Reactivity and prevalence of neutralizing antibodies against Japanese strains of bovine viral diarrhea virus subgenotypes

Bovine viral diarrhea virus (BVDV) field isolates show genetic and antigenic diversity. At least 14 subgenotypes of BVDV-1 and 4 of BVDV-2 have been identified in Artiodactyla worldwide. Of these, 6 subgenotypes of BVDV-1 and 1 of BVDV-2 have been isolated in Japan. Previously, we reported that each subgenotype virus expresses different antigenic characteristics. Here we investigated the reactivity of neutralizing antibodies against representative strains of Japanese BVDV subgenotypes using sera from 266 beef cattle to estimate the prevalence of this epidemic virus among cattle in Japan. Antibody titers at concentrations at least 4-fold higher than antibodies against other subgenotype viruses were considered subgenotype specific. Subgenotype-specific antibodies were detected from 117 (80.7%) of 145 sera samples (69.7% against BVDV-1a, 1.4% against BVDV-1b, 8.3% against BVDV-1c, and 1.4% against BVDV-2a). The results suggest that neutralization tests are useful in estimating currently epidemic subgenotypes of BVDV in the field.     

Effect of the methoxyiminoacetamide fungicide,SSF129, on respiratory activity in Botrytis cinerea

(E)-2-Methoxyimino-N-methyl-2-[2-(2,5-dimethylphenoxymethyl)phenyl]acetamide (SSF129) has been developed as a broad-spectrum systemic fungicide for control of cereal and fruit diseases. This compound inhibited NADH-oxidation by submitochondrial particles from mycelial cells of Botrytis cinerea, with an EC₅₀ value of 14.5 nM , due to blockage of electron transport through the cytochrome bc₁ complex in the mitochondrial respiratory chain. However, SSF129 did not suppress, but rather increased, oxygen consumption by mycelial cells of the fungus. This was because mycelial cells contain an alternative oxidase protein and the cells have the ability to rapidly switch electron flux from the main cytochrome pathway to the alternative pathway on blockage of the former by SSF129. The alternative pathway of the mycelia seems not to be operative when the cytochrome pathway is functional. Naturally occurring flavonoids inhibited the alternative oxidase of the mycelial cells in a dose-dependent manner, with EC₅₀ values of 68.4 µM for flavone and 63.7 µM for flavanone. These observations suggested that plant components play an important role in control of gray mould by SSF129. © 1999 Society of Chemical Industry     

Growth and root sucker ability of field-grown transgenic poplars overexpressing xyloglucanase

Xyloglucan is thought to be a key hemicellulose cross-linking adjacent cellulose microfibrils in plant cell walls. The growth traits of transgenic poplars (Populus alba) with decreased xyloglucan from overexpression of Aspergillus aculeatus xyloglucanase were characterized during a 4-year field trial. The field-trial site consisted of two blocks, a fertile soil block and a non-fertile soil block, determined by soil analysis. In the fertile block, the growth of aboveground biomass of the transgenic poplars was reduced to 24?C44?% compared to that of wild-type poplars, in contrast to the growth seen in chamber and greenhouse conditions. In the non-fertile block, the aboveground biomass of transgenic poplars was also smaller than that of the wild-type poplars. Because poplars reproduce asexually by root suckers, we also compared the formation of root suckers from transgenic and wild-type poplars. Root suckers formed less frequently from transgenic poplars than from wild-type poplars. The growth rates of root suckers from transgenic poplars were also slower than those from wild-type poplars. The results showed that constitutive degradation of xyloglucan impairs poplar growth and vegetative reproduction ability.     

Usefulness of intraoperative ultrasonography for two dogs with spinal disease

Intraoperative ultrasonography was performed during spinal surgery in two dogs. An extradural lesion was demonstrated by myelography in case 1, and an extradural tumor was found during surgery. Case 2 was diagnosed with a spinal subarachnoid cyst by myelography and magnetic resonance imaging. Ultrasonographically, pathologic conditions of the spinal canal or cord were adequately imaged during surgery in both cases. Intraoperative ultrasonography was a superior method that non-invasively revealed intradural and extradural conditions and thus constitutes a valuable diagnostic mean to ensure safe and precise spinal surgery.     

One-step Generation of Phenotype-expressing Triple-knockout Mice with Heritable Mutated Alleles by the CRISPR/Cas9 System

Triple-knockout mice generated by the one-step CRISPR/Cas9 system were examined for the effects of multiple gene modifications on each phenotype and individual gene function. Sixty embryos were transferred, and 9 pups were obtained; all 9 pups had mutations on 3 loci, and 7 pups showed mutations in all-alleles. F0 mice showed knockout phenotypes or no protein expression of target genes simultaneously, and these mutations were normally inherited in the next generation.     

Detection of microbial groups metabolizing a substrate in soil based on the [l4C] quinone profile

Analysis of [^l4C]respiratory Quinones synthesized in soil for 6 h after spiking with [U-¹⁴C]glucose, [U-^l4C]glycine, and [1,2-^l4C]acetate enabled to fingerprint the microorganisms metabolizing each substrate in soil and to determine the whole structure of the microbial communities at the same time. The [^l4C]- Quinones synthesized from [U-^l4C] glucose were the same as those from [U-^l4C] gIycinc in soil, suggesting that the same microbial groups metabolized glucose and glycine under the given conditions. No [^l4C]quinones from [1,2- ¹⁴C] acetate were detected in soil, indicating that the metabolism of acetate by microorganisms is negligible. The profiles of [^l4C]quinones from [U-^l4C]- glucose were compared between Nagoya University Farm soils subjected to 4 different fertilizing practices. The soils receiving farmyard manure contained [^l4C]menaquinones with highly hydrated isoprenoid units, which indicated the presence of Actinobacteria metabolizing glucose. The soil receiving only chemical fertilizers contained [¹⁴C]ubiquinone with 8 isoprenoid units (Q-8), indicating the presence of beta and gamma subdivisions of Proteobacteria. All the 4 soils were characterized by the high proportions of [¹⁴C] MK-6 and a mixture of [^l4C]MK-8(H₄) and [^l4C]MK-9. The Q-9 and Q-10(H₂) indicators of fungi, were not labeled under most of the conditions.     

Usefulness of myelography with multiple views in diagnosis of circumferential location of disc material in dogs with thoracolumber intervertebral disc herniation

The usefulness of myelography with multiple views (lateral, ventrodorsal, left and right oblique view) in the diagnosis of the exact circumferential location of herniated disc material around the spinal cord in 80 dogs diagnosed with thoracolumbar intervertebral disc herniation at surgery was assessed by comparison of clinical and surgical findings. The circumferential location of the compressing mass was diagnosed in 94% of dogs on myelography. The oblique view was of more benefit than the ventrodorsal view in diagnosing the circumferential distribution of the compressing mass. Only the oblique view contributed to a diagnosis of lateralization of the compressing mass in 45% of dogs. Fourteen percent of dogs had clinical lateralization contralateral to myelographic lateralization. The myelographic localization agreed with the surgical localization in 97% of dogs with regard to the exact location of herniated disc material. The presence of clinical lateralization contralateral to myelographic lateralization and a high proportion of agreement of myelographic and surgical localization documents that myelography with multiple views is useful and essential to accurately determine the circumferential location of disc material around the spinal cord.     

A new garlic growing system from bulbils through transplanting

Summary

We have developed a new garlic growing system from bulbils through transplanting. By this system, extremely small seed bulbils produced bulbs as large as those from cloves of bulbs. In the new system, the bulbils harvested in summer and stored at 18–20°C in winter are grown from the following spring to summer in paper pots, and then transplanted to the field. In Hokkaido, northern Japan, July transplanting produced larger bulbs than August or September transplanting, and the bulbs produced were almost the same size as those of traditional growing methods. One bulbil weighed 75.3±5.6 mg and 84.2±5.4 mg in cv. Furano and a clone, No. 205, respectively, while one clove weighed 5–7g. A clone, H500, produced bulbs of 79.9±3.0.g although its original bulbil weighed only 15.4±0.8.mg. Moreover, this clone produced a great number of bulbils, 250–300, per plant.     

Canine serum alkaline phosphatase isoenzymes detected by polyacrylamide gel disk electrophoresis

Serum alkaline phosphatase (ALP) isoenzymes were studied in normal dogs using a commercially available polyacrylamide gel disk electrophoresis kit (PAG/disk kit). Serum samples taken from the dogs were incubated with neuraminidase, after which most showed ALP isoenzymes as two characteristic stained bands. To determine the origin of each band, ALP isoenzymes of serum and tissue extracts (liver, intestine and bone) were characterized by heating, wheat germ agglutinin (WGA) and levamisole treatments. The results suggested that the band detected on the anode was liver ALP (LALP) and that the band detected on the cathode represented bone ALP (BALP), and both were corticosteroid-induced ALP (CALP). The percentage of each ALP isoenzyme to total ALP activity was estimated by densitometry. The percentage of BALP was the highest in young dogs (age<1 year, 64.7% ), and this value decreased with age. In contrast, the percentage of LALP in young dogs (22.2%) was much lower than that in middle-aged dogs (ages 1 year to 7 years, 59.3%) and old dogs (ages>7 years, 50.4%). The present results suggested that a commercially available PAG/disk kit is capable of detecting three serum ALP isoenzymes in dogs, and further that it may have clinical applications in the evaluation of ALP isoenzymes in veterinary medicine.