0.2 g larvae^–1 day^–1) during the second half. PUFAs were principally catabolized (468.6 ng larvae^–1 day^–1), primarily 22:6(n-3), 20:5(n-3) and 20:4(n-6) (221.8, 58.5 and 12.1 ng larvae^–1 day^–1, respectively). Saturated and monounsaturated fatty acids were also utilized (227.2 and 256.7 ng larvae^–1 day^–1, respectively), principally 16:0 and 18:1(n-9) that were both consumed at 149.8 and 156.7 ng larvae^–1 day^–1, respectively. The rank order of utilization of fatty acids (ng larvae^–1 day^–1) by D. dentex larvae from total lipids, PC and TAG coincided with the order of abundance of the different fatty acids in the respective lipid fractions. However, in PE, the most abundant fatty acid, DHA, was relatively conserved and 16:0, the second most abundant fatty acid, was catabolized to the greatest extent. D. dentex showed a pattern of lipid metabolism during early development similar to that of marine larval fish from temperate waters whose eggs contain high levels of total lipids, including an oil globule, and which preferentially utilize neutral lipids as the primary energy source.     

Induced Maturation of Striped Bass Morone saxatilis Exposed to 6, 9, and 12 Month Photothermal Regimes

Photoperiod and temperature manipulation were used to induce out-of-season spawning in striped bass Morone saxatilis Maturational status of 40 age V striped bass, held in five environmental chambers exposed to compressed 6 mo and 9 mo cycles and a 12 mo reference cycle, was monitored by monthly egg and gonadal (ultrasound) diameter measurements. The primary effect of shortened photothermal and reproductive cycles was a reduction in egg and ovary diameters, and the number of ovulating females. Egg diameters were directly related to cycle length, averaging 845 μm, 946 μm, and 1,073 μm for females held on the 6, 9, and 12 mo cycles, respectively. Ovary diameters of females held on the 6 mo cycle were smaller (19.6 mm) than those held on the 9 mo (27.4 mm) and 12 mo (24.5 mm) cycles. Ovary diameter was highly correlated with egg diameter ( r = 0.91). The number of ovulating females decreased with cycle length, totalling 38%, 88%, and 100% in the 6, 9, and 12 mo cycles, respectively. However, no differences in fecundity (58,452 eggslkg) or fertility (39%) were detected between photothermal treatments or tank-spawned and strigspawned females. When held for a second 6 mo cycle, none of the females spawned repetitively. Males held on the 6 mo cycle had smaller testicular diameters (16.2 mm) than those held on 9 mo (23.6 mm) and 12 mo (19.2 mm) cycles. Testicular diameter of spermiating males (15.7 nun) was significantly larger than that of non-spermiating males (6.9 mm) in all cycles. The low number of spawning females and small egg, ovarian, and testicular diameter offish held on the 6 mo cycle suggest that subjecting striped bass brood stock to an abbreviated cycle length (<9 mo) constrains maturation development and reproductive success.     

Effect of different schedules for broodstock exchange on the seed production of Nile tilapia Oreochromis niloticus (L.) in freshwater

This study compares the effect of three broodstock exchange treatments (exchange every month, two months and three months) versus no-exchange control treatment on year-round seed production and spawning synchrony of Nile tilapia Oreochromis niloticus stocked in 0.4 m³ fiberglass tanks within a recirculating water system. Adult males and females with mean body weights of 161.7 g and 127.6 g, respectively, were stocked at a rate of 4 fish/tank with a male to female sex ratio of 1:3. Water temperature at 29 ± 1.0 °C and photoperiod of 18 h/day were maintained throughout the experiment. The results showed that broodstock in the exchange treatments had higher seed production and spawning synchrony compared with those in the no-exchange treatment. The exchange treatments resulted in 70.5 to 97% increase in seeds kg female^–1 day^–1. The different spawning parameters of broodstock in the two- and three-month exchange treatments were higher than the one-month exchange treatment. Results suggest that broodstock exchange strategy has a long-term improvement effect on seed production and spawning synchrony, and exchanging all breeders every three months is more profitable in terms of better utilization of breeders and in reducing both the holding/conditioning facilities for broodstock and the labor cost.     

A new technique of feeding, repeated sampling of blood and continuous collection of urine in white sturgeon

A new technique combining oesophageal intubation, dorsal aorta cannulation, and urinary catheterization was developed to monitor concentration of nutrients in the blood circulation and their metabolites in the urine of 1–2 kg white sturgeon, Acipenser transmontanus. Three experiments were conducted to assess the technique based on: (1) quantitative delivery of nutrients; (2) stress levels post-operation; and (3) monitoring of nutrients in the blood and metabolites in the urine. In Experiment 1, recovery of intubated Cr₂O₃ was 105±5% (mean ± SEM, n=3) 3 h after intubation. In Experiment 2, plasma cortisol and glucose concentrations returned to respective basal levels of 8.2±1.8 ng ml^–1 and 74± mg dl^–1 (n=9) 48 h post-operation. In Experiment 3, sturgeon intubated with 1 g kg^–1 body weight of glucose or dextrin at 48 h post-operation showed a significantly different (p<0.05) peak plasma glucose level of 139±4 and 100±5 mg dl^–1 (n=5), respectively, at 4 h. Urinary glucose excretion was 1.8±0.9 mg kg^–1 h^–1 (n=5) 4–8 h after intubation with glucose. Our results show that the new technique allows quantitative delivery of nutrients, repeated sampling of blood, and continuous collection of urine in white sturgeon with minimum stress.