Protective immunity of sevenband grouper, Epinephelus septemfasciatus Thunberg, against experimental viral nervous necrosis

This paper describes the protective immune responses of sevenband grouper, Epinephelus septemfasciatus Thunberg, immunized with live piscine nodavirus, the causative agent of viral nervous necrosis (VNN), or the Escherichia coli – expressed recombinant coat protein. Nodavirus-neutralizing antibodies were detected at titres ranging from 1:158 to 1:1257 in serum of sevenband grouper which survived intramuscular injection with the virus, by a cell culture assay system. The virus-neutralizing ability of immune serum was also confirmed by injecting virus previously treated with serum into fish. This indicates establishment of acquired immunity in survivors and thus explains why survivors from natural infection are resistant to recurrence of the disease. Young sevenband grouper were immunized twice by intramuscular injections with the recombinant coat protein. Immunized fish produced neutralizing antibodies at high titres for at least 110 days and showed significantly lower mortalities in virus challenge tests. These results suggest the potential for vaccination against VNN in sevenband grouper, which is susceptible to piscine nodavirus at all life-stages.     

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.     

Decadal trends in abundance,size and condition of Antarctic toothfish in McMurdo Sound,Antarctica, 1972–2011

We report the analyses of a dataset spanning 39 years of near‐annual fishing for Dissostichus mawsoni in McMurdo Sound, Antarctica, 1972–2011. Data on total length, condition and catch per unit effort (CPUE) were derived from the > 5500 fish caught, the large majority of which were measured, tagged and released. Contrary to expectation, the length frequency of the McMurdo Sound catch was dominated by fish in the upper two‐thirds of the overall distribution exhibited in the industrial catch for the Ross Sea shelf. Fish length and condition increased from the early 1970s to the early 1990s and then decreased. Fish length positively correlated with Ross Sea ice extent in early spring, a relationship possibly caused by more ice encouraging larger fish to move farther south over the shelf and into the study area. Fish condition positively correlated with the amount of open water in the Ross Sea during the previous summer (Feb), perhaps reflecting greater availability of prey with the higher productivity that more open water brings. Decreasing fish size corresponds to the onset of the fishery, which targets the large individuals. CPUE was constant through 2001 and then decreased dramatically. We hypothesize that this decrease is related to the industrial fishery, which began in the 1996–97 austral summer, and concentrates effort over the ice‐free Ross Sea continental slope. As a result of limited prey choices and close coupling among mesopredators of the region, Antarctic toothfish included, the fishery appears to be dramatically altering the trophic structure of the Ross Sea.     

Present state of imposex in neptune whelk Neptunea arthritica inhabiting shallow waters around Hokkaido, Japan

ABSTRACT:   The state of imposex in Neptunea arthritica from seven sites along the coast of Hokkaido, Japan was examined in 2002 based on the criteria: (i) relative penis size index (RPSI); (ii) imposex frequency; (iii) stage frequency distribution of imposex in adult and immature whelks; and (iv) sex ratio. RPSI differed from site to site, although values from all sites were low (0.186–5.294). In particular, the RPSI values for four sites were very low (<1.0). In sites where immature whelks were also collected, the frequency of imposex was considerably lower in immature (7.7%–55.6%) than in adult whelks (50%–95.2%) except at one site. The imposex stage frequency distribution also differed among sites, and the trend in the adult whelks corresponded with their RPSI value. Female whelks showing severe imposex (Stage 3 and 3+) were restricted to large individuals.     

Evaluation of Cassia fistula meal as a replacement for soybean meal in practical diets of Oreochromis niloticus fingerlings

This study was undertaken to determine the replacement value of Cassia fistula seed meal (CFM) for soybean meal (SBM) in practical diets of Oreochromis niloticus fingerlings. Five practical diets (350 g kg^?1 crude protein) containing 0 g kg^?1 (control), 170 g kg^?1 (diet II), 340 g kg^?1 (diet III), 509 g kg^?1 (diet IV) and 670 g kg^?1 (diet V) substitution levels of CFM for SBM were formulated and fed to triplicate groups of O. niloticus fingerlings (mean initial weight of 10.22 ± 0.03 g) for 70 days. Fish mortality increased linearly with increase in inclusion levels of CFM in the diet. Growth and diet utilization efficiency were depressed in fish fed diets containing CFM at varying inclusion levels. Feed conversion ratio, specific growth rate and protein efficiency ratio of O. niloticus fed on diet containing 170 g kg^?1 substitution level of CFM were similar (P > 0.05) to the control diet. Digestibility of the different diets decreased with increase in inclusion levels of CFM. Fish fed diet containing 670 g kg^?1 CFM had significantly lower carcass protein. However, no significant differences were observed in carcass protein and lipid contents between fish fed the control diets and diet containing 170 g kg^?1 CFM. The most efficient diet in terms of cost per unit weight gain of fish was obtained in 170 g kg^?1 CFM dietary substitution.     

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.     

Effects of various levels of dietary protein on survival,molting frequency and growth of juvenile blue crabs (Callinectes sapidus)

An 18-week study was conducted to examine growth characteristics of the juvenile blue crab (Callinectes sapidus Rathbun) fed various levels of dietary protein. Hatchery raised individuals (157) of identical age (160 days) from the same gravid female were assigned to one of four dietary treatments: adult brine shrimp (Artemia salina Leach) and artificial pelleted diets containing crude protein levels of 23, 37, and 49%.Growth rate, measured as a function of mean weight and mean carapace width, indicated that brine shrimp-fed crabs were larger (P < 0.05) than crabs fed artificial pelleted diets. Also, blue crabs fed either 37 or 49% crude protein were larger than crabs fed the 23% crude protein diet. Molting frequency was greater in the brine shrimp-fed crabs than in those fed the artificial diets. No differences in growth were detected when comparing males vs. females fed the artificial diets. However, females demonstrated greater growth than males within the brine shrimp treatment. Protein concentration of freeze-dried whole blue crab carcasses was greater in brine shrimp-fed crabs than in crabs fed the artificial pelleted diets (P < 0.05).     

Evolutionary implications of two rainbow trout growth hormone genes

The primary structures of two rainbow trout growth hormone mRNAs (GH1 and GH2) have been deduced by direct sequencing of their respective cDNA clones and portions of the mRNA. Both GH1 and GH2 mRNA contain open reading frames comprised of 630 nucleotides and encode 210 amino acid residues of which 11 are variant. The translated regions of both mRNA are flanked by a short but rather conserved 5′-end, and a relatively long but highly diverged 3′-end. The differences at translated and 3′-untranslated regions suggest that the GH1 and GH2 mRNA originate from different loci. The GH1 and GH2 mRNA are likely transcribed from two distinct loci which were duplicated during tetraploidization of salmonid genome between 50 to 100 million years ago. The GH2 gene has been isolated and sequenced from a rainbow trout genomic library. This gene spans a region of approximately 4 kilobases. The trout GH gene is comprised of 6 exons and 5 introns, in contrast to 5 exons and 4 introns in mammals. The additional intron in the trout gene interrupts the translated regions that are analogous to the last exon of the mammalian counterpart. The alleged internally repeating sequences in mammalian GH, prolactin (Pr1) and placental lactogen (PL) are not observed in the predicted polypeptide sequence of trout GH. In addition, direct repeats that flank exons I, III and V of mammalian GH, Pr1 and PL genes are absent in trout gene. These findings indicate that the rainbow trout GH gene structure does not support the current hypothesis that internally repeated regions in GH, Pr1 and PL arose from a small primordial gene.