Development of an efficient method for recovery of Puumala and Puumala-related viruses by inoculation of Mongolian gerbils

Puumala (PUU) virus and PUU-related viruses are difficult to isolate in cell culture. To determine whether animal inoculation would be a better alternative for virus recovery, the Sotkamo strain of PUU virus was inoculated into several animal species. Newborn Mongolian gerbils (MGs), mice, and rats were infected with the Sotkamo strain by intracerebral (ic), intraperitoneal (ip), and subcutaneous (sc) inoculation. Antibodies to PUU appeared in MGs at 30 days post-infection (dpi), and in mice and rats at 15 dpi. Interestingly, virus appeared at 7 dpi in lung and brain of MGs inoculated via ic and ip routes. Virus was detected in all tested tissues of MGs at 15 dpi, with a peak level of 1.36 x 10 (5) focus forming units (FFU)/g in brain tissue. The virus titer declined with the onset of the antibody response and became undetectable by 75 dpi, when the antibody titer reached the maximum level. The appearance of the virus in mice and rats was delayed as compared to MGs, and the virus titer was apparently lower, at approximately 4 to 8 x 10(3) FFU/g, at 15 dpi. In addition, lung homogenates of antibody-positive Clethrionomys (C.) rufocanus (captured in Tobetsu, Hokkaido, Japan) were inoculated into MGs by the ic route. PUU-related viral RNA was detected at 16 dpi in the brains of MG inoculated with the lung homogenate, and antibodies were detected at 45 dpi. These findings indicate that newborn MG inoculation is an efficient method to recover PUU and PUU-related viruses.     

Complex vertebral malformation in a stillborn Holstein calf in Japan

A female stillborn Holstein calf with shortened cervical and thoracic regions, protrusion of the tongue, and bilateral symmetric flexural contraction of the anterior limbs was delivered on gestation day 281. Multiple hemivertebrae, fused and misshaped vertebrae, synostosis and scoliosis of cervical, thoracic and lumbar vertebral column were found in the affected calf by radiographic and computed tomographic (CT) analysis. Ten pairs of ribs were present and the sternum consisted of 9 sternebrae. Multiple morphologic abnormalities including fusion, malformation, and displacement, were found in the ribs and sternum. Cardiac anomalies, including atrial septal defect and hypertrophy of right ventricle, were observed. DNA-polymerase chain reaction (PCR) analysis demonstrated that amplified product from the liver DNA of the affected calf had identical pattern to that associated with complex vertebral malformation (CVM) of Holstein calves and that her dam was a heterozygous carrier of CVM. The affected calf was diagnosed as having CVM based on the DNA-PCR results and the characteristic findings, and was recorded as a first documentation of CVM confirmed in a Holstein calf in Japan.     

Myrmecofaunal change with bamboo invasion into broadleaf forests

Recently, it is documented that bamboo (Mousouchiku:Phyllostachys pubescens Mazel) invasion deteriorates ecological and/or landscape diversity. In order to investigate the ecological effects on forest-floor arthropods caused by bamboo invasion, we compared the myrmecofauna among broadleaf forests, bamboo-broadleaf mixed forests, and bamboo forests in Hiroshima City and its vicinity. Myrmecofauna in bamboo forests were more impoverished than in broadleaf forests. Myrmecofauna in the mixed forests, however, were similar to that in broadleaf forests in both species diversity and species composition. The change in myrmecofauna may occur relatively late after the initial bamboo invasion.     

Effects of fiber length and orientation on elasticity of fiber-reinforced plywood

Short carbon fibers, a reinforced material in wood veneer composites, were used to investigate the effects of fiber length and orientation of fibers on the elasticity of plywood. The technical feasibility, elasticity, and strength of the reinforced plywood with short carbon fiber were evaluated. In a short fiber reinforcement system, the fiber length does not directly influence the reinforcement in Cox's theory when the fiber length exceeded a certain length. When the length of short carbon fiber is beyond 3 mm, the high reinforced result was obtained in the experiment. However, if fiber length was too long, the reinforced result was less owing to the bridge between fibers and the increase of holes. The optimum fiber length must be considered. The orientation of fibers has a strong influence on the reinforcement. Unidirectional, perpendicular, and random orientation displayed different influence on the elasticity. Experimental results were discussed with Cox's method. Reinforced plywood with short carbon fibers in random orientation has a higher shear modulus and bending strength than the controls, in addition to other mechanical properties.     

Nuclear genome constitution and other characteristics of somatic hybrids between dihaploid Solanum acaule and tetraploid S. tuberosum

Eleven somatic hybrids (2n = 68 to 74) obtained between S. tuberosum ssp. tuberosum cv. Dejima (2n = 48) and ATDH-1 (2n = 24), an anther-culture-derived dihaploid of S. acaule (Yamada et al., 1997), were characterized by nuclear RFLP markers using 49 single-copy DNA probes distributed throughout the potato genome (2 to 6 probes per chromosome). One of the somatic hybrids, DA8-2, had 72 chromosomes and all the Dejima- and ATDH-1-specific markers (124 and 103 bands, respectively), suggesting the presence of a whole set of both parental chromosomes. The other somatic hybrids lost varying numbers of markers up to seventeen. The pattern of the loss of markers indicated the elimination of five chromosomes among four somatic hybrids. A nucleolar organizer region of chromosome 2 was often eliminated in the somatic hybrids. The somatic hybrids studied here had higher frequencies of multivalent formation than the S. tuberosum parent. They had reasonably good seed set when pollinated with S. tuberosum pollen. Hence, homoeologous recombination between S. acaule and S. tuberosum chromosomes is possible and useful traits from S. acaule may be transferred to the S. tuberosum gene pool. This revised version was published online in July 2006 with corrections to the Cover Date.     

Estimating the spawning season of black sea bream Acanthopagrus schlegelii in Hiroshima Bay,Japan, from temporal variation in egg density

Fisheries Science - We aimed to determine the spawning period of black sea bream Acanthopagrus schlegelii in Hiroshima Bay, Japan, based on the distribution of its eggs, to assist in the resource...     

Annual and seasonal changes in the assemblage of planktonic copepods and appendicularians in Funka Bay before and after intrusion of Coastal Oyashio Water

Fisheries Science - Annual and seasonal changes in the abundance and the assemblage of planktonic copepods and appendicularians were clarified in Funka Bay. Zooplankton samples were obtained by...     

Stock enhancement programme for black sea bream, Acanthopagrus schlegelii (Bleeker), in Hiroshima Bay, Japan: a review

This paper reviews the stock enhancement programme for black sea bream ( Acanthopagrus schlegelii ) in Hiroshima Bay. This bay is one of the biggest production areas for black sea bream in Japan, accounting for about 10% of the total catch of the species in this country in 2004. After intensive fishing pressure caused a drastic decline in the catch of the species in this bay in the 1970s, a stock enhancement programme was conducted in its northern part since 1982 to restore the depleted population. The number of black sea bream juveniles released in 1996 surpassed 9 million, representing the third main species stocked in Japan. Almost 1.4 million of these juveniles were released into Hiroshima Bay. The fast acclimatization of hatchery-reared juveniles released into the bay may have contributed to the recovery of landings in the late 1980s and 1990s. However, this recovery was accompanied by a reduction in the market price of black sea bream. Further studies to assess the effectiveness of the stock enhancement programme as well as the carrying capacity of Hiroshima Bay to maintain the stock of black sea bream at a stable, healthy level are desirable. The necessity of evaluating the secondary effects derived from using a reduced number of breeders as well as finding new markets are suggested.     

Changes in plasma prolactin and growth hormone concentrations during freshwater adaptation of juvenile chum salmon (Oncorhynchus keta) reared in seawater for a prolonged period

Freshwater adaptability of chum salmon was examined in juvenile fish reared in seawater for 4 months. The fish, weighing about 40g, were transferred directly to fresh water in October, when their cohorts are migrating in the North Pacific Ocean. Plasma sodium concentration decreased from 167 mM in seawater to about 130 mM during the first 24h, and increased gradually during 2–7 days after the transfer. No immunoreactive prolactin (PRL) was detected in the plasma of the seawater-adapted fish nor during the first 24h in fresh water. Significant levels of PRL were detected after 2–3 days. The maximal level (2.6 ng/ml) was observed after 5 days and became undetectable again after 7 days; no significant correlation was seen between the changes in plasma sodium and PRL levels during the transfer. Plasma growth hormone levels were relatively constant, except for a significant decrease 12h after the transfer. Although plasma thyroxine levels were highly variable during the experiment, a significant decrease and an increase were observed 12h and 5 days after the transfer, respectively. The present study indicates that juvenile chum salmon retain hyperosmoregulatory ability even after prolonged rearing in seawater. Examination of turnover rates, rather than changes in plasma levels, seems to be essential to clarify the osmoregulatory roles of the hormones.     

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.