Protective effect of Clostridium septicum alpha-toxoid vaccine against challenge with spores in guinea pigs

The protective effect of an alpha-toxoid vaccine of Clostridium septicum purified alpha-toxin was investigated in guinea pigs. Purified alpha-toxin was treated with formalin to make toxoid, and alpha-toxoid vaccine was prepared by mixing alpha-toxoid (4 to 64 microg/dose) with an aluminum phosphate gel as adjuvant. Guinea pigs were immunized twice with different doses of alpha-toxoid vaccine, and challenged with spores of C. septicum. The guinea pigs surviving after challenge had been immunized with 8 microg/dose or more of alpha-toxoid. All these animals produced titers of 20 units or higher of antitoxin at the challenge. The results suggest that C. septicum alpha-toxin plays an important role in protection against challenge with spores in guinea pigs.     

Allele frequency distribution of the canine dopamine receptor D4 gene exon III and I in 23 breeds

Various canine breeds are remarkably different from each other not only in their sizes and shapes but also in behavioral traits, suggesting that some of them are under genetic control. Although dopaminergic neurotransmission system is considered to affect animal behavior, little is known about related genes in canine. Relations between specific alleles in polymorphic regions of the dopamine receptor D4 gene (DRD4) and personality or psychiatric disorders have been reported in humans, and we first found polymorphism in exon III region of the gene in 4 canine breeds. In this study we surveyed allele frequency distribution in 23 breeds including a total of 1,535 unrelated individuals. In exon III, 8 alleles including a novel allele were identified. A group of breeds in which the alleles 447b, 498 and 549 were frequent tended toward high scores in aggression-related behavioral traits than that with frequent alleles 435 and 447a. Moreover, a polymorphism based on 24 bp insertion/deletion was found in exon I region for the first time in dogs. This information may be of use for candidate gene studies of behavioral variation in dogs.     

Spatiotemporal distribution of Flavobacterium psychrophilum and ayu Plecoglossus altivelis in rivers revealed by environmental DNA analysis

Outbreaks of bacterial cold-water disease (BCWD), caused by Flavobacterium psychrophilum, are widespread in Japan, especially among ayu Plecoglossus altivelis. There are few investigations of F. psychrophilum in river water, and its seasonal distribution has not been clarified. We aimed to identify the spatiotemporal dynamics of F. psychrophilum and ayu to provide information that is useful for establishing a countermeasure for BCWD. Quantitative analysis of environmental DNA (eDNA) was used to clarify the year-round dynamics of ayu and F. psychrophilum. We sampled river water from the Nagara and Ibi rivers in Japan, and conducted monthly water sampling and eDNA quantification. Changes in the eDNA concentration of ayu were consistent with the known life histories of the fish. There was a strong negative correlation between the eDNA concentration of F. psychrophilum and water temperature, suggesting a strong dependence of F. psychrophilum dynamics in the river on water temperature. Furthermore, relatively high eDNA concentrations were recorded for both organisms in early summer and fall, suggesting that ayu is infected with F. psychrophilum during these seasons when experiencing up- and downmigration, respectively.

     

云纹石斑鱼(♀)×七带石斑鱼(♂)杂交F1形态特征与其亲本子代的比较分析

为研究云纹石斑鱼(♀)×七带石斑鱼(♂)杂交F1的形态特征以及与其亲本子代的关系,运用常规测量方法和聚类分析法对云纹石斑鱼、七带石斑鱼及云纹石斑鱼(♀)×七带石斑鱼(♂)杂交F1 3个群体6月龄幼鱼的形态特征进行测量和分析,对3个群体的12项可量、可数性状进行指数分析。结果显示,杂交F1体型呈椭圆形,侧扁而粗壮,体侧具7条灰褐色横向条带,杂交F1在体色条纹上均具有云纹石斑鱼与七带石斑鱼的特征;可量、可数的性状中有6项性状偏向于云纹石斑鱼,5项性状偏向于七带石斑鱼,1项与二者相同;聚类分析结果表明,杂交F1与云纹石斑鱼聚为一类。研究表明,杂交F1在体色条纹上表现为二者的中间态,在形态上较接近于母本云纹石斑鱼。     

Natural hybridization between <Emphasis Type="Italic">Pinctada fucata</Emphasis> and <Emphasis Type="Italic">Pinctada maculata</Emphasis> inferred from internal transcribed spacer regions of nuclear ribosomal RNA genes

ABSTRACT:   Genetic evidence of the occurrence of natural hybridization between female Pinctada fucata and male Pinctada maculata among wild pearl oysters ( n  = 20) collected for use as the mother shell for private pearl farming in the Oshima Strait at Amami-o-shima, Kagoshima Prefecture, Japan, were obtained. A polymerase chain reaction-based species identification method for Pinctada was developed using polymorphisms in the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA (rRNA) gene. This method enabled the amplification of the ITS regions using a primer set specific for P. maculata and P. fucata . However, 10 of 20 individuals morphologically identified as P. fucata had sequences specific to both P. maculata and P. fucata in the ITS region. These putative hybrids showed sequences of a maternally inherited mitochondrial 16S rRNA gene, identical to that of P. fucata . Shells of the putative hybrids were difficult to discriminate from those of P. fucata exhibiting similar taxonomic traits. Moreover, the hybrids exhibited slower growth than P. fucata but faster growth than P. maculata .     

Antinecrotic and Antiapoptotic Effects of Different Concentrations of Copaiferalangsdorffii Essential Oil in Macrophages and Lymphocytes of Mice

Purpose： to evaluate the in vitro influence of Copaiferalangsdorffii essential oil on the antinecrotic activity in macrophages, and on the antiapoptotic activity in macrophages and lymphocytes of Swiss mice. Methods： lymphocytes and macrophages were obtained from ten 60-day-old male mice weighing 42-45 g, and then cultured in 24-well plates. Cells were treated with concentrations of 2%, 5%, 10% and 20% of oil from Copaiferalangsdorffii before or after induction of apoptosis with ultraviolet light. We used positive and negative controls. Hoechst staining was performed, and then images of cells under fluorescence microscope were captured and digitalized. We used Image J_1.46o to quantify the fluorescence intensity/brightness of condensation of the chromatin ofapoptotic cells. We used one-way ANOVA with Bonferroni＇s and Tukey＇s multiple comparison tests. Results are expressed as the means of relative densities after adjustment and 95% confidence intervals; P 〈 0.05 was considered statistically significant. Results： oil from Copaiferalangsdorffii at different concentrations has decreased necrosis and apoptosis of both lymphocytes and macrophages from mice, when compared to C＋ （all P 〈 0.05）. Discussion： Copaiferalangsdorffii is typically Brazilian and found in the Amazon region, with anti-inflammatory, antioxidant and antineoplastic effects due to its oil compounds, such as xyloglucans and kaurenoic acid. Its potential therapeutical usage was demonstrated in a national study. The present study showed that its oil has significant in vitro antiapoptotic and antinecrotic activity and it should be tested experimentally in vivo.     

Effects of momilactone on the protein expression in Arabidopsis germination

Although momilactone is known to play an important role in the allelopathy of rice (Oryza sativa L.), there is no information available about the mode of action of momilactone on the allelopathy of Arabidopsis. The present research describes the allelopathic activity of momilactone A and B on the germination of Arabidopsis thaliana and the effects of momilactone A and B on protein expression during germination. Momilactone A and B inhibited the germination of Arabidopsis at concentrations >30 and 10 μmol L^?1, respectively. The protein bands of 18, 19 and 21 kDa on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) were abundant in the momilactone A‐ and B‐treated Arabidopsis compared with the control. The protein bands of 18, 19 and 21 kDa were identified by the N‐terminal amino acid sequencing as cruciferin 2, cruciferina and cruciferin 3, respectively. Cruciferins and cruciferina are important storage proteins and play an important role as the initial source of nitrogen for seed germination. These results suggest that momilactone A and B may inhibit the germination of Arabidopsis seeds by inhibiting the degradation process of cruciferin and cruciferina.     

Importance of AC-rich Element on Pea Phenylalanine Ammonia-Lyase Gene 1 Promoter for Expression Induced by Nonpathogenic Attack

Regulatory elements in the promoter of phenylalanine ammonia-lyase gene 1 of pea (PSPAL1) in response to nonpathogenic attack were identified by in vivo footprinting analysis. The footprints determined AC-rich sequences, Box-I and Box-II, that were conserved at similar positions in the phenylpropanoid gene promoters from several plants. To reveal the functions of the AC-rich sequence in nonpathogen-responsiveness, we constructed Box-I-deletion PSPAL1 promoter (dB-1) with GUS reporter gene and transformed it into tobacco plant. The dB-1 had reduced basal expression and a complete loss of nonpathogen-responsiveness. These results indicate the essentiality of Box-I for PSPAL1 activation induced by nonpathogenic attack. Received 27 October 1999/ Accepted in revised form 25 November 1999     

A series of 19'-hexanoyloxyfucoxanthin derivatives from the sea mussel, Mytilus galloprovincialis, grown in the Black Sea, Ukraine

A series of carotenoids with 19- or 19'-hexanoyloxy moieties, 19'-hexanoyloxyfucoxanthin (1), 19'-hexanoyloxyfucoxanthinol (2), 19'-hexanoyloxyhalocynthiaxanthin (3), 19-hexanoyloxycrassostreaxanthin A (4), 19-hexanoyloxymytiloxanthin (5), and 19-hexanoyloxyallenicmytiloxanthin (6) were isolated from the edible part of the sea mussel, Mytilus galloprovincialis, grown in the Black Sea, Ukraine. Among them, 3, 4, and 6 were new compounds. These structures were charcterized by UV-vis, fast atom bombardment mass spectrometry, circular dichroism, and (1)H NMR spectroscopic data. They were assumed to be metabolites of 19'-hexanoyloxyfucoxanthin (1).