Flow cytometric determination of genome size in European sunbleak Leucaspius delineatus (Heckel, 1843)

The aim of this study was to compare DNA content in hepatocyte and erythrocyte nuclei of the European sunbleak, Leucaspius delineatus, in relation to nuclear and cell size by means of flow cytometry and fluorescence microscopy. The DNA standards, chicken and rainbow trout erythrocytes, were prepared in parallel with both cell types, with initial separation of liver cells in pepsin solution followed by cell filtering. Standards and investigated cells were stained with a mixture of propidium iodide, citric acid, and Nonidet P40 in the presence of RNAse, and fluorescence of at least 50,000 nuclei was analyzed by flow cytometry. Average cell size was determined by flow cytometry, using fresh cell suspension in relation to latex beads of known diameter. The size of nuclei was examined on the basis of digital micrographs obtained by fluorescence microscopy after nuclei staining with DAPI. The sunbleak’s erythrocyte nuclei contain 2.25 ± 0.06 pg of DNA, whereas the hepatocyte nuclei contain 2.46 ± 0.06 pg of DNA. This difference in DNA content was determined spectroscopically using isolated DNA from the two cell types. The modal diameters of the erythrocytes and hepatocytes were estimated to be 5.1 ± 0.2 and 22.3 ± 5.0 μm, respectively, and the corresponding modal dimensions of their nuclei (measured as surface area) were 15.2 and 21.4 μm², respectively. The nucleoplasmic index, as calculated from diameters estimated from surface area of nuclear profiles, was 2.51 for the erythrocytes compared with 0.08 for hepatocytes.     

Cytogenetic analysis of hybrids and hybrid triploids between the river puffer,Takifugu obscurus,and the tiger puffer,Takifugu rubripes

Cytogenetic analyses of the river puffer, Takifugu obscurus, the tiger puffer, Takifugu rubripes and hybrids produced between female T. obscurus and male T. rubripes and their hybrid triploids (produced by cold shock treatment at 4°C) were performed. T. obscurus had 2n = 44 chromosomes and 1.84 ± 0.019 pg DNA/cell, T. rubripes had 2n = 44 and 2.64 ± 0.015, the hybrids had 2n = 44 and 2.15 ± 0.010 and the hybrid triploids had 3n = 66 and 3.22 ± 0.010. The erythrocyte values of the hybrids were more similar to those for T. obscurus, whereas the hepatocyte, midgut and proximal tubule kidney cell values of the hybrids fell down between those for the parental species (p < .05). The erythrocyte, proximal tubule, hepatocyte and midgut epithelial cell sizes for the hybrid triploids were 1.5‐fold larger than those for the hybrids (p < .05). The thickness of retina and each layer in the hybrid triploids were 1.1‐fold larger than those of the hybrids (p < .05) and did not differ significantly among T. obscurus, T. rubripes and the hybrids (p > .05); however, the hybrid triploids had fewer cell nucleus outer layers than the hybrids (p < .05). Gonad development in the hybrids and hybrid triploids was less matured than in T. obscurus and T. rubripes. The metaphase nucleolus organizer regions (NORs) and the gill cells of T. obscurus, T. rubripes and the hybrids contained two satellite telocentrics, whereas the hybrid triploids contained three satellite telocentrics.     

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.     

Salt- and pH-induced functional changes in protein concentrate of edible green seaweed <Emphasis Type="Italic">Enteromorpha</Emphasis> species

Protein concentrates (PCs) were extracted from three edible green seaweed species of Enteromorpha (E. compressa, E. linza, and E. tubulosa) and were studied for their functional properties with respect to salt and pH. The protein content in the PC was found to be 60.35 ± 2.0, 53.83 ± 0.70, and 33.36 ± 1.04% in E. compressa, E. tubulosa, and E. linza respectively. The minimum nitrogen solubility was observed at pH 4.0 in all three PCs. The water-holding and oil-holding capacities in the three PCs ranged from 1.22 ± 0.06 to 1.53 ± 0.07 ml H₂O/g PC and from 1.05 ± 0.07 to 1.34 ± 0.10 ml oil/g PC respectively. Foaming capacity and stability were found to be pH-specific. They varied significantly with pH and NaCl concentration (P < 0.05). The inexpensive source of protein concentrate from Enteromorpha species could be incorporated into value-added food products.     

Tetraploid induction in tropical oysters,Crassostrea belcheri (Sowerby) and Crassostrea iredalei (Faustino)

Tetraploid induction has been conducted on temperate oysters but not on tropical oysters. In this study, different heat shocks (32, 35 and 38°C) and cold shocks (1, 4 and 7°C) were used to induce tetraploidy in two tropical oyster species, Crassostrea belcheri and Crassostrea iredalei, through meiosis I inhibition. Temperature shocks were applied on the newly fertilized eggs at 8–10 min post fertilization and terminated when second polar bodies began to form in the control eggs. The ploidy of the larvae and spat was determined via direct chromosome count. The percentage of larval survival until Day 20 was low (between 0.4% and 42.9%) for both temperature shocks and oyster species. No surviving larva was recorded for induction at 1, 4 and 38°C. Tetraploid spat was only recorded in C. iredalei but the percentage is low through heat shock induction of 32 and 35°C. This study shows that the tetraploid induction success rate was slightly higher in C. iredalei compared to C. belcheri. No surviving tetraploid spat were recorded for both oyster species through the cold shock method. This study shows that heat shock can be used to inhibit meiosis for the production of tetraploids but more experiments need to be conducted to determine the optimum temperature when dealing with tropical oysters.     

Growth and gut morphology of diploid and triploid juvenile Atlantic cod (Gadus morhua)

The objective of this paper was to compare the growth and gut morphology of juvenile diploid and triploid Atlantic cod (Gadus morhua) reared under similar conditions. Individually tagged 36‐week‐old diploid (mean weight 49.3 ± 13.8 g) and triploid (mean weight 43.6 ± 11.2) juvenile cod were measured at intervals during a 29‐week growth trial. Data for weight, length, condition factor (K), hepato‐somatic index (HSI), gonado‐somatic index (GSI), Relative Gut Length (RGL) and pyloric caeca number were collected and results were analysed in relation to ploidy status, gender and family contribution. At the end of the experiment, only one family (M2xF3) had many representatives with a relatively even distribution of sexes and ploidies. Diploid females were significantly heavier and had higher K than triploid females in the M2xF3 family (body weight 371.2 ± 120.2 vs. 298.4 ± 100.7 g; K 1.1 ± 0.1 vs. 0.93 ± 0.1), but no differences were found between diploid and triploid males. In the other families (pooled data), no differences in body weight were found between the ploidy groups. In general, triploids had a shorter intestine (RGL) and fewer pyloric caeca than their diploid siblings regardless of gender suggesting possible impairments in nutrient utilization and growth.     

Effects of different natural or prepared diets on growth and survival of juvenile spider crabs, <Emphasis Type="Italic">Maja brachydactyla</Emphasis> (Balss, 1922)

The effects of different diets (natural or pellets) on growth, survival, and moulting interval of juvenile spider crabs, weighing between 0.011–1.56 g and up to 17.6 mm in carapace length, were tested over a period of 90 days. During experiment I, five diets were tested: (1) frozen shrimp—Paleomonetes sp., (2) fresh mussels—Mytilus sp., (3) white fish fillets—Merlucius merlucius, (4) blue fish fillets—Sardina pilchardus, and (5) commercial crustacean pellets. Spider crabs fed fresh mussels grew larger (0.98 ± 0.69 g) and had higher growth rates (4.0 ± 0.7 %BWd⁻¹) compared to the other four diets. The crabs fed shrimp pellet and frozen shrimp grew to intermediate sizes and were smaller than the ones fed fresh mussels, but they were larger than spider crabs fed either blue or white fish fillets (0.46 ± 0.63 and 0.26 ± 0.13 g, respectively) compared to the ones fed white fish fillets (0.12 ± 0.04) and blue fish fillets (0.04 ± 0.02 g). The spider crabs fed blue fish fillets only lasted until day 60 of the experiment, after this day none of the 20 fed this diet were left. During experiment II, two diets were tested: (1) white and blue fish fillets and (2) commercial fish pellet. There were no differences in growth both in weight or carapace length (2.9 ± 1.8 and 2.1 ± 1.5 g in weight, and 18.9 ± 5.0 and 17.7 ± 3.3 mm, respectively) at the end of the experiment. Similarly, there were no differences in growth rates in weight between the two diets (1.2 ± 0.4 and 0.9 ± 0.3 %BWd⁻¹, respectively) or in carapace length (0.4 ± 0.1 and 0.4 ± 0.2 %BWd⁻¹, respectively). Fresh mussel appears to be a very good diet to culture the early stages of this species, while shrimp pellets also deliver acceptable results. On the contrary, frozen shrimp, fish fillets either from blue or white species (much higher lipid content in the blue species), and fish pellets were found to be bad diets for the culture of the early stages of M. brachydactyla.     

Hypoxia affects performance traits and body composition of juvenile hybrid striped bass (Morone chrysops × M. saxatilis)

Performance traits and body composition of juvenile hybrid striped bass (Morone chrysops × M. saxatilis) in response to hypoxia were evaluated in replicate tanks maintained at constant dissolved oxygen concentrations that averaged 23.0 ± 2.3%, 39.7 ± 3.0% and 105.5 ± 9.5% dissolved oxygen saturation. Fish were fed a commercially formulated feed daily to apparent satiation. Total feed intake and fish growth and yield increased linearly in response to increased dissolved oxygen concentration. Nutrient utilization was reduced significantly only at the greatest level of hypoxia. With the exception of whole body protein content, whole body compositional indices and nutrient retention efficiencies were linearly related to dissolved oxygen concentration. Results demonstrate that as hypoxia becomes more severe, juvenile hybrid striped bass feed intake is reduced, which affects growth and nutrient retention.     

Factors influencing the quality of half‐pearls (mabé) produced by the winged pearl oyster,Pteria penguin (Röding, 1758)

The winged pearl oyster, Pteria penguin, is cultured primarily to produce half‐pearls (mabé). The mabé quality is influenced by culture techniques, but there is limited information in this field. P. penguin with mean (±SE) dorso‐ventral height of 250 ± 6.5 mm were used to investigate the influence of culture period and nucleus position on mabé quality. Oysters were relaxed using 1‐propylene phenoxetol, and five nuclei were glued at different positions to the inner surfaces of the oyster shells; three on the more concave left valve and two on the right valve. Nucleated oysters were then cultured for 10 months under commercial pearl farming conditions at Savusavu in Fiji. Nacre deposited at the base and top of the nuclei was measured monthly, from the 6th to the 10th months of culture and the different qualities of mabé produced at different positions were scrutinized. Nacre thicknesses at the base and top of the resulting mabé were significantly different at different months (P < 0.05) and the rate of nacre deposition was highest during the warmer months. The different positions of nuclei on the valve greatly affected the quality of mabé formed. After a 10‐month culture period, around 1 mm of nacre covered the nuclei although the best quality mabé were obtained after 9 months.     

Enzyme-linked immunosorbent assay (ELISA) measurement of vitellogenin in plasma and liver histopathology in barfin plaice <Emphasis Type="Italic">Liopsetta pinnifasciata</Emphasis> from Amursky Bay,Sea of Japan

Vitellogenin (Vg) of the barfin plaice Liopsetta pinnifasciata was isolated and purified. In native polyacrylamide gel electrophoresis, Vg appeared as one band. After being subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE), Vg fraction produced several polypeptides with molecular masses of 180, 98, 70, 52, 41 and 37 kDa. MALDI–TOF mass spectrometry (MS) of the 180- and 98-kDa Vg polypeptides from the SDS–PAGE gel and de novo sequencing of their four peptide fragments based on MS/MS analysis confirmed that the purified proteins were vitellogenins, which shared high similarity with the Vgs of the barfin flounder Verasper moseri and Atlantic halibut Hippoglossus hippoglossus. The most part of the predicted sequences obtained from the L. pinnifasciata 180-kDa polypeptide has previously been found in the V. moseri vitellogenin type B, the sequences obtained from the 98-kDa polypeptide were found in V. moseri vitellogenin type A, so these findings allow us to propose that L. pinnifasciata has at least two different forms of Vg. Rabbit polyclonal antibodies against Vg were produced, and a quantitative enzyme-linked immunosorbent assay was developed. The concentration of Vg in barfin plaice from the moderately contaminated area of Amursky Bay in the Sea of Japan was detected based on the maturity stage of their gonads. In November 2008, the Vg concentration in the plasma of females with advanced oogenesis varied from 5.295 to 28.367 mg/ml (mean 16.38 ± 6.73 mg/ml, CV = 41.1%); in the plasma of males, the concentration ranged from non-detectable to 0.957 mg/ml (0.29 ± 0.42 mg/ml, CV = 127.9%). In October 2009, the Vg concentration in female plasma was lower than in November 2008 (2.21–13.87 mg/ml). High individual variability of plasma Vg was characteristic for maturing males (CV = 200.3%) and immature females (CV = 255.5%), and there was no significant difference between plasma Vg concentrations in males captured in November 2008 and October 2009 or in maturing males and immature females. Vacuolisation of hepatocytes was more typical for males with low plasma Vg concentrations and females with high plasma Vg concentrations. Necrosis and pyknosis of hepatocyte nuclei were more frequent in males with high Vg concentrations and in females with low plasma Vg concentrations.     

Effect of prebiotic xylooligosaccharides on growth performances and digestive enzyme activities of allogynogenetic crucian carp (<Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>)

The effect of prebiotic xylooligosaccharides (XOS) on the growth performance and digestive enzyme activities of the allogynogenetic crucian carp, Carassius auratus gibelio, was investigated. XOS was added to fish basal semi-purified diets at three concentrations by dry feed weight: diet 1, 50 mg kg⁻¹; diet 2, 100 mg kg⁻¹; diet 3, 200 mg kg⁻¹, respectively. Twelve aquaria (n = 20) with three replicates for each treatment group (diets 1–3) and control treated without XOS were used. Weights of all collected carp from each aquarium were determined at the initial phase and at the end of the experiment, and the carp survival was also determined by counting the individuals in each aquarium. After 45 days, there were significant differences (P < 0.05) in the relative gain rate (RGR), and daily weight gain (DWG) of diets 1–3 were compared with the control. However, the survival rate was not affected (P > 0.05) by the dietary treatments. For enzymatic analysis, dissection produced a crude mixture of intestine and hepatopancreas of each segment to measure. The protease activity in the intestine and hepatopancreas content of fish in diet 2 (487.37 ± 20.58 U g⁻¹ and 20.52 ± 1.93 U g⁻¹) were significantly different (P < 0.05) from that in the control (428.13 ± 23.26 U g⁻¹ and 12.81 ± 1.52 U g⁻¹) and diet 3 (428.00 ± 23.78 U g⁻¹ and 14.04 ± 1.59 U g⁻¹). Amylase activity in the intestine was significantly higher for diet 2 compared to diet 1 and the control. As for amylase in the hepatopancreas, assays showed higher activity in diet 2 (P < 0.05) compared to the rest.     

The use of alternative prey (crayfish, <Emphasis Type="Italic">Procambarus clarki</Emphasis>, and hake, <Emphasis Type="Italic">Merlucius gayi</Emphasis>) to culture <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier 1797)

The effects of two alternative prey (crayfish and hake) were tested on growth and survival of both juveniles and adults of Octopus vulgaris in two experiments. Octopuses fed the control (squid) were larger (3.0 ± 0.7 kg) than those fed crayfish (2.4 ± 0.6 kg) at the end of experiment I. Similarly, overall growth rates were higher for octopuses fed squid (1.7 ± 0.3 and 1.2 ± 0.2 %BW day⁻¹, respectively). Average feeding rates for the experiment were not different, being 6.5 ± 0.9 and 7.5 ± 0.9 %BW day⁻¹, respectively, for octopuses fed either squid or crayfish. Nevertheless, food conversions for the experiment were higher (42.4 ± 2.7%) for octopuses fed squid compared to the ones fed crayfish (23.9 ± 1.9 g). For experiment II, hake and crayfish were compared to squid; the final weight of octopuses fed squid, hake or crayfish was 1,183.0 ± 242.7 g, 1,175.6 ± 240.1 g and 922.3 ± 160.1 g, respectively. Overall growth rates for the experiment were 1.9 ± 0.2 %BW day⁻¹, 1.9 ± 0.3 %BW day⁻¹ and 1.1 ± 0.3 g, respectively. Final weight and growth rates were never different (P > 0.05) between octopuses fed squid and hake, but were always higher (P < 0.05) compared to the ones fed crayfish. Average feeding rates for experiment II were similar for the three diets, and of 4.6 ± 1.5, 4.2 ± 1.3 %BW day⁻¹ and 5.1 ± 0.9 %BW day⁻¹, respectively, for octopuses fed squid, hake or crayfish. Food conversions for experiment II were of 41.0 ± 9.6%, 40.5 ± 9.9% and 21.3 ± 7.4 g, respectively, for octopuses fed squid, hake or crayfish, and were always higher for octopuses fed squid and hake compared to crayfish. The results indicate that crayfish is not an adequate replacement for the usual prey to fatten octopus, even considering its much lower market price.     

Characterization of triploid hybrid groupers from interspecies hybridization (Epinephelus coioides ♀ × Epinephelus lanceolatus ♂)

Interspecies hybridization is widely used in aquaculture as a beneficial strategy. Diploid and triploid hybrids have been detected from the interspecies hybridization of Epinephelus coioides ♀ × Epinephelus lanceolatus ♂. This is the first report of triploidization through hybridization in grouper. Confirmation has been obtained through flow cytometry, karyotyping and erythrocyte nuclei measurement. The chromosome numbers of E. coioides, E. lanceolatus, diploid hybrid grouper are 48 and triploid hybrid grouper are 72. Measurements of erythrocyte nuclei indicate that triploid fish have a larger nuclear surface than the diploid groupers, and the average ratio of triploid to diploid surface area is 1.59. During the first 1.5 years, triploid hybrid groupers grow faster than diploid hybrid groupers or either parent species. The average growth rate of triploid hybrids is 1.61 times greater than that of diploid hybrids at 6 months of age and 1.43 times greater at 18 months of age. The triploid hybrid groupers are inferior in gonadal development, with no primary‐growth‐stage oocytes appearing in the gonads at 18 months of age. Morphological studies indicate that triploid hybrid groupers have distinctive differences in snout length, eye diameter, body trunk shape, and tail shape development compared with diploid hybrid groupers. Triploid hybrid groupers have an advantage in growth ability, and artificial breeding of triploid groupers might be of great potential use in the grouper aquaculture industry.     

Interspecific variations in age and size at settlement of 8 emperor fishes (Lethrinidae) at the southern Ryukyu Islands,Japan

Molecular and otolith analyses were conducted for 173 settlement-stage larvae of emperor fishes (family Lethrinidae) collected by light traps at Ishigaki Island, southern Japan, in July and August (summer season), to (1) present diagnostic DNA markers for identification of lethrinid species and (2) compare the size and age at settlement of each species. PCR–RFLP and direct nucleotide sequencing analyses identified 8 species. Size (standard length, SL) at settlement differed significantly between species; Lethrinus ornatus (mean SL ± SD, 12.8 ± 1.5 mm), L. obsoletus (14.2 ± 0.8 mm) and L. harak (15.8 ± 1.6 mm) settled at a smaller size than L. atkinsoni (17.0 ± 1.3 mm), L. genivittatus (17.3 ± 1.0 mm), L. olivaceus (18.1 ± 0.6 mm), L. nebulosus (18.6 ± 4.2 mm), and L. sp.2 reported by Lo Galbo et al. (J Mol Evol 54:754–762, 2002) (21.7 ± 1.4 mm). Age at settlement tends to increase with settlement size; L. obsoletus (mean age ± SD, 25.6 ± 1.2 days), L. atkinsoni (26.1 ± 2.1 days) and L. ornatus (26.3 ± 2.9 days) were younger at settlement than L. nebulosus (28.4 ± 2.1 days), L. harak (29.2 ± 1.7 days), L. olivaceus (29.5 ± 1.0 days), L. genivittatus (30.5 ± 1.7 days) and L. sp.2 (31.0 ± 2.0 days). Although our study showed interspecific variation in body size and age at settlement among 8 lethrinid species, further seasonal replication is necessary to clarify the general patterns.     

Composition of ovarian fluid in endangered Caspian brown trout, <Emphasis Type="Italic">Salmo trutta caspius</Emphasis>, and its effects on spermatozoa motility and fertilizing ability compared to freshwater and a saline medium

In this study, ovarian fluid composition and its effects on the motility and fertilizing ability of sperm were studied in endangered Caspian brown trout, Salmo trutta caspius, and were compared with a saline activation medium (125 mM NaCl, 30 mM Glycine, 20 mM Tris–HCl, pH = 9.0) and freshwater as the control. The ovarian fluid was composed of sodium 164.4 ± 4.4 mM l⁻¹, potassium 1.8 ± 0.1 mM l⁻¹, calcium 0.6 ± 0.1 mM l⁻¹, magnesium 0.4 ± 0.02 mM l⁻¹, chloride 127.4 ± 5.9 mM l⁻¹, total protein 389.5 ± 89.6 mg 100 ml⁻¹, cholesterol 9.3 ± 1.2 mg dl⁻¹, and glucose 3.3 ± 0.2 mM l⁻¹. The percentage of motile spermatozoa and the duration of sperm motility were significantly higher in ovarian fluid (62 ± 3%, 74.6 ± 0.8 s) than freshwater (35 ± 4%, 44 ± 1 s), but they did not differ significantly from saline medium (56 ± 3%, 74.3 ± 0.7 s) (P > 0.05). Higher eyeing rates were observed after the activation of sperm in ovarian fluid and saline solution than freshwater when 35,000 or 350,000 spermatozoa per egg were added into the activation media. However, no significant differences were observed at higher concentrations of spermatozoa per egg (730,000) (P > 0.05). Also, this study showed that the ovarian fluid composition can be considered as a species-specific character among salmonid fishes. As a conclusion, the results of this study recommend the use of ovarian fluid or the saline solution as an activation medium in the artificial reproduction of Caspian brown trout.     

Differences in uptake and killing of pathogenic and non‐pathogenic bacteria by haemocyte subpopulations of penaeid shrimp,Litopenaeus vannamei, (Boone)

Phagocytosis is an important function of both invertebrate and vertebrate blood cells. In this study, the phagocytic activity of haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei, (Boone), against pathogenic and non‐pathogenic particles was investigated in vitro. The haemocytes of penaeid shrimp were firstly separated by centrifugation on a continuous density gradient of iodixanol into four fractions with five subpopulations (sub), of which sub 1 (hyalinocytes) and sub 4 (semi‐granulocytes) have the main function in phagocytosis of both pathogenic and non‐pathogenic bacteria as well as fluorescent polystyrene beads. It was found that these haemocyte subpopulations engulfed virulent Vibrio campbellii and Vibrio harveyi at a higher rate than non‐virulent Escherichia coli and polystyrene beads. When these bacteria were mixed with shrimp haemocyte subpopulations and incubated for 180 min, the percentage of viable intracellular V. campbellii (25.5 ± 6.0%) recovered was significantly higher than the percentage recovered from V. harveyi (13.5 ± 1.1%). No viable intracellular E. coli was observed in this study. In contrast to V. harveyi and E. coli, V. campbellii containing endosomes did not acidify in time. Incubation of haemocyte subpopulations with the most virulent V. campbellii strain resulted in a significant drop in haemocyte viability (41.4 ± 6.3% in sub 1 and 30.2 ± 15.1% in sub 4) after 180 min post‐inoculation in comparison with the less virulent V. harveyi (84.1 ± 5.6% in sub 1 and 83.4 ± 4.1% in sub 4) and non‐virulent E. coli (92.7 ± 2.8% in sub 1 and 92.3 ± 5.6% in sub 4) and polystyrene beads (91.9 ± 1.6% in sub 1 and 84.4 ± 3.4% in sub 4). These findings may be a valuable tool for monitoring shrimp health and immunological studies.     

Long‐term low‐salinity stress affects growth,survival and osmotic related gamma‐aminobutyric acid regulation in the swimming crab Portunus trituberculatus

A 30‐day experiment was conducted to investigate the effects of long‐term low‐salinity stress on the growth performance and osmotic related chlorine ion channel ligand regulation: gamma‐aminobutyric acid (GABA) content and GABA_A receptor‐associated protein (GABARAP) expression in Portunus trituberculatus. The salinity levels of both the control group and the experimental group were 30 and 12 psu respectively. After rearing for 30 days, the specific growth rate and survival rate were compared between the two groups, and salinity 6 psu was used to test the salinity tolerance. The results were as follows: (a) Both the specific growth rate and survival rate were significant lower in the experimental group (p < 0.05) after 30 days; (b) After challenge with salinity 6 psu for 72 hr, the crabs of experimental group had a 100% survival rate, whereas the crabs of the control group were all dead within 48 hr; (c) The content of GABA and the gene expression level of GABARAP in experimental group were significant different from control group (p < 0.05) after challenge via salinity 6 psu. In the control group, the GABA content increased rapidly from 9.96 ± 2.09 to 42.00 ± 5.94 µg/g; however, in the experimental group, it only increased to 27.82 ± 2.55 µg/g; the gene expression of GABARAP in the experimental group increased to the maximum at 24 hr, then decreased and stabilized at 48 hr, suggesting that GABA and GABARAP were trigged during the early stage of low‐salinity stress resistance.     

Heritability of economically important traits in the Atlantic cod Gadus morhua L

During the development of breeding programme for Atlantic cod Gadus morhua L., in Iceland, genetic parameters were estimated for 1402 individuals, which were assigned with DNA profiling to 140 dams and 70 sires. The cod was reared in cages on the eastern and western coasts of Iceland from 2004 to 2005. At the average body weight of 1.8 kg, the estimated heritability (h² ± SE) for body weight, gutted weight and the condition factor (CF) were 0.31 ± 0.06, 0.34 ± 0.04 and 0.24 ± 0.06 respectively. Genetic correlation (r_G) in body weight between the two rearing locations was estimated as 0.95, which reflects a low G × E interaction. The estimated heritability for hepatosomatic index (HSI) and fillet yields was 0.061 ± 0.04 and 0.04 ± 0.04 respectively. The HSI and fillet yields were highly genetically correlated with body weight or 0.67 and 0.82 respectively. The genetic correlation between the CF and body weight was estimated as 0.31. There appears to be substantial amount of additive genetic variation for body weight suggesting that selection is likely to be successful. Low heritability for fillet yields and the HSI indicates less promise of genetic improvement. Assigning of parentage to individuals with DNA profiling was 80% successful.     

The effect of stocking density on yield,growth, and survival of Asian river catfish (<Emphasis Type="Italic">Pangasius</Emphasis><Emphasis Type="Italic">bocourti</Emphasis> Sauvage, 1880) cultured in cages

Asian river catfish (Pangasius bocourti Sauvage, 1880) were cultured at five different stocking densities in cages (submerged volume 1 m³) suspended in a dugout pond from August to November 2009. Pangasius bocourti fingerlings (mean weight 27.09 ± 0.54 g) were stocked at densities of 12, 25, 50, 100, and 200 fish m⁻³. At the end of 3 months, the harvest weights (gross yields) were, respectively, 2.05 ± 0.30, 5.20 ± 0.31, 10.60 ± 0.42, 19.98 ± 0.78, and 42.37 ± 0.41 kg m⁻³. The mean fish weights among the stocking densities of 25, 50, 100, and 200 fish m⁻³ were not significantly different, but were significantly higher than that of the 12 fish m⁻³ density. The specific growth rates among high stocking densities of 50, 100, and 200 fish m⁻³ were not significantly different; however, they were significantly higher than those of the low stocking densities of 12 and 25 fish m⁻³. Asian river catfish performed poorly at the lowest density. The results indicate an initial lower stocking threshold for Asian river catfish of above 5.20 kg m⁻³. The Asian river catfish cultured in small cages placed in a pond reached the desirable market size (>200 g) within a 90-day grow-out period. The results show that the maximum yield for Asian river catfish during a 3-month production cycle was not reached.     

Effect of ascorbic acid supplementation on zootechnical performance,haematological parameters and sperm quality of lebranche mullet Mugil liza

The feed provided to breeding fish is one of the main factors influencing the quality of fish gametes. This study was carried out to evaluate the influence of ascorbic acid on growth, haematological parameters and sperm quality of Lebranche mullet males (Mugil liza). Six diets with different levels of ascorbic acid (0; 53; 107; 216; 482 and 708 mg/kg) were tested in triplicate for 75 days. During spermiation (first gonadal maturation), 144 individuals (205.7 ± 11.5 g and 25.7 ± 0.4 cm) were randomly distributed in 18 experimental tanks. Growth parameters were evaluated at the beginning and end of the experiment. Fish blood was collected to analyse glucose, total protein and erythrocyte count (EC) (n = 9). Fish (n = 12) from each treatment were euthanized to determine hepatosomatic index (HSI) and gonadosomatic index (GSI). Semen was collected to evaluate spermatic density, cell membrane integrity and sperm motility. No difference (p > .05) was found on growth parameters, GSI, HSI and total protein. However, EC was lower in fish fed without ascorbic acid (the control group). Ascorbic acid supplementation provided positive effects on sperm characteristics. Fish from treatments with 53 and 107 mg/kg presented the best results for motility time (133.30 ± 4.25 and 135.00 ± 2.77 respectively). Treatments with 107, 216 and 708 mg/kg provided the best results for motility rate (92.0 ± 2.9%, 93.0 ± 5.8% and 93.0 ± 5.8% respectively). Supplementation with 107 and 216 mg/kg provided the best results for plasma membrane integrity (70.12 ± 9.10% and 76.3 ± 3.1% respectively). Lower spermatic density was observed in fish without ascorbic acid supplementation, although no difference was found in sperm density among the treatments with ascorbic acid (p < .05). Considering these results, supplementation of dietary ascorbic acid between 107 and 216 mg/kg optimizes the spermatic quality in males of lebranche mullet.