Meagre’s melatonin profiles under captivity: circadian rhythmicity and light sensitiveness

The present study reveals the first characterization of the plasma melatonin rhythms of the meagre (Argyrosomus regius) under aquaculture conditions. Melatonin levels were monitored during a 24 h cycle under a photoperiod of 16 L:8D and under constant darkness (DD), respectively to characterize the daily rhythm of this indoleamine and to test its endogenous origin. Besides, to identify which light intensities are perceived as night or day by this species, the degree of inhibition of nocturnal melatonin production caused by increasing intensities of light was tested (3.3, 5.3, 10.5, and 120 μW/cm²), applying 1 h light pulses at Mid-Dark. The result for melatonin daily rhythm in plasma showed a typical profile: concentration remained low during all daytime points, increasing greatly during dark points, with maximum values at 16:00 and 22:00 h, zeitgeber time. Under DD conditions, the plasma melatonin profile persisted, with a similar acrophase but with a lower amplitude between subjective day and night periods, indicating this rhythm as being endogenously driven. Moreover, meagre seemed to be very sensitive to dim levels of illumination during the night, since an intensity of just 3.3 μW/cm² inhibited melatonin production. However, only the pulse of 5.3 μW/cm² caused a melatonin drop till daytime concentrations. Thus, the threshold of light detection by the pineal organ was suggested as being located between 3.3 and 5.3 μW/cm². Such results are an added value for this species biology knowledge, and in consequence to its adaptation to aquaculture conditions, allowing the improvement of culture husbandry protocols.     

The effect of light intensity on development and hatching success of lingcod, Ophiodon elongatus (Girard), eggs

The influence of incubation light intensity on development and hatching success of the lingcod (Ophiodon elongatus Girard) was studied by determining time to hatch, per cent hatch (total and viable) and per cent of deformities for embryos incubated at three different light intensities: ～0, 1, and 563 lux. Photoperiod for the last two treatments was 16 h dark: 8 h light. Chemical parameters throughout incubation remained within acceptable ranges. Hatching in all treatments began 43 days post fertilization (353 °C days) and was complete on day 46 (377 °C days), with peak hatch for all treatments on day 44 (361 °C days). Per cent viable hatch for eggs incubated in the 1 lux treatment (88.6 ± 2.1%; mean ± SEM) was significantly greater than eggs incubated in the ～0 lux (59.6 ± 11.3%) and 563 lux (61.4 ± 9.2%) treatments. A significantly greater per cent of deformed embryos with curled bodies occurred at 563 lux (9.5 ± 2.6%) compared with the 1‐lux treatment (2.5 ± 0.6%). No significant differences for the other categories of deformities (ball, short, distended gut) were detected among treatments. Total deformities (all categories combined) for ～0 lux (16.0 ± 4.2%) and 563 lux (17.2 ± 3.3%) were significantly greater than total deformities for 1 lux (5.0 ± 1.4%).     

The Effects of light intensity on growth and survival of cuttlefish (sepia officinalis) hatchlings and Juveniles

The effects of three light intensities (100, 350 and 1200 lux) on cuttlefish hatchling rearing performance was studied in black tanks. A total of 270 cuttlefish with a mean wet weight (MWW) of 0.089 ± 0.012 g were used in the experiment, which was undertaken during the first 50 days after hatching (hatchling stage plus the transition to the juvenile stage). According to results of the present study, light intensity is an important factor for growth and survival consistency in cuttlefish rearing. All three light intensity groups displayed exponential growth. The effect of days, light intensity and their interaction only displayed differences (P < 0.05) between light groups in terms of mean wet weight. The 100 lux light intensity promoted the best absolute values of total biomass and total mortality. We believe that the higher mortality observed in 1200 lux reared cuttlefish during the first 10 days after hatching (DAH) was due to light intensity and individual adaptation to light conditions. Therefore, the 100 lux light intensity, obtained with daylight spectrum bulbs, is recommended for cuttlefish rearing during the first 50 DAH. This light setup promotes higher growth and survival rates and lower energetic costs, which are key aspects to consider in a cuttlefish hatchery.     

Effects of skeleton photoperiods on melatonin secretion in the rainbow trout, Oncorhynchus mykiss (Walbaum)

Abstract Plasma melatonin levels were measured at hourly intervals in rainbow trout, Oncorhynchus mykiss (Walbaum), maitained under three different skeleton photoperiods: 8L:2D:2L:12D;8L:7D:2L:7D; and 8L:12D:2L:2D. Blood samples were taken before, during and after the 2-h light pulses. Melatonin levels increased rapidly to mean scotophase (dark period) values of 261 ± 7 pg/ml after the first light-dark transition and had returned to mean photophase (light period) values of 51 ± 2 pg/ml 30min after the end of the scotophase. Light pulses during the early, mid- and late scotophase elicited similar reductions in melatonin levels, followed by rapid increases after the light pulse, reaching previous scotophase values within 90 min. The inability of any of the light pulses to modulate or truncate melatonin secretion under a LD 8:16 photoperiod provides further support for the hypothesis that melatonin secretion in the rainbow trout is a direct response to darkness, and is not under endogenous circadian control as in other vertebrates examined.     

Selective behaviour of a tench, Tinca tinca (L.), stock in different light–substrate combined conditions

Two physical factors light or substrate (fibre glass, artificial vegetation, gravel and mud) were determined in a 24 m long artificial channel to have importance on the distribution of 33 tench 2+, mean size 11.8±0.9 cm S.L. For this purpose in five trials tench could select between the half of the channel occupied by their preferred light intensity (inferior to 10 lux) combined with the less preferred substrate (fibre glass) or the half with the combination of one of the substrates considered and higher light conditions (40 lux). Fish preferred, with a 100% frequency, the optimal light-less preferred substrate when compared with the combinations fibre glass, gravel and mud. When artificial vegetation was the substrate considered, tench selected the combination preferred substrate – non-optimal light intensity with a 98.26±2.68% frequency. This result changed when increasing light intensity up to 150 lux, avoiding the shading effect. We demonstrated that light has a bigger influence than substrate in juvenile tench habitat selection under controlled conditions.     

Swimbladder inflation,growth and survival of yellowtail kingfish Seriola lalandi (Valenciennes, 1833) larvae under different temperature,light and oxygen conditions

Poor swimbladder inflation leads to low fish survival due to resulting spinal deformities and the inability to feed and develop normally. Failure of swimbladder inflation may be attributed to the inappropriate range of abiotic conditions. This study investigated the effects of temperature, light source and intensity, and oxygen conditions on initial swimbladder inflation, growth and survival of yellowtail kingfish Seriola lalandi larvae. The study consisted of four separate trials including low (21.5°C) and high (24.5°C) temperatures, natural and artificial light sources, low (1000 lux) and high (32 000 lux) light intensities, and ambient and supersaturated dissolved oxygen. Initial swimbladder inflation was only significantly affected by light source, with the highest inflation rate (97.5 ± 3.5%) under artificial light. Fish growth was improved at the higher temperature and at the higher light intensity. Survival was only significantly influenced by light intensity, with the highest survival (11.0 ± 2.3%) at the high intensity (32 000 lux). This study indicates that light source affects swimbladder inflation timing and high artificial light intensity improves larval fish growth and survival.     

Influence of light intensity and spectral composition of artificial light at night on melatonin rhythm and mRNA expression of gonadotropins in roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis>

In this study we investigated the influence of artificial light at night (ALAN) of different intensities (0, 1, 10, 100 lx) and different colours (blue, green, red) on the daily melatonin rhythm and mRNA expression of gonadotropins in roach Rutilus rutilus, a ubiquitous cyprinid, which occur in standing and moderately flowing freshwater habitats of central Europe. Melatonin concentrations were significantly lowered under nocturnal white light already at 1 lx. Low intensity blue, green and red ALAN lowered the melatonin levels significantly in comparison to a dark control. We conclude that ALAN can disturb melatonin rhythms in roach at very low intensities and at different wavelengths and thus light pollution in urban waters has the potential to impact biological rhythms in fish. However, mRNA expression of gonadotropins was not affected by ALAN during the period of the experiments. Thus, suspected implications of ALAN on reproduction of roach could not be substantiated.     

Effects of melatonin and green-wavelength LED light on the physiological stress and immunity of goldfish, <Emphasis Type="Italic">Carassius auratus</Emphasis>, exposed to high water temperature

This study investigated the effects of increasing water temperature (22–30 °C) on the physiological stress response and immunity of goldfish, Carassius auratus, and the ability of green light-emitting diode (LED) irradiation or melatonin injections to mitigate this temperature-induced stress. To evaluate the effects of either green-wavelength LED light or melatonin on stress in goldfish, we measured plasma triiodothyronine (T₃), thyroxine (T₄), and thyroid hormone receptor (TR) mRNA expression; plasma cortisol and glucose; and immunoglobulin M (IgM) and lysozyme mRNA expression. The thyroid hormone activities, TR mRNA expression, and plasma cortisol and glucose were higher in goldfish exposed to high-temperature water, but were lower after exposure to melatonin or green-wavelength LED light. Lysozyme mRNA expression and plasma IgM activity and protein expression were lower after exposure to high water temperatures and higher after melatonin or green-wavelength LED light treatments. Therefore, high water temperature induced stress and decreased immunity; however, green-wavelength LED light and melatonin treatments mitigated the effects of stress and enhanced immunity. The benefits of melatonin decreased with time, whereas those of green-wavelength LED treatment did not.     

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.     

Effect of light intensity and feed density on feeding behaviour,growth and survival of larval sablefish Anoplopoma fimbria

We studied the effects of light intensity on larval activity, feeding behaviour, growth and survival of a candidate species for aquaculture – sablefish (Anoplopoma fimbria). Of six light intensities ranging from 2 to 750 lux at the water surface, the greatest surviving biomass in small tanks was observed at 12–42 lux. In another experiment in larger tanks, behavioural observations showed that larvae fed poorly under light brighter than 800 lux but fed better at lower light intensities, depending on tank type. In a separate experiment, where live feed densities were varied between 2.5 and 17.5 rotifers per mL of tank water, surviving biomass and dry weight increased with higher feed densities. These experiments help refine methods for rearing larval sablefish by demonstrating effects of light intensity and feed density on larval performance.     

超低温冷冻对俄罗斯鲟精浆和精子酶活性的影响

研究了超低温冷冻保存(-196℃)对俄罗斯鲟(Acipenser gueldenstaedti)精浆和精子中总三磷酸腺苷酶(AT-Pase)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)、肌酸激酶(CK)等酶活性的影响,并分别测定了冷冻前后俄罗斯鲟精浆和精子中酶的活性。结果显示,经过超低温冷冻保存后,俄罗斯鲟精子活力下降,精子内各酶活性均显著降低,添加冷冻保护液组精子中总ATPase、SDH、LDH和CK的活性分别从(198.47±14.43)U/mL、(30.00±2.65)U/mL、(6 982.29±24.32)U/L和(1.94±0.05)U/mL下降至(110.19±2.32)U/mL、(16.33±2.08)U/mL、(5 122.93±195.07)U/L和(1.49±0.14)U/mL。未添加抗冻剂组则分别下降至(2.25±0.33)U/mL、(11.67±0.58)U/mL、(4 488.04±78.33)U/L和(1.16±0.02)U/mL;精浆中酶的活性均显著升高,添加冷冻保护液组精浆总ATPase、SDH、LDH和CK活性分别从(12.70±0.57)U/mL、(7.50±0.71)U/mL、(2017.26±116.81)U/L和(2.93±0.59)U/mL升高至(92.49±5.18)U/mL、(13.33±0.58)U/mL、(3 688.97±172.67)U/L和(4.39±0.24)U/mL,未添加抗冻剂组则分别上升至(200.27±12.97)U/mL、(24.67±3.06)U/mL、(6 124.40±329.14)U/L和(5.20±0.16)U/mL。结果表明,超低温冷冻对俄罗斯鲟精浆和精子中酶活性及精子活力均有较大影响。     

Effects of carbohydrate supplementation on water quality,microbial dynamics and growth performance of giant tiger prawn (Penaeus monodon)

Biofloc technology based 45-day indoor growth trial was conducted to evaluate the effect of carbohydrate (CHO), molasses supplementation on two developmental stages viz, juvenile (J) and sub-adult (SA) of Penaeus monodon in zero-water exchange system. P. monodon juveniles (1.56 ± 0.04 g) and sub-adults (14.32 ± 0.22 g) were stocked in fibre-reinforced plastic tanks (1,000 L) with soil base. Carbohydrate supplementation significantly reduced (p < 0.05) the total ammonia nitrogen (TAN), nitrite-N (NO₂-N) and nitrate-N (NO₃-N) over the time periods in sub-adult group. However, no significant difference in TAN and NO₂-N was observed in juvenile-based treatments. Supplementation of carbohydrate significantly increased (p < 0.05) the water and soil total heterotrophic bacterial (THB) count in both juvenile and sub-adults. Despite increase in total Vibrio count (TVC), lower TVC/THB % was noticed in the water column of J + CHO (0.82) and SA + CHO (0.73) compared to control groups, J ? CHO (1.48) and SA ? CHO (1.21). Supplementation of carbohydrate increased the final body weight in J + CHO (6.51 ± 0.44 g) and SA + CHO (22.52 ± 0.98 g) compared with respective controls, J ? CHO (5.05 ± 0.45 g) and SA ? CHO (20.00 ± 0.33 g). Similarly, significantly lower (p < 0.05) feed conversion ratio and better protein efficiency ratio were recorded in carbohydrate-supplemented juvenile and sub-adults treatment groups. The present study demonstrates that supplementation of carbohydrate reduced the nitrogenous metabolites and significantly enhanced the growth performance of juvenile and sub-adult of P. monodon.     

LED光色及注射褪黑素对虹鳟抗氧化酶活性及相关激素含量的影响

为探究光色对虹鳟抗氧化酶活性及相关激素含量的影响,对体质量(42.0±1.5) g的虹鳟进行了28 h的持续光照及注射褪黑素处理。实验设置白、红、绿、蓝4种光色,注射和不注射褪黑素两种处理方法,共8个处理组,分别在实验开始后的4、8、12、16、20、24和28 h取样,检测血清SOD和头肾CAT的活性以及血清皮质醇和褪黑素的含量。结果显示:①SOD和CAT活性以及皮质醇含量均呈现先下降后升高再下降后趋于平稳的趋势,在实验16 h处于较高水平,实验24～28 h趋于稳定。②SOD和CAT平均活性以及皮质醇平均含量均为红光下显著高于其他光色下,除CAT平均活性绿光与白光下差异不显著外,SOD活性和皮质醇含量均为绿光下显著低于其他光色下。③注射褪黑素使SOD和CAT活性以及皮质醇含量在28 h持续光照过程中呈下降趋势;相比较未注射处理组,注射褪黑素在各光色下均可显著降低SOD和CAT活性及皮质醇含量,提高褪黑素含量。研究表明,在28 h急性光照射下,虹鳟在16 h受到的胁迫最强烈,实验后期机体逐渐适应光环境;红光对虹鳟氧化胁迫最强烈,绿光对虹鳟基本不产生胁迫;注射褪黑素可缓解光照对虹鳟的胁迫作用。     

Protective role of antifreeze proteins on sterlet (<Emphasis Type="Italic">Acipenser ruthenus</Emphasis>) sperm during cryopreservation

The loss of sperm quality in sterlet (Acipenser ruthenus) due to freeze-thaw process in cryopreservation was investigated in the present study. Two antifreeze proteins (AFPI or AFPIII) were used at different concentrations of 0.1, 1, 10, and 100 μg/mL. We compared motility, curvilinear velocity, and plasma membrane integrity of fresh, cryopreserved sperm, and sperm cryopreserved in the presence of antifreeze proteins. Fresh sperm (control) had 85?±?4% motility and 160?±?2 μm/s curvilinear velocity, respectively. After cryopreservation, the motility of frozen-thawed sperm without addition of antifreeze proteins significantly decreased (44?±?9%), compared to the control. The highest motility of frozen-thawed sperm was obtained in cryopreserved sperm with addition of 1 μg/mL of AFPIII (58?±?14%). No significant differences were observed in curvilinear velocity between fresh sperm and cryopreserved sperm with/without addition of AFPI or AFPIII. The flow cytometry analysis revealed that fresh sperm contained 94.5?±?6% live cells, while the cryopreserved sperm only contained 26.6?±?14% live cells. Supplementation of antifreeze proteins has significantly improved the percentage of live cells in frozen-thawed sperm, except 0.1 μg/ml of AFPI group. No significant difference in percentage of live cells was detected in the sperm cryopreserved with 10 μg/mL of AFPI or AFPIII, compared to fresh sperm. Thus, addition of antifreeze proteins to cryopreservation medium could be considered to improve the post-thawed sperm quality of sterlet.     

Production of melatonin by the gilthead sea bream pineal: anin vivo andin vitro study

This study deals with the variations of plasma melatonin content and ofin vitro pineal melatonin secretion, in one year-old male sea bream. Plasma melatonin levels varied from almost undetectable levels during the day to 150–200 pg ml^–1 during the night, in animals maintained under a light/dark (12L/12D) cycle. The duration of the nocturnal surge corresponded to the duration of the night. A similar pattern of melatonin secretion was obtainedin vitro, with pineals cultured under a 24h L/D cycle. When the photoperiod cycle was advanced by 8h, the first nocturnal peak in melatonin secretion was more acute compared to the profile described above, but the rhythmic variations were immediately synchronized to the new L/D conditions. Under constant darkness (D/D), circadian variations in melatonin release were observed for no more than two 24h cycles. No melatonin was detected in the culture media from retinas cultured under L/D or under D/D. These results strongly suggest that the pineal is the major source of plasma melatonin in the gilthead sea bream, and that the 24h photoperiod cycle is a powerful synchronizer of the rhythmic production of melatonin by this organ.     

Effect of light intensity and photoperiod on Nile tilapia Oreochromis niloticus L. seed production

The objective of this study was to compare the effect of six light intensity/photoperiod combinations (2500 lux/18 h, 2500 lux/15 h, 2500 lux/12 h, 500 lux/18 h, 500 lux/15 h and 500 lux/12 h) on seed production in the Nile tilapia Oreochromis niloticus L. Each combination represented a treatment and was triplicated in 0.4‐m³ fibreglass tanks within a recirculating water system. Water temperature was maintained at 29.0 ± 1.0 °C. Males and females with mean body weights of 116.8 and 91.6 g, respectively, were stocked at a rate of eight fish/tank with a male to female ratio of 1:3. Seeds (eggs, sac fry and swim‐up fry) were collected every 2 weeks. The experiment lasted for a total of 120 days. The results showed that the 2500 lux/18 h treatment produced significantly greater (P < 0.05) total seed/tank (4944), seed kg^?1 female day^?1 (50.9), seed m^?2 day^?1 (40.3) and seed/ female day^?1 (6.7) than treatments with medium or short photoperiods (15 and 12 h day^?1 respectively) or lower light intensity (500 lux). The degree of spawning synchrony and percentage of the sac and swim‐up fry stages were significantly higher in the 2500 lux/18 h treatment than in the other treatments. Under the conditions tested in this study, seed production and spawning synchrony in the Nile tilapia may be improved by subjecting breeders to a light intensity of 2500 lux and a photoperiod of 18 h day^?1.     

The effect of low temperatures and photoperiods on growth and vertebra morphometry in Atlantic salmon

The aim of this study was to reveal possible interactive effects of temperature and photoperiod on somatic and skeletal growth, feed conversion, organ indexes and blood chemistry in Atlantic salmon postsmolts. A total of 1140 (initial mean weight 96.0 g ± 3.1 SEM) juvenile Atlantic salmon reared in seawater were in duplicates exposed to six different combinations of temperatures (4.3, 6.5 or 9.3 °C) and photoperiods (continuous light, LL or simulated natural photoperiod (69ºN), LDN) for 124 days. An interactive effect of photoperiod and temperature on somatic growth was found as the fish exposed to low temperature and continuous light regime (4LL) had a significantly higher growth (30 % gain in overall SGR) than the 4LDN group, corresponding to the effect of approx. 1.2 °C temperature increase. Fish in the 6 and 9 °C groups did not show any significant growth benefit of continuous light. Compared to the 4LDN group, the 4LL group showed higher total feed conversion efficiency, lower levels of blood Na⁺ and lower hepato-somatic and cardio-somatic indexes. In the skeleton, cervical vertebra were largest in the 4LL group, while the length of the head was largest in the 4LDN group, continuous light promotes growth at lower temperatures while supporting a normal development. It is suggested that a considerable growth benefit may be achieved by exposing juvenile Atlantic salmon to continuous light when reared at low (in this trial 4.3 °C) water temperature during winter.     

Effect of rearing density on the growth and welfare indices of juvenile spotted wolffish,Anarhichas minor (Olafsen)

The goal of this study was to determine the optimal stocking density for rearing juvenile spotted wolffish, Anarhichas minor (Olafsen), at two different sizes and assess the welfare status in relation to density. No major growth impairment was observed, although smaller fish (50–100 g) were significantly affected by density during the 120 days of the experiment, with final mean weights of 119.6 ± 11.6, 118.0 ± 5.8 and 88.7 ± 0.6 g for initial rearing densities of 10, 20 and 40 kg m^?2 respectively. No effect of rearing density was seen for larger fish (100–160 g) during the 90 days of the experiment, with final mean weights of 160.2 ± 5.9, 159.7 ± 3.7 and 163.7 ± 11.5 g at fixed rearing densities of 20, 30 and 40 kg m^?2 respectively. Our results suggest that the optimal rearing density of juvenile spotted wolffish is below 40 kg m^?2 for smaller size fish (～50–100 g) and probably ≥40 kg m^?2 for the larger fish (100–160 g). Furthermore, it appears that the range of rearing density used did not have a significant effect on a selection of stress indicators (Na⁺ and K⁺ concentration, haematocrit, hepatosomatic index, total amount of plasma proteins and liver and muscle water content) and immunity response (plasma lysozyme activity) of juvenile spotted wolffish, making it a very tolerant species to crowding.     

Critical swimming speed and maximum sustainable swimming speed of juvenile Pacific bluefin tuna, Thunnus orientalis

Tank wall collision is one of the major causes of mortality during the early-stage rearing of Pacific bluefin tuna, Thunnus orientalis (PBT). Therefore, to design a rearing environment that meets the needs of juvenile PBT, it is important to gather information about their swimming capabilities. We conducted experiments to examine the relative critical swimming speed (RCSS) and maximum sustainable swimming speed (MSSS) of early-stage PBT. The fish were kept in 3-tonne tanks and fed on artificial pellets every 2 h from dusk to dawn. We conducted two sets of experiments to measure swimming speed; the fish were introduced one at a time into a water funnel, and the water current velocity was gradually increased over time to estimate RCSS, or the water current was kept at a constant velocity to estimate MSSS. We measured the RCSS of 72 PBT juveniles (24–29 days after hatching (DAH); standard length (SL), 15.0 ± 2.3 mm) and the MSSS of 32 PBT juveniles (28–37 DAH; SL, 20.0 ± 5.1 mm) in the laboratory. The RCSS ranged from 4.7 to 20.3 SL/s (average, 12.4 ± 3.3 SL/s), and the MSSS was estimated to be approximately 4 SL/s. We speculate that introducing a water current in the rearing tank of no more than 4 SL/s could positively affect the survival of juvenile PBT.     

Endocrine (plasma cortisol and glucose) and behavioral (locomotor and self-feeding activity) circadian rhythms in Senegalese sole (Solea senegalensis Kaup 1858) exposed to light/dark cycles or constant light

The existence of daily rhythms under light/dark (LD) cycles in plasma cortisol, blood glucose and locomotor and self-feeding activities, as well as their persistence (circadian nature) under constant light (LL), was investigated in Senegalese sole (Solea senegalensis). For the cortisol and glucose rhythms study, 48 soles were equally distributed in 8 tanks and exposed to a 12:12 LD cycle and natural water temperature (experiment 1). After an acclimation period, blood was sampled every 3 h until a 24-h cycle was completed. Blood glucose levels were measured immediately after sampling, while plasma cortisol was measured later by ELISA. In experiment 2, the fish were exposed to LL for 11 days, and after this period, the same sampling procedure was repeated. For the study of locomotor and self-feeding rhythms (experiment 3), two groups of sole were used: one exposed to LD and the other to LL. Each group was distributed within 3 tanks equipped with infrared photocells for the record of locomotor activity, and self-feeders for feeding behavior characterization. The results revealed a marked oscillation in cortisol concentrations during the daily cycle under LD, with a peak (35.65 ± 3.14 ng/ml) in the afternoon (15:00 h) and very low levels during the night (5.30 ± 1.09 ng/ml). This cortisol rhythm persisted under LL conditions, with lower values (mean cortisol concentration = 7.12 ± 1.11 ng/ml) and with the peak shifted by 3 h. Both rhythms were confirmed by COSINOR analysis (p < 0.05). The synchronizing role of temperature and feeding schedule, in addition to light, is also discussed. Diel rhythms of glucose were not evident in LD or LL. As to locomotor and self-feeding activity, a very marked rhythm was observed under LD, with higher activity observed during the night, with acrophases located at 2:14 and 3:37 h, respectively. The statistical significance of daily rhythms was confirmed by COSINOR analysis. Under LL, both feeding and locomotor rhythms persisted, with an endogenous period (τ) around 22.5 h. In short, our findings described for the first time the existence of circadian cortisol and behavioral circadian rhythms in flat fish. Such results revealed the importance of taking into account the time of day when assessing stress responses and evaluating physiological indicators of stress in fish.