Lipid characteristics of coastal migratory Sarda orientalis tissues

To characterize the 22:6 n-3 (docosahexaenoic acid, DHA) levels of the oriental bonito Sarda orientalis, a coastal migratory tuna species, total lipids (TL) of white muscle, dark muscle, liver, pyloric cecum, gonad, and other viscera were separated into lipid classes, the constituents of TL were quantified, and the fatty acid composition of TL, triacylglycerols (TAG), phosphatidylethanolamine, and phosphatidylcholine was analyzed. The crude lipid contents of muscle and other organs were 0.8–3.2% and 2.3–9.1%, respectively. DHA was found in TL of various organs at levels ranging from 19.2% to 27.6% in muscle and 16.3% to 28.5% in other organs. The levels of DHA in muscle TAG (8.2–16.0%) were lower than or comparable to those in visceral TAG (6.9–24.0%). These findings did not coincide with those observed in active-migratory tuna species, which accumulate DHA in their muscle TAG during migration. These findings suggest that the DHA distribution of S. orientalis is different from that observed in active-migratory tuna species, and that the differences may be due to migration type of the fish.     

Faster growth before metamorphosis leads to a higher risk of pseudoalbinism in juveniles of the starry flounder <Emphasis Type="Italic">Platichthys stellatus</Emphasis>, as suggested by otolith back-calculation

The mechanism underlying malformation during flatfish metamorphosis is not clear. Here, the relationship between growth before metamorphosis and the occurrence of malformation after metamorphosis was examined in individuals reared in a single tank. Otoliths of starry flounder Platichthys stellatus larvae were labeled with alizarin complexone before metamorphosis [12 days after hatching (DAH)]. The positive linear regression between the lapillus otolith diameter and body length at 12 DAH enabled the estimation of individual larval size after completion of metamorphosis. At 12 DAH, the average body length of the typical pseudoalbino juveniles was significantly greater (5.37 mm) than that of the normal juveniles (5.13 mm) (P < 0.01, n = 100). In addition, of the larvae that were estimated to measure less than 4.8 mm at 12 DAH, approximately 80% metamorphosed into normal juveniles, whereas more than 60% of the larvae that measured more than 5.4 mm became pseudoalbino juveniles. These results indicate that the larvae that grow rapidly during the first 12 days are more likely to become pseudoalbino individuals after metamorphosis.     

Hen egg yolk and skinned krill as possible foods for rearing leptocephalus larvae of Anguilla japonica Temminck & Schlegel

Usual diets for rearing leptocephalus larvae of Japanese eel Anguilla japonica include eggs of the endangered spiny dogfish Squalus acanthias (SE). We investigated the effects of alternative food materials, hen egg yolk (HEY) and exoskeleton‐free (skinned) Antarctic krill (SAK), on the growth and survival of eel larvae. We found that feed comprising whole krill including exoskeleton (WAK) containing higher levels of fluoride (37.89 mg kg^?1) was acutely toxic to eel larvae exposed to this alone. In contrast, extract from SAK containing lower concentrations of fluoride (4.25 mg kg^?1) showed no apparent adverse effects. Growth of larvae fed a mixture of SE and SAK in a feed trial of 58 days [mean body weight (BW), 6.0 mg] was about twofold higher than that of larvae fed a mixture of SE and WAK (3.2 mg) (P < 0.01). A mixture of HEY and SAK also had some dietary benefits for eel larvae, enabling them to survive for up to 58 days and to grow significantly (mean BW, 2.4 mg), compared with their initial weight (mean BW, 0.2 mg) (P < 0.001). Although additional nutritional improvements are needed, the present results suggest that combination diet HEY and SAK may be a good alternative to SE as an effective diet for eel larvae.     

Changes in the immunostaining intensities of follicle-stimulating hormone and luteinizing hormone during ovarian maturation in the female Japanese flounder

The role of gonadotropin (GTH) in the reproduction of the Japanese flounder, Paralichthys olivaceus, was studied by assessing the changes in the apparent activity of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in the pituitary gland during gonadal maturation by immunohistochemical analyses. Corresponding changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α,20β-dihydroxy-4–pregnen-3-one (DHP) were also studied. Reared fish at the early spawning to termination stages were sampled from May to August and wild fish at the previtellogenic to termination stages were caught at 3- to 4-week intervals between April and September offshore from the northern mainland of Japan by gill nets. The gonadosomatic index of the reared fish decreased from the early spawning stage to the termination stage, while that of the wild fish increased significantly from the previtellogenic stage to the early spawning stage and decreased thereafter. In the reared fish, the immunostaining intensities of FSH and LH were high during the spawning period, accompanied by high plasma levels of T, E₂, and DHP. In the wild fish, the immunostaining intensities of FSH and LH were low during the previtellogenic stage but increased during the maturing and spawning stages. These results indicate that both FSH and LH are likely associated with oocyte maturation in the Japanese flounder.     

Egg collection from hatchery-reared broodstock of spotted halibut Verasper variegatus treated with LHRH analog

The objective of this study is to establish a method for acquiring large quantities of high-quality eggs for artificial fertilization from hatchery-reared broodstock of spotted halibut Verasper variegatus. We estimated the optimum conditions for implantation of luteinizing hormone-releasing hormone analog (LHRHa) cholesterol pellet (LHRHa-CP) to induce ovulation from two experiments which focused on the dose and the timing combined with monitoring of oocyte development by ovarian cannulation. From the results of cannulation, vitellogenesis of hatchery-reared broodstock was suggested to occur normally, although final oocyte maturation was not initiated in the first clutch of oocytes. In late December, oocytes in the most advanced clutch still underwent vitellogenesis, having diameter of about 0.82?mm. LHRHa-CP implantation during this period had no remarkable effects, except for administration at high dose (100???g/kg). In contrast, in mid January, when oocyte diameter reached about 0.95?mm, ovulation occurred in most individuals, even at low dose (20???g/kg). In mid February atretic oocytes became remarkable and LHRHa-CP implantation showed much lower performance in terms of egg quality. The diameter of growing oocytes converged to about 0.95?mm, which was that of fully grown postvitellogenic oocytes. Thus, oocyte diameter is suggested to be an effective indicator to estimate the timing of LHRHa-CP implantation.     

Development of cleaved amplified polymorphic sequence (CAPS)-based markers for identification of sweetpotato cultivars

To develop a simple and reliable method to identify sweetpotato cultivars, we designed cleaved amplified polymorphic sequence (CAPS)-based markers and used them to perform genotyping of Japanese sweetpotato cultivars. In order to screen the CAPS-based markers, 13 primer pairs were designed from the exon sequences of 11 sweetpotato genes to amplify fragments containing an intron. By digesting the amplified products with 8 restriction enzymes having different recognition sites, a total of 27 polymorphic marker fragments were obtained. Genotyping of 60 Japanese sweetpotato cultivars using these markers suggested that the markers can effectively distinguish sweetpotato cultivars. Among the genes used for primer design, the gene encoding the dihydroflavonol 4-reductase (DFR) showed the largest degree of polymorphism. To our knowledge, this is the first report on the development of CAPS-based markers in sweetpotato.     

Evaluation of a new eastern blotting technique for the analysis of ginsenoside Re in American ginseng berry pulp extracts

A new eastern blotting technique has been established for ginsenoside Re (G-Re) contained in American ginseng berry pulp extracts. G-Re in American ginseng berry pulp was extracted using 100% methanol, 100% ethanol, 50% aqueous methanol, and 50% aqueous ethanol. The combined crude extracts were applied onto a polyethersulfone membrane and developed using the methanol–water–acetic acid solvent system (45:55:1 v/v). Separated components were immunostained using anti-G-Re monoclonal antibody. G-Re was first specifically detected and then quantitatively analyzed using NIH Imaging software. We also confirmed that the most suitable solvent was 50% aqueous methanol for extracting G-Re from American ginseng berry pulp.     

3-Prenyl luteolin,a new prenylated flavone with melanin biosynthesis inhibitory activity from wood of Artocarpus heterophyllus

In our efforts to find new whitening agent from natural resources, we focused on wood of Artocarpus heterophyllus which shows anti-melanogenesis activity. By activity-guided fractionation of A. heterophyllus wood extract, a new prenylated flavonoid, 3-prenyl luteolin (1) was isolated. The IC₅₀ of mushroom tyrosinase inhibitotry activity of 1 was 76.3 µM. The results of the comparison with that of luteolin showed the prenyl substituent at C-3 position of 1 play an important role for revealing tyrosinase inhibition. In melanin formation inhibition on B16 melanoma cells, IC₅₀ of 1 was 56.7 µM with less cytotoxicity.     

Ontogenetic changes in water-use efficiency (δC) and leaf traits differ among tree species growing in a semiarid region of the Loess Plateau,China

On the Loess Plateau, China, several planted tree species such as Populus hopeinsis and Robinia pseudoacacia suffer occasional diebacks in the top shoots, reducing growth rates after maturation. However, this does not usually occur in other species, e.g., Ulmus pumila and Zizyphus jujuba. We compared stable carbon isotope ratios (δ¹³C) as indicators of leaf water-use efficiency (WUE), leaf mass per unit area (LMA), and leaf area- and mass-based nitrogen content (N_area and N_mass) by tree height (1, 5, and 10 m) in P. hopeinsis, R. pseudoacacia, U. pumila, and Z. jujuba. In P. hopeinsis and R. pseudoacacia, leaf δ¹³C and N_area were significantly lower in 10-m trees than in 1-m saplings, indicating that leaf WUE and photosynthetic rates of both species decreased with tree height. In contrast, δ¹³C in Z. jujuba varied little with tree height. The δ¹³C of 10-m-tall U. pumila trees was significantly higher than that of 1-m plants, demonstrating an increase in WUE with tree height. Decreasing WUE, leaf N_area, and N_mass with height increases in P. hopeinsis and R. pseudoacacia may be related to water and nutrient limitations for these species in semiarid regions. In contrast, stable or increasing δ¹³C with tree height in U. pumila and Z. jujuba may account (to some extent) for successful production of these species under identical environmental conditions. Diameter growth rate also decreased with maturity in P. hopeinsis and R. pseudoacacia, but increased or was stable for U. pumila and Z. jujuba. The differences in leaf WUE and LMA among species with tree maturity may be related to species’ growth patterns and susceptibility to drought stress, and are likely to be important new criteria for plantation species selection on the Loess Plateau.