Diel Rhythms of Serum Metabolites and Thyroid Hormones in Red Porgy Held in Different Photoperiod Regimes

Diel rhythms in serum glucose, lactate, cholesterol, triglycerides and thyroid hormones were studied in red porgy, Pagrus pagrus, held under different photoperiod regimes (constant darkness – DD, 8L:16D,12L:12D), at a constant temperature (17.1–18.7 °C) and fed with commercial pellets, by means of a self-feeder. A clear diurnal rhythm in feeding activity, regardless of the photoperiod regime was demonstrated. All serum components showed significant diel rhythms, although they were not necessarily consistent or circadian in periodicity. As well as this, differences in the average values among the varying treatments were observed. Fish held under the 12L:12D protocol displayed significantly higher average T₄, T₃ and lactate levels during the day rather than at night. Maximum glucose values occurred 8–12 h after dawn and maximum lactate concentrations at 06:00 or 14:00 h. Diel variations in cholesterol were evident only in the DD group with peak values inversely correlated with the rhythm of food intake. Triglycerides displayed a similar pattern of changes. Significant diel fluctuations in T₄ serum levels were observed only in fish exposed to the 12L:12D protocol, with peak values at dawn. A clear diurnal peak (10:00 h) in T₃ concentrations was observed in fish subjected to the 12L:12D regime, while fish exposed to the 8L:16D protocol showed two peaks: one in the photophase (10:00 h) and another in the scotophase (02:00 h). The light–dark alternation and the general activity rhythm of fish seem to be the main synchronizers of the diel rhythms observed in this study.     

Responses of circulating arginine vasotocin, isotocin, and melatonin to osmotic and disturbance stress in rainbow trout (Oncorhynchus mykiss)

In teleost fish exposed to stressors AVT has been implicated in activation of the hypothalamo-pituitary-interrenal axis. Mel has been shown to counteract several behavioural and endocrine consequences of stress in mammals. These studies were undertaken to investigate the effects of either disturbance or osmotic stress, applied either separately or simultaneously, on plasma AVT, IT and melatonin in rainbow trout (Oncorhynchus mykiss). Hormones were determined in plasma by high-performance liquid chromatography preceded by solid-phase extraction. The results showed that both forms of stress caused significant increase in plasma AVT concentration, although more pronounced elevation was observed in physically disturbed fish. Conversely, neither osmotic nor disturbance stress affected plasma IT concentration. The apparent difference in response to stress by the two close related neurohypophysial nonapeptides suggests independent mechanisms controlling their synthesis and/or release and supports the idea that only AVT plays a role in physiological response to stress. Plasma Mel level was depressed in fish subjected to disturbance stress and to both stresses applied simultaneously, an effect possibly associated with the elevation of plasma AVT concentration. Results are discussed in relation to physiological interactions between hormones.     

Food-entrainable circadian oscillator in goldfish: multiple daily feeding times and food-anticipatory activity

The existence of a food-entrainable circadian oscillator (FEO) was examined in goldfish Carassius auratus. Single goldfish were exposed to light–dark (LD) 12:12 (lights on 06:00–18:00 h) and fed at 12:00 h (phase I). The photoperiod regime was shifted to constant light (LL) (phase II). The fish were then fed at 06:00 or at 18:00 h (phase III), and all fish were fasted (phase IV). In both groups, three out of eight fish exhibited food-anticipatory activity (FAA) in phases II and III, the free-running period (τ) of locomotor activity was approximately 24.0 h, and circadian rhythms of locomotor activity started to free-run from the previous FAA in phase IV, indicating the existence of an FEO. Next, single goldfish were exposed to LD 12:12 and were fed twice daily at 12:00 and 00:00 h (phase I). The photoperiod regime was shifted to LL (phase II), and the fish were fasted (phase III). In phase II, FAA was observed in three out of six fish for both feeding times. In phase III, a circadian rhythm of locomotor activity was observed in five fish, and circadian rhythms started to free-run from the previous feeding time (either 12:00 or 00:00 h) in three fish. These results suggest that individual goldfish can remember two daily feeding times.     

Combined effects of photoperiod and temperature on growth and survival of African catfish (Clarias gariepinus,Burchell 1822) larvae under laboratory conditions

ABSTRACT

The effect of photoperiod (24L:00D, 12L:12D, and 00L:24D) and temperature (22 ± 1°C and 28 ± 1°C) on performance of Clarias gariepinus larvae was tested. Larvae weighing 3.2 ± 0.24 mg were cultured in aquaria at a stocking density of 20 fish L^?1 and fed twice a day on catfish starter diet (40% CP) at 10 % BW day^?1. Highest mean weight gain (31.00 mg), SGR (7.56% day^?1), and survival (83%) were achieved at photoperiod and temperature combination of 00L:24D and 28 ± 1°C. Percent survival of larvae differed significantly (p < .05) among treatments with optimal survival of (83%) in treatment combination of 28 ± 1°C and 00L:24D, while lowest survival (40%) in treatment combination of 22 ± 1°C and 24L:00D.     

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.     

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.     

Age-related changes in 2-[125I]-iodomelatonin binding sites in the brain of sea breams (Sparus aurata,L.)

The pineal organ of fish, through its 24h rhythmic release of melatonin, acts as a transducer of the photoperiod, influencing different physiological functions (e.g., reproduction, growth). The target sites for melatonin are poorly known in fish, especially marine species. A radioligand study was undertaken using the gilthead sea bream (Sparus aurata) maintained under natural temperature and photoperiod (at 28°N latitude). This species exhibits the property of changing sex during growth. Brains of one year-old males were collected at 16:00h and brains of three year-old females at 03:00, 10:00, 16:00 and 23:00h. Membrane homogenate receptor assays were run using 2-[¹²⁵I]iodomelatonin as a ligand. Binding sites were detected in brains of young and old fish. In the younger, the exhibited a B_max between 3.52 and 4.29 fmol mg protein^–1 and a K_D between 358–380 pmol l^–1. In the older fish, the K_D varied according to a daily pattern: values were three times higher at 03:00 and 10:00h (500–600 pmol l^–1) than at 16:00 and 23:00h (150–300 pmol l^–1). The number of sites also were higher at 03:00 and 10:00h (180–200 fmol mg protein^–1) than at 16:00 and 23:00h (95–110 fmol mg protein^–1). Melatonin and iodomelatonin displaced 2-[¹²⁵I]iodomelatonin binding in a dose dependent manner, the second being more potent than the first. Binding was also inhibited by GTP. The results provide the first evidence for the presence of membrane melatonin binding sites in the brain of an exclusively marine fish. They suggest that their number and affinity varies during growth and throughout a light/dark cycle. Future experiments will aim to precise the anatomical location and role of these binding sites.     

Effect of temperature and photoperiod on the growth, feed consumption, and biochemical content of juvenile green abalone, Haliotis fulgens, fed on a balanced diet

Juvenile green abalone, Haliotis fulgens (31.3 ± 0.1 mm, 3.7 ± 0.04 g live weight) were reared in laboratory for six months in order to determine their survival, growth, tissue composition, feed consumption (C), feed conversion ratio (FCR) and protein efficiency ratio (PER) under two temperatures (20 °C and 25 °C) and three photoperiods (00:24, 12:12 and 24:00 light:dark hours). Survival was ca. 100% at 20 °C, and between 68% and 75% at 25 °C. The highest gross growth rate (109 ± 4.3 μm d^− 1, 69 ± 3.9 mg d^− 1) was observed in abalone from the combination 20 °C-00:24 L:D. Slowest GGR (38 ± 4.1 μm d^− 1, 26 ± 1.5 mg d^− 1) was observed in the combination 25 °C-24:00 L:D. Organisms from 25 °C exhibited signs of the withering syndrome at the end of the experiment. Gross energy content (4.4 to 4.6 kcal g^− 1 tissue dry wt, TDW) and crude protein (60 to 68% TDW) were not significantly affected by either temperature or photoperiod, but organisms from 20 °C exhibited lower moisture content (86.8 to 88.6%) and higher tissue:shell ratio (0.34-0.40) than those from 25 °C (88.9 to 92.1%; 0.22-0.31 respectively). Feed consumption markedly increased at night, decreased with age, and was higher at 25 °C than 20 °C, irrespective of photoperiod. Highest C was observed under continuous darkness (0.66 to 0.95% BW d^− 1 at 20 °C and 0.84 to 1.25% BW d^− 1 at 25 °C), and was lowest under continuous light (0.50 to 0.82% BW d^− 1 at 20 °C and 0.71 to 1.02% BW d^− 1 at 25 °C). FCR and PER were both affected by temperature but not by photoperiod. Higher PER (2.2 to 3.4) and lower FCR values (0.69 to 1.05) were observed at 20 °C, when compared to 25 °C (PER 1.35 to 2.09, FCR 1.10 to 1.86). Sex ratios were ca. 1:1 in the 00:24 and 12:12 L:D photoperiods, yet ca. 50% of abalone from the 24:00 L:D photoperiod were immature at the end of the experiment. It is concluded that H. fulgens can be best cultured at 20 °C and 00:24 or 12:12 L:D regimes, while sustained temperatures at or above 25 °C may result in cumulative stress, altered physiological rhythms, and delayed maturation.     

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.     

The effects of continuous photoperiod (24L:0D) on growth of juvenile barramundi (<Emphasis Type="Italic">Lates calcarifer</Emphasis>)

The efficacy of photoperiod manipulation to influence growth and developmental processes is well documented in a range of temperate aquaculture species. However, the application of such techniques with tropical species requires further investigation. This preliminary 20-day study investigated the influence of continuous photoperiod on growth of barramundi (Lates calcarifer). In addition, diel plasma melatonin profiles provided a physiological measure of how the endocrine system of barramundi responded to continuous photoperiod. Juvenile barramundi (1.33 ± 0.02 g) were held in recirculation systems under 12-h light: 12-h dark (12L:12D) or 24-h light (24L:0D) with a light intensity of 1,000 lux throughout the water column. Fish from both treatments grew to more than 14 times their original weight, with final weight (24L:0D = 21.59 ± 0.85 g; 12L:12D = 19.12 ± 0.55 g), total length (24L:0D = 12.67 ± 0.14 cm; 12L:12D = 11.96 ± 0.13 cm) and specific growth rate (24L:0D = 9.60 ± 0.05% bw day⁻¹; 12L:12D = 9.14 ± 0.06% bw day⁻¹) being significantly higher for fish grown on 24L:0D compared with 12L:12D. There were no significant differences in feed intake (24L:0D = 226.46 ± 6.27 g; 12L:12D = 219.02 ± 5.73 g) or feed conversion ratio (24L:0D = 0.71 ± 0.06; 12L:12D = 0.80 ± 0.07) between light treatments. Barramundi held under 12L:12D exhibited diel melatonin secretion, which peaked mid-dark phase (171.83 ± 4.81 pg ml⁻¹) followed by a gradual decrease in base levels at the onset of illumination (68.61 ± 8.77 pg ml⁻¹). When juvenile barramundi were subjected to 24L:0D, the amplitude of peak melatonin secretion was significantly suppressed during the subjective mid-dark phase (129.71 ± 2.36 pg ml⁻¹). This preliminary study confirmed that barramundi respond to photoperiod manipulation in a similar manner to many temperate fish species, thus demonstrating the future potential use of artificial lighting to improve growth in this species commercially.     

Stress-induced changes in the affinity and abundance of cytosolic cortisol-binding sites in the liver of rainbow trout,Oncorhynchus mykiss (Walbaum), are not accompanied by changes in measurable nuclear binding

Plasma cortisol levels and the number (N_max) and affinity (K_d) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml^–1 within 24h and although levels declined to 8.0 ± 3.0 ng ml^–1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml^–1). There was a 60% reduction in cytosolic N_max in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 ± 15.2 fmol mg^–1 protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the K_d of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in K_d was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in N_max in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that high-affinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.     

Diel feeding rhythms of greenback flounder Rhombosolea tapirina (Günther 1862): the role of light–dark cycles and food deprivation

The role of light–dark (LD) cycles and food deprivation in the regulation of diel feeding rhythms in greenback flounder Rhombosolea tapirina (Günther 1862) was examined. A computerized monitoring system was developed to record uneaten pellets, and food intake was estimated as the difference between pellets presented and those uneaten. Three groups of five fish each (75–281 g) were exposed sequentially to the LD cycles: LD 12:12, LD 12:12 (scotophase advanced by 9 h), LD 6:18, DD, LD 6:6:6:6, LL and LD 12:12 (return to the initial cycle). Illumination was 120 lux at the water surface (complete darkness at scotophase), and water temperature was 15 ± 1 °C. Fish fed mainly during the light hours under LD 12:12 and resynchronized to the scotophase-advanced LD 12:12 regime after about 12 h. Feeding started at the onset of light and extended to the early scotophase under LD 6:18. A circadian feeding rhythm was detected in fish under DD and LD 6:6:6:6 regimes. Under the LL regime, two groups of fish displayed arrhythmic feeding patterns and did not resynchronize to LD 12:12 for at least 13 days. In contrast, the third group of fish exhibited a circadian feeding rhythm under the LL regime and immediately re-entrained to LD 12:12. Fish were deprived of food for a single period of 26–57 h once feeding patterns had been established under LD 12:12, LD 12 : 12 (scotophase advanced by 9 h) and DD regimes, and feed delivery was resumed during the scotophase or subjective night; the timing of feeding rhythms was shifted by the reintroduction of feed, but progressively resynchronized to the LD cycles. The results suggest that the greenback flounder is a diurnal feeder, that a LD 12:12 cycle is a potent environmental cue to entrain circadian feeding rhythms, and that a biological clock is involved in the timing of feeding.     

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.     

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.     

Inhibition of sexual maturation in tank reared haddock (Melanogrammus aeglefinus) through the use of constant light photoperiods

The haddock (Melanogrammus aeglefinus) is believed to be a potential candidate for aquaculture in the Atlantic coastal countries including the UK, Norway and Canada. However, under culture conditions, haddock will sexually mature prior to the attainment of a suitable harvest weight. Therefore, a long term tank based experiment was performed where three populations of haddock (hatched spring 2002, approximately 150 fish per population) were exposed to either a simulated natural photoperiod (SNP) or SNP until January or July 2003 and thereafter continuous illumination (Jan LL or July LL respectively) with individual growth rate and maturation status being recorded until July 2004 (27 months post-hatch). While the SNP treated population matured in the spring of 2004 (two years post-hatch) with 88% of the population being observed to release gametes, no mature individuals were observed in either of the LL treated populations and furthermore there was no evidence of gonadal development or elevation in sex steroids (testosterone, estradiol-17β or calcium). The application of LL appeared to directly stimulate the growth rate of haddock (e.g. 14 to 27% increase in weight thermal growth coefficient) and improve food conversion rates. At the end of the trial there was an approximate 50% increase in wet weight (SNP: 647 ± 53 g, Jan LL: 982 ± 34 g and July LL 985 ± 33 g), however no significant difference in weight in relation to the length of LL exposure was observed. Plasma melatonin analysis revealed that the natural diel rhythm had been inhibited in both LL treated populations. Along with highlighting the similarity in reproductive entrainment between haddock and other gadoids, these results demonstrate how photoperiod manipulation could be used as a management strategy to improve growth performance in farmed haddock stocks.     

Effect of the dusk photoperiod change from light to dark on the incubation period of eggs of the spotted rose snapper, Lutjanus guttatus (Steindachner)

Spotted rose snapper, Lutjanus guttatus (Steindachner), eggs were incubated under different photoperiods to examine the effect of photoperiod on incubation. The eggs from two fish were incubated under five artificial photoperiods: constant dark (D), constant light (L) from 06:00 hours and 6, 10 and 14 h of light from 06:00 hours. The eggs from seven other fish were incubated under a natural photoperiod. Different spawning times (21:00 – 01:00 hours) and different photoperiods combined to give the start of the dusk photoperiod change after 11–23 h of incubation. Constant light or applying the dusk photoperiod change after ≥20 h of incubation appeared to extend the hatching period. The mean hatching period for groups of eggs incubated in darkness or that received the dusk photoperiod change after ≤19 h of incubation (n=8 different groups) was 2 h 15±10 min, which was significantly lower (P<0.05) than the mean hatching period of 4 h±37 min for groups that did not receive the dusk photoperiod change or that received the dusk photoperiod change after ≥20 h of incubation (n=9 groups). However, despite these differences, the majority of the eggs hatched during a 2–3 h period from 17 to 20 h of incubation, and a sigmoid regression (r²=0.9) explained the relationship between percentage hatch and hours of incubation for all photoperiod groups.     

Neuropeptides isotocin and arginine vasotocin in urophysis of three fish species

In this study, for the first time, both neuropeptides isotocin (IT) and arginine vasotocin (AVT) have been identified and measured in urophysis, the neurohaemal organ of the caudal neurosecretory system of teleost fish. So far, AVT, but not IT, was quantified by radioimmunoassay (RIA) in urophysis of several fish species. We have used high-performance liquid chromatographic assay with fluorescence detection (HPLC-FL) preceded by solid-phase extraction (SPE) and liquid chromatography-electrospray ionization triple-quadrupole tandem mass spectrometry (LC-ESI MS/MS) technique to determine both neuropeptides in urophysis of three fish species. The efficiency of peptide’s SPE extraction was 79–85 %. In HPLC-FL method, the limits of detection (LOD) and quantification (LOQ) were estimated as 1.0 and 3.4 pmol/mL for IT and 0.25 and 2.20 pmol/mL for AVT. In LC–MS/MS method, LOD and LOQ were estimated as 0.4 and 1.2 pmol/mL for IT and 0.06 and 0.2 pmol/mL for AVT. The chromatographic methods are good alternative for RIA, because enable to measure both nonapeptides simultaneously in one sample. In round goby (Neogobius melanostomus), three-spined stickleback (Gasterosteus aculeatus) and sea bream (Sparus aurata), urophysial IT concentrations ranged between 0.056 and 0.678 pmol/mg tissue and AVT concentrations ranged between 0.0008 (or even below detection threshold) and 0.084 pmol/mg tissue.     

南海北部陆坡金鼻眶灯鱼秋季摄食节律

中层鱼是深海食物网的重要组成部分,其摄食节律的研究对探讨以中层鱼为核心的能量流动与物质循环具有重要意义。本研究利用2014年10月南海北部陆坡海域拖网调查数据,对中层鱼优势种金鼻眶灯鱼(Diaphus chrysorhynchus)秋季的摄食节律进行了研究。结果表明:(1)金鼻眶灯鱼取样体长范围为28~110 mm,集中于50~59 mm体长组,各时间段摄食等级与体长间均不存在正相关;(2)金鼻眶灯鱼在18:00左右上浮摄食,次日05:30左右达到摄食高峰,为连续摄食,5个取样时间段平均摄食等级范围为1.50~2.57,呈增长趋势;(3)差异性分析表明,金鼻眶灯鱼傍晚与夜间或清晨间摄食等级差异性显著,表现在18:30~19:30时间段与01:00~02:00、04:45~05:50或06:30~07:30三个时间段间(P=0.041;P=0.000;P=0.007),另03:30~04:30与04:45~05:50时间段间摄食等级也存在极显著差异(P=0.005);(4)金鼻眶灯鱼各时间段摄食等级频率呈波浪形变化,4级频率在06:30~07:30时间段达到最高峰;0级频率在黄昏18:30~19:30时最高,01:00~02:00降为0,03:30~04:30上升到黄昏时的一半,此后连续下降。总体而言,金鼻眶灯鱼傍晚至次日清晨连续摄食,连续排遗,摄食集中于清晨。综上所述,金鼻眶灯鱼是一种昼夜垂直迁移、夜间连续摄食与排遗的浮游动物食性鱼类,其在南海陆坡水域能量流与物质流中起着承上启下的作用。     

Photoperiodic mechanisms and rhythms of reproduction in the female rainbow trout

The present work investigates the importance of circadian and circannual rhythms in the photoperiodic control of reproduction in the rainbow trout. Maintenance of groups of 20–30 female trout under continuous light (LL), constant long (18L:6D) or short (6L:18D) days and conditions of constant temperature (8.5–9.0°C) and feeding rates (0.5% body weight, day^–1), starting in February, produced markedly different spawning periodicities during the first and subsequent years of treatment. At the end of the first year, spawning was advanced by up to 2 months in the fish under LL and 18L:6D and delayed by up to 5 months in those under 6L:18D when compared with the December spawning of control fish under ambient light. Continued exposure of the fish to the same photoperiod regimes produced cycles of spawning and peak levels of testosterone, oestradiol-17 and calcium (as an index of vitellogenin) at intervals of 150–170 days under LL and 18L:6D and 320–420 days under 6L:18D. The functional importance of these spawning cycles and their relationship to circannual rhythms and the control of reproduction is discussed.Exposure of fish to skeleton (6L:4D:2L:12D, 6L:6D:2L:10D and 6L:8D:2L:8D) and resonance (6L:42D, 6L:48D and 6L:54D) procedures produced ranges of spawning times up to two months in advance of control fish. Results with the resonance regimes, where fish received only a half the light-dark cycles and a quarter of the total daylight hours of those on ambient light cycles show that trout do not measure daylength and time by counting daily cycles or by hour-glass mechanisms. Collectively, these data support the proposition that rhythmic processes of photosensitivity are involved in the modulation of reproductive development in the rainbow trout and that circadian and circannual rhythms cooperate in the timing and entrainment of this cycle.     

Effect of stocking density and photoperiod on growth and survival in cultured early juvenile pot‐bellied seahorses Hippocampus abdominalis Lesson, 1827

The effect of stocking density and photoperiod on Hippocampus abdominalis was examined in this study. Stocking densities of 45, 30, 15 and 5 seahorses 3 L^?1 were tested on newborns. Growth and survival were independent of stocking density. A second stocking‐density experiment aimed to remove the effect of an early mortality experienced in experiment 1 using older juveniles at 25, 15 and 5 seahorses 3 L^?1. There were no differences in the parameters measured. Also, the effect of photoperiod was investigated on seahorses cultured under 24:00, 16:08 and 08:16 (L:D) photoperiods. A single Artemia meal was delivered at 10:00 hours. Survival and body growth in 16:08 and 08:16 hours were higher than in 24:00 hours. In a second experiment, seahorses were cultured in the photoperiods and conditions described for experiment 1, except they were fed twice the amount in two meals delivered at 10:00 and 16:30 hours. The seahorses in 16:08 hours showed better growth than the other treatments, but there were no differences in survival. These results suggest that early juvenile H. abdominalis can be cultured at higher stocking densities than previously reported, without compromising growth and survival, and when feeding was not limiting, grew better in an extended photoperiod (16:08) but not in 24:00 hours.