Segregation of microsatellite loci in second generation hybrid striped bass (<Emphasis Type="Italic">Morone saxatilis</Emphasis> × <Emphasis Type="Italic">Morone chrysops</Emphasis>)

Within the framework of a larger project aimed to assess the potential of second generation hybrid striped bass for German aquaculture the genotypic segregation of five microsatellite loci was analysed in two progeny lots (n = 74 and 76, respectively). There was no consistent correlation between microsatellite genotypes and phenotypic category (white bass, hybrid, or striped bass). None of the individuals expressed neither only white bass nor only striped bass genotypes at all five loci. On the other hand, only hybrid genotypes at all five loci were detected in three individuals of lot 1 and four individuals of lot 2. Single loci tests for conformity of microsatellite genotypic segregation with Mendelian rules revealed significant deviations (P < 0.05) in four cases for lot 1 and in three cases for lot 2. If pooled over all five loci, both lots displayed highly significant deviations (P < 0.01) with an excess of hybrid genotypes and a deficiency of white bass genotypes. It is concluded that stabilizing selection performed on hybrid genotypes might be a suitable approach for practical aquaculture in Europe if the goal is to become independent of first generation hybrid fry supply and/or if establishing domesticated brood stocks of both parental species is impossible. However, more detailed studies on the characteristics and performance of multiple hybrid generations are needed.     

Control of lipid oxidation and meat color deterioration in skipjack tuna muscle during ice storage

The present study investigated the effects of lipid oxidation on quality deterioration in the ordinary and dark muscles of skipjack tuna Katuwonus pelamis during the early stages of ice storage for 72 h. The lipid hydroperoxide content of the dark muscle was significantly higher (P < 0.01) than that of the ordinary muscle throughout 72 h of ice storage. The metmyoglobin content of the ordinary muscle gradually increased, and was accompanied with darkening in the fish meat color. On the other hand, the addition of sodium ascorbate or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox^®) to the ordinary muscle of skipjack tuna significantly inhibited the formation of lipid hydroperoxide as well as metmyoglobin formation. Thus, a decrease in α-tocopherol content in the ordinary muscle with antioxidant addition was not observed during ice storage period. In conclusion, the rate of lipid oxidation of skipjack tuna ordinary muscle is closely related to metmyoglobin formation, and the addition of antioxidants to fish meat is effective at inhibiting lipid oxidation as well as myoglobin oxidation in post-mortem meat.     

Production of androgenetic amago salmon<Emphasis Type="Italic"> Oncorhynchus masou ishikawae</Emphasis> with dispermy fertilization

With the aim of improving artificial androgenesis in teleost fishes, we tested two methods for producing androgenetic diploids of amago salmon (Oncorhynchus masou ishikawae), namely, fertilization of gamma-ray irradiated eggs with fused spermatozoa (sperm-fusion method) and the fertilization of irradiated eggs with untreated sperm followed by the blocking of cell division (mitosis-inhibition method). Our results showed that the optimal condition for sperm fusion was to treat the sperm with 50% polyethylene glycol (molecular weight 7500) for 100 s. The efficiency of the two methods of androgenesis was compared in terms of fertilization rate, hatching rate, and larval survival after hatching. The rate of fertilization was lower with the sperm-fusion method than with the mitosis-inhibition method, but the reverse was true for the hatching rate. The survival rate of hatched larvae was the same with the two methods. Androgenesis was confirmed with a recessive albino color marker, and all viable offspring were found to be heterozygous based on analysis of the microsatellite markers. Our results suggest that androgenesis with the sperm-fusion method is a promising approach with potential applications in both aquaculture breeding programs and the preservation of endangered freshwater fishes.     

Weaning of the wedge sole <Emphasis Type="Italic">Dicologoglossa cuneata</Emphasis> (Moreau): influence of initial size on survival and growth

The weaning phase can be decisive in fish-culture viability. In this work, the relationship between the initial size and weaning success has been studied in wedge sole (Dicologoglossa cuneata). For each age (30, 50, and 70 days after hatching, DAH), two to three sizes were selected, and all were put on the same feeding schedule for 20 days. Each batch (three replicates) was sampled at 1, 10, and 20 days. Specific growth rate (SGR) and survival were compared at the end of the co-feeding period, after 10 days on dry feed only. The best results for survival and growth were found with the smallest larvae, and vice versa. The SGRs and survival rates recorded during the co-feeding period were higher (0.8–15.6 day⁻¹ and 68.3–97.8%) than those from the dry-food phase (0.9–4.7 day⁻¹ and 56.3–66.7%). Successful weaning (survival = 65% and SGR = 9.3 day⁻¹) is possible with 30 DAH larvae (7.6–8.1 mm and 3.9–4.6 mg). In conclusion, the most effective weaning would be possible at 30 DAH, implying significant Artemia savings (25–50%).     

Effects of three culture densities on growth and survival of <Emphasis Type="Italic">Octopus vulgaris</Emphasis> (Cuvier, 1797)

The purpoase of this research was to test the effects of three culture densities on the growth and survival of Octopus vulgaris. A total of 141 sub-adult octopuses (1,175.4 ± 194.9 g) were randomly distributed in nine tanks of 2,000 l each (3.6 m × 1.1 m, and 50 cm water depth). Three tanks were stocked with a low initial density of 4 kg/m³, while three other tanks were stocked with an initial density of 8 kg/m³, and the remaining three tanks were stocked at an initial density of 15 kg/m³. Octopuses were all fed frozen squid (Loligo gahi) at 5% body weight per day (%BW/day). The experiment lasted for 70 days. Water temperature varied between 20 ± 2°C, and salinity varied between 36 ± 1 ppt. During the entire experiment, dissolved oxygen was always >75%, and ammonia was always lower than 0.1 mg/l. No differences in growth or growth rates (between 0.9 and 1.1%BW/day for the three densities) were found. Nevertheless, mortality was significantly lower for the low density compared to the other two densities tested. Maximum densities in the culture tanks (>25 kg/m³) were attained in the higher culture densities after 56 days of the experiment.     

Growth comparison of Asian Nile and red tilapia strains in controlled and uncontrolled farm conditions

Growth of several genetically improved Nile tilapia (GIFT or Genetically Improved Farmed Tilapia, FaST or Freshwater Aquaculture Center Selected Tilapia, SEAFDEC-selected) and domesticated red tilapia (BFS or Binangonan Freshwater Station, NIFI-red or National Inland Fisheries Institute red, HL or Hacienda Luisita) stocks were compared in controlled (tank) and uncontrolled farm conditions (lake-based cages) with unselected NIFI or Chitralada Nile tilapia as control. Specific growth rate differed significantly (P = 0.009) in tank-reared Nile tilapia stocks where GIFT grew best at 1.358%/day followed by FaST (1.307%/day), control stock NIFI (1.257%/day) and SEAFDEC-selected (1.202%/day). Genetic effect explained 84.4% of the variance in growth of Nile tilapia in tanks. Although Nile tilapia growth in cages followed the same trend where GIFT grew best at 1.570%/day, no significant stock differences (P = 0.479) were noted. Meanwhile, red tilapia reared in either tanks or cages showed no significant stock differences in terms of growth. However, survival of the red tilapia stocks in cages differed significantly with HL having the highest percentage survival at 93.3%. The different growth responses of the Nile tilapia stocks especially under controlled (tank) farm conditions were largely due to genetic factors (stock differences).Under uncontrolled farm conditions, environmental factors were generally observed to have also affected the survival and to some extent, the growth of Asian Nile and red tilapia stocks.     

Effect of trawling with traditional and ‘T90’ trawl codends on fish size and on different quality parameters of cod <Emphasis Type="Italic">Gadus morhua</Emphasis> and haddock <Emphasis Type="Italic">Melanogrammus aeglefinus</Emphasis>

The effect of trawling on fish size and on different quality parameters of cod (Gadus morhua) and haddock (Melanogrammus aeglefinus) was evaluated after conducting 16 valid hauls using two trawls in a double rig fitted with a traditional and a novel ‘T90’ codend, respectively. The total catch volume during the fishing period was 47.6 metric tons, with an average catch per codend of 1.5 (range 0.5–2.9) tons. The mean haul duration was 5 h. The catch was assessed according to fish size, mortality, external damage, initial white muscle pH and development of rigor mortis. Fillet quality (colour, blood spots, gaping) was assessed after 1 week of freeze-storage. Our results showed there was no difference between the two types of nets in terms of catch volume, but significantly slightly bigger fish were caught with T90 than with the traditional trawl net (p < 0.05). Haddock caught with the traditional trawl net had more external injuries related to the trawl gear than haddock caught with the T90 gear (p < 0.05). The gaping frequency for cod caught with the traditional trawl net tended to be higher than cod caught with the T90 gear, but the difference was not significant (p = 0.07). No other differences in fish quality between fish caught in the trawl nets were observed.     

Relative potencies of natural estrogens on vitellogenin and choriogenin levels in the Indian freshwater spotted snakehead, <Emphasis Type="Italic">Channa punctata</Emphasis>: in vivo and in vitro studies

The relative efficacies of three natural estrogens viz., estrone (E₁), estradiol-17β (E₂) and estriol (E₃) to induce synthesis of vitellogenin (Vg) and choriogenin (Chg) were assessed in primary hepatocyte cultures of the Indian freshwater spotted snakehead, Channa punctata. Hepatocytes were isolated from the spotted snakehead liver by a non-enzymatic protocol. Optimum culture conditions were standardized for ensuring their viability and functioning. Isolated hepatocytes were cultured for 48 h for monolayer formation and then exposed to various concentrations (0.001–10 μM) of the three estrogens. Competitive homologous ELISAs, developed and validated for spotted snakehead Vg and Chg were employed to determine the amounts of these two proteins secreted into the culture medium after 48 h of incubation. The results reveal that although all the three estrogens were effective in inducing the production of Vg and Chg in a dose-dependent manner, there were differences in their relative potencies. Of three estrogens, E₁ was the least potent and could induce synthesis of Vg and Chg only at a minimum concentration of 0.5 μM; whereas significant levels of both the proteins were quantified in culture medium by exposing the hepatocytes to E₂ or E₃ even at a concentration of 0.001 μM. All three estrogens were effective in inducing synthesis of Vg and Chg in vivo also. These results suggest the possibility of employing the above in vitro experimental design to monitor the presence of estrogens/estrogen-like chemicals in natural waters, which could interfere with the estrogen receptor system of fish. This study further points to the possibility of using Chg, in addition to Vg, as a parameter for screening various chemicals for their estrogenic activity.     

Cloning,expression, and induction by 17-β estradiol (E<Subscript>2</Subscript>) of a vitellogenin gene in the white cloud mountain minnow <Emphasis Type="Italic">Tanichthys albonubes</Emphasis>

Vitellogenins (Vtgs), the precursors for the yolk proteins, are very important for the embryonic development of teleosts, and have also been studied extensively as biomarkers for environmental estrogenic mimics. The cDNA for a Vtg was isolated from the liver of the female white cloud mountain minnow (Tanichthys albonubes) by 3′- and 5′-RACE methods. It is 4,171 bp in full length, and encodes a putative protein of 1,326 amino acids. This putative Vtg, designated as wcmmVtg, contains complete portions of LVI and PV, but lacks the C-terminal half of LVII and thus belongs to type I vitellogenin. In addition to the liver of the female fish, wcmmVtg was also shown to be expressed in the ovary. During ovarian development, the mRNA expression of wcmmVtg in both the liver and ovary was continuously increased from the previtellogenic to late vitellogenic stages, but then decreased significantly at post-spawning stage. In the male fish, expression of wcmmVtg mRNA was induced in the liver by treatment with E₂ (10 and 100 ng/l) for 14 days. These results suggest that the Vtg originated from the ovary of the white cloud mountain minnow may also contribute to the accumulation of yolk proteins during oocyte growth, and that the male white cloud mountain minnow is sensitive to the estrogenic treatment in terms of Vtg mRNA expression, which could also be applied to monitor the environmental estrogenic mimics.     

Identification of oyster-derived hypotensive peptide acting as angiotensin-I-converting enzyme inhibitor

Angiotensin-I-converting enzyme (ACE) plays a crucial role in the crisis of hypertension. Some peptides that originate from protease hydrolysates are known to suppress ACE activity in vitro and in vivo. Here, we investigated whether trypsin hydrolysate of oyster Crassostrea gigas showed hypotensive activity and ACE inhibition. The hydrolysate significantly suppressed systolic blood pressure and ACE activity in spontaneously hypertensive rats following a one-shot oral administration and a long-term feeding experiment lasting 9 weeks. Each hydrolysate from oyster tissue showed ACE inhibitory activity, indicating the hypotensive effect was due to synergism. One potent ACE inhibitory peptide, Asp-Leu-Thr-Asp-Tyr, was identified from the hydrolysate of the striate muscle, and the peptide exhibited hypotensive activity in vivo. Protease digestion analysis suggested that Asp-Tyr could be the real effector of this penta-peptide in vivo.