Serological survey of Neospora caninum infection among dogs in Japan through species-specific ELISA

A seroepidemiological survey of Neospora caninum infection among dogs in Japan was conducted using species-specific enzyme-linked immunosorbent assay with recombinant surface antigen (Nc-SAG1t). Among 1,206 dogs examined, 126 dogs (10.4%) from 30 prefectures from Hokkaido to Okinawa were positive to N. caninum infections, which were more frequently detected in females than males. Siberian Huskies showed the highest positive rate compared with the other breeds. Dogs with pyometra and diabetes mellitus showed the higher positive rates than dogs with other diseases or without diseases.     

Two linked genes on rice chromosome 2 for F1 pollen sterility in a hybrid between Oryza sativa and O. glumaepatula

Hybrid incompatibility plays an important role in establishment of post-zygotic reproductive isolation. To unveil genetic basis of hybrid incompatibilities between diverged species of genus Oryza AA genome species, we conducted genetic dissection of hybrid sterility loci, S22(t), which had been identified in backcross progeny derived from Oryza sativa ssp. japonica (recurrent parent) and South American wild rice O. glumaepatula near the end of the short arm of chromosome 2. The S22(t) region was found to be composed of two loci, designated S22A and S22B, that independently induce F₁ pollen sterility. Pollen grains containing either of the sterile alleles (S22A-glum^s or S22B-glum^s) were sterile if produced on a heterozygous plant. No transmission of the S22A-glum^s allele via pollen was observed, whereas a low frequency of transmission of S22B-glum^s was observed. Cytological analysis showed that the sterile pollen grains caused by S22A could reach the bicellular or tricellular stage, and the nearly-sterile pollen grains caused by S22B could reach the tricellular stage. Our genetic analysis showed repulsion linkage effect is possible to induce strong reproductive barrier by high pollen sterility based on recombination value and transmission ratio of hybrid sterility gene to the progeny was influenced by frequency of competitors on fertilization.     

Migration of pine wood nematodes in the tissues of <Emphasis Type="Italic">Pinus thunbergii</Emphasis>

The distribution of pine wood nematodes (Bursaphelenchus xylophilus, PWNs) in Japanese black pine (Pinus thunbergii) tissues was investigated by staining with fluorescein isothiocyanate-conjugated wheat germ agglutinin. After PWNs were inoculated to current-year stems of pine seedlings, their distribution at about 5 cm below the inoculation site was confined only to cortical resin canals 1 day after inoculation, and then spread to other tissues, including resin canals of short branches. When PWNs were inoculated onto cross or tangentially cut surfaces of stem segments, maximal PWN migration speed was estimated to be faster through cortical resin canals and xylem axial resin canals vertically (>6.7 and <2.3 mm/h, respectively) than through cortical tissues both vertically and horizontally (<1.2 and <0.2 mm/h). To examine whether PWNs in cortical resin canals could invade surrounding tissues, segments in which PWNs resided only in cortical resin canals were prepared by removing the top portion 6 h after inoculation. Additional incubation of such segments caused extended PWN distribution to xylem axial resin canals and then to other tissues. A similar experiment with top portions of girdled segments removed 12 h after inoculation also showed extended PWN distribution from xylem axial resin canals and pith to cortical resin canals and then to other tissues. These results provided direct evidence that PWNs have the ability to migrate from cortical resin canals and xylem axial resin canals to other tissues.     

茉莉花中芳樟醇生成相关酶的精制

从酶学角度探讨了茉莉花重要香气成份芳樟醇的 生成机理⒚以双瓣茉莉花为材料,经过调制丙酮粉,缓冲液粗提,２０％ ～８０％ 饱和度硫铵分部, Ｃ Ｍ  Ｔｏｙｏｐｅａｒｌ ６５０ Ｍ 和 Ｆ Ｐ Ｌ Ｃ Ｃ Ｍ  Ｔｏｙｏｐｅａｒｌ ６５０ Ｓ两次柱层析,得到了 Ｆ Ｃ 蛋白分部液⒚同时经 Ｇ Ｃ Ｍ Ｓ 定性和 Ｇ Ｃ 定量分析,明确 Ｆ Ｃ 部分是催 化茉莉花粗 香气前 体物 质生 成芳 樟醇 的主 要部分⒚进 一步 经等 电电 泳、β Ｄ葡萄 糖苷 酶活 性染色 等电 电泳 和 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 凝胶电泳,初步判断 β Ｄ葡萄糖苷酶参与了芳樟醇的生成,且该酶的等电点在 ８．７～９．０,其单体蛋白的分子量在 ３５～３８ ｋ Ｄ 之间 ⒚     

Fluorescein-labeled wheat germ agglutinin stains the pine wood nematode, <Emphasis Type="Italic">Bursaphelenchus </Emphasis><Emphasis Type="Italic">xylophilus</Emphasis>

Pine wilt disease, caused by the pine wood nematode (PWN, Bursaphelenchus xylophilus), is a major threat to pine forests throughout East Asia. Nonetheless, its mechanism of invasion has not yet been described in detail. To better understand the pathology of this disease, it is important to examine the distribution of PWNs within pine tissue during the course of disease development. We attempted to stain nematodes with fluorescein-conjugated wheat germ agglutinin (F-WGA) as a means to locate and track the spread of PWNs. Although PWNs proliferated on Botrytis cinerea fungus were successfully stained only on their vulvas and spicule holes, PWNs extracted from inoculated Pinus thunbergii seedlings were stained on their surface. Stainability, or the percentage of PWNs stained with F-WGA over more than half of their surface, was about 20% by 1 day after inoculation, but increased to 80% at 10 days. The stainability of PWNs extracted from a 5-cm main stem segment that included the inoculation site was less than that of PWNs extracted from other parts of the main stem farther away (i.e., those that had dispersed). These results suggest that stainability is related to dispersal activity in time. Thus, to raise the stainability of PWNs at shorter timeframes after inoculation, PWNs with more than 80% stainability were re-inoculated into pine seedlings. This resulted in more than 70% stainability from 1 to 6 days after inoculation. In F-WGA stained thin paraffin sections of pine tissue of re-inoculated seedlings, PWNs brightly fluoresced under epifluorescence and were easily detected against the dark background of pine tissue. This staining technique with F-WGA is an excellent tool for detecting PWNs in pine tissue.     

Faster growth before metamorphosis leads to a higher risk of pseudoalbinism in juveniles of the starry flounder <Emphasis Type="Italic">Platichthys stellatus</Emphasis>, as suggested by otolith back-calculation

The mechanism underlying malformation during flatfish metamorphosis is not clear. Here, the relationship between growth before metamorphosis and the occurrence of malformation after metamorphosis was examined in individuals reared in a single tank. Otoliths of starry flounder Platichthys stellatus larvae were labeled with alizarin complexone before metamorphosis [12 days after hatching (DAH)]. The positive linear regression between the lapillus otolith diameter and body length at 12 DAH enabled the estimation of individual larval size after completion of metamorphosis. At 12 DAH, the average body length of the typical pseudoalbino juveniles was significantly greater (5.37 mm) than that of the normal juveniles (5.13 mm) (P < 0.01, n = 100). In addition, of the larvae that were estimated to measure less than 4.8 mm at 12 DAH, approximately 80% metamorphosed into normal juveniles, whereas more than 60% of the larvae that measured more than 5.4 mm became pseudoalbino juveniles. These results indicate that the larvae that grow rapidly during the first 12 days are more likely to become pseudoalbino individuals after metamorphosis.     

Interactions of picrodendrins and related terpenoids with ionotropic GABA receptors of mammals and insects

Different structural features govern the interaction of picrodendrins and related terpenoids with rat and with housefly GABA receptors. This supports previous studies which suggest that there are significant differences between the structures of the binding sites in these two receptors.     

Evaluation of Loads and Sources of Heavy Metals in Tama River, Tokyo

The loads and sources of heavy metals (Cd, Co, Cu, Mn, Mo, Ni, Pb, V, and Zn) in the midstream of Tama River were evaluated on the basis of their chemical analyses in ordinary and stormwater runoff from July 2007 to November 2008. Tama River is one of the three major rivers flowing into Tokyo Bay. The total annual water discharges differed largely for 2007 and 2008, depending on the scales of typhoon rainfalls and other heavy rainfalls in each year. The concentrations of the metals other than Mo in the river did not change markedly at a flow rate of less than approximately 200 m³ s^?1, but at higher flow rates, the concentrations of all the metals increased linearly with the increase in the log of flow rate (r ²?=?0.94–0.99). The annual loads of heavy metals for 2007 and 2008 were estimated using regression equations between the above parameters and the hourly flow rate data for each year. For the metals other than Mo, the contribution of the loads at higher flow rates (>200 m³ s^?1) was much larger than that at lower flow rates (<200 m³ s^?1), showing the importance of the particulate loads (primarily crustal materials) during storm runoff following typhoon rainfalls and other heavy rainfalls. In contrast, the loads of Mo at lower flow rates accounted for major portions (56–78%) of the total loads, because of a relatively small contribution of particulate load during storm runoff. The contribution of the loads of Mo, Zn, Cd, Cu, and Ni at lower flow rates to the annual loads was larger than that of other metals. It was found that the concentrations of these metals in ordinary runoff are strongly affected by the discharge of treated water from sewage treatment plants which are located along the catchment. Thus, treated water from sewage treatment plants may be the primary source contributing to the present pollution of Mo, Zn, Cd, Cu, and Ni in Tokyo Bay.     

Indicated detection of two unapproved transgenic rice lines contaminating vermicelli products

We analyzed the DNA fragments extracted from four rice vermicelli products. The Bacillus thuringiensis (Bt) rice line, which has a construct similar to the GM Shanyou 63 line, was detected in some vermicelli products by identification of the junction region sequence between rice Act1 promoter and the Cry1Ac gene, and that between Cry1Ac and nos. In addition, we also detected a different Bt rice line by means of the junction region sequence between the maize ubiquitin promoter and cry1Ab gene and that between the cauliflower mosaic virus 35S promoter and the hygromycin phosphotransferase in some vermicelli products. Accordingly, we for the first time have detected the two transgenic Bt rice lines contaminating rice vermicelli samples. Furthermore, we developed a duplex real-time polymerase chain reaction (PCR) method for the simultaneous detection of both Bt rice lines.