Putative virulence-related genes in Vibrio anguillarum identified by random genome sequencing

The genome of Vibrio anguillarum strain H775-3 was partially determined by a random sequencing procedure. A total of 2,300 clones, 2,100 from a plasmid library and 200 from a cosmid library, were sequenced and subjected to homology search by the BLAST algorithm. The total length of the sequenced clones is 1.5 Mbp. The nucleotide sequences were classified into 17 broad functional categories. Forty putative virulence-related genes were identified, 36 of which are novel in V. anguillarum, including a repeat in toxin gene cluster, haemolysin genes, enterobactin gene, protease genes, lipopolysaccharide biosynthesis genes, capsule biosynthesis gene, flagellar genes and pilus genes.     

Inhibition of hirame rhabdovirus growth by RNA aptamers

RNA aptamers are artificial nucleic acids that specifically bind to a wide variety of targets. They are an effective tool for pharmaceutical research and development of antiviral agents. Here, we describe four Hirame rhabdovirus (HIRRV)‐RNA aptamers (H1, H2, H3 and H4) that we obtained from an in vitro process called the systematic evolution of ligands by exponential enrichment (SELEX). The HIRRV‐RNA aptamers specifically bind to HIRRV. Hirame natural embryo (HINAE) cells treated with virus and the RNA aptamer showed a decrease in appearance of cytopathic effect when compared with control (treated only with virus). Rhodovulum sulfidophilum was transformed with genes for the RNA aptamers, and the aptamers were detected in the culture medium, indicating that they were secreted from the cells. Thus, the recombinant R. sulfidophilum might be a powerful tool for the prevention of HIRRV in aquaculture.     

Recreational fishery as a conservation tool for endemic Dolly Varden <Emphasis Type="Italic">Salvelinus malma miyabei</Emphasis> in Lake Shikaribetsu,Japan

Recreational angling is a popular leisure activity globally and can be a threat or useful conservation tool depending on the management strategy. Miyabe charr Salvelinus malma miyabei is an endemic fish that inhabits Lake Shikaribetsu, and a management program was established to harmonise recreational fishery and conservation. To examine the suitability of this program, population assessment was conducted with the cooperation of anglers. The population size in 2014 as assessed by tag-and-release angling was estimated to be 105,300 [95% confidence interval (CI) 37,300–178,600], much higher than the reported estimate in 1995. Further, angling mortality was estimated to be quite low. Moreover, no decreasing population trend was detected on analysing 8 years of anglers’ catch data. Consequently, angling has facilitated stock assessments of Miyabe charr under the current program. This case is a good example of recreational angling acting as a conservation tool under appropriate management.     

Evaluation of stock management procedures for walleye pollock in northern waters of the Sea of Japan using a simulation

To evaluate walleye pollock stock management procedures in the northern waters of the Sea of Japan, 30-year population dynamics, including uncertainties, were forecast. Errors in current stock size estimation, variability in future recruitment and changes in future fishing mortalities were incorporated. Results of virtual population analysis (VPA) from resampled catch-at-age data with bootstrap methods was used as the current stock size estimation with uncertainty. Performances of each scenario were evaluated using conservation, utilization, stability and reliability factors. Twenty-two management scenarios and continuing the current fishing mortality were evaluated. Scenarios with minor regulation changes and continuing the current fishing mortality showed poor stock conservation performances. Scenarios with minor regulation changes produced good short-term but poor long-term utilization. Stabilities were poor in continuing the current fishing mortality and fishing ban scenarios. Reliability in all scenarios after 30 years was smaller than in continuing the current fishing mortality; however, small differences among scenarios were observed. The simulation results indicated that multilateral assessment is needed to evaluate the management candidates. Uncertainty caused by recruitment variability mostly affected future population dynamics. The role of simulations in the production of effective scientific advice is discussed.     

Effect of bleeding on the quality of amberjack Seriola dumerili and red sea bream Pagrus major muscle tissues during iced storage

The effect of bleeding on white muscle quality in amberjack and red sea bream was evaluated by measuring ATP-related compounds, volatile basic nitrogen (VBN), and the trimethylamine (TMA) content. The freshness was evaluated by the K value, and the degree of spoilage was elucidated by VBN and TMA. ATP was rapidly converted to ADP and AMP in the muscle and IMP was the main product of ATP degradation during iced storage. For both species, the IMP content was higher in the muscles of fish that were bled than in those unbled during iced storage. Conversely, the K value and the levels of hypoxanthine (Hx) and VBN were higher in the muscle tissue of unbled fish than in the bled tissue of both species. Similarly, the TMA content was higher in the unbled muscle tissues of both species after a week of storage.     

Effects of glutathione treatments during sperm washing and in vitro fertilization on the in vitro early development of embryos of Japanese Black cattle

This study was conducted to examine the effects of adding glutathione (1 mM) to media used for sperm washing and in vitro fertilization (IVF) on the improvement of early development of embryos produced using cryopreserved spermatozoa of the less IVF-competent bull (the one considered unqualified as spermatozoa supplier for the production of bovine blastocysts using IVF). The cryopreserved spermatozoa of this bull were characterized by normal motility and lower ATP content and blastocyst productivity than those of IVF-competent bulls. The addition of glutathione to the sperm washing medium was more effective in improving the productivity of blastocysts and ATP content than the addition of glutathione to the IVF medium or no glutathione addition at all (control). These results suggest that this simple method may be used to improve the potential of cryopreserved spermatozoa of less IVF-competent bulls to fertilize oocytes in vitro and to induce normal embryonic development after fertilization.     

Age estimation of the wild population of Japanese mantis shrimp <Emphasis Type="Italic">Oratosquilla oratoria</Emphasis> (Crustacea: Stomatopoda) in Tokyo Bay,Japan, using lipofuscin as an age marker

ABSTRACT:   In this study, the age composition of the Japanese mantis shrimp, Oratosquilla oratoria , in Tokyo Bay, central Japan, was investigated using lipofuscin, an autofluorescent pigment, as an age marker. Lipofuscin in a histological section of protocerebral bridge cell mass (PBCM) in the brain was identified by confocal microscopy, and its concentration was quantified by image analysis. Modal analysis of the lipofuscin concentration showed four regularly-spaced modes that could each be regarded as a distinct age group. This implied a constant lipofuscin accumulation in PBCM at a 6.5 × 10⁻²% volume fraction per year; it also implied the existence of individuals that are at least 4 years old. The lipofuscin concentration was found to be a more appropriate index than body length for estimating the age of O. oratoria , because the modal analysis on the body–length histogram failed to detect apparent age groups. This was probably a result of the declining growth rate with age and the individual variations in growth through molting leading to considerable overlap in the size between different age groups. The lipofuscin analysis suggested that fast-growing individuals in each cohort are recruited to the fishery, and most individuals attain the exploitable size between 2 and 3 years of age.     

Microbial communities in various waters used for fish larval rearing

A major concern in larvae production is a mass mortality caused by fish diseases. In larvae production, pumped‐up natural seawater filtered through a sand filter system is used for fish rearing, and microalgae and rotifer cultures. Here, we investigated the community structures of eukaryotic microbes, as well as total bacteria and vibrios, in various processed ‘waters’ used in a larvae production site. We observed that ultraviolet irradiation of seawater was effective to reduce not only total bacteria and vibrios but also eukaryotic microbes. Moreover, the community structures of total bacteria and vibrios in rearing waters for fish larvae were different from those in rotifer cultures fed with Chlorella, but rather similar to those in natural seawater and microalgae cultures. These results suggest that the bacterial community in rearing waters may originate mainly from natural seawater and then be selected by microalgae in rearing water. Overall, this study provides useful information for avoiding the risk of fish disease outbreaks in a larvae production site.     

Universal PCR primers for ribosomal protein gene introns of fish

Human ribosomal protein (RP) gene sequences with respect to intron/exon structures and corresponding cDNA or genomic data of fish species were obtained from the GenBank database. Based on conserved exon sequences, 128 primer pairs for 41 genes were designed for exon-primed intron-crossing (EPIC) polymerase chain reaction (PCR). In reference to the draft genome sequences of the Pacific bluefin tuna (Thunnus orientalis), 12 primer pairs expected to amplify introns of the bluefin tuna with lengths of 500–1000 bp were selected and applied to six distantly related fish species belonging to the Orders Clupeiformes, Tetraodontiformes, Pleuronectiformes, Perciformes, Scorpaeniformes, and Anguilliformes. PCR amplification was observed for at least four species in each primer pair, and all fragments were larger than those expected for intronless amplification. Single fragment amplification was observed for at least seven primer pairs per species. Fragment sizes of the bluefin tuna for nine primer pairs corresponded to those expected from the genomic data. Thus, our primer pairs are potentially applicable to a wide variety of fish species and serve as an initial step for isolating single-copy nuclear DNA sequences.     

Characterization of ethylene biosynthesis and its regulation during fruit ripening in kiwifruit,Actinidia chinensis ‘Sanuki Gold’

Ethylene biosynthesis in kiwifruit, Actinidia chinensis ‘Sanuki Gold’ was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwifruit. Treatment of fruit with more than 5 μL L^?1 of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days.