Nondestructive observation of bovine milk by NMR spectroscopy: analysis of existing States of compounds and detection of new compounds

In this study were successfully observed the one- (1H, 13C) and two-dimensional (1H-13C, 1H-15N, 1H-31P) NMR spectra of milk directly without any pretreatment. The signals of each NMR spectrum were assigned, and their existing states were also analyzed. Lactose existed in a free state in milk. The signals due to the butyric acid chain can be assigned among the other fatty acid chains. Monounsaturated fatty acid (oleic acid chains) and polyunsaturated fatty acid chains (linoleic and linolenic acid) were assigned by their characteristic signals. The signals from citrate, N-acetylcarbohydrates, and lecithin could be observed directly in the 1H-13C HSQC NMR spectra; the assignment of their signals was made through the 1H-13C, 1H-15N, and 1H-31P HMBC spectra of extracted milk. Signals from creatine and N-acetylcarbohydrates were detected for the first time.     

Origin of waxy common millet, Panicum miliaceum L. in Japan

We developed PCR-based markers for genotyping Waxy genes in common millet (Panicum miliaceum L.) and proposed a new hypothesis on the origin and dispersal of waxy type of Japanese landraces. I₂-K staining indicated that almost all the Japanese landraces had waxy endosperm. Based on the PCR and dCAPs analyses of two waxy loci on different genomes we concluded that there were at least two possible lineages in Asian continent that had independent dispersal pathways into Japan. Most Japanese common millet had the similar genotypes with those in Korean Peninsula but a few particularly from northern part of Japan had common genotypes in northeastern China and Primorskaya Province of Russia.     

The W‐ and Z‐linked EE0.6 sequences used for molecular sexing of captive Japanese crested ibis on Sado Island

The Japanese crested ibis Nipponia nippon is a critically threatened bird. Accurate sexing is necessary to perform effective management of captive breeding toward a national project for a tentative release of the Japanese crested ibis on Sado Island. A PCR‐based sexing method targeting a 0.6 kb EcoRI fragment (EE0.6) sequence on W chromosome with AWS03 and USP3 primers has been developed for the Japanese crested ibis. However, the primers were selected from the EE0.6 sequences from bird species other than the Japanese crested ibis. In this study, we determined the W‐ and Z‐linked EE0.6 sequences in the Japanese crested ibis, and clarified Japanese crested ibis sequence mismatch in the binding sites of the primers. Further, we found no polymorphism in the primer binding sites among five founder birds for the Sado captive Japanese crested ibis population. These findings validated the PCR‐based sexing method with the AWS03 and USP3 as accurate molecular sexing methods of captive Japanese crested ibis on the Sado Island. Additionally, we designed a primer set for a novel PCR‐based sexing, based on the EE0.6 sequences obtained in this study. This novel sexing method may be useful for future ecological research following the release of Japanese crested ibis on Sado Island. This is the first report to show the EE0.6 sequences in Japanese crested ibis.     

Nationwide Cyprinid herpesvirus 3 contamination in natural rivers of Japan

Cyprinid herpesvirus 3 (CyHV-3) disease is a significant threat for common and koi carp cultivators and for freshwater ecosystems. To determine the prevalence of CyHV-3 in Japanese rivers, a nationwide survey of all national class-A rivers was undertaken in the Summer of 2008. The virus was concentrated from river water samples using the cation-coated filter method. CyHV-3 DNA was detected in 90 rivers, representing 90% of 103 successfully analysed rivers. More than 100,000 copies of CyHV-3 DNA per litre of sample were detected in four rivers, higher than that reported during the Yura River outbreak in 2007. For CyHV-3-positive rivers, the log CyHV-3 density was negatively correlated with the water temperature on the sampling date and positively correlated with the suspended solids and dissolved oxygen, which are annually averaged for each river. Our results demonstrate that virus detection using molecular biology techniques is a powerful tool for monitoring the presence of CyHV-3 in natural environments.     

日本对虾杆状核病毒性传染病诊断技术

本文提出了分别用暗视野显微技术和透射电子显微技术快速确诊日本对虾杆状核病毒性传染病的方法。在暗视野显微镜下观察到的血淋巴中的病毒粒子大小约为0.5μm;胃表层上皮被病毒感染的细胞核明显地呈白色均质小体,直径10～15μm,球形或椭圆形,坏死细胞被囊化为棕色小块(约20～50μm)。病虾血淋巴和胃经负染制样在透射电子显微镜下观察病毒颗粒大小为400×150nm;核衣壳大小为390×85nm。     

Keratoconus in a cynomolgus monkey

In a seven-year-old male cynomolgus monkey, erythema of the upper eyelid and forehead and corneal opacity, edema and conical protrusion in the eye were observed. At necropsy, ophthalmological and serological examinations revealed binocular corneal opacity and conical protrusion and a high IgE level, respectively. Thinning of the epithelium and stroma of the cornea were noted histopathologically. At the center of the corneal epithelium, the number of epithelial cells was reduced, their cytoplasm was poorer and the basal cells were flatter than at the periphery. Bowman's membrane was folded with partial loss or breakage. Collagen fibers were compacted or disarranged, and the keratocytes were increased in the stroma, with focal pyknosis or loss of the endothelium and folding of Descemet's membrane. Electron microscopical examination revealed atrophy of the corneal epithelial basal cells. This is the first report of a case of keratoconus in a cynomolgus monkey.     

Detection of a bovine group C rotavirus from adult cows with diarrhea and reduced milk production

Only two strains (Shintoku and porcine-like WD534tc) of group C rotavirus (GCR) from cattle have been reported to date. A GCR designated the Yamagata strain was the only pathogen detected in an outbreak of adult cow diarrhea accompanied by a decrease in milk production. The nucleotide sequences of the VP6 and VP7 genes from strain Yamagata were determined. Comparative sequence analysis showed that the sequence identities between strains Yamagata and Shintoku were markedly high in both VP6 gene (98.1%) and VP7 gene (93.5%), and that these strains belonged to the same clusters which were distinguished from GCRs from different host species in phylogenetic trees of these genes. These results suggested strongly that cattle species is one of the natural hosts of GCR infection, and that GCRs are a cause of adult cow diarrhea.     

Effect of the litter layer on runoff and evapotranspiration using the paired watershed method

The litter layer on a forest floor can influence both short-term runoff and long-term water balance through modification of various hydrological processes. In this study, we have quantified the watershed-scale effects of the litter layer on runoff and evapotranspiration using a paired watershed method. The removal of the litter layer in a forested watershed with an area of 1.19 ha was conducted annually over the latter half of a 6-year experimental period. An adjacent forested watershed with an area of 1.42 ha was preserved as a control. Our results indicated that litter removal increased the 3-year runoff by 80.3 mm during the post-treatment period. Furthermore, when the peak flow range in the control watershed was 0.4–1.0 mm/h and >1.0 mm/h, peak runoff during flood events was about 1.5 and 1.4 times greater than that observed before litter layer removal, respectively. These data suggest that litter layer removal can decrease litter layer interception and, hence, increase peak flow, particularly during relatively large flood runoff events.     

Deer-proof fence prevents regeneration of Picea jezoensis var. hondoensis through seed predation by increased woodmouse populations

The indirect consequence of sika deer (Cervus nippon) grazing on the regeneration of Picea jezoensis var. hondoensis through its effect on the small rodent population was assessed by evaluating seed predation by mice inside and outside the deer-proof fence on Mt Ohdaigahara, central Japan. Following construction of the deer-proof fence, Sasa nipponica, which covers the forest floor and is the main forage of deer, grew thicker inside the fence due to the absence of deer grazing than outside of the fence. Higher numbers of two species of woodmice (Apodemus speciosus and A. argenteus) were found inside the fence, possibly because the thick S. nipponica growth provides better cover. Inside the fence, more seeds of P. jezoensis var. hondoensis were eaten by the mice than outside the fence. These results suggest that the mouse populations inside the fence increased due to changes in S. nipponica cover, which subsequently increased seed predation by the mice, possibly preventing regeneration of P. jezoensis var. hondoensis.     

Studies on keeping-quality of vegetables and fruits. III. Changes in sugar and organic acid contents accompanying the chilling-injury of eggplant fruits

The mechanism of chiiling-injury in fruits of eggplants (Solanum melongena L. cultivar ‘Hyonaga’) was investigated by determining the sugar and organic acid content during cold storage and after exposure to low temperature for various periods. Fructose, glucose and maltose were detected, and totalled more than 96% of total sugar on the initial day. Total sugar content increased to 30% higher than the initial day after 2 days of cold storage, and after that decreased gradually, while in 20° C-stored fruits it decreased sharply. Fructose, glucose and maltose increased rapidly after transfer from 1° C to 20° C. Succinic, oxalic, malic and citric acid were detected before storage. Malic and citric acid were found to be the main organic acids. The ratio of malic/citric acid in fruits stored at 1°C decreased during storage, while after storage at 20°C it increased. The ratio in fruits after transfer from 1°C to 20°C increased rapidly during the time course. The ratio of malic/citric acid ranged from 6.0 to 6.8 in healthy fruits, while it was below 5.9 or above 7.3 in chilled fruits. It is suggested that according to this ratio the degree of chilling-injury of eggplant fruits might be foreseen.