Receptor assay-guided isolation of anti-GABAergic insecticidal alkaloids from a fungal culture

The gamma-aminobutyric acid (GABA) receptor bears sites of action for insecticides. To discover GABA receptor-directed insecticides in natural products, fungal culture extracts were screened for their ability to inhibit specific binding of the radiolabeled noncompetitive antagonist [3H]1-(4-ethynylphenyl)-4-n-propyl-2,6,7-trioxabicyclo[2.2.2]octane to housefly head membranes. The screening efforts led to the isolation of two alkaloids from Aspergillus terreus: PF1198A (alantrypinone) and PF1198B (serantrypinone), which had IC50 values of 0.34 and 2.1 microM, respectively, in this assay. These compounds were ca. 47-61-fold selective for housefly vs rat GABA receptors. Both compounds showed insecticidal activity against Myzus persicae in the range of 100-500 ppm. Binding assay-guided screening should provide significant opportunities for the identification of novel and selective insecticides.     

Detection of candidate polymorphisms around the QTL for fat area ratio to rib eye area on BTA7 using whole‐genome resequencing in Japanese Black cattle

In our previous study, we performed genome‐wide association study (GWAS) to identify the genomic region associated with Fat area ratio to rib eye area (FAR) and detected a candidate in BTA7 at 10–30 Mbp. The present study aims to comprehensively detect all polymorphisms in the candidate region using whole‐genome resequencing data. Based on whole‐genome resequencing of eight animals, we detected 127,090 polymorphisms within the region. Of these, 31,945 were located within the genes. We further narrowed the polymorphisms to 6,044 with more than five allele differences between the high and low FAR groups that were located within 179 genes. We subsequently investigated the functions of these genes and selected 170 polymorphisms in eight genes as possible candidate polymorphisms. We focused on SLC27A6 K81M as a putative candidate polymorphism. We genotyped the SNP in a Japanese Black population (n = 904) to investigate the effect on FAR. Analysis of variance revealed that SLC27A6 K81M had a lower p‐value (p = .0009) than the most significant SNP in GWAS (p = .0049). Although only SLC27A6 K81M was verified in the present study, subsequent verification of the remaining candidate genes and polymorphisms could lead to the identification of genes and polymorphisms responsible for FAR.     

The dietary effects of a fermented vegetable product on glutathione peroxidase activity and lipid peroxidation of Japanese flounder Paralichthys olivaceus

ABSTRACT:   To study the dietary effects of fermented vegetable product (FVP) on the protection against lipid peroxidation of various tissues in Japanese flounder, Paralichthys olivaceus . The fish were fed on experimental diets with or without FVP for 4 weeks. The glutathione concentrations in serum or liver had a tendency to increase on FVP-feeding fish. The FVP-feeding fish showed higher glutathione peroxidase (GPx) activity of liver than the control fish, and the GPx activity was increased by the administration of 6 mg/kg body weight/day FVP ( P  < 0.05). Conversely, fish fed on FVP containing diets exhibited significantly ( P  < 0.05) lower lipid peroxidants of serum and liver than the control fish. The FVP is suggested to suppress lipid peroxidation in the administrated fish, which led to enhancement of antioxidant effect against cultured fish.     

Age and growth of yellowstriped butterfish, <Emphasis Type="Italic">Labracoglossa argentiventris</Emphasis>, around Izu Oshima Island

ABSTRACT:   Age and growth of the yellowstriped butterfish, Labracoglossa argentiventris , around Izu Oshima Island were studied using a total of 1450 fish. Age was determined by counting the edge of the opaque zones as a ring mark on sectioned sagittal otoliths. Formation of the first ring was observed during spring or summer, corresponding to 1.5 years after hatching. Thereafter, one ring was formed each year in the same season as the previous year. The growth of the butterfish was rapid until 2 years of age. The maximum likelihood method was applied to the age and length data for estimating parameters in von Bertalanffy, Gompertz, and Logistic growth models. The selected model, based on the Akaike Information Criterion, was the von Bertalanffy growth model, which indicated differential asymptotic length and variance by sex.     

Simultaneous estimation of instantaneous mortality coefficients and rate of effective survivors to number of released fish using multiple sets of tagging experiments

ABSTRACT:   A method is proposed for estimating two instantaneous mortality coefficients, and rate of effective survivors to number of tagged fish released, using exact times at liberty from multiple sets of tagging experiments. Effective survivors are defined here by the product of survivors immediately after releasing and reporting rate of recaptures. A least-squares method was applied to estimate the parameters using statistical models expressing the numbers of effective survivors, which were calculated by a principle similar to virtual population analysis. In order to investigate the statistical performance of the method, simulation trials using operating models were carried out. The method was applied to data of tagging experiments for alfonsino Beryx splendens from 1957 to 1969, off Kanagawa Prefecture. In most of the results of simulation trials, the method yielded estimates of natural mortality coefficient of which expected values were within ± 12% of true values. Estimates of natural mortality coefficient, fishing coefficient and the rate of effective survivors for alfonsino are 0.57 (per year, 95% confidence limits [CL] 0.40–0.69), 0.094 (per year, 0.03–0.25) and 0.39 (0.12–1.00), respectively. Modifications for the generalization of the method and notes on its applications are discussed.     

The effect of ovary storage and in vitro maturation on mRNA levels in bovine oocytes; a possible impact of maternal ATP1A1 on blastocyst development in slaughterhouse-derived oocytes

Since BSE testing of slaughtered cattle is obligatory in Japan, storage of ovaries at 15-20 C overnight in phosphate buffered saline has become a routine protocol in in vitro production (IVP) of cattle embryos. Ovary storage is known to reduce developmental competence of oocytes; however, its effects on oocyte gene expression have not been clarified yet. This study compared oocytes collected from stored slaughterhouse-derived ovaries with those collected by Ovum Pick-Up (OPU) in terms of the expression of 20 selected genes to determine if ovary storage affects cellular processes at the molecular level. Expression of mRNA in oocytes was assayed before and after in vitro maturation (IVM) by real-time quantitative PCR. Maternal mRNA levels of genes were investigated in 2-cell stage embryos obtained from slaughterhouse oocytes to assess their roles for blastocyst formation. In immature OPU oocytes, genes related to metabolism (GAPDH), transporters (GLUT8, ATP1A1) and stress resistance protein (HSP70) showed significantly higher expression compared with oocytes derived from stored ovaries. During IVM, the expression of GDF9, GLUT8, CTNNB1 and PMSB1 was significantly decreased irrespective of oocyte source. Two-cell stage embryos cleaving at 22-25 h after in vitro fertilization (IVF) showed a significantly higher blastocyst formation rate and ATP1A1 gene expression level compared with those cleaving at 27-30 h after IVF. Our results reveal that storage of ovaries alters mRNA levels in oocytes. Correlation of Na/K ATPase ATP1A1 expression in IVP embryos at the 2-cell and 8-cell stages with their developmental ability to the blastocyst stage may suggest the importance of maternal mRNA of this gene during blastulation in embryos derived from slaughterhouse oocytes.     

Fine mapping the number of corpora lutea quantitative trait loci on SSC3: Analysis of the porcine follicle‐stimulating hormone receptor gene

In females, follicle‐stimulating hormone (FSH) targets a FSH receptor (FSHR) expressed only on granulose cells, inducing maturation of the ovarian follicles. We hypothesized that genetic variants in the FSHR gene influence litter size by affecting the number of corpora lutea. We fine‐mapped a region of Sus Scrofa chromosome 3 that contains quantitative trait loci for corpora lutea. Polymorphisms were detected in the exons and 5′ flanking region of the porcine FSHR gene, a positional candidate for the statistically most significant of the quantitative trait loci. Finally, 248 F₂ animals from a Duroc and Meishan cross were genotyped for three FSHR SNPs at positions 74, 532 and 1166, and these were correlated with the phenotypes of litter size and corpus luteum number. Three haplotypes were identified: M1 (G/G/C), M2 (C/A/T) and D (C/A/C). In the F₂ population, the M1 haplotype was associated with a greater number of corpora lutea (P < 0.01) and also seemed to be associated with increased litter size, although the association was not significant (P = 0.2571). Some polymorphisms resulting in amino acid substitutions in these genes were excluded from the polymorphisms possibly responsible for the number of corpora lutea.     

多因素影响下拖拉机侧向稳定性模型实验

针对拖拉机斜坡直线行驶工况,基于拖拉机比例模型和3D打印技术,建立了模型拖拉机轮胎-地面载荷实验测试系统。以斜坡上侧车轮-地面载荷为主要参考量,提出了针对拖拉机前、后轮的侧向稳定评价指标(拖拉机前、后轮的斜坡上侧车轮载荷分配系数)。采用田口实验设计方法,选择前后轮轮胎类型、前配重质量、前后轮距和机具位置6个影响因素作为控制因子,以E级和F级随机路面作为噪声因子,设计了6因子混合水平的田口实验方案,并对实验结果进行信噪比和均值的方差分析。实验结果表明,对拖拉机斜坡上侧前、后轮侧向稳定性影响最大的控制因子分别是前配重质量和后轮距;得出基于前、后轮侧向稳定性评价指标的拖拉机最优配置,为拖拉机的稳定性优化设计提供了一定参考,也为拖拉机防侧翻预警控制提供了理论基础。     

Clarification of relationship between single-nucleotide polymorphism panels of Shiga toxin-producing Escherichia coli O157:H7/H- strains

Eighty strains of enterohemorrhagic Escherichia coli O157:H7/H- were analyzed by three single-nucleotide polymorphism (SNP) panels using whole-genome sequencing data. The partial concordance of SNP types among the different SNP panels was observed on minimum spanning trees reconstructed with SNP data. As for lineage I/II strains, some of the clade 7 strains belonged to one unique SNP type as determined by three panels, suggesting that clade 7 should be divided into at least two genotypes, namely, the unique type and the rest. In addition, clade 8 contained two unique genotypes, which was consistent with the previous prediction. Similarly, for lineage II, clade 12 should be divided into three genotype strains. In contrast, many strains of several clades belonging to lineage I were clustered into the same node on each minimum spanning tree upon testing with the three SNP panels. Previous studies reported that lineage I diverged more recently than lineages I/II and II. Such low diversity in lineage I in this study may have arisen because this lineage has not accumulated SNPs because of its relatively recent divergence. Based on the concordance observed in this study, some of the previously published O157 genotype distribution data were successfully interpreted to clarify the clade distribution, which was well supported by previous literature.