Onset of individual growth difference in larviculture of Pacific bluefin tuna <Emphasis Type="Italic">Thunnus orientalis</Emphasis> using fertilized eggs obtained from one female

We conducted a rearing experiment on Pacific bluefin tuna (PBT) larvae, which originated from one female broodstock, and determined the growth history of the larvae to clarify when the growth difference occurs. We investigated the otolith microstructure of the PBT larvae to examine the individual growth history and to predict the age and body size at the onset of individual growth difference. Since total length (TL) of fish can be back-calculated from otolith radius, we back-calculated TLs of 100 fish of 19 days post hatch (dph) divided into three size groups (small, intermediate, large). Growth difference was recognized from 3 dph (mouth opening), and the difference became larger thereafter. Growth of large-size fish was assumed to be promoted by the feeding conditions of PBT larvae.     

First record of wild ephyrae of the giant jellyfish Nemopilema nomurai

To understand the causes and to control harmful blooms of the giant Nomura??s jellyfish Nemopilema nomurai, it is essential to study the seed population (benthic polyps) and its environment. To locate the habitat of polyps, the first step is to find ephyrae shortly after detachment from polyps. We found five ephyrae of N. nomurai of 1?C2?mm diameter for the first time from plankton samples collected at two sites, one specimen in the northwestern East China Sea (32°12.3??N, 123°12??E) and the other four specimens in the Yellow Sea (34°05.8??N, 121°50.0??E), on 22 and 26 May 2011. From the developmental state and water temperature of ca. 16?°C at the sampling stations, the collected ephyrae were estimated to have detached from polyps in early May of the year. Considering the increase of water temperature which induces strobilation and the direction of transport in the northwestern East China Sea and in the Yellow Sea, the locality of the seed polyps of the present specimens is probably the sea-floor close to the Changjiang River mouth and along the coast of Jiangsu Province.     

Factors Influencing Early Survival and Growth of Laboratory‐reared Pacific Bluefin Tuna,Thunnus orientalis,Larvae

The collection of fertilized eggs for mass culture of Pacific bluefin tuna (PBT) relies on the spontaneous spawning of broodstock in captivity, and the spawning season of the broodstock is generally from mid‐May to September. The diameter of fertilized eggs from the broodstock decreases during the spawning season. To investigate the influence of three potential factors, the egg diameter (larger or smaller), the rearing water temperature (25 or 28 C), and the aeration rate during the night (strong or weak), on early survival and on growth of PBT larvae, replicate trials were conducted to assess these three factors until 7 d after hatching (d.a.h.). At 7 d.a.h., survival rates of larvae reared with strong nighttime aeration were found by a three‐way ANOVA to be significantly higher than for larvae reared with weak aeration. Furthermore, growth rates of PBT larvae hatched from larger eggs were significantly faster than those from smaller eggs. However, a significant difference in the survival rate was not detected for the factors, egg diameter and rearing water temperature. The results indicate that rearing with strong nighttime aeration significantly improved the early survival of PBT larvae and the egg diameter (a proxy for egg quality) and influenced growth.     

Study on the Mechanism of the Blood-Glucose-Lowering Effect of Collagen Peptides from Sturgeon By-Products

In a previous study, we found that the collagen peptides prepared from the by-products of Bester sturgeon had an inhibitory effect on elevated blood glucose levels in a glucose tolerance test with ICR mice. In the present study, we examine the mechanism of the effect of sturgeon collagen peptides (SCPs) in detail. When glucose was orally administered to mice along with the SCPs, it was found that the glucose remained in the stomach for a longer time. In the above tests, the amount of glucose excreted in the feces of mice also increased. On the contrary, it was revealed that the SCPs have a dipeptidyl-peptidase-IV (DPP-IV) inhibitory ability in an in vitro test. In subsequent oral and intravenous glucose administration tests, glucagon-like peptide-1 (GLP-1) and insulin levels in the blood of mice were maintained at high levels. These results suggested the following three mechanisms: SCPs slow the rate of transportation of glucose from the stomach into the small intestine, resulting in delayed glucose absorption; SCPs suppress the absorption of glucose in the small intestine and excrete it from the body; SCPs inhibit DPP-IV in the blood and maintain a high GLP-1 level in blood, which in turn stimulates insulin secretion.     

Effects of manufacturing factors on the viscosity of a polysaccharide solution extracted from Gagome Kjellmaniella crassifolia

A highly viscous polysaccharide solution was extracted from Gagome Kjellmaniella crassifolia in 20°C water. The eluted sugar concentration was 0.16%, and the extracted carbohydrates consisted of fucoidan, laminaran, and alginate at an approximately ratio of 8.2:0.8:1.0. An increase in the extraction temperature resulted in a less viscous solution, even though the amount of eluted sugar was higher than that obtained at a lower temperature. The most viscous extraction solution was obtained at a neutral pH, with a more acidic or alkaline extraction solution being a less viscous. When the highly viscous polysaccharide solution was heated, the viscosity decreased markedly with increasing temperature. The viscosity of the polysaccharide solution increased after dialysis against water and decreased with the addition of either KCl or NaCl. However, the viscosity was recovered to previous levels by following re-dialysis against water. The removal of divalent cations by EDTA and the re-addition using CaCl₂ or MgCl₂ also caused reversible changes to the viscosity. These characterizations will be useful for widespread applications of viscous K. crassifolia polysaccharides.     

Occurrence of toxic <Emphasis Type="Italic">Dinophysis acuminata</Emphasis> (Dinophyceae) in Uranouchi Inlet,Japan

In the Uranouchi Inlet, which has a commercial fishery of littleneck clam Ruditapes philippinarum, potentially toxigenic species of Dinophysis acuminata showed an annual occurrence except for the summer of 2000–2002. Relatively high abundances of D. acuminata were observed in April 2000 and November 2002. Some samples of D. acuminata cells, which were collected from the inlet in autumn 2002, contained pectenotoxin-2 (0.9–2.8 pg/cell) and okadaic acid (1.9–2.8 pg/cell). Cell length and hypotheca width of D. acuminata found in the Uranouchi Inlet were 33.7–45.0 and 21.0–31.6 μm, respectively. The length-width ratios ranged 1.21–1.71. Sequences of the 5.8S ribosomal RNA gene and flanking internal transcribed spacers, 1 and 2 of cells of D. acuminata from the inlet fell into a D. acuminata complex clade that included, D. acuminata-D. sacculus-D. acuta reported by others. In the inlet, occurrences of D. acuminata were detected within a broad water temperature range between 13.0 and 26.9°C, and those showing more than 25 cells/L were within 16.4–17.3°C for temperature and 27.6 and 34.1 for salinity. This is the first report that cells of Dinophysis acuminata from southern Japanese coastal waters contained okadaic acid and pectenotoxin-2.     

Suspected panosteitis in a crossbred calf

A male crossbred calf developed a limp and pain upon deep pressure on the right hind limb and the right forelimb. The radiographic findings of affected limbs and pathological findings of bone biopsy were similar to those observed in canine panosteitis. This is the first case of suspected panosteitis reported in cattle.     

The PFA-AMeX method achieves a good balance between the morphology of tissues and the quality of RNA content in DNA microarray analysis with laser-capture microdissection samples

Recently, large-scale gene expression profiling is often performed using RNA extracted from unfixed frozen or formalin-fixed paraffin embedded (FFPE) samples. However, both types of samples have drawbacks in terms of the morphological preservation and RNA quality. In the present study, we investigated 30 human prostate tissues using the PFA-AMeX method (fixation using paraformaldehyde (PFA) followed by embedding in paraffin by AMeX) with a DNA microarray combined with laser-capture microdissection. Morphologically, in contrast to the case of atypical adenomatous hyperplasia, loss of basal cells in prostate adenocarcinomas was as obvious in PFA-AMeX samples as in FFPE samples. As for quality, the loss of rRNA peaks 18S and 28S on the capillary electropherograms from both FFPE and PFA-AMeX samples showed that the RNA was degraded equally during processing. However, qRT-PCR with 3’ and 5’ primer sets designed against human beta-actin revealed that, although RNA degradation occurred in both methods, it occurred more mildly in the PFA-AMeX samples. In conclusion, the PFA-AMeX method is good with respect to morphology and RNA quality, which makes it a promising tool for DNA microarrays combined with laser-capture microdissection, and if the appropriate RNA quality criteria are used, the capture of credible GeneChip data is well over 80% efficient, at least in human prostate specimens.     

Overcoming the self-incompatibility of Lilium longiflorum Thunb. by application of flower-organ extract or temperature treatment of pollen

Summary The application of flower organ extracts to stigmas and the temperature treatment of pollen were tried to overcome self-incompatibility of Lilium longiflorum cv. Georgia.Substances in stigma, style, ovary and anther were extracted with ethanol and fractionated with ethylacetate into the acidic, basic and aqueous fractions. The extracs melted in a small volume of distilled water were applied to stigmas prior to self-pollination. Hinomoto stigma extract, self-pollinated and cross-pollinated Georgia stigma extracts of high concentrations and Georgia anther extract of high concentration were effective in overcoming the self-incompatibility and resulted in a high percentage of fruit set and many normal seeds. Extracts from Hinomoto ovary, style and anther were ineffective, except a basic fraction, which was very slightly effective.Pollen was treated with 40°C for 60 or 90 minutes and 50°C for 30 or 60 minutes, and a half of each lot was followed by –20°C for 24 h, prior to self-pollination. All treatments were effective, especially at 40°C for 60 minutes or 50°C for 30 minutes, and 40°C for 90 minutes or 50°C for 60 minutes followed by –20°C for 24 h.     

Microbial community profiles in intercellular fluid of rice

Plants harbor microorganisms that are thought to stimulate plant defense systems or promote plant growth. Individual species in these intercellular microbial communities are often not sufficiently abundant to be easily described, although some endophytic microorganisms amenable to culture have been characterized. To better understand the microbial population of plants, we collected intercellular fluid (IF) from leaf blades and sheaths of rice and subsequently isolated DNA from the IF. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S and 18S rDNA fragments amplified from IF DNA by PCR indicated that these band patterns were distinguishable from those of a leaf surface-wash fluid (SF). Analysis of a set of rDNA fragments amplified from IF DNA of rice with different genotypes, paddies or growth stages for the primary survey of overall microbial community in the IF suggested that this approach is suitable for analyzing microbial diversity in the IF from various plant samples. Actually, comparative analysis of amplified rDNA fragments of rice and other five plant species indicated that the microbial diversity in IF is likely to vary substantially among plant species. We can also use sequence analysis of 16S rDNA fragments amplified from rice IF DNA to identify species including unculturable bacteria and proteobacteria and Xanthomonas and 18S rDNA fragments to identify Tilletiaria anomala, Tilletia iowensis, Ustilago maydis and unculturable eukaryotes. Thus, IF DNA analysis seems to be a good tool to further study the microbial ecology of plants.