Molecular characteristics of horse phospholipase C zeta (PLCζ)

A sperm‐specific phospholipase C (PLC), PLCzeta (PLCζ), is thought to underlie the initiation of calcium ([Ca²⁺]_i) oscillations that induce egg activation in mammals. In large domestic species, only bovine, porcine and recently equine PLCζ have been cloned, and the physiological functions of these molecules have not been fully characterized. Here, we evaluated the physiological functions of equine PLCζ (ePLCζ) in mouse oocytes. ePLCζ was cloned from testis using RT‐PCR. The expression of ePLCζ messenger RNA was confirmed in testis but not in other tissues. Microinjection of ePLCζ complementary RNA (cRNA) into mouse oocytes induced long‐lasting [Ca²⁺]_i oscillations, and most of the injected oocytes formed pronuclei (PN). The injection of cRNAs encoding horse, mouse, human and cow PLCζ into mouse oocytes showed that ePLCζ had the highest [Ca²⁺]_i oscillation‐inducing activity among the species tested. Mutation of D202R, which renders the protein inactive, abrogated the activity of ePLCζ. The nuclear translocation ability of ePLCζ was defective when expressed in mouse oocytes. Taken together, our findings show for the first time that ePLCζ has highest activity of the mammalian species studied to date. Our findings will be useful for the improvement of reproductive technologies in the horse.     

Molecular analysis of β-proteobacterial ammonia oxidizer populations in surface layers of a submerged paddy soil microcosm

The structure of the β-proteobacterial autotrophic ammonia-oxidizing bacterial (AOB) communities in a microcosm of submerged paddy soil was determined by denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments amplified using AOB-selective primers. Shift in the community composition was observed 4 weeks after submergence. The communities from the surface layers (0–1, 2–3 mm) of the soil microcosm were different from those of the subsurface layers (6–9, > 15 mm) and DGGE bands specific to each layer were detected. The majority of the retrieved sequences were Nitrosospira-like, whereas no Nitrosomonas-like sequences were obtained. The 16S rDNA primer set also amplified sequences that were not related to the known Nitrosospira-Nitrosomonas group, although they showed a close relationship with other groups of β-proteobacteria. The results suggest that Nitrosospira-like populations are dominant AOB populations in the submerged paddy soil, and that the oxic layer of submerged paddy soil harbours the specific AOB.     

A simple method for a mini-preparation of fungal DNA

A simple method was established to prepare DNA from fungal mycelia cultured on an agar plate. The fungi tested successfully with this method contained Zygomycetes, Ascomycetes, Basidiomycetes, and Oomycetes. This method did not require any time-consuming steps to crush or digest mycelia or fractionation in a phenol–chloroform mixture. The DNA was easily extracted by immersing and dispersing the mycelial plugs in a specific buffer (200 mM Tris-HCl, 50 mM ethylenediaminetetraacetic acid, 200 mM NaCl, 1% n-lauroylsarcosine, pH 8.0), then concentrated by ethanol precipitation. The total time to complete the whole procedure was less than 1 h. The quality and quantity were sufficient for polymerase chain reaction amplification and Southern blot analysis.     

Longitudinal shrinkage variations within trees of sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) cultivars

The longitudinal shrinkage variations within trees and the relationship with density, microfibril angle, and modulus of elasticity were examined for five sugi cultivars selected for different within-tree distributions of density, microfibril angle, and modulus of elasticity. The cultivars showed significant differences in longitudinal shrinkage and in its within-tree distribution. The within-tree distributions were categorized into two types: (1) large values of longitudinal shrinkage near the pith that decreased with height and from pith to bark, (2) small values of longitudinal shrinkage near the pith that increased slightly from pith to bark. There were strong relationships between longitudinal shrinkage and microfibril angle, and modulus of elasticity, with large values of longitudinal shrinkage associated with large microfibril angle and low modulus of elasticity. Sugi exhibited large variation in longitudinal shrinkage within stem and among cultivars, with the variation strongly affected by microfibril angle. Part of this article was presented at the 56th Annual Meeting of the Japan Wood Research Society, Akita, Japan, August 2006     

Avian haemosporidian parasites infection in wild birds inhabiting Minami-daito Island of the Northwest Pacific, Japan

Haemosporidian parasites infection among wild birds inhabiting Minami-daito Island was studied. Blood films from 183 birds representing 4 species of 4 families were examined microscopically. Avian haemosporidian parasites were detected in 3 species with an overall prevalence of 59.6%. None of the 30 Daito scops owls (Otus scops interpositus) examined were infected. Either Haemoproteus sp. or Plasmodium sp. infection was found in 14 of 31 (45.2%) Borodino islands white-eyes (Zosterops japonicus daitoensis). Plasmodium spp. were found in 94 of 102 (92.2%) bull-headed shrikes (Lanius bucephalus) and 1 of 20 (5%) tree sparrows (Passer montanus).     

Genetic analysis of phosphoprotein and matrix protein of rabies viruses isolated in Brazil

To investigate the genetic characteristics of phosphoprotein (P) and matrix protein (M) genes of variable rabies virus (RV) prevalent in Brazil, the authors genetically characterized the P and M genes from 30 Brazilian RV field isolates. Phylogenetic analysis based on the P and M genes revealed the presence of six RV variants that consisted primarily of three insectivorous bats, the vampire bat, dog and fox in Brazil. Specific amino acid substitutions corresponding to these phylogenetic lineages were observed, with Asp(42) and Glu(62) in the P protein found to be characteristic of Brazilian chiroptera- and carnivora-related RVs, respectively. Amino acid sequence motifs predicted to associate with a viral function in the P and M proteins were conserved among Brazilian RV variants.     

Effect of cross-sectional dimensions on bow and surface checking of sugi (Cryptomeria japonica) boxed-heart square timber dried by conventional kiln drying

The effect of cross-sectional dimensions on bow and surface checking were investigated, using the boxed-heart square timber of two sugi cultivars with dimensions 80, 120, and 140 mm and length 1.9 m taken at two different heights above the ground. The smaller cross-sectional timber tended to have larger bow, less surface checking, and larger dimensional shrinkage. However, the drying defects were different between the cultivars and sampling heights, depending on the shrinkage properties of the juvenile wood and the heartwood proportion in the core part of the stem. The bow was larger in the smaller cross-sectional timber in which the longitudinal shrinkage was large in the juvenile wood. Surface checking was more prominent in larger cross-sectional timber containing sapwood in its outer part, which suggested the surface checking was induced by drying stress, owing to large moisture gradients between the heartwood and sapwood. The cross-dimensional shrinkage of the timber was larger in timber with larger tangential shrinkage.