Establishment of a potency test by ELISA for a rabies vaccine for animal use in Japan

The ELISA we developed was able to determine the antigen content and was suitable for a potency test, and we described a relative potency assay method which determines the potency of test vaccines by comparing the ELISA value of a test vaccine to that of a reference vaccine. In the present study, we standardized the reference vaccine used for determining the potencies of test vaccines, and established a potency test by ELISA. We evaluated the proposed reference vaccine by the neutralizing antibody responses in dogs after vaccination, by the challenge protection test in guinea pigs (GP potency test), which is the earlier official potency test used in Japan, and by the NIH potency test, which is widely used throughout the world. The results showed that a 4-fold dilution of the proposed reference vaccine induced sufficient immunity in dogs. A 3-fold dilution of the proposed reference vaccine passed the GP potency test. The international units (IU) calibrated by the NIH potency test were 3.7 IU/dose. From the results and the WHO recommendation that veterinary rabies vaccines should have a potency of at least 1.0 IU/dose, we determined to dilute the proposed reference vaccine by 3 fold and regarded it as the reference vaccine. Finally, we confirmed that there is a good agreement between the results of the potency test by ELISA and the results of the GP potency test. The establishment of the potency test by ELISA has made it possible to monitor the potency in the production process and has contributed to the stable production of the vaccine.     

Aloe ring spot,a new disease of aloe caused by <Emphasis Type="Italic">Haematonectria haematococca</Emphasis> (Berk. &; Broome) Samuels &; Nirenberg (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis> sp.)

A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates. Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot.     

Strain differences in whole-body protein turnover in the chicken embryo.

1. Whether or not there is a strain difference in embryonic whole-body protein turnover rates was tested using the chicken embryos of Rhode Island Red carrying a sex-linked dwarf gene (dwarf), White Leghorn (layer), and White Cornish X White Plymouth Rock (broiler) strains on day 12 of incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine either intraperitoneally or intravenously on day 12 of incubation in order to investigate the effect of the route of isotope administration. The results showed that the values for fractional and absolute synthesis rates were approximately 13% higher by intravenous injection than by intraperitoneal injection. 3. Whole-body protein turnover, both in terms of fractional and absolute rates, was significantly faster in dwarf than in broiler embryos, with intermediate values in layer embryos, although no growth differences were observed on day 12. 4. Difference in egg weight, measured before incubation, did not affect protein turnover. 5. It was concluded that the strain difference manifested in whole-body protein turnover of the chicken embryo would probably be a reflection of differences in genetic background.     

Cloning, characterization and expression of cDNA containing major histocompatibility complex class I, IIα and IIβ genes of Japanese flounder Paralichthys olivaceus

ABSTRACT:   The nucleotide sequences of Japanese flounder Paralichthys olivaceus , major histocompatibility complex (MHC) cDNA, classical MHC class Iα, non-classical MHC class Iβ, MHC class IIα and IIβ, were determined. The domain structures and antigen binding motifs of vertebrate MHC are conserved in the Japanese flounder MHC. A phylogenetic analysis supports the classification of these genes into class I and class II MHC. Classical MHC class Iα was ubiquitously expressed, whereas the non-classical MHC class Iβ was expressed mainly in lymphoid organs, gills, intestine and stomach. The MHC classes IIα and IIβ were also ubiquitously expressed.     

Feeding activities of Coptotermes formosanus Shiraki and Reticulitermes speratus (Kolbe) as affected by moisture content of wood

The relationship between the moisture content (MC) of wood blocks and the feeding activities of two Japanese subterranean termites, Coptotermes formosanus Shiraki and Reticulitermes speratus (Kolbe), was investigated with two choice tests. When three wood blocks with low MC (6%–12%), middle MC (79%–103%), and high MC (140%–182%) were exposed to workers of C. formosanus and R. speratus in a choice test, the feeding preferences were found in the middle MC blocks for C. formosanus, and the middle and high MC blocks for R. speratus, although wood blocks of low MC were also attacked. In a second choice test, wood blocks consisting of five pieces with water-impregnated (MC: 133%–191%) top, middle, or bottom piece were exposed to workers of both species. Higher consumptions were generally obtained in water-impregnated wood pieces and bottom pieces.     

Termite repellent sesquiterpenoids from Callitris glaucophylla heartwood

Fractions of methanol and ethanol extracts from the heartwood of white cypress pine (Callitris glaucophylla Thompson et Johnson) were investigated for their repellent activity against subterranean termite Coptotermes formosanus Shiraki worker using a two-choice semicircular filter paper test at 0.5% (w/w) concentration. Fraction CY-E2 composed of (−)-citronellic acid, guaiol, α-, β-, and γ-eudesmol isomers as well as an unknown compound, showed the highest statistically significant repellency (97.8% ± 2.2 SEM) of all fractions tested. Bioactivity-guided fractionations using high-performance liquid chromatography led to the isolation of two, oxygenated eudesmane-type sesquiterpenes with α-methylene moieties, both termite-repellent compounds. These compounds were subsequently identified as ilicic acid methyl ester (IAME) and costic acid by means of spectroscopic analyses, electron impact mass spectrometry, and nuclear magnetic resonance spectroscopy. We report the isolation of both IAME and costic acid from C. glaucophylla heartwood for the first time. Part of this paper was presented at the 47th Symposium on the Chemistry of Terpenes, Essential Oils, and Aromatics, Tokyo, November 2003     

Differences in spectral trajectory with stand volume development between Japanese larch and Japanese oak in Hokkaido,Japan

Our objective was to investigate the differences in the spectral trajectory of the LANDSAT Thematic Mapper with stand volume development between Japanese larch (Larix kaempferi) and Japanese oak (Quercus mongolica var. crispula) in the experimental forest of Kyushu University, Hokkaido, Japan. We used nonlinear regression and a categorical variable to investigate the relationship between spectral reflectance factor and stand volume for each forest type. This research showed that the reflectance factor of Japanese larch and Japanese oak reached saturation and that both forest types had different spectral trajectories with stand development. In addition, we found that middle of development stage (stand volume was between 100 and 200 m³/ha) of each forest type had similarity in reflectance factor.     

Hydration behavior and compressive strength of cement mixed with exploded wood fiber strand obtained by the water-vapor explosion process

Poor compatibility was found between exploded wood fiber strand (WFS) and cement due to the excessive presence of water-soluble degraded polysaccharides in extractives of exploded WFS obtained from weathered wood waste treated by the water-vapor explosion process (WVEP). This study presents some comparative results from a continuing investigation on the compressive strengths of exploded WFS–cement mixtures. Based on results previously obtained with the hydration test, the relation between hydration behavior and compressive strength of the mixture was explored. In addition, the effect of the curing age on compressive strength development of the mixture with selected additive chemicals was examined. The results supported the results of early studies with hydration tests indicating that adding MgCl₂ to the mixtures of exploded WFS mixed with quick-curing cement or ordinary Portland cement and a composite of MgCl₂ + CaO added to the mixture of exploded WFS and furnace-slag cement effectively improved the hydration behaviors; it greatly enhanced the compressive strengths of mixtures as well. Compressive strengths were strongly correlated to maximum hydration temperatures (T_max) of wood–cement mixtures influenced by the cement type, wood wastes (treated or not with WVEP), additive chemicals, and their content levels. The results also indicated that adding selected chemicals had no significant effect on compressive strength among the mixtures of exploded WFS mixed, respectively, with three types of cement at a curing age of 180 days. X-ray diffraction, scanning electron microscopy, and energy dispersive X-ray spectroscopy were used to identify the hydration products and to probe the element distribution of the mixture in the wood–cement interface zone from a fractured surface.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002     

Screening method for wood extractives: direct cellulose thin-layer chromatography plate

A method for screening wood extractives was developed using cellulose thin-layer chromatography plate (Cell-TLC) separation and was directly applied to bioassays. Wood meal (60 mesh) from nangka (Artocarpus heterophyllus Lamk) heartwood was extracted with hot methanol. The crude extract was separated using a Cell-TLC plate (50 × 50mm). Two broad bands with R_f values of 0.46 and 0.96 were found, and the bands showed completely different effects against the pest termite Coptotermes formosanus Shiraki and the decay fungus Fomitopsis palustris. The band with R_f 0.46 was preferentially consumed by workers of C. formosanus, and it did not show any growth inhibition against F. palustris when the Cell-TLC plate was directly exposed to the organism. In contrast, the band with R_f 0.96 appeared to repel strongly the feeding by C. formosanus and inhibited the growth of F. palustris. It was concluded that the Cell-TLC system was applicable for screening wood extracts consisting of many compounds.