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Here, we explored cold‐shock‐induced haploid androgenesis in Japanese pufferfish (Takifugu rubripes) and analysed chromosomes by fluorescence in situ hybridization (FISH), single nucleotide polymorphism (SNP) in sex determination locus and microsatellite genotypes to verify all‐male inheritance. We also observed early embryonic development to have an insight into the cytological mechanism of cold‐shock‐induced gynogenesis. The chromosome number of control group was diploid (2n = 44), while in cold‐shock group haploid (1n = 22) and triploid (3n = 66) embryos were detected. Cellular DNA content flow cytometry showed that the rate of haploid induction was 90%. One signal of nucleolar organizing region (NOR) was detected by silver nitrate staining and FISH using rDNA probe in androgenetic haploids from cold‐shock group. SNP analysis revealed that about half of haploid embryos examined had the G genotype indicating male‐specific Y chromosome. Microsatellite genotyping showed that 26 out of 29 haploid embryos from cold‐shock group exclusively inherited paternally derived genotypes. Cytological observation revealed that both second polar body and egg nucleus were located on the blastodisc surface at 60 min post fertilization in cold‐shock group, suggesting simultaneous extrusion of both nuclei.

A 74‐day feeding trial was conducted to investigate the efficacy of n‐6 polyunsaturated fatty acid (n‐6 PUFA) in diets for tiger puffer. Six experimental diets with graded levels of n‐6 PUFA (from 6.01 to 37.98 g/kg) replacing saturated fatty acids (SFAs) were used in this study. Increasing levels of n‐6 PUFA resulted in decreased weight gain (from 472.6% to 349.4%), feed efficiency and crude protein content in fish, but increased lipid accumulation in whole fish and liver. Liver samples from fish fed the lowest and highest level of n‐6 PUFA were subjected to transcriptomic assay. Analysis of the differentially expressed genes showed that high dietary levels of n‐6 PUFA tended to induce glycolysis and triacylglycerol biosynthesis, but reduce synthesis of amino acid and protein, and cause impairment to liver health. The mitochondrial DNA copy number, activity of mitochondrial respiratory chain complex I and glycogen content in the liver, which indicate energy supply status, significantly decreased with increasing levels of dietary n‐6 PUFA. In conclusion, high n‐6 PUFA/SFA ratios in diet led to reduced energy supply (by approximately 50%) and increased hepatic lipid accumulation (by 55%), which probably consequently negatively affect protein synthesis, liver health and growth of tiger puffer.

Cytogenetic analyses of the river puffer, Takifugu obscurus, the tiger puffer, Takifugu rubripes and hybrids produced between female T. obscurus and male T. rubripes and their hybrid triploids (produced by cold shock treatment at 4°C) were performed. T. obscurus had 2n = 44 chromosomes and 1.84 ± 0.019 pg DNA/cell, T. rubripes had 2n = 44 and 2.64 ± 0.015, the hybrids had 2n = 44 and 2.15 ± 0.010 and the hybrid triploids had 3n = 66 and 3.22 ± 0.010. The erythrocyte values of the hybrids were more similar to those for T. obscurus, whereas the hepatocyte, midgut and proximal tubule kidney cell values of the hybrids fell down between those for the parental species (p .05); however, the hybrid triploids had fewer cell nucleus outer layers than the hybrids (p

Although jellyfish blooms are serious nuisances for fisheries and other industries, the utilization and nutritional value of by-catch jellyfish has drawn public attention. Here, we evaluated the efficacy of feeding tiger puffer Takifugu rubripes with moon jellyfish Aurelia sp. using 20-day rearing experiments. Feeding on jellyfish had no positive effect on growth performance, although it significantly reduced neutral lipids and increased proportions of polar lipids, n−3 and n−6 highly unsaturated fatty acids (HUFAs), especially arachidonic and docosahexaenoic acids, and taurine. Supplemental feeding on jellyfish plus pellets elevated activity and responsiveness in the fish, with no effect on burst swimming speed or prevention of aggressive biting by siblings. The present research suggests that feeding on jellyfish improves body composition and some behaviours in tiger puffers. As cultured fish tend to accumulate excessive amounts of lipids from lipid-rich pellets that can induce an unhealthy condition, and behavioural deficits of hatchery-reared fish reduce survival after release in the wild, we propose the use of jellyfish as a supplemental diet in the nursery production of tiger puffer. Moreover, feeding on jellyfish can overcome deficiencies in alternative soy-material feeds by supplying phospholipid, HUFAs, or taurine.

Summary The present study was conducted to evaluate the effects of different dietary chitosan oligosaccharide (COS) levels on digestive enzyme activity, intestinal bacteria flora, and growth performance of the tiger puffer (Takifugu rubripes). Some 600 fish (initial body weight 129.2 ± 3.1 g) were randomly allocated into twelve 2,000-L blue cylinder aquaria with 1,500-L of sea water and divided into four groups (n = 3 aquaria/diet). The fish were fed experimental diets supplemented with different levels of COS (0% [control, L1], 0.05% [LC1], 0.1% [LC2], and 0.2% [LC3]) for 8 weeks. During the experiment, the water temperature was 18 ± 1°C, salinity 28 ± 0.1, pH 8.0 ± 0.1 (mean ± SD, n = 672); airstones were positioned at the bottom of the tank, dissolved oxygen content was above 7.0 mg/L, with a photoperiod of 12 hr light: 12 hr dark. The results showed that dietary COS significantly increased growth performance, intestinal-somatic index (ISI), and intestinal protease as well as lipase activity (p .05). High-throughput sequencing analysis showed that COS affected the richness and diversity of intestinal microbial species. The Shannon index of intestinal bacterial flora was significantly higher in the control group than in other groups (p

In spring of 2011, an epidemic outbreak of torafugu with high mortality occurred in an aquafarm with marine industrial recirculation aquaculture system (MIRAS) in Yantai, Shandong Province, China. The diseased fish showed anorexia, haemorrhaging and festering fin and skin and swelling internal organs. Forty-five dominant bacterial isolates were obtained from the diseased fish, and were found to belong to 12 species according to 16S rRNA gene sequences. One strain from each species was selected to test the pathogenicity, and five strains were showed to be virulent to zebrafish. Whereas Enterovibrio nigricans Fr42 was highly virulent with the LD50 of 7.8 × 104 CFU g−1, Photobacterium swingsii Fr23, Vibrio owensii Fr40, V. harveyi Fr51 and V. rotiferianus Fr71 were moderately virulent with the LD50 of 1.7 × 106 to 8.4 × 106 CFU g−1. Both the bacteria and their extracellular products of the five strains were found to show phospholipase, caseinase, gelatinase, amylase and/or lipase activities. The production of N-acyl homoserine lactones (AHLs) of the five strains was detected by three different AHLs biosensors, and three of them were found to produce AHLs by at least one kind of biosensor. This is the first study describing various opportunistic bacterial pathogens of fish cultured in MIRAS in China.

A method for the production of sterile fish fillet was developed to create a raw fish that can be refrigerated for more than 21 days. Immediately after being killed, tiger puffer fish (Takifugu rubripes) were skinned, gutted and filleted using an aseptic method, and the prepared fillets were immersed in boiling water for 10 s after vacuum packing and were rapidly chilled. This processing method left the fish raw inside. Eleven trials of the production on 11 individuals succeeded in the production of 22 sterile fillets. The products showed no bacterial detection even after storage for 21 days at 4C. The contents of inosine 5′-monophosphate (IMP) of the refrigerated sterile fillets remained at ca. 2,200 to 2,400 nmol/g, while IMP of nonsterile fillets became undetectable. The refrigeration increased the levels of glutamic acid and alanine in the sterile fillets. The products can be safely consumed as a sliced raw fish, sashimi. Practical Applications Raw fish, especially sashimi, is a high-priced delicacy in Japan. However, raw fish is highly perishable, and its shelf life is limited to several days after killing. The method developed for preparing sterile fish fillet is applicable to all kind of fish, and meets the demands of the fish industry for fresh products with extended shelf life. The refrigeration of sterile raw fillet can save energy costs that are high in freezing and frozen storage. Sterile tiger puffer fish fillets can be refrigerated for a long time and can be used as a new ingredient in Western food restaurants that previously have not been able to utilize tiger puffer fish. Tiger puffer fish is served only in restaurants specializing in tiger puffer fish since it is very expensive and perishable.

Takifugu rubripesis teleost fish widely used in comparative genomics to understand the human system better due to its similarities both in number of genes and structure of genes. In this work we survey the fugu genome, and, using sensitive computational approaches, we identify the repertoire of putative protein kinases and classify them into groups and subfamilies. The fugu genome encodes 519 protein kinase-like sequences and this number of putative protein kinases is comparable closely to that of human. However, in spite of its similarities to human kinases at the group level, there are differences at the subfamily level as noted in the case of KIS and DYRK subfamilies which contribute to differences which are specific to the adaptation of the organism. Also, certain unique domain combination of galectin domain and YkA domain suggests alternate mechanisms for immune response and binding to lipoproteins. Lastly, an overall similarity with the MAPK pathway of humans suggests its importance to understand signaling mechanisms in humans. Overall the fugu serves as a good model organism to understand roles of human kinases as far as kinases such as LRRK and IRAK and their associated pathways are concerned.

A 2 × 3 factorial feeding trial was conducted to determine effects of dietary Ca/P ratio and dietary microbial phytase on growth, mineral digestibility and vertebral mineralization in tiger puffer. The treatments consisted of three levels of Ca/P ratios (0.5, 1.0 and 1.5) combined either with phytase (2000 FTU kg−1 diet) or without supplementation, respectively. The Ca/P ratios were achieved by supplementing calcium at 0, 6 and 12 g kg−1 combined with the same level of inorganic P at 5 g kg−1. After a 50-day feeding trial, puffer fish fed the diet at low Ca/P ratio (0.5) together with phytase had significantly higher growth rate and feed intake (FI) than other groups. Both dietary Ca/P ratio and phytase supplement were independent effects on plasma minerals and alkaline phosphatase. Interactive effect between both dietary treatments was observed on P and Zn contents in vertebrae and whole body. P and Zn digestibilities tended to increase with increased Ca/P ratio from 0.5 to 1.0, especially when phytase was supplemented. In conclusion, fish fed a diet with highest Ca/P ratio (1.5) showed the poorest growth performance and nutrients utilization. Dietary Ca/P ratio of 0.5 (without Ca supplement) with 2000 FTU phytase per kg would be the optimum combination in the diet of tiger puffer.

To confirm the toxicity or the tetrodotoxin (TTX) content of cultured puffer fish in China, 565 specimens of cultured puffer fish (Takifugu rubripes, Takifugu fasciatus or Takifugu obscurus, Takifugu flavidus and Takifugu bimaculatus) were collected from five provinces from November 2007 to November 2008. The toxicity of the skin, muscle, liver and testis/ovary was examined using methods based on a mouse assay for TTX. The results indicated that the skin, muscle, liver and testis of specimens were ‘non-toxic’ in both of the mouse assays [

Until recently, the Human Genome Project held centre stage in the press releases concerning sequencing programmes. However, in October 2001, it was announced that the Japanese puffer fish (Takifugu rubripes, Fugu) was the second vertebrate organism to be sequenced to draft quality. Briefly, the spotlight was on fish genomes. There are currently two other fish species undergoing intensive sequencing, the green spotted puffer fish (Tetraodon nigroviridis) and the zebrafish (Danio rerio). But this trio are, in many ways, atypical representations of the current state of fish genomic research. The aim of this brief review is to demonstrate the complexity of fish as a group of vertebrates and to publicize the ‘lesser-known’ species, all of which have something to offer.

Summary The present study determined whether puffer Takifugu rubripes and T. xanthopterus larvae use selective tidal stream transport (STST) for migration into the nursery area. The influence of the tidal cycle on the vertical distribution of Thkifugu larvae was investigated during a 24 h sampling period at one location off Shimabara Peninsula in Ariake Bay. Samples were collected in three depth layers, from near the sea floor to near the surface (5, 20 and 30 m depth). The change in vertical distribution in relation to tidal phase was not observed. This data did not support STST hypothesis. Diel vertical migration was observed irrespective of tidal phase, where larvae migrated to the middle layer during the night, and sank to the bottom layer during the day, however, larvae hardly emerged into the surface layer during the study period. In Ariake Bay, the residual current leads to a layered vertically stratified structure, in which surface water flows towards the mouth and the middle-bottom water flows toward inner part of the Bay. It is suggested that Takifugu larvae use not STST but residual currents for transport into the nursery ground, namely, undergoing nocturnal diel vertical migration in the water column between the middle layer and the bottom layer where the net flow is northward.