Quantitative loop-mediated isothermal amplification method for the detection of <Emphasis Type="Italic">Vibrio nigripulchritudo</Emphasis> in shrimp     

Jean Fall  Gunimala Chakraborty  Tomoya Kono  Minoru Maeda  Yoshihiro Suzuki  Toshiaki Itami  Masahiro Sakai 《Fisheries Science》2011,77(1):129-134

Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10² copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/μl. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T _t) versus the log of bacterial number. A high correlation coefficient (R ² = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.  相似文献   

Control of lipid oxidation and meat color deterioration in skipjack tuna muscle during ice storage     

Jeong-Ho Sohn  Toshiaki Ohshima 《Fisheries Science》2010,76(4):703-710

The present study investigated the effects of lipid oxidation on quality deterioration in the ordinary and dark muscles of skipjack tuna Katuwonus pelamis during the early stages of ice storage for 72 h. The lipid hydroperoxide content of the dark muscle was significantly higher (P < 0.01) than that of the ordinary muscle throughout 72 h of ice storage. The metmyoglobin content of the ordinary muscle gradually increased, and was accompanied with darkening in the fish meat color. On the other hand, the addition of sodium ascorbate or 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox^®) to the ordinary muscle of skipjack tuna significantly inhibited the formation of lipid hydroperoxide as well as metmyoglobin formation. Thus, a decrease in α-tocopherol content in the ordinary muscle with antioxidant addition was not observed during ice storage period. In conclusion, the rate of lipid oxidation of skipjack tuna ordinary muscle is closely related to metmyoglobin formation, and the addition of antioxidants to fish meat is effective at inhibiting lipid oxidation as well as myoglobin oxidation in post-mortem meat.  相似文献   

Extractive components and taste aspects of fermented fish pastes and bean pastes prepared using different koji molds as starters     

Giri  Anupam  Osako  Kazufumi  Ohshima  Toshiaki 《Fisheries Science》2009,75(2):481-489

We have evaluated four commercially available mold starters (SP-01, NY, M1, and “for kome miso”) for fermented fish pastes with the aim of determining the effect of different koji molds on the nutritional and sensory characteristics of the fermented product. Comparative studies among different fermented fish and bean pastes were also conducted. Protein contents were considerably higher in the fermented fish pastes (22.7–24.1%) than the fermented bean pastes (8.2–12.8%). Free amino acids, oligopeptides, organic acids, mineral content, and sensory scores of the fermented fish pastes were superior to those of the bean pastes in terms of nutritional aspects. Sensory evaluation of fermented fish and bean pastes clearly indicated that different koji molds used as starters had little effect on the taste of the fermented fish pastes. However, in terms of nutritional and sensory characteristics, fermented fish pastes from trash fishes were found to be both commercially valuable and a potential gain to food processing firms among the many new generic fermented food products becoming available.  相似文献   

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo     

Md. Shaheed Reza  Nanami Mizusawa  Ayaka Kumano  Chiharu Oikawa  Daisuke Ouchi  Atsushi Kobiyama  Yuichiro Yamada  Yuri Ikeda  Daisuke Ikeda  Kazuho Ikeo  Shigeru Sato  Takehiko Ogata  Toshiaki Kudo  Mitsuru Jimbo  Ko Yasumoto  Kazutoshi Yoshitake  Shugo Watabe 《Fisheries Science》2018,84(3):563-577

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.  相似文献   

Molecular physiological aspects of chalking mechanism in rice grains under high-temperature stress     

Toshiaki Mitsui  Hiromoto Yamakawa  Tohru Kobata 《Plant Production Science》2016,19(1):22-29

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Vitellogenesis and choriogenesis in fishes     

Akihiko Hara  Naoshi Hiramatsu  Toshiaki Fujita 《Fisheries Science》2016,82(2):187-202

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Preparation of Photocrosslinked Fish Elastin Polypeptide/Microfibrillated Cellulose Composite Gels with Elastic Properties for Biomaterial Applications     

Shinya Yano  Megumi Mori  Naozumi Teramoto  Makoto Iisaka  Natsumi Suzuki  Masanari Noto  Yasuko Kaimoto  Masashi Kakimoto  Michio Yamada  Eri Shiratsuchi  Toshiaki Shimasaki  Mitsuhiro Shibata 《Marine drugs》2015,13(1):338-353

Photocrosslinked hydrogels reinforced by microfibrillated cellulose (MFC) were prepared from a methacrylate-functionalized fish elastin polypeptide and MFC dispersed in dimethylsulfoxide (DMSO). First, a water-soluble elastin peptide with a molecular weight of ca. 500 g/mol from the fish bulbus arteriosus was polymerized by N,N′-dicyclohexylcarbodiimide (DCC), a condensation reagent, and then modified with 2-isocyanatoethyl methacrylate (MOI) to yield a photocrosslinkable fish elastin polypeptide. The product was dissolved in DMSO and irradiated with UV light in the presence of a radical photoinitiator. We obtained hydrogels successfully by substitution of DMSO with water. The composite gel with MFC was prepared by UV irradiation of the photocrosslinkable elastin polypeptide mixed with dispersed MFC in DMSO, followed by substitution of DMSO with water. The tensile test of the composite gels revealed that the addition of MFC improved the tensile properties, and the shape of the stress–strain curve of the composite gel became more similar to the typical shape of an elastic material with an increase of MFC content. The rheology measurement showed that the elastic modulus of the composite gel increased with an increase of MFC content. The cell proliferation test on the composite gel showed no toxicity.  相似文献   

Molecular phylogenetic relationship of Tetraodon pufferfish based on mitochondrial DNA analysis     

Yoji Igarashi  Hiroyuki Doi  Yusuke Yamanoue  Shigeharu Kinoshita  Toshiaki Ishibashi  Hideki Ushio  Shuichi Asakawa  Mutsumi Nishida  Shugo Watabe 《Fisheries Science》2013,79(2):243-250

Pufferfishes belonging to the genus Tetraodon form one of the largest groups in the family Tetraodontidae. This group consists of more than 20 species distributed across freshwater, as well as brackish and coastal waters, of Africa and Southeast Asia. However, their phylogenetic and evolutionary relationships are still ambiguous. In the present study, mitochondrial genes encoding 16S rRNA and cytochrome b were sequenced for 17 Tetraodon species collected from various areas mentioned above. Supermatrix analysis based on the complete sequences of the two genes from Tetraodon species was performed using a tree with 50 tetraodontid species (including 7 Tetraodon species) as a backbone. The obtained phylogenetic tree classified Tetraodon species into three groups, correlating well with their habitats, such as Asian freshwater, Asian brackish water, and African freshwater. We also showed that salinity tolerance of Asian freshwater species T. cochinchinensis was apparently lower than that of Asian brackish water species T. nigroviridis, suggesting that the speciation of Tetraodon species in the Asian freshwater group was caused by the molecular evolution associated with osmotic regulation.  相似文献   

Cloning and expression of a surface immunogenic protein in Streptococcus dysgalactiae isolated from fish and its application in enzyme‐linked immunosorbent assays to diagnose S. dysgalactiae infections in fish     

I Nishiki  T Minami  T Itami  T Yoshida 《Journal of fish diseases》2014,37(12):1031-1039

Lancefield group C Streptococcus dysgalactiae (GCSD) causes severe necrotic lesions in the caudal peduncle in the genus Seriola farmed in Japan. To develop a sero‐diagnostic method for GCSD infection in farmed fish, we attempted to identify a surface immunogenic protein that induces an antibody after infection with GCSD by immunoblot analysis using sera collected from infected fish. A protein obtained from sodium dodecyl sulfate (SDS) extracts of GCSD was identified as S. dysgalactiae surface immunogenic protein (Sd‐Sip). Sd‐Sip exhibited more than 94% homology with a surface antigen or a hypothetical protein from S. dysgalactiae mammalian isolates at the nucleotide sequence level. Expression of the recombinant Sd‐Sip (rSd‐Sip) was confirmed by immunoblot analysis, that is, its reactivity to GCSD‐infected sera. Antibody detection ELISA using rSd‐Sip and their usefulness for diagnosis of GCSD infection were examined. GCSD‐infected sera collected from farmed amberjack, Seriola dumerili (Risso), showed strong reaction with immobilized rSd‐Sip. Meanwhile, sera immunized by other pathogenic bacteria of fish were showed ELISA values similar to those of non‐infected sera. These results of this study suggest that the antibody detection ELISA using rSd‐Sip is an effective diagnostic method for GCSD infection in fish.  相似文献   

Profiling of phenylpropanoid monomers in developing xylem tissue of transgenic aspen (Populus tremuloides)     

Shiro Suzuki  Norikazu Sakakibara  Laigeng Li  Toshiaki Umezawa  Vincent L. Chiang 《Journal of Wood Science》2010,56(1):71-76

Here we describe alterations in the cinnamate/monolignol pathway in three transgenic aspen lines: one with downregulated expression of 4-coumarate:CoA ligase (4CL), one with upregulated expression of coniferaldehyde 5-hydroxylase (CAld5H), and a 4CL downregulated/CAld5H upregulated line. Compared with the wild type, the 4CL downregulated line showed significantly increased levels of p-hydroxycinnamic acids such as p-coumaric, ferulic, and sinapic acids. In contrast, the CAld5H upregulated line had increased content of p-coumaryl and 5-hydroxyconiferyl alcohols. In the 4CL downregulated line, it was likely that most hydroxycinnamic acids were glycosylated. These results strongly suggest that the downregulation of 4CL and upregulation of CAld5H disrupt the metabolic flow through the cinnamate/monolignol pathway and thus alter the amount and structure of its final product, lignin.  相似文献