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1.
In this study, the embryonic and larval development stages of one of the most important ornamental fish serpae tetra (Hyphessobrycon eques) are described. The early life stage is documented from fertilization until the beginning of the juvenile period. The fertilized eggs (the average diameter = 938.55 ± 35.20 µm) were incubated at a water temperature of 26 ± 0.5°C. The cleavage finished in 1:10 hr (=h) and the early blastula stage occurred at 1:26 hr post fertilization (hpf). The gastrulation started at 3:05 hpf, and 50% epiboly was observed at 3:25 hpf. Segmentation stage was monitored at 7:26 hpf. Embryonic developmental stage was completed and hatching occurred 20–21 hpf. The total length (TL) of newly hatched larvae was 2.64 ± 0.21 mm. The larval development of serpae tetra was divided into four different periods: Yolk‐sac larva (1–4 DAH, TL = 2.77 ± 0.09 mm ‐ 3.85 ± 0.11 mm), preflexion larva (5–12 DAH), flexion larva (13–15 DAH, TL = 5.78 ± 0.46 mm on the 15th day) and post‐flexion larva (16–30 DAH, TL = 10.7 ± 0.27 mm on the 28th–30th days). The mouth and anus are closed at 1 DAH. The mouth and anus opened at 4 DAH. Exogenous feeding started on the 4th day. The first gulping of the swim bladder was on days 3. The larva begins to swim freely, and the yolk sac was completely consumed at 4 DAH. Histological structures of the eye and brain of new hatched larva were clearly identified at 1 day after hatching (DAH). According to histological findings, the digestive system (stomach, intestine) started to develop and the liver could be seen on the ventral side of the swim bladder at 5 DAH. No histological difference was observed between the anterior intestine and the posterior intestine at 15–16 DAH. The larval metamorphosis was completed, and the larvae transformed into juveniles at 28–30 DAH.  相似文献   

2.
The present experiment was aimed at studying the conditioning, maturation and natural spawning of orange‐spotted grouper, Epinephelus coioides, in a recirculatory aquaculture system (RAS). Thirty fish (n = 30; 3.35 ± 0.05 kg) were stocked in a circular tank of 125 m3 capacity fitted with an RAS for conditioning and broodstock development. After 15 days, 15 fish were implanted with 17 α methyl testosterone and letrozole at the rate of 5 mg and 0.2 mg/kg body weight, respectively, for conversion from female to male. The gonadal development started after 1 month, and by 90th day, 63.53 ± 3.78% and 2.07 ± 0.84% of the oocytes attained a size of 400–500 μm and 500–600 μm respectively. Natural spawning commenced in the RAS from 4th month onward after stocking and spawning continued round the year. The spawning pair showed courtship behaviour with a typical vertical burst of swimming just before release of gametes. The total number of eggs spawned during 1 year was 47.23 million with spawning frequency varying form 5 to 13 times per month. The association of spawning events with new moon day (lunar cycle) weakened as time progressed. The mean monthly fertilization and hatching rates varied from 77.80 ± 3.34% to 83.70 ± 1.76% and 82.80 ± 4.21% to 88.33 ± 1.39% respectively. The study proved that RAS is an efficient system that provides a stable, controllable and conducive environment for year‐round natural breeding of orange‐spotted grouper.  相似文献   

3.
Despite intensive stocking programmes, wild Maraena whitefish Coregonus maraena (Bloch, 1779)‐stocks are in danger of extinction in the Baltic sea region. Current aquaculture rearing methods in recirculating aquaculture systems (RAS) are lacking efficient protocols for transitioning larvae from endogenous to exogenous feeding. In this 34‐day‐experiment the offspring of wild fish was used. Maraena whitefish larvae were weaned at three different temperatures (15.9°C, 17.9°C, 19.7°C) and three feeding regimes, resulting in nine treatments. The first group received pure live feed (freshly hatched Artemia sp. nauplii) for 10 days and a 1:1‐mixture of live and dry feed for 3 days. The second group received the live and dry feed mixture for 13 days and the third group directly received a commercial dry feed. All nine treatments were stocked in triplicate with 500 larvae per tank (19 ind. L?1). Feeding started 4 days post hatch. Survival was highest in the pure dry feed groups and lowest in the pure live feed groups. In contrast to growth, which was highest in the live feed groups and lowest in the dry feed group. Higher temperature increased growth in the live feed groups but had no effect on survival in all groups. These results will enhance the weaning of C. maraena in recirculation aquaculture.  相似文献   

4.
Constant and oscillating egg incubation temperatures on embryonic development and early larval morphology were studied in longfin yellowtail (Seriola rivoliana Valenciennes). We investigated the effects of constant temperatures from 16 to 32°C on embryo development and larval morphology at hatch, and whether oscillating temperature during embryogenesis could lead to larval morphological variations. After hatching, larval morphology and development during yolk sac (YS) utilization were examined in larvae at constant temperatures and larvae at 25°C that had oscillating temperature during egg incubation. Hatching rates were > 75%, only decreasing to ~ 50% at 30°C. At constant temperatures, the largest larvae occurred at 22 and 24°C. The oscillating temperature did not affect the timing of embryo development but resulted in larger and smaller larvae with a smaller and bigger YS, respectively, with a similar hatching time. Therefore, a growth response occurred in embryos during a window of development before hatching, depending on the adaptive response to temperature (spawn‐specific). After hatching, most of the YS was absorbed within 24 hr in all treatments, and the growth of the larval head was a priority with an optimal development at 26°C. There was compensatory growth in smaller larvae resulting in similar sizes after YS utilization, but larvae showed variations in body structure that could be important in further aquaculture research.  相似文献   

5.
The embryonic and larval development of black skirt tetra, Gymnocorymbus ternetzi, are described under controlled laboratory conditions. In addition, major histomorphological changes and the allometric growth patterns during larval development have been described. The laboratory‐reared broodstock, that is 1 year of age, were spawned. Hatching occurred 20–21 h after spawning at 24 ± 0.5°C. The cleavage was finished in 2 h and the early blastula stage occurred at 2:04 hours after spawning. The gastrulation started at 3:20 hours and 30% epiboly was observed at 3:34 hours after spawning. Eight‐somite stage was observed at 08:33 hours. And embryonic developmental stage was completed at 21 h after spawning. The newly hatched larvae were 1442 ± 14.3 μm in mean total length (TL). The mouth opened at 3 days after hatching (DAH). The yolk sac had been totally absorbed and the larvae started to swim actively within 3–4 days. Notochord flexion began at 11 DAH. The metamorphosis was completed and the larvae transformed into juveniles at 32 DAH. In this paper, the full developmental sequence from egg to juvenile of G. ternetzi is described for the first time.  相似文献   

6.
Knowledge of broodstock manipulation, gametogenesis, artificial fertilization and larval nutrition is a prerequisite to reach a large‐scale production of the polychaete Hediste diversicolor. In this study, the characteristics of oogenesis and spermatogenesis and embryonic and larval development were observed. Moreover, the effect of temperature on embryonic development and effect of diets and sediment on earlier juvenile development were tested. Mature oocytes were characterized by a spherical shape with a diameter of about 200 μm. They showed radial symmetry with a large centric nucleus surrounded by four distinctive layers. Embryo cleavage started between 4 and 10 hr after fertilization, and developed to the swimming trochophore stage after 5–6 days after fertilization at 11°C. The rate of embryonic development increased with a temperature increase from 6.1 to 21.2°C. The trochophore larvae increased in length from day 9 and commenced differentiation into 3‐setiger stage larvae at day 12. The length growth of larvae until 7‐setiger stage was 0.43 mm/day when fed with fish feed, while those fed shellfish diet and smolt sludge both grew around 0.21–0.23 mm/day. The results suggest that fish feed is a superior diet compared to shellfish diets and smolt sludge for the early larval stage of H. diversicolor.  相似文献   

7.
Natural spawning, early development and larviculture of the ornate goby Istigobius ornatus in captivity were studied for the first time. I. ornatus spawned 46 times from 31 October 2013 to 31 October 2014. Fecundity ranged from 246 to 10,214 eggs per clutch, with an average hatching rate of 77.8% ± 9.9% (M ± SEM). Fertilized eggs (1.31–1.54 × 0.46–0.50 mm in diameter) were adhesive demersal and oval‐shaped. Embryonic development lasted 84 hr at 27.5 ± 0.5°C. Newly hatched larvae [2.12 ± 0.04 mm in total length (TL)] transformed to the juvenile stage completely when TL was 7.79 mm. Effects of different water temperatures (24, 28 and 32°C) and salinities (10, 15, 20, 25, 30, 35 and 40 g/L) on per cent survivals (%) and survival activity indices (SAIs) were tested. Survival was not significantly different under different temperatures; SAIs was significantly higher at 28°C. Larvae showed the significantly higher survival and SAIs at salinities 10–30 g/L than at 35 and 40 g/L. Effect of different prey densities on survival was significantly higher in 7 days post hatch larvae fed 20 and 30 rotifers/ml. These findings could guide future programs in captive breeding technology development and commercial production of other marine ornamental gobies.  相似文献   

8.
The industrial aquaculture pond system has gradually replaced the use of traditional earthen pond, as it causes less pollution and is more economical. In this study, an industrial ecological purification recirculating aquaculture system consisting of the water source pond, high‐density culture ponds, a deposit pond, and ecological purification ponds for channel catfish cultivation was established. Twelve water samples from different ponds were sequenced, and the bacterial communities were analysed. The abundances of Cyanobacteria and Merismopedia varied in different functional ponds of the system. The water quality was stable after two months of cultivation at 1.89 ± 0.22 mg/L total nitrogen, 1.1 ± 0.08 mg/L NH4‐N and 0.43 ± 0.1 mg/L total phosphorus. The fish weight increased in a nearly linear manner, reaching 237.63 ± 23.8 per fish at day 120. An analysis of the environmental parameters, water quality and fish weight suggested that the system had an effective water purification process. Canonical correspondence analysis showed that the community was affected at the genus and phylum levels by different environmental parameters. We identified several dominant beneficial bacteria with nutrient removal abilities. Overall, our results demonstrated that the ecological purification recirculating aquaculture system had notable effects on water quality improvement and promoted changes in bacterial populations. These results provide important information on the microbial ecology of pond industrial eco‐aquaculture systems.  相似文献   

9.
In the present study, we report the first successful instance of controlled reproduction in Scatophagus argus, which has recently emerged as a new aquaculture resource. The controlled reproduction process for S. argus was optimized with regard to salinity acclimation. Gonadal maturation was affected by salinity in both sexes. Levels of plasma 17β‐estradiol (E2) and 11‐ketotestosterone (11‐kT) were salinity dependent and increased significantly with the duration of acclimation. Plasma levels of gonadal steroids were higher in fish held at 25‰ salinity. The highest gonadosomatic indices (GSI), 15.1 ± 1.6 in the female and 6.4 ± 1.2 in the male, were also observed at 25‰ salinity. Nevertheless, the optimal salinity for S. argus embryonic development and larval culture was 15‰. Thus, the salinity requirement for gonadal maturation and early development are quite different. The use of advanced reproductive technologies combining salinity acclimation and stimulation of luteinizing hormone‐releasing hormone analog (LHRH‐A2) resulted in a fertilization rate of 83.2%–91.3% and embryonic survival rates of over 90%. Embryos of S. argus at the 2‐cell, blastula, gastrula and pharyngula stages were observed. Most embryos hatched after 21.0 hr of incubation at 28.0 ± 1.0°C. The development of larvae into juveniles was completed at 40–45 days posthatch (dph). In this study, we provide information about the controlled reproduction of S. argus and identify the optimal environmental parameters for S. argus embryonic and larval culture, with the aim of developing reliable reproductive techniques for its mass production.  相似文献   

10.
Probiotic influence on fish immune response and digestive capacity is extensively discussed in aquaculture. In this experiment, a feeding trial was carried out for 100 days to evaluate the cross‐effects of probiotic supplementation and rearing temperature (17, 20 and 23°C) in juvenile seabass (Dicentrarchus labrax). The experimental diet was supplemented with a commercial probiotic blend (Biomin AquaStar Growout) at 3 g/kg diet (5.23 × 10CFU/kg diet), and tested against a non‐supplemented diet (control). Growth performance and innate immune responses were analysed at 70 and 100 days of feeding, whereas digestive enzyme activities were determined at 100 days of feeding. At the end of the feeding trial, fish were subjected to a handling stress and cumulative mortality was recorded. Digestive enzyme activities were influenced by temperature, with α‐amylase and lipase activities peaking at the higher temperature (23°C) and trypsin at the lower temperature (17°C). Immune parameters showed a significant temperature versus feeding duration effect, with complement system (ACH50) and peroxidase peaking at 70 and 100 days of feeding, respectively. Poststress cumulative mortality was higher at the lowest temperature (17°C), especially in fish fed the control diet. In conclusion, water temperature was the main variable affecting the studied parameters, whereas the dietary probiotic supplementation had influence on the chymotrypsin activity and survival rate in seabass reared at 17°C.  相似文献   

11.
《Aquaculture Research》2017,48(11):5463-5471
Hatchery production of great scallop, Pecten maximus, remains unpredictable, notably due to poor larval survival. Large‐scale flow‐through systems up to 3500 L have been developed to avoid the use of antibiotics in static systems. Alternatively, small‐scale flow‐through systems have been successfully applied for oysters but they proved to be unsuitable to rear scallop larvae. By focusing on physical factors presumed to limit P. maximus larval development, this study aimed to optimize great scallop larvae rearing parameters under controlled conditions. First, the influence of aeration on larval performances, energetic metabolism and antioxidant defences were studied both in static and flow‐through systems. Aeration depressed larval food intake, regardless of the intensities of flow tested (100 ml/min, 155 ml/min and 270 ml/min). On the other hand, antioxidant enzyme activities remained constant or decreased, suggesting that antioxidant defences were inactivated. The increase in citrate synthase activity suggested an increase in metabolic rate possibly due to a turbulent stressful environment. All larvae exposed to such turbulence died before reaching metamorphosis, whereas those reared without aeration survived well (≈ 95%). The effects of water renewal were thereafter studied in 50‐L flow‐through flat‐bottomed tanks. No differences in survival (20.4 ± 0.5%), growth (3.8 ± 0.2 μm/d), competence (5.6 ± 0.2%), energetic metabolism level and antioxidant enzyme activities were observed when comparing 12.5 and 25 L/hr water renewal. Whereas air bubbling leads to detrimental effects, flow‐through in small flat‐bottomed tanks appears to be a suitable technique for scallop larvae rearing.  相似文献   

12.
Y. Wang  M. Li  K. Filer  Y. Xue  Q. Ai  K. Mai 《Aquaculture Nutrition》2017,23(5):1113-1120
This trial was conducted to evaluate the effects of replacing dietary fish oil with Schizochytrium meal for Pacific white shrimp (Litopenaeus vannamei) larvae (initial body weight 4.21 ± 0.10 mg). Six test microdiets were formulated using Schizochytrium meal to replace 0 g/kg, 250 g/kg, 500 g/kg, 750 g/kg, 1000 g/kg or 1500 g/kg fish oil DHA. No significant differences were observed in survival, growth, final body length and activities of digestive enzyme among shrimp fed different diets (p > .05). No significant differences were observed in C20:5n‐3 (EPA) in muscle samples (p > .05). C18:3n‐3 and C20:4n‐6 in muscle increased as Schizochytrium meal replacement level increased (p < .05). No significant differences were observed in C22:6n‐3 (DHA) and n‐3 fatty acids among shrimp fed diets that algae meal replaced 0 g/kg ‐ 1000 g/kg of fish oil. Shrimp fed diet R150 had higher DHA content than other groups and had higher n‐3 fatty acids than that of shrimp fed diets R50, R75 and R100 (p < .05). C18:2n‐6, PUFA and n‐6 fatty acids in muscle increased, while n‐3/n‐6 ratio decreased with increasing algae meal replacement level from 0 g/kg to 1000 g/kg (p < .05). In conclusion, Schizochytrium meal could replace 1500 g/kg fish oil DHA in the microdiets without negatively affecting shrimp larvae survival, growth and activities of digestive enzyme.  相似文献   

13.
Hybridization was used to take advantage of desirable traits in offspring. In the present study, we applied the cryopreserved Epinephelus lanceolatus sperm into interspecific hybridization with E. moara. Successful hybridization between these two species was achieved and cultured in 23–24°C seawater (34‰). There was no difference in survival rate between hybrid (E. moara ♀ × E. lanceolatus ♂) and non‐hybrid (E. moara ♀ × Emoara ♂) at 24 hrs post hatch (HPH), but less hybrid (14.35 ± 8.02%) hatched than non‐hybrid (93.60 ± 1.65%), which might be due to irregularity cell cleavage and skeletal deformities from the formation of embryonic body to the later embryonic development. Similar phenomenon was found in hybrid embryos from fertilization with fresh sperm, indicating that species variations between parents, rather than cryopreserved sperm, resulted in deformities in embryos. Mean hatch time of the hybrid was 1 hr faster than that of E. moara. The hybrid was 1.95 ± 0.06 mm in total length when newly hatched and reached 38.00 mm at 58 days post hatch (DPH), which showed faster growth than E. moara (recorded in the previous study). Considering its faster growth, E. moara × E. lanceolatus hybrid was a potential breeding production in aquaculture. Over 212,000,000 larvae have been produced and launched in the market since 2015. The results of this study also shed some lights on further comparative studies in grouper hybrid performance.  相似文献   

14.
15.
The tongue sole Cynoglossus semilaevis, an inshore fish in China, has showed great potential in aquaculture recently. However, poor survival was recorded during the period of weaning from live Artemia to artificial diets. In this paper, the influence of co‐feeding larvae with live and inert diet on weaning performance was described. The C. semilaevis larvae were reared at 21 ± 1 °C and fed four different feeding regimes from 6 days post‐hatching (dph): A, Artemia (10 individuals mL?1); B, Artemia (5 individuals mL?1); C, mixed diet (10 Artemia individuals mL?1 and 12 mg L?1 inert diet); and D, mixed diet (5 Artemia individuals mL?1 and 12 mg L?1 inert diet). Rotifers were also supplied in all cases during the first days of feeding. Mixed diets of commercial formulated feed and live prey (rotifers and Artemia) allowed larvae to complete metamorphosis, achieving similar specific growth rate (SGR) (18.5 ± 1.4% and 18.7 ± 1.6%) and survival (40 ± 7.6% and 48.5 ± 6.8%) compared with larvae fed on live feed alone (SGR of 18.3 ± 1.2%, 19.3 ± 1.9% and survival of 41.2 ± 11.3%, 38 ± 4.9%). However, in metamorphosed fish, when live feed was withdrawn on 31 dph, there was significant difference (P < 0.05) in survival and growth among treatments. Metamorphosed fish, previously fed mixture diets during larval stages, had similar survival (62.1 ± 7.6% and 62.8 ± 3.9% for regimes C and D, respectively) but higher than that obtained for fish that previously fed on live feed (49.3 ± 2% and 42.1 ± 3.9% for regimes A and B, respectively) after weaning (day 60). The SGR of weaned fish previously fed live feed was similar (3.1 ± 0.6% and 2.92 ± 0.6% for regimes A and B, respectively) but lower than that recorded for fish that was fed from day 6 to day 30 on the mixed diet (4.5 ± 1.1% and 4.9 ± 0.3% for regimes C and D, respectively). It is suggested that weaning of C. semilaevis from early development would appear to be feasible and larval co‐feeding improves growth and survival.  相似文献   

16.
Aeromonads are ubiquitous in aquatic environments and have been implicated in fish and human infections. In this study, we isolated, studied antimicrobial susceptibility patterns and screened the existence of 15 virulence genes in aeromonads from two famously consumed fish species—seven marine Tiger Grouper (Epinephelus fuscoguttatus) and eight freshwater Marble Goby (Oxyeleotris marmoratus) from the aquaculture hatchery in Sabah, Malaysia. A total of 30 aeromonads (17 A. caviae, 9 A. rivuli, 4 A. dhakensis) were identified using PCR targeting GCAT gene, rpoD‐restriction fragment length polymorphism and multi‐locus phylogenetic analysis. All 30 strains were resistant to amoxicillin and cephalothin and five strains were multidrug‐resistant. Nine virulence genes (lip, ela, eno, fla, aerA, hylA, dam, alt and ser) present in A. dhakensis, suggesting the virulence potential of this species as a fish pathogen. This study offers as a baseline for future studies in monitoring and managing these two fish in aquaculture industry.  相似文献   

17.
This study aimed to evaluate the performance of Pyrrhulina larvae, Pyrrhulina brevis, under different photoperiods and feeding frequencies. A completely randomized design was used, with four replicates in a 5 × 2 factorial, with five photoperiods (6L:18D, 12L:12D, 16L:8D, 20L:4D, 24L:0D) and two feeding frequencies (two or four times a day). Four hundred larvae with an initial length of 4.6 ± 0.09 mm were randomly distributed in 40, 1‐L containers at a density of 10 larvae/L. Feeding was performed with Artemia nauplii in the ratio of 150 Artemia nauplii/larva/feeding. After 15 days, all larvae were euthanized for measurements of length (mm), final weight (mg), survival rate (%) and larvae uniformity in weight an length (%). The data were subjected to analysis of variance and subsequently, in case of significance, a Tukey test was performed to compare means. There was no interaction between photoperiod and feeding frequency on studied variables (P > 0.05). Likewise, there was no influence of feed rate on the same variables (P > 0.05). There was only influence of photoperiod, where the larvae subjected to longest photoperiods (24L:0D and 20L:4D) showed improved weight and length (P < 0.05). Therefore, a photoperiod of 20L:4E, with feeding twice a day is recommended.  相似文献   

18.
The study amalgamated earthworm and agro‐industrial wastes through vermicomposting and then evaluated the potential of the bedding (mixture of Eisenia fetida and vermicompost) to replace fishmeal in semi‐intensive farming of Oreochromis niloticus. The bedding was used to substitute fishmeal at inclusion rates of 100, 60, 30 and 0% (D100, D60, D30 and control D0). In triplicates, the four homogeneous diets were fed to quadruplicate groups of 30 g O. niloticus for 112 days. There was no significant difference (p > .05) in mortalities, average length gain and FCR among all tests. Nevertheless, diet D0 had significantly (p < .05) superior amino acid profile, low fibre content and fish carcass crude protein (63.2 ± 0.72% dry matter). Subsequently, D30 and D0 produced fish with significantly higher (p < .05) mean weight gain (256.03 ± 0.4 g) and biomass (369,136 g) respectively. On to the contrary, diet D100 had significantly higher (p < .05) crude lipids content (9.4 ± 0.6% dry matter), economic returns and profit index than the control diet due to the comparatively low cost of producing the earthworm bedding. This simple biotechnology can commercially be upscaled to sustainably produce cheap and nutritious fish feed capable of increasing yields and maximizing profits.  相似文献   

19.
Copepods are crucial source of live feeds in the aquaculture industry. In particular, several species of the genus Acartia are considered optimal prey for fish larvae. The species Acartia bilobata has excellent potential for marine larvae culture, as it is easy for mass culture. This study investigated the effects of various algal diets on the egg production and egg‐hatching rate of A. bilobata. The results indicated that the single‐species diet Isochrysis galbana was the most supportive diet for A. bilobata egg production and female life span in all treatments (egg production: 23.85 ± 0.70 eggs female?1 day?1 and female life span: 18.00 ± 1.45 days). Nannochloropsis oculata and Tetraselmis chui treatments gave markedly lower egg production and female life span as both single‐species and multiple‐species diets. For the egg hatching‐rate experiment, except for the T. chui treatment, which yielded a considerably lower hatching rate than the other diets, the hatching rate was only slightly affected by the algal diets. These results confirm that A. bilobata, a tropical brackish‐water copepod species, develops rapidly at 28°C and can produce a large number of eggs; therefore, it has considerable potential for larvae culture.  相似文献   

20.
The concept of nutritional programming raises the interesting possibility of directing specific metabolic pathways or functions in juvenile fish, for example, to improve the use of substitutes to fishmeal and oil, and hence to promote sustainability in aquaculture. The aim of the study was to determine effects of early nutritional stimuli of gilthead seabream larvae and check if nutritional programming of gilthead sea bream is possible between 16 days post hatching (dph) and 26 dph. A trial was conducted to determine the effects of early nutritional stimuli of gilthead seabream larvae. Five experimental microdiets (pellet size <250 μm) were formulated containing five different proportions of a marine lipid source rich in long‐chain polyunsaturated fatty acids (LC‐PUFA) and two vegetable lipid sources rich in linolenic and linoleic acids. The results of this study demonstrate that dietary n‐3 LC‐PUFA levels increased larval growth and survival affecting Δ6 desaturase gen (fads2) expression and retinal neurons density. However, the high mortalities obtained along on‐growing in fish fed low n‐3 LC‐PUFA at 16 dph constrained the feasibility of nutritional programming of gilthead seabream during this late developmental window and needs to be further investigated.  相似文献   

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