麦冬块根提取物对稻瘟病菌丝及苗期稻瘟病发生的抑制效果

采用不同浓度来自70%甲醇溶液的麦冬块根提取物对2种产自日本的稻瘟病菌生理小种菌株MAFF305480和MAFF101002的菌丝生长及其接种诱发水稻幼苗稻瘟病发生的抑制效果进行评价。试验表明：含有麦冬块根提取物的原液1/10,1/20,1/40以及1/80浓度都能明显抑制稻瘟病菌小种菌株(MAFF305480和MAFF101002)菌丝生长, 抑制率均56.4%以上, 对接种诱发稻瘟病的水稻幼苗处理发现, 幼苗植株发病级数为明显下降, 抑制率均38.4%以上, 并且都随着处理浓度提高抑制效果明显提高。本研究结果表明,麦冬块根提取物中含有抑制稻瘟病菌生长的化学物质, 并且能直接抑制稻瘟病菌发生和生长。     

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。     

Development of Peyer's patch and cecal tonsil in gut-associated lymphoid tissues in the chicken embryo

It is well known that chicken B cells develop in the bursa of Fabricius (BF), which is categorized as gut-associated lymphoid tissue (GALT). Chicken GALT also includes Peyer's patch (PP) and cecal tonsil (CT). The relationship between these tissues in GALT during B cell development is currently unknown. In this study, we conducted comparative examination of PP, CT and BF development during embryogenesis using immunohistochemical staining. On day 13 of embryogenesis (E13), accumulation of MHC class II(+) cells was observed in the intestine. Thereafter, Bu-1(+) cells and IgM(+) cells appeared, and their number continuously increased at the same sites where MHC class II(+) cells were present. Similar results were obtained in the CT. The locations of embryonic PP were limited to two sites; near the Meckel's diverticulum and the ileocecal junction. Anlage of bursal follicles first appeared at E13 and developed thereafter. Immigration of Bu-1(+) cells to bursal follicles began at E13, and the number of Bu-1(+) cell subsequently increased. When the follicle of BF was eliminated from the embryo by treatment with testosterone, development of PP and CT were observed. We concluded therefore that the development of PP and CT start during late embryogenesis at the same time as the follicle of BF, and that appearance of surface IgM(+) cells in PP and CT is independent form the development of the follicle of BF.     

Clarification of adsorption and movement by predicting ammonia nitrogen concentrations in paddy percolation water

We noted that ammonia nitrogen was not adsorbed by the cultivated layers of highly permeable paddy fields during the initial fertilization period, but reached the lower layers relatively early. In our study, we considered an exponential equation from an aqua-environmental perspective with the goal of obtaining good growth of rice plants in order to estimate the concentrations and integrated volume of ammonia nitrogen accompanying paddy percolation. Using this exponential equation, we were able to derive a relation between time and concentrations of paddy percolation water, and hypothesized that if percolation rates were less than 10 mm/day, percolation would have no effect on rice growth, while simultaneously helping to maintain the good water quality of the extra-paddy environment. We also clarified the differences between the potential ammonia nitrogen adsorption volume derived from the CEC value and the integrated amount of ammonia nitrogen water in soil, and considered the causes from the perspectives of solute movement and water movement. Electronic Publication     

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.     

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 blastocyst production rate and incidence of chromosomal abnormalities by developmental stage in in vitro produced porcine embryos

The present study was conducted to determine the relationship between embryonic development speed at different stages (the cleaved stage at 52 h and the blastocyst stage at 6 days post insemination) and incidences of chromosome abnormalities in in vitro produced porcine embryos. Porcine oocytes were collected from 3-6-mm ovarian follicles obtained at a slaughterhouse and matured in modified NCSU-37 medium for 44-46 h. Following in vitro fertilization with a final concentration of 1 x 10(5) sperm/ml for 3 h, all oocytes were cultured in vitro for 52 h. Day-2 (52 h after insemination) embryos were classified according to their cleaved stages into 2-cell, 3- to 4-cell, 5- to 8-cell, and >8-cell stages; these were cultured separately for additional 4 days (Day 6). The resultant Day-6 blastocysts were classified according to the morphological diameter into 3 grades: Grade A, expanded blastocysts; Grade B, expanding blastocysts; and Grade C, early blastocysts. They were then analyzed chromosomally. The 3- to 4-cell and 5- to 8-cell embryos had significantly high blastocyst development rates (46.1 and 36.9%, respectively), and these blastocysts contained significantly more cells (40.2 and 42.4 cells, respectively) than those derived from 2-cell embryos and >8-cell embryos (28.6 and 26.5 cells, respectively). The incidence of chromosomal abnormalities was significantly higher in the blastocysts derived from 2-cell and >8-cell stage embryos than in the blastocysts derived from the other stage embryos. Furthermore, the grade A blastocysts had the lowest incidence of chromosomal abnormalities (35.3%) and contained the most cells (48.7 cells). Porcine in vitro production (IVP) yielded a high blastocyst rate and an excellent embryo quality when 3- to 4-cell and 5- to 8-cell stage embryos were selected on Day 2 after insemination. The same criteria yielded a higher quality of expanded blastocysts based on the stage of embryo development and morphology.     

Pathology of cutaneous fowlpox with amyloidosis in layer hens inoculated with fowlpox vaccine

Cutaneous fowlpox occurring in vaccinated layer hens was investigated pathologically and microbiologically. Anorexia, decrease of egg production, increased mortality, yellow scabs on faces, and alopecia of feathered skins with yellow scabs were observed in affected hens. Histologically, proliferative and necrotic dermatitis with eosinophilic ring-shaped cytoplasmic inclusions (Bollinger bodies) and clumps of gram-positive cocci (Staphylococcus hyicus) were noted in the affected birds. Fowlpox lesions were primarily observed in the feathered skins. Proliferation of feather follicle epidermal cells, with cytoplasmic inclusions and degeneration of the feather, and bacterial clumps in the feather follicles were noted in the affected skins. Ultrastructurally, characteristic fowlpox viral particles were observed in the cytoplasmic inclusions of hyperplastic epidermal cells. Amyloid deposition was observed in the Disse space of the liver, splenic sinus, and lamina propria of the bronchiolar, bronchial, and tracheal areas. Amyloidosis could be one factor inducing the fowlpox infection in vaccinated chickens.     

Comparison of two quantitative assays for xenotropic murine leukemia virus-related virus

Xenotropic murine leukemia virus-related virus (XMRV), a novel gammaretrovirus in humans, was found in patients with prostate cancer (PC) and chronic fatigue syndrome (CFS). However, there has been controversy whether XMRV is directly associated with human diseases. In this study, we developed a LacZ marker rescue assay using human embryonic kidney 293T cells and a focus assay using a feline fibroblastic sarcoma-positive leukemia-negative QN10S cells. XMRV induced prominent foci in QN10S cells and the viral titer determined by the focus assay was as high as that by the LacZ marker rescue assay. Because the focus assay is simple and sensitive, it will be useful for monitoring infectious XMRVs in CFS and PC patients and virological studies for XMRV.