Detection of Borna disease virus in a pregnant mare and her fetus

A pregnant mare showing pyrexia, reduced appetite, ataxia and paresis was euthanized and examined for the presence of Borna disease virus (BDV). Her brain, showing multiple neuronal degeneration and necrosis with hemorrhage, and the histologically normal brain of the fetus were both positive for BDV RNA. The BDV nucleotide sequences were identical in the mare and fetus in the second open reading frame (ORF). This is the first report of the possible vertical transmission of BDV in a horse.     

Annual variation of potential predation impacts on larval and juvenile marbled sole <Emphasis Type="Italic">Pseudopleuronectes yokohamae</Emphasis> by sand shrimp <Emphasis Type="Italic">Crangon uritai</Emphasis> in Hakodate Bay,Hokkaido

An investigation was conducted to evaluate the annual variation in potential predation impact (PPI) on larval and juvenile Pseudopleuronectes yokohamae by Crangon uritai in Hakodate Bay using predator-prey size relationships. Laboratory experiments were designed to estimate the favorable prey size of C. uritai through back-calculation of body length (BL) of P. yokohamae from sagittal otolith diameter observed in the stomachs of C. uritai. The most favorable prey-predator size ratio (BL of P. yokohamae-total length (TL) of C. uritai) class was 0.15–0.19, and ranged from 0.12–0.31. There was a significant positive correlation between the BL increase of P. yokohamae and the bottom water temperature in the field, such that BLs stagnated during the cold year of 1999 from April to June, and increased during the warm year of 2002. In contrast, no significant correlation was found between the TL increase of C. uritai and the bottom water temperature. Moreover, there were no significant differences in the mean TL of C. uritai between months (April–June) or years (1998–2002). Variation of PPI depended on the relationships between the growth rates of P. yokohamae and bottom water temperature. Therefore, the warm year of 2002 resulted in higher potential predation impact (PPI≥20), and it was at least 20 days shorter than that of the cold year of 1999. These results suggest that increased bottom water temperature in the nursery area was one of the most important factors for cumulative predation loss.     

Characterization of fast skeletal myosin from white croaker in comparison with that from walleye pollack

ABSTRACT:   Enzymatic and structural properties of white croaker fast skeletal muscle myosin were determined and compared with those of walleye pollack counterpart. Ca²⁺-ATPase activity of white croaker myosin was decreased to approximately 70% of the original activity during 1 day of storage at 0°C and pH 7.0 in 0.5 M KCl and 0.1 mM dithiothreitol, whereas that of walleye pollack was decreased to approximately 20% under the same condition. The activation energy ( E _a) for inactivation of white croaker myosin calculated by the Arrhenius plot for inactivation rate constant (K_D) was 1.2-fold higher than that of walleye pollack. While Ca²⁺-ATPase showed a similar KCl-dependency for the two species, the maximal activity was observed at pH 6.2 and 6.3 for white croaker and walleye pollack, respectively. Actin-activated myosin Mg²⁺-ATPase activity of white croaker was approximately half that of walleye pollack at 0.05 M KCl and pH 7.0, although the two myosins showed a similar affinity to F-actin with K _m of 1.7 and 1.4, respectively. Limited proteolysis with α-chymotrypsin cleaved heat-denatured white croaker myosin mainly at heavy meromyosin/light meromyosin (HMM/LMM) junction, whereas walleye pollack myosin was cleaved at several sites in LMM as well as at the HMM/LMM junction.     

Isolation and characterization of a novel 45 kDa calponin-like protein from anterior byssus retractor muscle of the mussel Mytilus galloprovincialis

SUMMARY: A calponin-like protein of 45 kDa was isolated from mussel anterior byssus retractor muscle (ABRM) and its inhibitory effects on actomyosin Mg²⁺-ATPase was demonstrated. The 2-D electrophoresis for ABRM myofibrils gave a spot of 45 kDa protein in addition to myofibrillar proteins such as myosin and actin. The 45 kDa protein, which was more basic and showed a slightly higher molecular weight than actin, was isolated by ion-exchange chromatography and subjected to chymotryptic digestion. N-terminal amino acid sequencing of polypeptide fragments produced gave two sequences, ASQKGMTSFGAVRHH and GMDRALISKMGSKYDSGL, both of which showed a high homology to those of vertebrate calponins and invertebrate calponin-related proteins. Furthermore, the 45 kDa protein strongly reacted with commercially available antibody raised against chicken smooth muscle calponin, demonstrating that the mussel ABRM 45 kDa protein is a new member of the calponin family. Then, actomyosin Mg²⁺-ATPase activity of ABRM was measured in the presence and absence of the 45 kDa protein. The 45 kDa protein clearly inhibited actomyosin Mg²⁺-ATPase activity in a dose-dependent manner as in the case of other vertebrate calponins. These results indicate that the 45 kDa calponin-like protein is involved in the thin filament-associated regulation of molluscan smooth muscle contraction, possibly of a unique contraction called catch.     

Physiological difference between free and triglyceride-type conjugated linoleic acid on the immune function of C57BL/6N mice

Previous studies have shown the physiological significance of dietary conjugated linoleic acid (CLA) in various experimental animals and in human beings. One of the important problems to better elucidate is the difference between triglyceride (TG) and free (FFA) dietary CLA. Here, using splenocytes, this study assesses how TG- and FFA-CLA modulate immunoglobulin and various cytokine productions. In this study, C57BL/6N mice were fed an experimental diet containing 0% CLA, 0.1 or 1% FFA-CLA, or 0.1 or 1% TG-CLA for 3 weeks. The production of immunoglobulin tended to be up-regulated by 1% FFA-CLA. As a result of protein array analysis using the supernatant from splenocytes cultured with no CLA, 1% FFA-CLA, and TG-CLA, some cytokine production was shown to be remarkably regulated by dietary FFA- and TG-CLA. A total of 32 cytokines were examined, and 11-14 produced cytokines that were 2-fold up-regulated as compared with control for FFA- or TG-CLA, respectively. Especially, the production of IL-9 and MCP-5 and other cytokines was remarkably up-regulated by both FFA- and TG-CLA. In addition, seven cytokines were 2-fold down-regulated by TG-CLA. These data show that there is a slight but significant difference between the functionalities of FFA- and TG-CLA.     

Effects of n-3 highly unsaturated fatty acid content in Artemia on survival and development of laboratory-reared horsehair crab Erimacrus isenbeckii larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) content in Artemia sp. on survival, the number of days required to reach each larval stage, and the growth and morphogenesis of larvae of the horsehair crab Erimacrus isenbeckii were examined. Artemia enriched with materials of six types [condensed Nannochloropsis sp., ethyl oleate, two enrichment levels of eicosapentaenoic acid (EPA), and those of docosahexaenoic acid], and unenriched Artemia were fed to larvae. Newly hatched larvae were reared in groups of 50 individuals in two 2-l beakers until they reached the first crab stage. No significant difference was found in survival rate, growth, or morphogenesis of larvae, but fewer days tended to be required to reach the first crab stage in the group enriched with high EPA compared to the other groups. Results show that, although the amount of n-3 HUFA in unenriched Artemia might meet the demands of horsehair crab larvae for their survival, growth, and morphogenesis, Artemia should be enriched with high EPA to improve the rate of larval development.     

Ionic liquids as formaldehyde-free wood adhesives

Several ionic liquids promote depolymerization of wood components, i.e., polysaccharides and lignin, into low molecular weight compounds, some of which further re-polymerize into resin-like compounds. In this study, the depolymerization/re-polymerization of wood components in ionic liquids was applied to preparation of plywoods from Japanese cedar (Cryptomeria japonica) veneers by employing ionic liquids as adhesives. The adhesive solution was prepared by mixing an ionic liquid (pyridine hydrochloride ([Py][Cl]), imidazole hydrochloride ([IM][Cl]), or 1-ethylpyridinium chloride ([EtPy][Cl])) with water and d-glucose in various weight ratios. Tensile shear test of the three-ply plywoods prepared from the veneers and the adhesive solution through hot-pressing indicated that the plywood bonded with the [IM][Cl]-based solution ([IM][Cl]/water/glucose ratio: 9/3/2) exhibited the highest strength. Scanning electron microscope observation on the plywoods suggested that the ionic liquids softened the cell walls of the probably plywood through the depolymerization/re-polymerization reactions and the cell walls were compressed during the hot-pressing process. Entwining of the compressed cell walls and van der Waals force enhanced by the compression were considered to be origins of the adhesion of the veneers.     

Comparison of the nutritional transition date distributions of marbled sole larvae and juveniles in Hakodate Bay, Hokkaido

To examine survival processes in marbled sole Pseudopleuronectes yokohamae larvae, nutritional transition date distributions (NTDDs) were compared between larvae and large juveniles (LJs) in Hakodate Bay from 2001 to 2003. NTDDs were used instead of hatch date distributions because the onset of increment formation coincides with the timing of the transition from endogenous to exogenous nutritional sources for marbled sole. LJs were defined as being large enough to be safe from predation by the sand shrimp Crangon uritai. In 2001 and 2002, the ranges of the NTDDs of LJs largely overlapped with those of the larvae. However, in 2003, the early cohort (those whose otolith rings formed before 17 March) disappeared from the NTDDs during and after April, and the NTDDs of LJs were heavily biased toward a later period compared to those of the larvae. In 2003, Oyashio coastal water (<3°C and ≤33.0 salinity) unexpectedly intruded into the bay in late March and lowered the water temperature from 6.3°C on 6 March to 4.1°C on 25 March. One possible cause of mortality in the early cohort of 2003 may be feeding failure caused by the extremely low water temperature.     

Rapid identification of eels <Emphasis Type="Italic">Anguilla japonica</Emphasis> and <Emphasis Type="Italic">Anguilla anguilla</Emphasis> by polymerase chain reaction with single nucleotide polymorphism-based specific probes

ABSTRACT:   Standard molecular techniques, such as sequencing and restriction fragment length polymorphism analysis after polymerase chain reaction (PCR) amplification are relatively complicated, and species identification can take a long time when using such techniques. We established a quick method, using PCR with species-specific TaqMan Minor Groove Binder (MGB) probes based on single nucleotide polymorphism (SNP) to distinguish the two eel species Anguilla japonica and Anguilla anguilla . This method can be used in processed products. Partial sequences of the mitochondrial 16S rRNA gene were compared between A. japonica and A. anguilla to design a primer pair common to both A. japonica and A. anguilla and probes specific to A. japonica and A. anguilla . Different fluorescence intensities were produced in two PCR microtubes each containing A. japonica - and A. anguilla -specific probes for one target sample. We observed the fluorescence intensity of PCR products in microtubes under ultraviolet transillumination, with similar results to those obtained by real-time PCR. Therefore, SNP-based PCR is a powerful tool for identifying materials of processed foods from either A. japonica or A. anguilla .