Random sequencing of genomic DNA of <Emphasis Type="Italic">Photobacterium damselae</Emphasis> subsp. <Emphasis Type="Italic">piscicida</Emphasis>

ABSTRACT:   Random sequencing of the genomic DNA of Photobacterium damselae subsp. piscicida was conducted. The sequences were assembled into 930 contigs. The total length of these contigs accounts for 37.5% of the genome of P. damselae subsp. piscicida. The contigs contain 2055 open reading frames (ORFs) that have homology with genes in the GenBank database. Furthermore, some of the ORFs have homology with reported virulence related genes, and are classified as encoding colonization factors, exotoxin, and lipopolysaccharide (LPS). Thirty-seven ORFs have homology with colonization factors. In those colonization factors, 27, three, and four ORFs have homology with polar flagellar-related genes, capsule-related genes, and others (accessory colonization factors, superoxide dismutase (Cu-Zn), MshA biogenesis protein, and heme receptor genes), respectively. Five ORFs have homology with exotoxin-related genes (2 hemolysin, phospholipase, hyaluronidase, lysophospholipase L2 genes). Further, six ORFs have homology with LPS-related genes.     

Seasonal variation in docosahexaenoic acid content in horse mackerel caught in the East China Sea

ABSTRACT:   The fatty acid composition in horse mackerel caught off Nagasaki, off Tsushima Island, and in the middle of the East China Sea was investigated. The ratios of monoenoic and polyenoic acids to the total fatty acids in the Nagasaki and bull trawl catch specimens had a negative correlation. The docosahexaenoic acid (DHA; 22:6n-3) ratios of total fatty acids in summer-caught specimens were lower than those in winter-caught specimens, and this tendency was significantly clearer in smaller-sized fish, while DHA levels in tissues varied little throughout the year. Consistently high levels of DHA were found in various fishing areas, which suggests that horse mackerel offers a stable source of DHA.     

Effect of chinese parsley Coriandrum sativum and chitosan on inhibiting the accumulation of cadmium in cultured rainbow trout Oncorhynchus mykiss

ABSTRACT:   Toxic heavy metals such as cadmium (Cd) accumulate in cultured fish from solid-form feed mixtures. In this paper, two natural ingredients, supposed to inhibit accumulation of heavy metals in the fish body, were investigated to develop a functional feed mixture for healthy fish. Three test diets: (i) ♯1, regular diet as negative control; (ii) ♯2, ♯1 + 2% lyophilized Chinese parsley Coriandrum sativum (CP); and (iii) ♯3, ♯1 + 3% chitosan (CT), were each fed to 50 rainbow trout Oncorhynchus mykiss once a day, at the constant rate of 1.0–1.2% of body weight for 12 weeks. No undesirable effect on fish growth was observed by an addition of either CP or CT to regular feed. Then, 10 mg Cd was added to 1 kg each of the three test diets and they were named as: (i) ♯4, ♯1 + Cd; (ii) ♯5, ♯2 + Cd; and (iii) ♯6, ♯3 + Cd, respectively. The fish given positive a control feed (diet ♯4) for 3 weeks accumulated Cd in the liver and kidney at approximately 0.52 mg/kg. However, Cd in ordinary muscle was found to be just above the detection limit. Upon accumulation of Cd, three groups of 50 fish were each fed test diets ♯4, ♯5, and ♯6 for 12 weeks at the same feed rate described above. The concentrations of Cd accumulated in the liver, kidney, and ordinary muscle of the fish were measured every 3 weeks. Contrary to the positive control fish fed diet ♯4, those fish fed test diets ♯5 and ♯6 accumulated 20–30% and 25–40% less Cd in the liver, respectively. Test diets were a little less effective for inhibition of Cd uptake in the kidney than that observed in the liver. These results suggest that addition of CP or CT to fish feed might be used to decrease toxic heavy metal accumulation in fish.     

French consumers’ marginal willingness to pay for the pairing of Japan’s fall chum salmon and rice wine (sake)

The pairing of foods and beverages (especially alcoholic beverages) is an effective marketing approach to promote the value of Japanese gastronomy to overseas consumers because it requires little additional cost. In this study, the research target was French consumers, and the evaluation targets were grilled fall-season wild chum salmon from Japan (mainly from Hokkaido) and Japanese rice wine (sake). The difference in the consumers’ willingness to pay (WTP) was evaluated using the contingent valuation method. In addition, we analyzed the determinants by applying Tobit regression. Our results revealed that there was a higher WTP when Japan’s (Hokkaido’s) grilled fall salmon and sake were presented as a set than when they were presented individually. The geographical identity of the origin of food and beverages can engender a positive premium to consumers through the synergistic effect of the origin of the brand.

     

Vertical Changes in Bacterial Communities in Percolating Water of a Japanese Paddy Field as Revealed by PCR-DGGE

Percolating water was sampled from the plow layer and subsoil layer in a Japanese paddy field, and the bacterial communities were compared together with floodwater by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) targeting a partial 16S rRNA gene and subsequent sequencing. The number of DGGE bands ranged from 16 to 28 with no significant differences among the sampling sites and times. Only 2 bands were common for the three sources of water samples. DGGE bands specific for the floodwater samples and percolating water samples from the plow layer were identified, while percolating water samples from the subsoil layer did not show specific bands but displayed common bands to those of the floodwater samples (7 bands) and percolating water samples from the plow layer (1 band). Cluster analysis of the DGGE banding patterns showed a distinct clustering in the samples of percolating water from the plow layer and a closer relationship between the others. These results suggest that the bacterial communities in percolating water changed during downward movement through the plow layer and subsoil layer. Sequences of the DGGE bands specific for the samples of percolating water from the plow layer showed a close relationship with anaerobic bacteria such as iron-reducers or uncultured bacterial DNA isolated from environments that are considered to be less oxic. On the other hand, the sequences of the bands specific for the samples of floodwater and percolating water from the subsoil layer showed a close relationship with uncultured bacterial DNA isolated from freshwater environments.     

Phosphorylation of ovalbumin by dry-heating in the presence of pyrophosphate: effect on protein structure and some properties

Ovalbumin (OVA) was phosphorylated by dry-heating in the presence of pyrophosphate at pH 4.0 and 85 degrees C for 1 and 5 days, and the physicochemical and structural properties of phosphorylated OVA were investigated. The phosphorus content of OVA increased to 1.01% by phosphorylation, and the electrophoretic mobility of PP-OVA also increased. Although the solubility of dry-heated OVA decreased, the decrease was slightly depressed by phosphorylation. The circular dichroism spectra showed that the change of the secondary structure in the OVA molecule, as measured by alpha-helix content, was mild by phosphorylation. The exchange reaction between the sulfhydryl and disulfide groups was enhanced and the surface hydrophobicity of OVA increased by phosphorylation. The tryptophan fluorescence intensity of OVA decreased by phosphorylation, suggesting that the conformational change occurred in the OVA molecule by phosphorylation. Although the differential scanning calorimetry thermograms of OVA showed a lowering of the denaturation temperature from 78.3 to 70.1 degrees C by phosphorylation, the stability of OVA against heat-induced insolubility at pH 7.0 was improved. The results indicated molten (partially unfolded) conformations of OVA formed by dry-heating in the presence of pyrophosphate.     

Enzyme-linked immunosorbent assay based on a polyclonal antibody for the detection of the insecticide fenitrothion. Evaluation of antiserum and application to the analysis of water samples.

For development of an indirect competitive enzyme-linked immunosorbent assay (ELISA) for the organophosphorus insecticide fenitrothion, the specificity of the antiserum R-3 generated with the bifunctional hapten, LysMNPA (2-[[[(3-methyl-4-nitrophenyl)oxy]methylcarbonyl]amino]-6-(2,4-dinitrophenyl)aminohexanoic acid) and the application to the residual analysis of some water samples were evaluated. At optimized ELISA conditions, the quantitative working range was from 1 to 39 ng/mL with a limit of detection of 0.3 ng/mL and an IC(50) value of 6 ng/mL. Cross-reactivity to structurally similar organophosphorus compounds and related chemicals was determined. The antiserum R-3 showed significant cross-reactivity with fenitrooxon and 3-methyl-4-nitrophenol, which have a 3-methyl-4-nitrophenoxy group as common structures, but showed relatively low cross-reactivity with other compounds. Each water sample (river water, tap water, purified water, and bottled water) had a matrix effect and was investigated by adding Tween 20 in the assay buffer. These four kinds of water samples were fortified with fenitrothion at several concentration levels and were directly analyzed with only dilution with an equal volume of antiserum solution. The mean recovery was 105.9%, and the mean coefficient of variation was 10.9%. The results suggested that the developed ELISA would be very suitable for a preliminary screening for fenitrothion in water samples at such low levels.     

Detection of nifH Sequences in Sugarcane (Saccharum officinarum L.) and Pineapple (Ananas comosus [L.] Merr.)

To detect N₂-fixing bacteria in a plant without using culture methods, nifH gene segments were amplified with degenerate primers from DNA extracted from stems of sugarcane ( Saccharum officinarum L.) and leaves of pineapple ( Ananas comosus [L.] Merr.). Sequences of the nifH clones were homologous to those of bacteria in the genera Bradyrhizobium, Serratia , and Klebsiella. The bacteria with nifH sequences homologous to those of the three genera may be promising candidates for predominant endophytic diazotrophs in sugarcane and pineapple, because these sequences were commonly detected in samples from different host plants, which were collected at different locations. On the other hand, no nifH sequence related to Gluconacetobacter diazotrophicus , which is an endophytic N₂-fixing bacterium and had been previously isolated from sugarcane, was detected in sugarcane. This indicates the absence or the presence of few G. diazotrophicus in the stems of the sugarcane plants used in the current study.