Post-molt processes of cuticle formation and calcification in the Japanese mitten crab <Emphasis Type="Italic">Eriocheir japonicus</Emphasis>

This study examines the following in the Japanese mitten crab: (1) the structure of the exoskeleton with special reference to its calcification; (2) the progression of post-molt cuticle formation and calcification. In the crab, the structure and calcification state of the exoskeleton at the molt and during the inter-molt stage were similar to those of other crustaceans. During the inter-molt, the exoskeleton consisted of four cuticle layers; the outermost epicuticle, the exocuticle, the endocuticle and the innermost membrane layer. Intense calcification was observed in the exo- and endocuticle. At the molt, the synthesis of the epi- and exocuticle was already complete, and the addition of the endocuticle began after the molt. Calcification of the exocuticle initiated soon after the molt, but there was a delay between endocuticle matrix synthesis and calcification. Histology showed that the process of calcification was similar to that in other crustaceans. However, calcium concentrations within the exoskeleton continued to increase and never reached the levels of the inter-molt stage at the end of the experiment. This suggests that the Japanese mitten crab is relatively slow to calcify compared to other crustaceans.     

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.     

Guanine-cytosine contents of the host and symbiont cDNA in a symbiotic coral

ABSTRACT:   Hermatypic (reef-building) corals harbor dinoflagellate endo-symbionts Symbiodinium spp. In studying gene expression in such symbiotic corals, problems arise regarding how to distinguish the coral and symbiont mRNA, and how to estimate their fractions in the mRNA population of the holobiont (symbiotic complex of the coral and Symbiodinium cells). In this study, these issues were addressed using juveniles of hermatypic coral Acropora tenuis in symbiosis with Symbiodinium cells of strain PL-TS-1. First, the guanine-cytosine (GC) contents were determined in expressed sequence tags (EST) from PL-TS-1 cells cultured in vitro and symbiont-free larvae of A. tenuis , and their average GC contents were found to be significantly different. The average GC content of the EST from the holobiont was much closer to that of A. tenuis larvae, suggesting that the majority (>90%) of mRNA isolated from the holobiont originated in the host. In protein-coding sequences, little overlap was observed between the GC-content distributions of PL-TS-1 cells and A. tenuis larvae. All of the coding sequences ( n  = 59) found in the A. tenuis EST had GC contents below 0.5, whereas the GC content exceeded 0.5 in the majority (43/44) of coding sequences from the nuclear genome of PL-TS-1 cells.     

Microsatellite marker isolation and cultured strain identification in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.     

Relationship between zooplankton abundance and the early marine life history of juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis> in eastern Hokkaido,Japan

Interannual variations in abundance, timing of outmigration from rivers, growth rate and condition of juvenile chum salmon (Oncorhynchus keta) were studied in the Nemuro Strait (eastern Hokkaido, Japan) during 1999–2002 to establish a possible relationship to zooplankton abundance. The otolith microstructure of juveniles was examined each year in late June to determine their time and size at sea entry (i.e., outmigration), and to estimate the early marine growth rates. Salmon outmigration peaked in mid- or late May, which coincided, in three of the four study years, with the peak release of juveniles into rivers within the study area. Abundance, growth rate and condition of fish were higher in 2001, when—compared to other years—smaller fish experienced higher growth rates, coinciding with greater zooplankton abundance for that year. Our results suggest that high zooplankton abundance positively influenced juvenile chum salmon growth and the condition of the fish during their early marine life despite their small size at sea entry.     

Development of an in vitro culture system for producing eel larvae from immature ovarian follicles in Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To standardize conditions during the final maturation and ovulation of ovarian follicles from Japanese eel, we have developed a culture system for the production of fertilizable eggs from post-vitellogenic ovarian follicles in vitro. Post-vitellogenic ovarian follicles were incubated in culture medium supplemented with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) with or without bovine serum albumin (BSA) to assess the effects of protein concentration. Eggs that ovulated during incubation were fertilized, and the remaining follicles were incubated in prostaglandin F_2α (PGF_2α) for a further 3 or 6 h before fertilization. Male eels were injected repeatedly with human chorionic gonadotropin. The quality of eggs obtained under the different culture conditions was evaluated after artificial fertilization in terms of hatching success. Hatching rates tended to decrease with increasing concentrations of BSA in the incubation medium in a dose-dependent manner. The addition of PGF_2α drastically increased the number of eggs that ovulated, but the rate of hatching was greatly decreased compared with eggs obtained earlier by DHP incubation alone. The larvae obtained from artificially fertilized eggs produced in vitro survived for 14 days without feeding. We conclude that in vitro culture systems thus have a great potential for the acquisition of good quality eggs under tightly controlled artificial conditions, culminating in the production of eel larvae.     

Effect of N-acetyl cysteine and glycine supplementation on growth performance,glutathione synthesis,and antioxidative ability of grass carp, <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

An 8-week feeding trial was conducted to evaluate the effect of N-acetyl cysteine (NAC) and glycine supplementation on growth performance, glutathione (GSH) synthesis, and antioxidative ability of grass carp, Ctenopharyngodon idella. Four practical diets were formulated: control, control + 0.2% NAC, control + 0.5% glycine, and control + 0.2% NAC + 0.5% glycine. Each diet was randomly assigned to quadruplicate groups of 30 fish (approximately 8.8 g). Weight gain and specific growth rate were significantly increased with the supplementation of NAC. Supplementation of NAC plus glycine significantly increased the feed efficiency. Glutathione peroxidase (GPx) and γ-glutamine cysteine synthase (γ-GCS) in plasma were significantly increased with the supplementation of NAC plus glycine. GSH in plasma increased and malondialdehyde (MDA) decreased in fish fed diets supplemented with NAC. Respiratory burst, superoxide dismutase (SOD), and catalase (CAT) activity were not affected by NAC or glycine. These results clearly indicated that NAC improved the growth performance and restored GSH of grass carp, supplemented NAC together with glycine enhanced GSH synthesis, and improved the antioxidative ability of grass carp.     

Effects of mangrove deforestation on fish assemblage at Pak Phanang Bay,southern Thailand

Daytime sampling using a seine net was conducted at Pak Phanang Bay (Nakhon Si Thammarat Province, Thailand) in February and July 2006, to determine differences in fish assemblage structures between a mangrove site and an adjacent site completely cleared of mangroves. The overall numbers of fish species and individuals were significantly higher at the mangrove site than the cleared site in both months. Although benthic crustacean feeders showed more species and individual numbers at the mangrove site in both months, the opposite was found for zooplankton feeders. A cluster analysis, based on the abundance of each species, demonstrated that the fish assemblage structures were distinctly different between the two sites. In addition, significant differences in length frequency distributions for each of the four most abundant species were found between the sites in February and/or July. Small individuals of Scatophagus argus, Ambassis nalua, and Tetraodon nigroviridis were more common at the mangrove site, and of Chelon subviridis at the cleared site. These results suggest that mangrove deforestation exerts marked effects on fish assemblages.     

The development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish in the salmon aquaculture industry

Norway leads the world aquaculture production of Atlantic salmon Salmo salar and farmed Norwegian Atlantic salmon is currently consumed around the globe. However, sea lice infestation is a major problem faced by the salmon aquaculture industry in Norway and elsewhere. The use of wild-caught cleaner fish, mainly wrasses, has been recommended over the other available methods as the most economical and environmentally friendly option to control sea lice infestation in salmon farming. Here, we review the development of the Norwegian wrasse fishery and the use of wrasses as cleaner fish. In this document, we address the sea lice problem and introduce the main wrasse species employed as cleaner fish, document the cleaning behaviour of wrasses, present the development of a new wrasse fishery associated with the salmon aquaculture industry, and finally, we identify the main challenges associated with the intensive use of wild-caught cleaner wrasses and provide some insight for future directions of the wrasse fishery and further development of aquaculture techniques to supply salmon facilities with domesticated cleaner fish.