Age Difference in Morphology and Immunohistology inthe Thymus and Spleen in Crl:CD (SD) Rats

We investigated chronological changes in immunohistochemical phenotyping in the thymus and spleen in Crl:CD rats up to the age of about one year. In the thymus, T cells increased markedly from 3 to 4 weeks of age. Proliferating cells also increased markedly at these points. B cells tended towards an increase with age. In the spleen, white pulp increased until 9 weeks of age and remained fairly stable thereafter. In the periarteriolar lymphoid sheath and marginal zone, T cells gradually increased until 9 weeks of age and became almost flat thereafter. In the lymph follicle, T cells increased with age. B cells tended towards an increase with age in all areas of the spleen. It was concluded that development of the thymus was most marked from 3 to 4 weeks of age and that both the thymus and spleen had matured by 9 weeks of age.     

Response to mitogens of chicken splenocytes determined by three methods of measurement

The aim of the present study was to examine whether the 3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5‐diphenyl tetrazolium bromide (MTT) colorimetric assay can be applied to measurement of mitogen‐induced chicken splenocyte activation. Activation was also measured by a ³H‐thymidine uptake assay and a viable cell count assay. Optimal concentrations of mitogens and incubation periods required for maximal responses to mitogens differed between the MTT assay and the viable cell count and thymidine uptake assays. This probably reflects differences in the activities measured by the MTT assay which detects mitochondrial enzyme activity, and the thymidine uptake and viable cell count assays which detect cellular proliferation activity. The validity of the MTT assay was supported by the observation that the mitogen‐induced increase in succinate dehydrogenase activity paralleled the level of mitogen‐induced MTT formazan production. Mitogen concentrations inducing maximal formazan formation in chicken splenocytes were higher than those for chicken peripheral blood lymphocytes reported previously. Results of the present study indicate that mitogen‐induced chicken splenocyte activation could be measured by the MTT colorimetric assay, although mitogen concentrations and incubation periods required for maximal splenocyte activation differed between the MTT assay and the other two assays used in this study.     

Disease Resistance of Tissue-cultured Seedlings of Rhododendron after Treatment with Streptomyces sp. R-5

An endophytic actinomycete, Streptomyces sp. R-5, which had been isolated from a field-grown rhododendron plant, was used to protect rhododendron seedlings in tissue culture from Pestalotia disease caused by Pestalotiopsis sydowiana. R-5 had intense antagonistic activity against P. sydowiana without adversely affecting the seedlings in glass flasks. A suspension of R-5 was spread on the surface of the multiplication medium in glass flasks in which seedlings were growing. Ten days later, the 4th upper leaf of seedlings was inoculated with P. sydowiana and incubated for 14 days. In controls untreated with R-5, substrate mycelia of P. sydowiana grew on all leaves and stems above and below the 4th leaf within 2–3 days of inoculation. Such growth resulted in the wilting death of 54% of seedlings by 14 days. In contrast, only the inoculated leaves turned brown in ca. 90% of seedlings growing on medium treated with R-5. None of these seedlings died. Thus, treatment of the medium surface with R-5 efficiently protects the seedlings from infection by P. sydowiana. Scanning electron microscopy revealed that substrate mycelia of R-5 grew on and beneath the cuticle of leaves of the treated seedlings. Fluorescent microscopy showed that R-5 was also inside the leaves. Received 8 June 2001/ Accepted in revised form 4 July 2001     

Experimental studies on rain splash erosion of forest soils after clearing in Okinawa using an artificial rainfall apparatus

The main island of Okinawa, in southwestern Japan, is characterized by a subtropical climate, coral reefs, and reddish soils, which contain Acrisols and Cambisols (FAO/UNESCO classifications). Recently, due to soil erosion on hilly lands, the coral reefs have been damaged by an inflow of reddish soil. Forest clearing is thought to one of the factors increasing rain splash erosion rates on hilly lands, because it is believed that clearing disturbs the forest floor and causes the supply of litter to stop. After clearing, A0, A and B horizons will be denuded, one after the other, by rain splash. In this study, we measured rain splash erosion rates of undisturbed samples of A0, A and B horizons using an artificial rainfall apparatus. The results of experiments on rain splash erosion clarified several aspects of the erosion process in a clearing site. First, it was found that the rain splash erosion rates were higher in fresh litter than in decomposed litter, in decomposed litter than in the A horizon, and in the A horizon than in the B horizon. Thus, the erosion rate increases with soil depth. Secondly, surface-gleyed red and yellow soils are mostly vulnerable to rain splash erosion. The erosion rates of these soils are two to three times higher than those of the red soil and the yellow soils. Thirdly, the erosion rates are affected mainly by bulk density and saturated hydraulic conductivity, not by clay ratio and dispersion ratio. Lastly, the difference between erosion rates of surface-gleyed red and yellow soils and other soils will be even greater if a soil crust has formed, because the formation of a soil crust increases the rain splash erosion rate.     

Production of Diabetic Offspring Using Cryopreserved Epididymal Sperm by In Vitro Fertilization and Intrafallopian Insemination Techniques in Transgenic Pigs

Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains that model human diseases. However, production of numerous cloned disease model pigs by SCNT for large-scale experiments is impractical due to its complexity and inefficiency. In the present study, we aimed to establish an efficient procedure for proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and treated with insulin to allow for normal growth to maturity, at which point epididymal sperm could be collected for cryopreservation. In vitro fertilization and intrafallopian insemination using the cryopreserved epididymal sperm resulted in diabetes model transgenic offspring. These results suggest that artificial reproductive technology using cryopreserved epididymal sperm could be a practical option for proliferation of genetically modified disease model pigs.     

Recent races of spinach downy mildew pathogen <Emphasis Type="Italic">Peronospora farinosa</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan

Isolates of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, collected at seven sites in Japan from December 2013 to March 2016, were identified as belonging to races 8, 10, 12 and 13, and one was unauthorized. Races 10, 12, 13 and the unauthorized were first found in Japan.     

Effect of nitrogen deposition on CH4 uptake in forest soils in Hokkaido,Japan

Abstract

It has been well documented by short-term artificial experiments that the CH⁴ uptake is inhibited by N input, especially NH^{4 p+}-N input. To investigate the effect of the natural N input by throughfall and other factors on the CH⁴ uptake in forest soils, we measured the CH⁴ uptake rates for 6 months during the snow-free period of the year and N input by throughfall throughout the year at 10 sites in Hokkaido, Japan, from 1997 to 2002. Water filled pore space (WFPS) and pH values in the soils varied widely among the sites (38-93% and 3.9-6.2, respectively). The rates of NH^{4 p+}-N and NH^{3 p-}-N inputs ranged from 1.3 to 6.9 kg N ha^p-1 year^p-1 and from 0.8 to 2.9 kg N ha^p-1 year^p-1, respectively. The NH^{4 p+}-N input was generally higher than the NH^{3 p-}-N input. Total N input by throughfall amounted to 2.3-9.4 kg N ha^p-1 year^p-1. The highest CH⁴ uptake rate occurred within the period from July to September (41-215 μg CH⁴ m^p-2 h^p-1) each year at most sites. CH⁴ uptake rate was relatively low (~50 μg CH⁴ M-2 h^p-1) at northern sites, while a high CH⁴ uptake rate was observed throughout the year 100 (? CH⁴ m^p-2 h^p-1) at southern sites. The mean CH⁴ uptake rates were significantly different among the sites. Cumulative CH⁴ uptake ranged from 1.4 to 6.6 kg CH⁴ ha^p-1 [184 d]^p-1 with a mean values of 3.22 ± 1.36 kg CH⁴ ha^p-1 [184 d]^p-1. Cumulative CH⁴ uptake increased with increasing temperature and decreased with an increase in precipitation (Rain), NH^{4 p+}-N input (TF^NH4) WFPS, soil total C (TC), and total N (TN). There was a quadratic relationship between the CH⁴ uptake and NH^{3 p-}-N input (TF^NO3), soil pH, and C / N ratio in soil. A regression equation was obtained as follows to predict the CH⁴ uptake in forest soils: Cumulative CH⁴ uptake = 0.47 / Rain + 0.38 / TF^NH4 + 0.34 / TC - 0.30 / TF^N03 (R ^p2 = 0.74, p = 0.0001). This equation indicates that atmospheric N input into forest soils is one of the main factors that control cumulative CH⁴ uptake with precipitation, total carbon content in soil in Hokkaido, Japan.