Population structure of rice blast isolates resistant to scytalone dehydratase inhibitors in Mie Prefecture and implications for their origin

In Mie Prefecture in Japan, rice blast isolates resistant to melanin biosynthesis inhibitors targeting scytalone dehydratase (SDH) were first observed in 2005. To analyze the distribution of the resistant isolates, 527 isolates were collected from wide areas in this prefecture during 2006 and 2007. Almost half of the isolates collected (233 of 527 isolates) carried a point mutation in the SDH gene conferring the resistant phenotype. To compare population structures of resistant and sensitive isolates, we analyzed the isolates with repetitive-element-based PCR DNA fingerprinting using a single primer complementary to a sequence in the terminal inverted repeat of transposable element Pot2. A majority of the resistant isolates were classified into a single DNA fingerprint haplotype, Mie1. Despite its prevalence in the resistant isolates, Mie1 was not found in the sensitive isolates. Furthermore, in a dendrogram constructed from the DNA fingerprint data, Mie1 and six other haplotypes formed a cluster composed of resistant isolates alone. These results suggest that the resistant isolates that belonged to the Mie1 haplotype had migrated from regions outside Mie Prefecture and selectively propagated in a short period in this prefecture.     

Detection and analysis of genetic variations in GOB locus of Plasmopara viticola by DNA sequence analysis

The population genetic structure of Plasmopara viticola in Japan was analyzed using grapevine downy mildew specimens collected from two islands, Honshu and Hokkaido. By simple sequence repeat analysis with the GOB microsatellite marker and DNA sequencing, an accurate copy number of the (CT) _n (CTAT) _n repeat was determined. Consequently, we found a large number of genetic variations in (CT) _n and (CTAT) _n repeats. Also, a single nucleotide polymorphism in the (CTAT) _n repeat of the GOB locus was detected. These genetic variations may serve as a valuable tool to understand the population structure of P. viticola.     

A case of congenital hyperostosis in a newborn piglet

One female newborn piglet showed prominent thickening of both forelimbs and died soon after birth. Histopathologically, thin and woven trabeculae of bone was extending out from the edge of cortical bone in the affected forelimbs, and diagnosed as congenital hyperostosis. The extent of radially proliferated trabeculae was most prominent in radioulna. Many round- to spindle-shaped cells were observed in periosteum, which were considered to be osteoblasts. Around the periosteum, the mesenchymal proliferation was extensive with abundant mucus, and cartilaginous metaplastic changes were observed mainly around the radioulna and humerus. Dilatation of vessels with fibrin deposition in vessel walls was often observed, which were considered to reflect the localized circulatory disturbance.     

Stimulation of Adrenal Chromaffin Cell Proliferation by Hypercalcemia Induced by Intravenous Infusion of Calcium Gluconate in Rats

Increased incidence of adrenal pheochromocytoma is frequently encountered in rat carcinogenicity studies. In some of the studies, the finding is judged to be due to a rat-specific mechanism of carcinogenesis caused by a disturbance of calcium homeostasis. However, direct evidence that the proliferation of chromaffin cells in the adrenal medulla is induced solely by hypercalcemia is not available. In this study, calcium gluconate was intravenously infused for 7 days to rat chromaffin cells by a tail cuff method, and cumulative labeling with bromodeoxyuridine (BrdU) was carried out to evaluate the proliferative activity. The serum calcium concentration was dose-dependently increased, and a high calcium concentration was stably sustained from day 2 to 7. In the adrenal medulla, BrdU-positive chromaffin cells increased in the calcium gluconate-treated animals, and the BrdU-labeling index increased in a dose-dependent manner. In addition, an increased BrdU-labeling index of chromaffin cells was shown to correlate with the serum calcium concentration. Our results demonstrate that hypercalcemia directly enhances the proliferative activity of chromaffin cells and that the proliferative activity is correlated with the serum calcium concentration.     

High-grade myofibroblastic sarcoma of inguinal region in a dog

A subcutaneous tumor in the left inguinal region was present in an 11-year-old female bloodhound. Histopathologically, the tumor showed invasive growth and extensive necroses, and it was composed of spindle-shaped, elongated, and stellate neoplastic cells accompanied by occasional giant cells arranged in fascicular, herringbone, or irregular storiform patterns with abundant production of collagen fibers. The cytoplasm of most tumor cells was positive for vimentin, alpha-smooth muscle actin, and calponin, but was negative for desmin, smoothelin, and S-100. Furthermore, most of the tumor cells were negative for Iba1 while some tumor cells were weakly positive. Thus, this tumor was diagnosed as a high-grade myofibroblastic sarcoma according to the diagnostic criteria for human myofibroblastic sarcomas.     

Transcriptional profiling of inflammatory cytokine genes in African buffaloes (Syncerus caffer) infected with Theileria parva

Theileria parva (T. parva) causes East Coast fever (ECF), which is of huge economic importance to Eastern and Southern African countries. In a previous bovine model, inflammatory cytokines were closely associated with disease progression in animals experimentally infected with T. parva. The African Cape buffalo (Syncerus caffer), the natural reservoir for T. parva, is completely resistant to ECF despite a persistently high parasitaemia following infection with T. parva. Characterizing basic immunological interactions in the host is critical to understanding the mechanism underlying disease resistance in the African Cape buffalo. In this study, the expression level of several cytokines was analyzed in T. parva-infected buffaloes. There were no significant differences in the expression profiles of inflammatory cytokines between the infected and uninfected animals despite a remarkably high parasitaemia in the former. However, the expression level of IL-10 was significantly upregulated in the infected animals. These results indicate a correlation between diminished inflammatory cytokines response and disease resistance in the buffalo.     

Epizootic canine distemper virus infection among wild mammals

In the spring of 2007, seven raccoon dogs and a weasel were captured near the city of Tanabe in Wakayama prefecture, Japan. The causative agent of the animals' death 1-2 days after capture was identified as canine distemper virus (CDV) by virus isolation, immunostaining with an anti-CDV polyclonal antibody, and a commercially available CDV antigen-detection kit. Sequence analysis of hemagglutinin genes indicated the isolated viruses belong to genotype Asia-1 and possess the substitution from tyrosine (Y) to histidine (H) at position 549 that is associated with the spread of CDV to non-canine hosts. A serosurvey for CDV was then conducted among wild animals in the region. The animals assayed consisted of 104 raccoons, 41 wild boars, 19 raccoon dogs, five Sika deer, two badgers, one weasel, one marten, one Siberian weasel and one fox. Virus-neutralization (VN) tests showed that, except for fox and weasel, all of the species assayed had VN antibodies to CDV. Interestingly, 11 of the 41 wild boars (27%) and two of the five Sika deer assayed possessed VN antibodies to CDV. These findings indicate that CDV infection was widespread among wild mammals during this epizootic.     

Development of microsatellite markers and evaluation of genetic diversity of the Amazonian ornamental fish Pterophyllum scalare

The objective of the present work was to develop species-specific microsatellite markers for P. scalare and to analyze the diversity and genetic structure of a wild population, from the Amazon River, and three commercial stocks (common, marble, and clown morphological variants), from farmers in Vieras-Minas Gerais. Through microsatellite-enriched genetic libraries, 11 microsatellite markers with adequate amplification patterns were characterized. Population genetic analysis identified eight polymorphic loci that generated 66 alleles ranging from two alleles (PSCA1B3) to nine (PSCA2H1). The polymorphic information content ranged from 0.031 to 0.827. High genetic differentiation was observed between the wild population and the stocks, and moderate differentiation between the three stocks. Deviation in the Hardy-Weinberg equilibrium was observed in one locus in the wild population, in five loci in the common morphological variant, in two in the marble, and in two in the clown morphological variant. Bayesian analysis of genetic structure revealed the existence of two clusters, one represented by the natural population and the other by the stocks. The developed microsatellite markers serve as a tool for the analysis of diversity and genetic structure and conservation studies of P. scalare.

     

Identification and validation of QTLs for cold tolerance at the booting stage and other agronomic traits in a rice cross of a Japanese tolerant variety,Hananomai, and a NERICA parent,WAB56-104

In Africa, cold temperatures occur in the highlands of East and Southern Africa and in some areas of the Sahel region of West Africa leading to substantial rice yield losses. Cold tolerance (CT) at booting stage on basis of spikelet fertility after cold water irrigation was evaluated using F₂ population derived from a cross between temperate japonica, Hananomai, and tropical japonica, WAB56-104. Two Quantitative trait loci (QTLs) for CT were detected on chromosome 8 and 10 with enhanced effects on the trait coming from Hananomai and WAB56-104 allele, respectively. The QTLs explained 30% and 33% of phenotypic variation in spikelet fertility, respectively. CT was negatively correlated with panicle number (r = ?0.35, p < 0.01) and positively correlated with panicle weight (r = 0.61, p < 0.001). Selected BC₁F₄ and BC₁F₅ genotypes having homozygous alleles for both CT QTLs exhibited higher spikelet fertility under cold stress. The identified QTLs will be useful in the development of cold-tolerant varieties for production in high altitude areas through marker-assisted selection.     

Partial migration of juvenile temperate seabass <Emphasis Type="Italic">Lateolabrax japonicus</Emphasis>: a versatile survival strategy

Partial migration describes intrapopulation variation in the migratory behavior, i.e. some individuals from a population migrate to low-salinity river areas, while others remain in coastal areas. This paper reviews the partial migration pattern of juvenile temperate seabass Lateolabrax japonicus, which is a migration pattern not commonly seen in Japan. Seabass spawn offshore, and eggs and larvae are transported to coastal areas. Some of these juveniles then ascend rivers, while others remain in coastal areas. Juveniles efficiently use physical structures in their habitat; they use tidal currents to ascend rivers in macrotidal estuaries, while they use the salt wedge in microtidal estuaries. Once juveniles ascend the river, they can feed on the abundant prey and attain more rapid growth than those remaining in coastal areas. As estuaries are highly productive areas, they play significant roles as nurseries for juveniles of various fishes. However, compared with coastal areas, the relative area of estuaries is considerably smaller and its environmental conditions are more variable. For example, nearly 40% of adult seabass in Tango Bay were estimated to use estuarine areas as a nursery, while the other 60% use coastal areas during their juvenile stage. Using both estuaries and coastal areas through partial migration during the juvenile stage is concluded to contribute to the stabilization and yield of seabass populations.