Abundance and use of cavity trees at the tree- and stand-levels in natural and plantation forests in Hokkaido, Japan

The conservation of cavity trees in plantations is important for maintaining biodiversity because cavity trees are one of the forest attributes most strongly affected by forestry operations. We surveyed the occurrence of cavities at the tree-level and the abundance of cavity trees and the number of uses by secondary cavity users at the stand-level in natural and conifer plantation forests in Hokkaido, Japan. We classified cavities into two types by their formation processes: excavated and non-excavated. Excavated cavities were more common in dead trees than in live trees. Non-excavated cavities showed high occurrence in dead and live broad-leaved trees. The abundance of non-excavated cavity trees was two times greater than that of excavated cavity trees in natural forests, but, in plantations, excavated cavity trees were dominant. The abundance of non-excavated cavity trees was lower in thinned plantations than in not-thinned plantations. The number of cavity uses increased with the number of cavity trees, indicating the importance of maintaining cavity trees. Conifer plantations are established by removing native broad-leaved trees, which results in a lower abundance of non-excavated cavity trees in these locations. The conservation of broad-leaved trees is important for maintaining the current and future resource of non-excavated cavity trees in plantation forests, as is the conservation of dead trees, which showed a high occurrence of excavated cavities.     

Characterization of monoclonal antibodies directed against the canine distemper virus nucleocapsid protein

We have established four monoclonal antibodies (MAbs) against the nucleocapsid protein (NP) of canine distemper virus (CDV). A competitive binding assay has revealed that the MAbs are directed against two antigenic domains. An immunofluorescence assay using a series of deletion clones of the NP and an immunoprecipitation assay using the NP have revealed that two of the MAbs recognize the C-terminal region of the NP while the other two recognize the tertiary structure of the N-terminal domain. These MAbs reacted with all eight strains of CDV used in this study, but showed different reactivities against measles virus and rinderpest virus.     

Gentian Kobu-sho-associated virus: a tentative, novel double-stranded RNA virus that is relevant to gentian Kobu-sho syndrome

A virus-like dsRNA of about 23 kbp was detected in gentian plants showing Kobu-sho syndrome including stunting, shortened internodes, and tumors on stems, nodes and roots. Nucleotide sequence analysis has suggested that this dsRNA is related to Pestivirus species but not to any other plant viruses. It was protected from externally added RNase, suggesting that the dsRNA is encapsidated. The dsRNA was co-extracted in a crude homogenate of glutaraldehyde-fixed tissue with the virus-like particles that have been associated previously with Kobu-sho syndrome in gentian (Usugi et al. Jpn J Phytopathol 76:21–24, 2010). The RNA sequence was detected in more than 99 % of Kobu-sho gentian individuals but in less than 20 % of apparently healthy gentian individuals from fields affected with Kobu-sho. Thus, we propose naming the virus Gentian Kobu-sho-associated virus.     

Sea urchin coelomocyte arylsulfatase: a modulator of the echinoderm clotting pathway

Sea urchin petalloid coelomocytes effectuate the clotting pathway by undergoing a rapid and dynamic cellular transformation that leads to cellular adhesion and wounds closure. We have identified high levels of activity of arylsulfatase (Ars) associated with coelomocytes of the sea urchin Lytechinus variegatus (Lamarck, 1816). Ars activity was extracted from clotted coelomocytes with EDTA and showed high levels of activity up to a 1:100 dilution. Clot formation from isolated coelomic fluid was significantly inhibited by the ARS inhibitor, p-nitrophenyl phosphate. Ars activity was collected by 80% ethanol precipitation, a diagnostic test previously used in Ars isolation. Cellular extraction studies in the presence and absence of the non-ionic detergent Triton X-100 indicated that some Ars activity was present intracellularly, possibly in intracellular membrane-bound compartments, however the majority of Ars activity was extracted from the extracellular coelomocyte membrane. Polyclonal anti-sea urchin embryo Ars antibodies recognized a single protein band with an approximate molecular weight of 75 kDa on western blots. Immunofluorescence using the anti-sea urchin Ars antibody revealed an intracellular and extracellular staining of Ars in both petalloid and filopodial coelomocytes. Taken together, these data indicate that coelomocyte Ars might be involved in cell-to-cell crosslinking of surface sulfated polysaccharides vital for clot formation.     

Comparison of ultrasound- and electrostimulation-guided nerve blocks of brachial plexus in dogs

Objective

To compare the effectiveness of ultrasound- and electrostimulation-guided nerve blocks of the brachial plexus and to determine whether ultrasound guidance is feasible in conscious dogs.

Inheritance of resistance to brown leaf spot disease in gentians

Brown leaf spot caused by Mycochaetophora gentianae is a serious disease on gentian in Japan. Previous studies revealed that Gentiana triflora cultivars were susceptible to M. gentianae and that G. scabra cultivars and their interspecific hybrid cultivars were resistant. We subsequently analyzed the mode of inheritance of resistance to M. gentianae in gentians using several populations derived from crosses between G. scabra and G. triflora. Parental G. scabra and G. triflora lines, F₁ and BC₁ plants were inoculated with a conidial suspension and assessed for susceptibility to M. gentianae. All tested F₁ plants were resistant to M. gentianae, indicating that the resistance of G. scabra was inherited dominantly. Segregation ratios in backcross progenies using G. scabra line OK fit the Mendelian ratio of 1:1, showing the involvement of a single dominant locus. These results indicate that the resistance to M. gentianae in G. scabra line OK is controlled by a single dominant allele designated as gentian brown leaf spot resistance (GBLS-1). Finding this allele should facilitate future breeding efforts to develop resistant gentian cultivars.     

Scale-independent significance of river and riparian zones on three sympatric Myotis species in an agricultural landscape

Conserving biodiversity in agricultural landscapes is an urgent issue. The effective conservation of biodiversity requires plans based on species’ habitat preferences at multiple spatial scales. We examined how the foraging habitat selection of bats varied with grain size (50, 150, 250, and 350 m) and how habitat selection in the home range differed from that in the foraging habitat in an agricultural landscape. Focusing on three sympatric Myotis species (Myotis petax, Myotis gracilis, and Myotis frater), we radio-tracked 10 individuals of each species for 121 nights in Hokkaido, northern Japan. Rivers and broadleaved riparian forests were commonly preferred at multiple scales, although the preferred land-cover type was dependent on both extent and grain size for most species. The best grain sizes for predicting the foraging-habitat use of M. petax and M. gracilis were 50 and 150 m. By contrast, M. frater showed no tendency across the grain sizes. Our results indicate that it is necessary to consider both extent and grain size to understand the habitat selection of bats. Our findings also suggest that focusing primarily on preferred land-cover types at multiple scales is effective for conservation planning, given the limited resources in terms of time, manpower, and finances. Although arable did not negatively affect the selection of foraging habitats and home ranges in these bat species, preventing the loss of rivers and forests should be prioritized over preventing arable land expansion.     

Validation of a direct time-resolved fluoroimmunoassay for progesterone in milk from dairy and beef cows

The aim of this study was to validate a direct time-resolved fluoroimmunoassay (TR-FIA) for quantifying progesterone concentrations in milk during the bovine oestrous cycle. Holstein-Friesian and suckled and non-suckled Japanese Black cows were used to demonstrate the relationship between milk and plasma progesterone concentrations and to monitor progesterone profiles in milk and plasma during the oestrous cycle. The minimum detection level of the assay was 1.53ng/mL. Progesterone concentrations in milk and plasma changed in a similar manner throughout the oestrous cycle in dairy and beef cows, and milk and plasma progesterone profiles were significantly correlated (P<0.001). The study confirmed that a direct TR-FIA can be used to monitor the oestrous cycle in cattle and to quantify progesterone concentrations in whole milk.     

Pond area and distance from continuous forests affect amphibian egg distributions in urban green spaces: A case study in Sapporo,Japan

We examined factors that affect egg distributions of amphibians, which are currently declining globally, in urban areas. We counted the number of egg masses of two amphibian species (Hynobius retardatus and Rana pirica) in ponds in urban green spaces during two years (2011 and 2012) and investigated the effects of local and landscape factors on them. Forest area in green spaces and pond area had positive effects, and distance from continuous forests in the suburbs and the rate of pond shore protection had negative effects on the number of eggs. Among these factors, distance from continuous forests and pond area had strong effects on egg distributions. These results suggest the importance of water habitat size and the need for colonization by amphibians from surrounding continuous forests. To conserve urban amphibian assemblages, the preservation of ponds with sufficient area and continuous forests close to urban habitats would be important.     

Phylogeography based on the nuclear ribosomal DNA internal transcribed spacer region of native Miscanthus sinensis (Poaceae) populations in Japan

Nucleotide sequence variations were investigated with respect to the geographical distribution patterns of Miscanthus sinensis populations that were sampled from 26 Japanese national parks and three populations of the Ryukyu Islands. Twelve homozygous sequences in the nuclear ribosomal DNA internal transcribed spacer region were detected. The populations of M. sinensis in mainland Japan mainly were composed of a monophyletic group with a symapomorphic character, whereas those in the Ryukyu Islands included a polyphyletic group. Only an internal transcribed spacer haplotype with a plesiomorphic character was found in both mainland Japan and the Ryukyu Islands. Thus, no clear geographical isolation was observed in this species. These facts might be caused by the ability of M. sinensis as a pioneer plant to have a high migration potential and high gene flow by outcrossing. On the basis of the results of this study (nuclear ribosomal DNA) and the previous study (chloroplast DNA), a phylogeographical history of M. sinensis was presumed where the ancestral and derived lineages were distributed in the southern and northern regions of Japan, respectively, without strict geographical isolation.