Reproduction,growth and stress response in adult red sea bream,Pagrus major (Temminck & Schlegel) exposed to different photoperiods at spawning season

Adult red sea bream, Pagrus major (body weight, 1.0–2.0 kg) was exposed to three photoperiods [12 h light:12 h dark (12L:12D), 16 h light:8 h dark (16L:8D) and 24 h light:0 h dark (24L:0D)] from 2 months before spawning till the end of the spawning season to investigate growth, spawning and stress response. During the spawning season, tanks were checked every morning for spawned eggs. The growth performance in fish under 24L:0D was stimulated with significantly higher feed intake than those under other photoperiods (P<0.05). The number of eggs and gonadal histology confirmed that three and five females out of six in each of duplicate tanks of the 16L:8D treatment spawned. In contrast, only two out of six females in one tank of the 24L:0D treatment spawned, and no spawns were observed in the 12L:12D treatment. At the end of the spawning period, both 17β‐estradiol and testosterone levels were significantly higher in fish exposed to 16L:8D followed by 12L:12D and 24L:0D photoperiods (P<0.05). Photoperiod manipulation did not cause significant stress response in fish (P>0.05). The results suggest that stimulating the growth performance of red sea bream at reproductive stage with a 24L:0D photoperiod is possible if the fish are subjected to this photoperiod long before the onset of the spawning season.     

Induced maturation and spawning of domestic and wild striped bass, Morone saxatilis (Walbaum), broodstock with implanted GnRH analogue and injected hCG

Abstract. A domestic striped bass. Morone saxatilis (Walbaum), broodstock was established by rearing fish to sexual maturity in ponds. A method was developed to reproduce the domestic females, and also wild females too immature to be successfully induced to spawn with injected human chorionic gonadotropin (hCG). The fish were implanted with pellets containing 100–150μg of a synthetic analogue of mammalian gonadotropin reieasing-hormone, [D-Ala⁶- Pro⁹-NEt]-LHRH (mGnRHa), in a matrix of cholesterol (CH) and cellulose. They were implanted with one fast hormone-release (80% CH) pellet and one slow hormone-release (95% CH) pellet and allowed to mature for 1–3 days, until they entered the process of final oocyte maturation and were induced to ovulate or spawn with an hCG injection. The secondary hCG injection was found to be necessary to speed the maturation of the wild fish; they otherwise would succumb to the stresses of capture, handling and confinement before they could be spawned. The total mGnRHa dosages used ranged from 33 to 111μg mGnRHa/kg body weight, and the hCG doses were either 165 or 330 IU hCG/kg body weight. Using the combined mGnRHa implant-hCG injection technique, fry production rates were comparable to those obtained using fully mature wild females taken directly from their spawning grounds.     

Domestication of Silver Perch,Bidyanus bidyanus,Broodfish

Abstract

Most breeding programs for the Australian native freshwater silver perch, Bidyanus bidyanus, are based on fish originally captured from natural populations. However, over the last 20 years, there has been a dramatic decline in the distribution and abundance of silver perch in the wild and the species now has the conservation status of “vulnerable.” Unless broodfish can be domesticated a serious shortage is imminent. The aims of this study were to determine the age and size at sexual maturity and the reproductive performance of silver perch reared in captivity. Fingerlings (mean weight, 4.0 g) from a hatchery were stocked into an aerated 0.32-ha earthen pond at a density of 22,000 fish/ ha for 12 months, harvested, and restocked at 10,000 fish/ha for a further 2 years. The fish were fed a formulated diet containing 35% crude protein. No fish were mature at 1 year of age; at 2 years, most males (93%; total length, 304.0 mm, weight 412.7 g), but no females were mature; and at 3 years, most females (97%; 316.4 mm, 549.6 g) were mature. Three-year-old fish injected with 200 IU/kg human chorionic gonado-trophin spawned viable gametes. High fecundity (mean 139,286 eggs/ kg), fertilization rates (84.5%) and hatch rates (76.8%) demonstrate that broodfish can be domesticated and the life cycle of silver perch can be closed. The use of domesticated broodfish will eliminate reliance on wild populations and provide a basis for future hatchery production and selective breeding programs.     

No significant effect of additive ratios of docosahexaenoic acid to eicosapentaenoic acid on the survival and growth of cobia (Rachycentron canadum) juvenile

An experiment was conducted in the laboratory to investigate the effects of additive ratios of docosahexaenoic acid (DHA) to eicosapentaenoic acid (EPA) on the growth and survival of cobia (Rachycentron canadum) juveniles from August to October 2005. Three hundred and eighty cobia juveniles (56 days of age, body weight 6.9 ± 0.1 g, body length 9.2 ± 0.1 cm) were selected and 20 of them were freely taken for initial sample analysis in the week 0. Additional 360 juveniles were randomly assigned into eight groups with triplicate, total 24 tanks with 15 fish each. Cobia juveniles were reared in glass‐steel tanks (200‐L volume per tank) using filtered seawater with temperature 26–30.5 °C, salinity 25.4–33.0 g L^?1 and pH 7.8–8.0. Cobia juveniles were fed for 8 weeks using seven treatment diets (D‐1 to D‐7) with the same amount of DHA and EPA (15.0 ± 1.2 g kg^?1 of dried diet), but varying ratios of DHA to EPA (0.9, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, respectively) and a control diet (D‐0, DHA + EPA = 8.0 g kg^?1 of dried diet, DHA/EPA = 1.3). Five juveniles per tank were randomly taken for sample analysis at the end of weeks 4 and 8, respectively. The highest protein efficiency rate (PER; 1.5 in mean), average body weight (BW; 73.3 g per fish in mean) and the lowest feed conversion ratio (FCR; 1.6 in mean) were obtained in cobia juveniles fed the control diet at the end of week 8. These parameters were significantly different (P < 0.05) among juveniles fed the control and treatment diets; however, no significant difference (P > 0.05) was found among juveniles fed the treatment diets evaluated in this study. It was concluded that the survival and growth of cobia juveniles were not greatly influenced by additive ratios of DHA to EPA in our experimental conditions.     

Preliminary study on broodstock rearing,induced breeding and grow‐out culture of the sea cucumber Holothuria scabra in Sri Lanka

Experiments were conducted to identify suitable methods for broodstock rearing, induced breeding and grow‐out culture of Holothuria scabra in Sri Lanka. Two hundred and seventy‐two brooders (500–600 g) collected from off Mannar were individually packed in oxygen‐filled polythene bags with and without sea water and transported to a sea cucumber hatchery at Kalpitiya. Lagoon pens, sand‐filled fibreglass tanks and bare tanks were used in triplicates to maintain brooders. Spawning was initiated using air dry, water jet and thermal‐stimulation methods. Hatchery produced juveniles with an average weight of 11 ± 5 g were reared (2 individuals m^?2) in lagoon pens, mud ponds and fibreglass tanks in triplicates. The significantly high evisceration rate was observed when brooders were transported without sea water (t‐test, P < 0.05). Brooders maintained in bare tanks showed a significant weight reduction than the brooders in sand‐filled tanks and lagoon pens (anova , P < 0.05, d.f. = 2). Thermal stimulation (ambient temperature ± 3–5°C) was found to be the most successful method of spawning initiation of H. scabra. The mean (±SD) percentage males and females participated for spawning per trial was 9.2 (±10) and 4.6 (±5.6) respectively. On an average, 1.16 millions of eggs (±1.03 SD, n = 5) were obtained per spawning trial. H. scabra juveniles reared in tanks showed significantly lower growth rate than the juveniles in pens and ponds (anova , P < 0.05). Lagoon pens and sand‐filled tanks are suitable to maintain brooders and lagoon pens can be successfully used for mass rearing of juveniles.     

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone analogue for ovulation induction in black sea bass Centropristis striata (Linnaeus, 1758)

Mature black sea bass, Centropristis striata L. (200–800 g), were captured in coastal South Carolina during the spawning season and administered hormones for ovulation induction and strip spawning. During both study years, control groups of females were incorporated into the study design and administered sham injections containing physiological saline solution. In 2004, females received a single intramuscular injection of human chorionic gonadotropin (hCG) (330 IU kg⁻¹) (n=8) or two injections of hCG at 24‐h intervals (n=8). In 2005, females received a single injection of hCG (n=10) or an analogue of luteinizing hormone releasing hormone (LHRHa) (n=10). In 2004, all fish administered a single dose of hCG ovulated at least once. Six fish ovulated on two consecutive days and one fish ovulated on 3 days consecutively. In contrast, six of eight fish receiving two doses of hCG ovulated once, five ovulated on 2 days successively and three fish ovulated 3 days in succession. Of the fish that spawned, no differences were found in any reproductive parameters. In 2005, all fish administered hCG or LHRHa ovulated at least once. Three fish administered hCG ovulated twice, four fish ovulated on three consecutive days and one fish 4 days successively. All fish administered LHRHa spawned at least twice, six fish ovulated thrice and three fish ovulated 4 days, successively. A significant difference in fertility was found between hCG (75.6±11.4%) and LHRHa (55.6±27.4%). The results of this study indicate that both hCG and LHRHa are effective for ovulation induction in prespawning black sea bass.     

Hormone-induced ovulation and spawning of captive and wild broodfish of the catadromous Australian bass, Macquaria novemaculeata (Steindachner), (Percichthyidae)

Australian bass, Macquaria novemacideata (Steindachner),mature but do not spawn in fresh or brackish water ponds. Ovulation and spawning of captive (n = 158)and wild Australian bass (n = 123) was induced in the normal breeding season by single injections of human chorionic gonadotropin (hCG). Doses of 100-4000IU kg-“ hCG induced ovulation and the optimum dosage was 500 IU kg” hCG. The breeding season was from mid-May to late August for wild fish,and extended into September for captive fish. There was a tendency for mean fertilization and hatching success to decline over the breeding season. Greater fertilization and hatching success was obtained from fish which spawned naturally than from stripped fish. Fish spawned after 34,2 ± 0.4 h (mean ± SE, n = 74). Ovulating fish that failed to spawn were stripped after 40.2 ± 0.3 h (n = 76). The timing of stripping and fertilization was an important factor determining hatching success. There was no apparent difference in latency periods or the number of eggs spawned between captive and wild fish. However, the mean number of eggs obtained from naturally spawned fish was higher than for stripped fish. The techniques described in this paper will assist the largescale production of Australian bass by increasing the quality and quantity of larvae from hCG-induced spawnings.     

Does the wild‐caught Chinese mitten crab megalopae perform better than the hatchery‐produced seed during the juvenile culture?

Although wild Yangtze Eriocheir sinensis juveniles (coin‐sized seed) have better culture performance than the pond‐reared juveniles during the adult crab culture stage, the culture performance of wild‐caught megalopae (WC) remains unknown. Therefore, this study was designed to compare the growth, survival rate, crab yield, feed conversion rate (FCR), precocity, final weight and size distribution between WC and hatchery‐produced megalopae (HP) during the juvenile culture stage. Wild‐caught megalopae had significantly higher body weight, weight gain rate (WGR) and specific growth rate (SGR) during 0–4 weeks but lower WGR and SGR than HP during 4–8 weeks (p < .05). Although WC had a significantly lower survival rate and yield of normal juvenile crabs than HP, total yields of normal juveniles and precocious crabs were similar. Wild‐caught megalopae had a significantly higher precocious rate and yield of precocious crabs than HP (p < .05). Regardless of sex, no significant differences were found in the gonadosomatic index (GSI), hepatosomatic index (HSI) and final weight of normal juveniles and precocious crabs between WC and HP. As for the size distribution of normal crabs, WC had a higher percentage of small female juveniles (<3 g) but a lower proportion of middle size ranges (3–5.99 g and 6–8.99 g) than HP for females (p < .05), and there was no significant difference for males. In conclusion, wild megalopae of the Yangtze E. sinensis had worse culture performance than the hatchery‐produced seeds; therefore, wild megalopae would not be recommended for commercially juvenile culture of E. sinensis.     

Analysis of growth and first reproduction of hatchery‐reared juvenile Cortez oyster (Crassostrea corteziensis) in northwestern Mexico: proposal of a minimal fishing size

This study presents complimentary data of early growth and age/size of first reproduction of hatchery‐reared Crassostrea corteziensis juveniles cultivated in the field for 9 months. Samples of oysters and gonads were collected monthly to determine absolute growth and growth rates, as well as overall gonad development. At the end, adult oysters ranging 75.6 ± 0.62 mm shell height (SH) yielded a mean growth rate of 0.308 mm day⁻¹, which is among the highest reported for the species. Gametogenesis began in March–April and spawning occurred during August and September. Males outnumbered females at smaller sizes, but female were more abundant at larger sizes. From planting time in the field, the age/size of first maturity was 3.25 months for males (42 mm SH) and 4.5 months for females (54 mm SH), with a mean population age of 4.75 months for males (size range 55–59 mm) and 5.25 months for females (60–64 mm). Based on these results, and the time needed for hatchery care prior planting juveniles in the field (2.25 months), we propose limiting fishing of wild oysters to sizes >65 mm height to allow them to breed at least once and contribute to restoring wild populations.     

Induced spawning of sea bass, Lates calcarifer, and rabbitfish, Siganus guttatus, after implantation of pelleted LHRH analogue

Captive Lates calcarifer broodstock at Tigbauan, Iloilo (Philippines) were implanted with cholesterol-based pellets of the LHRH analogue D-Trp⁶-desGly¹⁰-LHRH ethylamide or D-hArg(Et₂)⁶,Pro⁹-NHet-LHRH at doses between 9.0 and 23.5 μg/kg body weight. In May, one of ten LHRH-treated females released partially hydrated ova into the tank 4 days after implantation. In July, at least one (and probably four) of five LHRH-treated females spawned in the tank 2 days after implantation; 2.6 million hatchlings were collected. In August, both LHRH-treated females spawned in the tank 2 days after implantation; 978 000 hatchlings were collected. None of the sham-operated control fish spawned in any of the experiments.Captive Siganus guttatus broodstock implanted with silastic-based pellets of the LHRH analogue D-Nal (2)⁶ LHRH spawned 1–2 days earlier than sham-operated controls.     

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

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

Mass Production of Striped Patao Eugerres brasilianus Juveniles in Cuba

Abstract.— Two spawning and larval rearing trials with striped patao Eugerres brasilianus were conducted at Tunas de Zaza, Cuba, to develop a method for experimental-scale mass production of juvenile fish. Mature striped patao were captured from coastal lagoons in September 1988 and January 1989. Sexually mature females and males were induced to spawn by treatment with human chorionic gonadotropin (HCG). After spawning, eggs were collected and placed in 1.2-m³ cylindroconical fiberglass incubators at densities of 150 and 200 eggs/L. About 1 h before hatching, eggs were transferred to 5–m³ concrete tanks. Larvae were fed a mixture of marine microalgae, rotifers, copepods, and Artemia nauplii; artificial food was supplied at 30 d post-hatch as a shrimp diet with 25% protein and 0.1–0.4 mm particles. Fourteen of 20 females spawned after 2 or 3 HCG injections (total dose = 6–10 IU/g body weight). Fecundity was 467–2,167 eggs/g body weight. Egg incubation time in seawater (38–39 ppt salinity) was 15–17 h at 29–30 C and 21–23 h at 25–26 C, with 98–100% hatching rate. The oil droplet and yolk sac were consumed by 3 to 5 d post-hatch and first food (rotifers) was eaten during second day. High larval mortalities began after the first month. Growth in 39- ppt seawater was faster (0.41 ± 0.01 md/d) at 29.1 ± 0.3 C than at 25.2 ± 0.2 C (0.27 ± 0.008 mm/d). Overall survival was 6–12.5% with tinal densities of 5–10 juveniles/L after 48–60 d. A total of 132,000 juveniles was harvested from the two rearing cycles.     

Reproductive ability of second generation ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate the reproductive ability of second generation (F₂) koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. Only diploid F₂ males and females were fertile and used in crosses. A significant increase was recorded in male fertility in F₂ versus F₁. In contrast with an earlier study in which only one fertile F₁ male was found, about 20% of F₂ males produced sperm. The observed reproductive ability of F₂ hybrids was similar to that demonstrated by the only fertile F₁ male and F₁ females. F₂ males produced diploid spermatozoa and generated triploids when crossed with koi females. All triploid fish in these progenies were males indicating that F₂ males had a sex chromosome constitution of XY. F₂ females produced diploid eggs and generated mostly triploids when crossed with koi males. In progenies obtained by crosses of F₂ males with F₁ and F₂ females, most of the surviving juveniles (63%–100%) were diploid; a minority of juveniles were aneuploid (ploidy ranged from 2.1n to 3.6n). Diploid fish in these progenies were presumably the result of spontaneous androgenesis and gynogenesis, by the same mechanisms observed earlier in progenies obtained by crossing the F₁ male with F₁ females.     

Distribution patterns of saprolegniosis cutaneous lesions in wild and farmed brown trout (Salmo trutta L.) obtained using a geographic information system (GIS)

A retrospective study was conducted using 250 clinical records of brown trout (Salmo trutta L.) with saprolegniosis by Saprolegnia parasitica, which had been collected from 8 rivers and 1 fish farm in the province of León (Spain). A geographic information system (GIS) was used to obtain skin lesion distribution patterns in males and females. Lesions in wild brown trout affected 15.31 ± 13.33% of the body surface, with a mean of 12.76 ± 6.56 lesions per fish. In addition, 51.23% of wild trout presented lesions with necrosis of the skin or fins. The pattern obtained when not distinguishing between sexes indicated that saprolegniosis lesions are mainly located above the lateral line and most frequently affect the dorsal cephalic region, the adipose fin, the peduncle and the caudal fin. However, differences were observed between males and females. Farmed trout presented a lower percentage of affected body surface (2.06 ± 4.36) and a lower number of lesions with and without necrosis because they received preventive treatment for saprolegniosis.     

Efficacy of Vaccination Against Streptococcus iniae During Artificial Spawning of the Red-Tail Black Shark (Epalzeorhynchos bicolor,fam. Cyprinidae)

Artificial spawning may precipitate high mortalities of red-tail black shark (Epalzeorhynchos bicolor) broodstock from Streptococcus iniae infection. Two vaccine formulations (aluminum or oil adjuvant) were evaluated for effectiveness. Fish (mean weight 21.2 ± 5.7 g) were harvested, acclimatized, and vaccinated by intracoelomic injection. After 21 days, fish were moved to a biosecure laboratory and, one day later, simultaneously spawned and challenged by bath or by intracoelomic injection with S. iniae. Both formulations increased relative% survival (RPS) in unspawned (RPS [formulation]: 80 [aluminum]; 54 [oil]) and spawned fish (87–95 [aluminum]; 73–75 [oil]), suggesting that vaccination increases survival of red-tail black shark broodstock after spawning.     

Reproductive ability of triploid ornamental (koi) carp (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids and characteristics of their offspring

The purpose of this study was to investigate reproductive ability of backcross triploid koi (Cyprinus carpio L.) × goldfish (Carassius auratus L.) hybrids. These triploids have been obtained by crossing of F₁ hybrid females producing diploid eggs with males of parental species. Triploid hybrid females, when crossed with goldfish or koi males, produced mostly aneuploid fish with ploidy range from approximately 2.2n–3.2n with a mean value 2.5n; some fish in crosses of triploid females with koi males were tetraploid (4.0n). Since analysed fish had in their genomes one haploid set from parental males, the data indicate that triploid hybrid females mostly produced aneuploid eggs with ploidy range from approximately 1.2n–2.2n and a mean ploidy around 1.5n while some eggs were triploid. Triploid hybrid males were completely sterile and have not released any sperm after hormonal injection. Despite their low viability, some aneuploid fish obtained from triploid hybrid females were raised in indoor recirculating systems until the age of 2 years and their reproductive ability has been evaluated. One aneuploid female with ploidy 2.1n produced larvae with ploidy range from 2.9n to 3.4n with a mean ploidy of 3.1n when crossed with a koi male; about 60% of obtained larvae had ploidy from 3.0n to 3.2n. These data indicate that this female produced mostly eggs with unreduced ploidy level.     

Effect of different broodstock densities on the reproductive performance of Nile tilapia, Oreochromis niloticus (L.), in a recycling system

The present study was conducted to evaluate the effect of different stocking densities on the seed production of Nile tilapia, Oreochromis niloticus (L.), under intensive recycling hatchery system conditions. Males and females with mean body weights of 163.2 and 105.0 g, respectively, were stocked at three broodstock densities (4, 8 and 12 fish m^–2) at a male:female ratio of 1:3 in 1 × 1 × 0.43 m (W × L × H) fibreglass tanks. The tanks were illuminated at 2500 lux for 18 h day^–1 and the water temperature was maintained at 29 ± 1 °C. Effluent from spawning tanks was recycled through a biological filter with 10–15% replacement of new water per day. The experiment lasted for 126 days. The results showed that breeders stocked at 4 fish m^–2 had significantly higher (P < 0.05) mean values for total seed production, seed kg^–1 female day^–1, seed female^–1 day^–1, seed m^–2 day^–1 and spawning synchrony than at 8 and 12 fish m^–2 broodstock densities. The mean percentage of seeds in the yolk-sac and swim-up fry stages was highest at 4 fish m^–2 broodstock density. However, the recovery rate was not affected by broodstock density. It is recommended that further research should be conducted to determine whether weight m^–2 , number m^–2 or age of broodstock should be the basis for stocking broodstock.     

Hormone-induced ovulation, natural spawning and larviculture of Brazilian flounder Paralichthys orbignyanus (Valenciennes, 1839)

Mature Brazilian flounders Paralichthys orbignyanus were captured in coastal southern Brazil and their reproduction in captivity was studied. Brazilian flounder will spawn naturally in captivity when the water temperature is around 23 °C and 14 h of light is provided daily. Females were induced for ovulation and hand stripping using human chorionic gonadotropin, luteinizing hormone‐releasing hormone analogue or carp pituitary extract. There was no need to inject males, as running milt was observed during the spawning season. Fertilization and hatching rates were above 80% independent of the hormone used. Notochord length at hatching was 2.18±0.07 mm for larvae hatching from naturally spawned eggs. Larvae were reared in salt water (30–35 g L^?1) at 24 °C and under continuous illumination. Larviculture was with green water (Tetraselmis tetrathele 50 × 10⁴ cells mL^?1). Rotifers (10–20 ind mL^?1) were offered as first food 3 days after hatching and gradually replaced by Artemia nauplii (0.5–10 ind mL^?1). Larvae settled to the bottom 20 days after hatching and completed metamorphosis within a week after that. The total length for newly metamorphosed juveniles was 12.9±2.2 mm and the mean survival was 44.8%. The results demonstrate the feasibility of producing Brazilian flounder fingerlings for stock enhancement or grow‐out purposes.     

Effect of dietary prebiotic inulin on histology,immuno‐haematological and biochemical parameters of Asian seabass (Lates calcarifer)

The present study was carried out to investigate the effect of supplementation of inulin on histology, immuno‐haematological and biochemical parameters of Asian seabass (Lates calcarifer) fingerlings with an initial average body weight of 7.14 ± 0.05 g. Inulin was supplemented at four different concentrations (control) 0, 5, 10, 15, and 20 g/kg in the L. calcarifer diet containing 400 g/kg protein and 90 g/kg lipid. At the end of the 60 days feeding trial, the absorptive surface area of the intestine and glycogen deposition in liver were increased in fish fed inulin supplemented diets. The immune parameters lysozyme, alternative complement pathway, superoxide dismutase and nitroblue tetrazolium assay showed significant (p < 0.05) difference between control and treatment groups. Haematological parameters showed that red blood cells, white blood cells, haemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration levels increased significantly (p < 0.05) in the fish fed with inulin supplemented diets. The analysis of biochemical parameters revealed that glucose, urea, cholesterol, triglyceride, aspartate aminotransferase, alkaline phosphatase, alanine aminotransferase, and lactate dehydrogenase showed significant differences (p < 0.05) between control and treatments groups. No significant difference (p > 0.05) was observed for total protein among different treatments. The results of the study revealed that 15–20 g/kg inulin supplementation has a beneficial effect in the histology, immuno‐haematological, and biochemical parameters in L. calcarifer juveniles.     

Effect of dietary protein and energy levels on growth,oxygen consumption,haemolymph and digestive gland carbohydrates,nitrogen excretion and osmotic pressure of Litopenaeus vannamei (Boone) and L. setiferus (Linne) juveniles (Crustacea,Decapoda; Penaeidae)

The influence of protein and energy levels on growth rate, survival, pre- and post-prandial oxygen consumption, ammonia excretion, haemolymph glucose (HG), glycogen in digestive gland and osmotic pressure (OP) in white shrimp Litopenaeus vannamei (Boone) and L. setiferus (Linne) juveniles was studied. Diets containing a high-quality protein at a protein/energy (P/E) ratio of 16, 26, 31 and 36 mg kJ⁻¹ were fed at 20% of shrimp body weight of two sizes: < 1 g and > 1 g. Both species showed a optimum P/E ratio of 36 mg kJ⁻¹ (33–44% protein and 6–23% carbohydrate) in juveniles < 1 g. For shrimp > 1 g, L. setiferus showed a higher growth rate in the diet with 16 mg kJ⁻¹ (27% protein; 32% carbohydrate) and L. vannamei between 26 and 36 mg kJ⁻¹ (33–44% protein and 6–23% carbohydrate). In both experiments, the growth rate of L. vannamei was 2–3 times that observed in L. setiferus. Routine oxygen consumption and apparent heat increment (AHI) of L. setiferus juveniles was two times higher than that observed in L. vannamei juveniles, which could indicate that L. setiferus has a higher metabolic rate. The O/N ratio varied according to protein level, with higher values (O/N = 180) with a 16-mg kJ⁻¹ diet and lower values (O/N = 73) with a 36-mg kJ⁻¹ diet in L. setiferus juveniles. A similar variation in O/N ratio was obtained in L. vannamei fed with all diets with an interval between 22 and 50. An inverse relation between ammonia excretion and HG, and digestive gland glycogen (DGG) in relation to an increase in the P/E ratio indicate that both shrimp species are well adapted to use carbohydrates and/or proteins from their diet. The higher values of hyper-osmotic capacity (hyper-OC) were observed in L. setiferus < 1 g fed with 36 mg kJ⁻¹ and the lowest in L. vannamei < 1 g fed with 31 mg kJ⁻¹. Intermediate values of hyper-OC were observed in both species fed all diets indicating that osmotic factors of juveniles < 1 g of both species are more affected by the P/E ratio than juveniles > 1 g. All results showed that juveniles > 1 g of both species are less dependent of P/E ratio than juveniles < 1 g. Litopenaeus vannamei is a most tolerant shrimp species with a high capacity to use a wide range of dietary P/E ratios for growth, which may be due to its lower energy requirements. Litopenaeus setiferus showed a lower capacity to accept different P/E ratios but the optimum P/E ratio obtained with this species shows that L. setiferus accept diets with a high carbohydrate level as well. These results demonstrate that there are nutritional and physiological differences that explain the differences that have been observed when both species were cultured in commercial ponds.