Analysis of the triple gene block sequence in an important melon pathogen,Melon yellowing-associated virus

A novel viral disease of melon, caused by Melon yellowing-associated virus (MYaV), has become one of the most serious problems for melon production in Brazil. Despite its importance, little is known about the genome sequence of the virus. In this study, the triple gene block genomic region including the 3′-end region of an isolate of MYaV was elucidated (ca. 3.1 kb). Sweet potato chlorotic fleck virus, a newly suggested carlavirus species, was the most closely related virus with 62% nucleotide identity. The presence of two heptanucleotide-motifs for subgenomic RNA synthesis was confirmed in this sequence.     

Aurantiochytrium sp. meal can replace fish oil in practical diets for the juvenile Pacific white shrimp

This study evaluated the use of microalgae (Aurantiochytrium sp.) meal as a substitute for fish oil in the diet of juvenile Pacific white shrimp (Litopenaeus vannamei) reared in a clear‐water system. Dietary treatments at five replacement levels (0%, 25%, 50%, 75%, 100%) were performed in triplicate. After 46 days, only a slight difference in shrimp final weight was observed among treatments (0.61 g). An increase in final weight was observed with replacement of up to 50% fish oil for microalgae meal, while the optimal percentage of replacement estimated was 44.7%. Feed conversion rate (FCR) of animals fed a diet of Aurantiochytrium sp.meal to replace up to 50% fish oil decreased, and the optimal percentage of replacement estimated was 49.3%. The fatty acids profile in shrimp muscle tissue demonstrated an increase in docosahexaenoic acid (DHA) from 10.03% to 14.28% with increased replacement of fish oil by microalgae meal in the diet. Therefore, the partial replacement of fish oil by microalgae meal resulted in improved shrimp growth and FCR, and total replacement of dietary fish oil had no negative effects on these parameters. In addition, inclusion of microalgae meal raises the level of DHA in shrimp muscle.     

利用内部信息的农用自动引导行走车的研究(第4报)——适应农业环境的控制系统

研究的行走控制系统由软、硬两部分组成，通过建立系统的控制模型，开发了利用非线性理论进行线性变换的直线行走控制方法和利用预测控制进行曲线行走控制的方法，通过实验确定了控制系统的参数；改造设计了行走车的行走与操向部分。实现了计算机的行走控制。     

Vitrification procedure decreases inositol 1,4,5‐trisphophate receptor expression,resulting in low fertility of pig oocytes

Although cryopreservation of mammalian oocytes is an important technology, it is well known that unfertilized oocytes, especially in pigs, are highly sensitive to low temperature and that cryopreserved oocytes show low fertility and developmental ability. The aim of the present study was to clarify why porcine in vitro matured (IVM) oocytes at the metaphase II (MII) stage showed low fertility and developmental ability after vitrification. In vitro matured cumulus oocyte complexes (COCs) were vitrified with Cryotop and then evaluated for fertility through in vitro fertilization (IVF). Although sperm‐penetrated oocytes were observed to some extent (30–40%), the rate of pronuclear formation was low (9%) and none of them progressed to the two‐cell stage. The results suggest that activation ability of cryopreserved oocytes was decreased by vitrification. We examined the localization and expression level of the type 1 inositol 1,4,5 trisphosphate receptor (IP₃R1), the channel responsible for Ca²⁺ release during IVF in porcine oocytes. Localization of IP₃R1 close to the plasma membrane and total expression level of IP₃R1 protein were both decreased by vitrification. In conclusion, our present study indicates that vitrified‐warmed porcine COCs showed a high survival rate but low fertility after IVF. This low fertility seems to be due to the decrease in IP₃R1 by the vitrification procedure.     

Oocyte maturation and active motility of spermatozoa are triggered by retinoic acid in pen shell <Emphasis Type="Italic">Atrina pectinata</Emphasis>

For recovery of the declining population of pen shells in the wild, the production of pen shell juveniles for transplantation or aquaculture is underway in Japan. For more stable juvenile production, artificial fertilization methods for pen shells are needed, but methods to induce oocyte maturation (meiosis resumption) used in other bivalves, which make oocytes fertilizable, were ineffective for pen shells. Here, we report evidence showing that retinoic acid (RA) has strong activity in inducing oocyte maturation and activating sperm motility in pen shells. Treatment of fully developed oocytes with 1.0 μM all-trans-RA (at-RA) induced germinal vesicle breakdown, a typical morphological sign of oocyte maturation, but 1.0 μM at-retinol and at-retinal, 2 mM ammonia, and 1.0 μM serotonin were ineffective. Treatment with at-RA for 30 min was sufficient for oocyte maturation and was more potent than its isomers, 9-cis- and 13-cis-RA. Parallel results were obtained for sperm motility activation. Oocyte responsiveness to at-RA increased during the final stage of ovary development. Artificial fertilization was successful only with the oocytes treated with at-RA, and fertilized eggs developed to D-shaped (veliger) larvae without apparent morphological abnormalities. These results indicate the possible application of RA for the artificial fertilization of pen shells.     

Chronological analysis of canine parvovirus type 2 isolates in Japan

Fifty-five canine parvovirus type 2 (CPV) samples, 12 fecal specimens and 43 cell culture isolates, were examined for their genetic characteristics of VP2 gene. They were collected from the diseased dogs at various districts of Japan during 27 years from 1980 to 2006. A fragment of VP2 gene was analyzed by restriction fragment length polymorphism assay and DNA sequencing. The original antigenic type 2 of CPV (CPV-2) was no longer found in the samples since 1984, and two antigenic variants CPV-2a and CPV-2b replaced CPV-2 as predominant types for about 5 years from 1982. A new genetic variant of prototype CPV-2a with non-synonymous substitution at the VP2 amino acid residue 297 from Ser to Ala was first detected in 1987. New CPV-2b with the same amino acid substitution at position 297 as new CPV-2a was also detected from the samples collected in 1997. Since then new CPV-2b has been the predominant CPV over the field of Japan. Several additional amino acid substitutions were detected in the VP2 gene of some recent CPV strains. Neither CPV-2c(a), CPV-2c(b), nor "Glu-426" of the antigenic variants previously found outside the country was detected in any samples tested. Reactivity of new CPV-2a and 2b variants against antibodies produced by the current vaccine products was determined by a cross hemagglutination-inhibition test. The recent field CPV isolates reacted more efficiently to the antibodies produced in dogs vaccinated with the new CPV-2b vaccine strain than the conventional CPV-2 vaccine strain.     

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.     

A complement-dependent neutralizing monoclonal antibody against glycoprotein II of pseudorabies virus

A monoclonal antibody (MAb), 1.21, was produced against pseudorabies virus (PRV). It exhibited virus neutralization activity only in the presence of complement. Immunoblot analysis after sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of virions revealed that MAb 1.21 bound with the 230 kilodalton (kD) virus protein only under non-reducing conditions. This protein was purified by immuno-affinity chromatography using MAb 1.21 and was found to be composed of three subunits, 60, 70 and 120 kD polypeptides when analyzed by SDS-PAGE under reducing conditions. This protein is probably glycoprotein II of PRV.     

Two cases of feline malignant craniopharyngioma

Tumors at the cranial base in 2 cats (a 9 1/2-year-old, castrated male Chinchilla and a 7-year-old, castrated male American Shorthair) were diagnosed as malignant craniopharyngioma. The tumor lesion was histopathologically divided into four parts: 1) a small acinus part, in which relatively large cells with a pale cytoplasm composed small acini; 2) a duct part, in which small cuboidal cells composed ducts; 3) a cyst part, in which there were large cysts lined with flat cells; and 4) a pavement part, in which large multi-angular-shaped cells proliferated in a pavement pattern. The epithelial cells of some parts were positive for keratin by immunohistochemistry. Histopathologic findings of the present feline cases were identical to those of malignant craniopharyngioma in other animal species.     

Synthesis and structure-activity study of fungicidal anilinopyrimidines leading to mepanipyrim (KIF-3535) as an anti-Botrytis agent

A series of 2-anilinopyrimidines was prepared and their fungicidal activities against Botrytis cinerea Pers were examined. The activity fell sharply with any substitution on the anilinobenzene ring. Substituents at the 5-position of the pyrimidine ring greatly reduced the activity. Substituents such as chloro, methoxy, methylamino, methyl or 1-propynyl were well tolerated at the 4- and 6-positions of the pyrimidine ring. Among these substituents, the combination of methyl and 1-propynyl groups was the most favourable. 2-Anilino-4-methyl-6-(1-propynyl)pyrimidine (KIF-3535), which showed excellent activity and no significant phytotoxicity, was finally selected for development and has been given the common name mepanipyrim.