Characterization of Aquimarina hainanensis isolated from diseased mud crab Scylla serrata larvae in a hatchery

Mass mortality due to necrosis signs occurred in hatchery-reared zoea stage larvae of the mud crab Scylla serrata in Okinawa, Japan, and a causative bacterium was isolated. In this study, we identified and characterized the bacterium by genome analysis, biochemical properties and pathogenicity. The bacterium was a Gram-negative, non-motile, long rod, forming yellow colonies on a marine agar plate. It grew at 20–33°C (not at 37°C) and degraded chitin and gelatin. Phylogenetic analysis of the 16S rRNA gene sequence identified the bacterium as Aquimarina hainanensis. Genome sequence data obtained from Illumina MiSeq generated 29 contigs with 3.56 Mbp in total length and a G + C content of 32.5%. The predicted 16 chitinase genes, as putative virulence factors, had certain homologies with those of genus Aquimarina. Experimental infection with the bacterium conducted on larvae of four crustacean species, brine shrimp Artemia franciscana, freshwater shrimp Caridina multidentata, swimming crab Portunus trituberculatus and mud crab S. serrata, revealed that this bacterium was highly virulent to these species. The present study suggests that the bacterium caused mass mortality in mud crab seed production was A. hainanensis and can be widely pathogenic to crustaceans.     

Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

Efficiency of stagnant water larviculture using disinfected amictic eggs of the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> in Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

There is a need to develop low-cost methods for larviculture that emphasize disease prevention. We evaluated stagnant water larviculture in flounder Paralichthys olivaceus using rotifer amictic eggs treated with glutaraldehyde. Growth and survival of the larvae were similar to in larvae reared by standard running water larviculture with daily feeding of rotifers (control). In the experimental group, the n-3 highly unsaturated fatty acid content of rotifers in the rearing water was higher than in the enriched rotifers fed to the control group. We also observed a similar pattern for the fatty acid composition of flounder larvae. The viable bacterial counts associated with rotifers in stagnant water were lower than those associated with enriched rotifers, and there were fewer bacterial genera. However, the viable bacterial count was the same or higher in the experimental rearing water compared with that in the control group. The α-Proteobacteria and Cytophaga–Flavobacterium–Bacteroidetes group dominated the bacterial community structure after larval hatching. To our knowledge, this is the first time that stagnant water larviculture, using only disinfected amictic eggs, has been successfully demonstrated on a mass production scale. This method appears to offer benefits including improved nutrition, more favorable bacterial communities, and lower cost.     

A review of seed production and stock enhancement for commercially important portunid crabs in Japan

We reviewed the present status of seed production for stock enhancement and evaluated the impact of stocking on commercial catches in Japan, of portunid crabs particularly Portunus trituberculatus and Scylla paramamosain. The mean survival rate from hatching to first-stage crabs was around 10%, and 20–30% of the larval culture trials conducted in recent years could not harvest juveniles during seed production. To achieve reliable mass seed production technologies, measures for controlling disease and the nutritional condition of larvae in seed production tanks are required to be developed. The main spawning season extends from April/May to July. Reflecting their life cycle characteristics such as high growth rates, crabs recruit to the fishery after September and largely contribute to the commercial landings until December in the hatching year. The main release season of juveniles is from June to July. Consequently, released juveniles are expected to contribute to commercial landings in the release year. Analyses of catch and release statistics in two small bays estimated the yield from released individuals (YPR, yield per release) at 2.4 g for P. trituberculatus and 3.3–7.7 g for S. paramamosain, which were similar to values reported from tagging surveys. On a major regional basis, catch and release histories of P. trituberculatus highlighted the impact of hatchery releases on commercial landings as 33.6 g YPR in the Seto Inland Sea, where catches and releases have been greatest. Although the YPR estimates were different between small bays and major regional seas, Japanese stock enhancement programmes should have had an impact on portunid crab production, dependent on the magnitude of the releases.     

Stem water transport of Lithocarpus edulis, an evergreen oak with radial-porous wood

The stem water conducting system of an evergreen broad-leaved oak, Lithocarpus edulis (Mak.) Nakai, was investigated. Evergreen broad-leaved oaks (Lithocarpus, Castanopsis, Cyclobalanopsis, Quercus) belonging to the Quercoideae are a major component of Asian monsoon forests, and are characterized by the possession of radial-porous wood. A characteristic of radial-porous wood is the development of aggregate rays between radially oriented files of vessels. We measured the distribution of vessel lumen diameters in a stem cross section and calculated the theoretical water conductivity of the wood. The radial profile of the heat pulse velocity (HPV) was measured for an intact whole tree under field conditions and compared with the theoretical distribution of water conductivity. Soft X-ray photographs of frozen stem sections indicated that most of the vessel lumina were filled with water, including those of vessels more than 20 years old. Even when vessels were relatively wide (lumen diameters > 100 microm), cavitation was negligible. The rate of water uptake from the cut stem base correlated closely with HPV (r = 0.96), and HPV closely reflected the mean volume flow per stem sectional area (SFVS) around the sensor probes. However, the ray tissue sharply inhibited heat transfer, and the positioning of the probes strongly affected the absolute value of HPV. It was also found that HPV more closely reflected the mean sap flow velocity in the vessels than did SFVS.     

Structure of xylan from culms of bamboo grass (Sasa senanensis Rehd.)

Xylan prepared from culms of kumaizasa (Sasa senanensis Rehd.), a representative species of bamboo grass, was hydrolyzed with-xylanase ofStreptomyces olivaceoviridis E-86. Four arabinoxylo-oligosaccharides and two glucuronoxylo-oligosaccharides were isolated from the enzymatic hydrolysate of the xylan by chromatography on a charcoal column, a Dowex 1-x8 column, a Toyo-pearl HW-40S column, and a LiChrospher 100 NH₂ column and on preparative paper chromatography. The results of the structural analyses of the saccharides showed that the isolated oligosaccharides had the structures of 3²--l-arabinofuranosyl-xylobiose, 3²--l-arabinofuranosyl-xylotriose, 3²--[-d-xylopyranosyl-(1 2)-l-arabinofuranosyl]-xylobiose, 3³--[-d-xylopyranosyl-(1 2)-l-arabinofuranosyl]-xylotriose, 2³--4-O-methyl-d-glucuronosyl-xylotriose, and 2³--d-glucuronosyl-xylotriose. From the structural analysis of the oligosaccharides derived from the xylan, kumaizasa xylan was concluded to be a kind of arabinoglucuronoxylan having not only stubs of singlel-arabinose and singled-glucuronic acid but also stubs of disaccharide units such as-d-xylopyranosyl-(1 2)-l-arabinofuranose.     

Stable carbon isotope ratios of water-extractable organic carbon affected by application of rice straw and rice straw compost during a long-term rice experiment in Yamagata,Japan

ABSTRACT

Hot-water- and water-extractable organic matter were obtained from soil samples collected from a rice paddy 31 years after the start of a long-term rice experiment in Yamagata, Japan. Specifically, hot-water-extractable organic carbon and nitrogen (HWEOC and HWEON) were obtained by extraction at 80°C for 16 h, and water-extractable organic carbon and nitrogen (WEOC and WEON) were obtained by extraction at room temperature. The soil samples were collected from surface (0–15 cm) and subsurface (15–25 cm) layers of five plots that had been treated with inorganic fertilizers alone or with inorganic fertilizers plus organic matter, as follows: PK, NPK, NPK plus rice straw (RS), NPK plus rice straw compost (CM1), and NPK plus a high dose of rice straw compost (CM3). The soil/water ratio was 1:10 for both extraction temperatures. We found that the organic carbon and total nitrogen contents of the bulk soils were highly correlated with the extractable organic carbon and nitrogen contents regardless of extraction temperature, and the extractable organic carbon and nitrogen contents were higher in the plots that were treated with inorganic fertilizers plus organic matter than in the PK and NPK plots. The HWEOC and WEOC δ¹³C values ranged from ?28.2% to ?26.4% and were similar to the values for the applied rice straw and rice straw compost. There were no correlations between the HWEOC or WEOC δ¹³C values and the amounts of HWEOC or WEOC. The δ¹³C values of the bulk soils ranged from ?25.7% to ?23.2% and were lower for the RS and CM plots than for the PK and NPK plots. These results indicate that HWEOC and WEOC originated mainly from rice plants and the applied organic matter rather than from the indigenous soil organic matter. The significant positive correlations between the amounts of HWEOC and HWEON and the amount of available nitrogen (P < 0.001) imply that extractable organic matter can be used as an index for soil fertility in this long-term experiment. We concluded that the applied organic matter decomposed more rapidly than the indigenous soil organic matter and affected WEOC δ¹³C values and amounts.     

Methane emissions from different vegetation zones in a Qinghai-Tibetan Plateau wetland

We measured methane (CH₄) emissions in the Luanhaizi wetland, a typical alpine wetland on the Qinghai-Tibetan Plateau, China, during the plant growth season (early July to mid-September) in 2002. Our aim was to quantify the spatial and temporal variation of CH₄ flux and to elucidate key factors in this variation. Static chamber measurements of CH₄ flux were made in four vegetation zones along a gradient of water depth. There were three emergent-plant zones (Hippuris-dominated; Scirpus-dominated; and Carex-dominated) and one submerged-plant zone (Potamogeton-dominated). The smallest CH₄ flux (seasonal mean=33.1 mg CH₄ m⁻² d⁻¹) was observed in the Potamogeton-dominated zone, which occupied about 74% of the total area of the wetland. The greatest CH₄ flux (seasonal mean=214 mg CH₄ m⁻² d⁻¹) was observed in the Hippuris-dominated zone, in the second-deepest water area. CH₄ flux from three zones (excluding the Carex-dominated zone) showed a marked diurnal change and decreased dramatically under dark conditions. Light intensity had a major influence on the temporal variation in CH₄ flux, at least in three of the zones. Methane fluxes from all zones increased during the growing season with increasing aboveground biomass. CH₄ flux from the Scirpus-dominated zone was significantly lower than in the other emergent-plant zones despite the large biomass, because the root and rhizome intake ports for CH₄ transport in the dominant species were distributed in shallower and more oxidative soil than occupied in the other zones. Spatial and temporal variation in CH₄ flux from the alpine wetland was determined by the vegetation zone. Among the dominant species in each zone, there were variations in the density and biomass of shoots, gas-transport system, and root-rhizome architecture. The CH₄ flux from a typical alpine wetland on the Qinghai-Tibetan Plateau was as high as those of other boreal and alpine wetlands.     

Fishery biology of mud crabs <Emphasis Type="Italic">Scylla</Emphasis> spp. at Iriomote Island,Japan: species composition,catch, growth and size at sexual maturity

The fishery biology of mud crabs Scylla spp. was examined using baited traps and gill nets from September 2001 to August 2005 at Iriomote Island, Japan. To elucidate the growth of the crabs, artificially produced S. serrata juveniles were released and recaptured at the study site. The sizes at which 50% of females and males of S. serrata reached sexual maturity (SM₅₀) were estimated as an external carapace width (ECW) based on the morphology of the abdomen and the chela respectively. Two species, S. serrata and S. olivacea, were identified in the area with S. serrata being the dominant species (>95% of the catch). Changes in the mean ECW and the results of the release and recapture experiments suggested that the recruitment of young crabs to the fishery occurred from December/January to April/May. The SM₅₀ of females and males occurred at 132.4 and 150.7 mm ECW respectively. The body size composition of S. serrata revealed that immature crabs comprised approximately 40 and 65% of the catch for females and males respectively. To maintain a sustainable fishery for S. serrata, a minimum landing size based on the SM₅₀ estimates should be implemented as a fishing regulation.     

High-yield SRI in West Java by decomposing straw in waterlogged paddy field

System of Rice Intensification (SRI) often achieved higher yield than conventional practice. We identified the high-yielding farmers from the yield records of 1909 paddy fields belonging to an organic farmers’ association. Farmers whose yields were from 8.4 to 10.4 t ha^?1 were interviewed and their fields surveyed. Their yields had increased by an estimated average of 40% following the adoption of SRI practices. They applied 2–12 t ha^?1 of compost. Compared to the conventional practice, they shortened seedling age at transplanting from 27.4 to 17.6 days and reduced the number of seedlings per hill from 4–6 to 2–3, while hill spacing remained unchanged. Instead of intermittent irrigation which is recommended in standard SRI, they kept shallow flooding of 1–2 cm. Although they applied a lot of compost, no correlation was found between the amount of compost application and the yields. Instead, high-yielding farmers returned rice straw into waterlogged paddy after harvest, which presumably is an ideal condition for biological nitrogen fixation. This may occur around rice straw during decomposition under waterlogged condition and might supplement the negative nitrogen balance, thereby enabling the high yield as compared with conventional practices where the fresh rice straw is removed and/or burned.