Analysis of a pair of END+ and END? viruses derived from the same bovine viral diarrhea virus stock reveals the amino acid determinants in Npro responsible for inhibition of type I interferon production

The Exaltation of Newcastle disease virus (END) phenomenon is induced by the inhibition of type I interferon in pestivirus-infected cells in vitro, via proteasomal degradation of cellular interferon regulatory factor (IRF)-3 with the property of the viral autoprotease protein N^pro. Reportedly, the amino acid residues in the zinc-binding TRASH motif of N^pro determine the difference in characteristics between END-phenomenon-positive (END⁺) and END-phenomenon-negative (END⁻) classical swine fever viruses (CSFVs). However, the basic mechanism underlying this function in bovine viral diarrhea virus (BVDV) has not been elucidated from the genomic differences between END⁺ and END⁻ viruses using reverse genetics till date. In the present study, comparison of complete genome sequences of a pair of END⁺ and END⁻ viruses isolated from the same virus stock revealed that there were only four amino acid substitutions (D136G, I2623V, D3148G and D3502Y) between two viruses. Based on these differences, viruses with and without mutations at these positions were generated using reverse genetics. The END assay, measurements of induced type I interferon and IRF-3 detection in cells infected with these viruses revealed that the aspartic acid at position 136 in the zinc-binding TRASH motif of N^pro was required to inhibit the production of type I interferon via the degradation of cellular IRF-3, consistently with CSFV.     

Analysis of the equine ovarian structure during the first twelve months of life by three-dimensional internal structure microscopy

A three-dimensional internal structure microscopy (3D-ISM) can clarify the anatomical arrangement of internal structures of equine ovaries. In this study, morphological changes of the equine ovary over the first 12 months of life were investigated by 3D-ISM in 59 fillies and by histological analysis in 2 fillies. The weight and volume of the paired ovaries initially decreased from 0 to 1 months to 2 to 3 months of age and then significantly increased at 8 to 12 months of age. The ovulation fossa was first observed around the 3rd month and became evident after the 6th month. The number of follicles with a diameter of ≥10 mm and the diameter of the largest follicle increased gradually after 6 months of age. On a volume basis, the medulla accounted for nearly 90% of the whole ovary at 0 to 1 months of age, but significantly decreased from 2 to 3 months of age. The volume of the cortex increased progressively after birth and reached approximately 60% of the total volume at 8 to 12 months of age. This significant development of the cortex coincided with the increased number and size of large follicles observed from 6 months of age. These results suggest that the development of the cortex plays a role in the maturation of the follicles and the equine ovary undergoes substantial morphological changes postnatally until puberty.     

Fatal winter dysentery with severe anemia in an adult cow

An adult dairy cow fatally affected with winter dysentery was investigated pathologically and virologically. The cow had severe anemia and diarrhea with massive blood. Pathologically, the loss of surface epithelial cells and necrosis of crypt epithelial cells in the large intestine were observed. Bovine coronavirus (BCV) antigen was observed in necrotic crypt epithelial cells of the large intestine. Virus particles were found in the necrotic epithelial cells of the large intestine. Virologically, BCV was isolated from the feces of the dead cow. The dead cow had no serum antibody against BCV although the co-habitants did. These suggest that severe infection of BCV in the cow without the BCV antibody accompanied by severe hemorrhagic anemia resulted in the cow's death.     

Malachite-green-removing properties of a bacterial strain isolated from fish ponds in Thailand

Malachite green (MG) has been focused on as a biotreatment target and its biological properties have also been an issue in food fish aquaculture. An MG-removing bacterium was isolated from aquaculture fish pond sediment samples in Thailand. The isolate, strain T-5-2, is a Gram-negative, aerobic rod-shaped bacterium, and has been identified as a member of the Pseudomonas putida group. Proton nuclear magnetic resonance spectroscopy (¹H-NMR) analysis of a broth culture medium containing MG showed that the concentration of MG decreased markedly and that other molecules, including leucomalachite green (LMG), were generated. Moreover, liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis showed that the MG concentration in the broth culture medium continuously decreased. This analysis also demonstrated that the concentration of LMG initially increased and then gradually decreased. Furthermore, gas chromatography–mass spectrometry (GC–MS) analysis showed 4-(dimethylamino)benzophenone (4DABP) as a degradation component of MG, which was confirmed by ¹H-NMR and LC–MS/MS analysis. These findings suggest that this bacterial strain can remove MG in broth culture and degrade it to certain metabolites, including LMG and 4DABP. This study is the first detailed evaluation by the combination of LC–MS/MS, GC–MS, and ¹H-NMR analyses of an MG-removing bacterium isolated from Thai aquaculture fish ponds.     

Factors affecting the seed germination and seedling emergence of redflower ragleaf (Crassocephalum crepidioides)

Redflower ragleaf (Crassocephalum crepidioides) is a weed, as well as a minor vegetable, in tropical and subtropical regions of the world. The influence of environmental factors and seed conditions on the germination and emergence of redflower ragleaf have been evaluated in order to help understand its distribution and to develop effective management strategies. The seeds germinated at a constant temperature in the range of 10–30°C and reached a maximum at 15–20°C. The highest germination rate was recorded at an alternating temperature of 20/15°C (day/night). The seeds germinated over a wide pH range (2–12), with the highest germination rate at between 4 and 10. Germination under saturated and flooded conditions was also high. The germination of seeds from opened (mature) capitula was significantly higher than from partially opened or unopened capitula. The germination of seeds without a pappus was significantly higher than for seeds with a pappus. The germination rate of 1 year old seeds decreased drastically when compared to that of freshly harvested seeds. The seedling emergence rate was ～63% for those seeds placed on the soil surface, but no seedling emerged from a depth of ≥1 cm. These results indicate that redflower ragleaf seeds can germinate in various environmental conditions, but that the percentage that germinates will be different in different environments. Regeneration could be effectively prevented by at least a 1 cm soil covering or by destroying the plant before the capitula open. In contrast, freshly harvested seeds from opened capitula should be sown on the soil surface when redflower ragleaf is to be cultivated as a vegetable.     

Adverse and beneficial effects of long-term high-concentration ascorbic acid supplementation in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

The present study examined the effects of high-concentration ascorbic acid (AsA) supplementation over a long period on growth, plasma components, nonspecific immune responses, and thermal tolerance in rainbow trout Oncorhynchus mykiss. Four commercial diets supplemented with 0, 100, 1000, and 5000 mg AsA per kg of diet (designated AsA0, AsA100, AsA1000, and AsA5000, respectively) were fed to rainbow trout (initial weight 1.85 g) for 100 days. AsA contents in liver increased with increasing dietary AsA levels. Feeding period, growth performance, and plasma components did not differ significantly between the AsA groups. On the other hand, the phagocytic assay [nitro-blue tetrazolium (NBT) assay] was significantly higher in fish fed AsA1000 and AsA5000 than those fed a diet without AsA (AsA0) at the end of feeding trial. Superoxide dismutase (SOD) activity in the intestines of fish fed AsA5000 was also significantly higher than that of fish fed AsA0. In the thermal tolerance test, fish fed AsA5000 only showed significantly lower cumulative mortality compared with the AsA0 group. In conclusion, high-concentration AsA supplementation such as AsA5000 over a long period does not induce any adverse effects and can enhance disease and stress resistance in rainbow trout.     

Phylogeography of the Japanese eight-barbel loach <Emphasis Type="Italic">Lefua echigonia</Emphasis> from the Yamagata area of the Tohoku district,Japan

The Japanese eight-barbel loach Lefua echigonia, which is a freshwater fish native to Japan, is distributed from the Tohoku to Kinki districts and is divided into six regional populations according to mtDNA analysis. In this study, we investigated L. echigonia collected from several locations in Yamagata Prefecture and neighboring prefectures using mtDNA control region sequences and confirmed the spatial distribution pattern among the new regional population (Yamagata population). The new population was limited to the Mogami river system in the inland area of Yamagata Prefecture and is distinguished from other regional populations by high sequence divergences.     

The influence of auxin type on the array of somaclonal variants generated from somatic embryogenesis of eggplant, Solanum melongena L.

Somaclonal variation is a possible source of variation in plant breeding. To apply this approach to eggplant breeding, somaclonal variations were observed among plants regenerated through somatic embryogenesis induced by 1-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Variations including leaf shape, plant height, flower number per cluster, fruit shape, anther number per flower and pollen fertility were compared among ? 300 plants (R₀). Although these variations were confirmed among plants regenerated using both auxins, the frequencies of somaclonal variations in leaf shape, plant height, fruit shape and pollen fertility in the NAA experiment were higher than those in the 2,4-D experiment. Variations in flower number and anther number were also confirmed among plants from both experiments, although no significant differences in their frequency were observed. Subsequently, the inheritance of variations (leaf shape, fruit shape, and flower number) observed in R₀ was investigated from generation to generation (R₁). Variations in leaf shape and fruit shape were inherited while those in flower number were not. From these results we concluded that, although a high frequency of somaclonal variations were observed among plants from both methods, embryogenesis with NAA was more efficient than 2,4-D in eggplant.     

Seasonal variations in major components of Crassostrea gigas from Seto Inland Sea

Fisheries Science - The proximate composition and nutritional content of oysters define their quality and commercial value. Cultivation site and harvest date may impact these characteristics. We...