Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) using recombinant toxin for detection of antibodies against Pasteurella multocida toxin

To facilitate the control of progressive atrophic rhinitis (PAR) of swine caused by toxigenic Pasteurella multocida, an enzyme-linked immunosorbent assay (ELISA) and a serum neutralization test (NT) have recently been developed to detect antibodies against the P. multocida dermonecrotic toxin (PmDNT). However, the NT is a cumbersome and time-consuming technique. To overcome these drawbacks, we developed an indirect ELISA, using recombinant PmDNT expressed in Escherichia coli, for the detection of antibodies to PmDNT in serum samples from pigs. The practical usefulness of this ELISA was compared with the NT using serum samples obtained from experimentally infected and naturally infected pigs. In the pigs experimentally inoculated with vaccine including PmDNT toxoid, the ELISA and neutralization antibodies were detected at almost the same time, and a good correlation was demonstrated between both tests (P<0.01, R(2)=0.807). Therefore, the ELISA can be used to evaluate the immune reaction of pigs after vaccination with P. multocida toxoid. In a survey conducted on a field herd with a history of clinical AR, the seropositivity by ELISA in pigs of age 4.5-6 months was increased even though the NT was negative, and the correlation was low between the results obtained with the two tests (P<0.01, R(2)=0.38). Therefore, the results indicated that this ELISA might be a useful alternative to the NT currently used to detect the antibody to PmDNT after vaccination or infection with P. multocida.     

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.     

Detection of Mycoplasma hyopneumoniae in lung and nasal swab samples from pigs by nested PCR and culture methods

We examined nasal swab and lung homogenate samples collected from pigs experimentally and naturally infected with Mycoplasma hyopneumoniae for the detection of M. hyopneumoniae by the nested PCR (nPCR) and culture methods. In the 23 experimentally infected pigs, M. hyopneumoniae was commonly detected in nasal swabs by the nPCR and culture methods at 4 weeks after inoculation, and there was a significant correlation (P<0.01) between the titers of viable organisms in nasal swabs and in lung homogenates in the experimentally inoculated pigs. In the naturally infected pigs, on the other hand, discrepancies in detection were found between nasal swab and lung homogenate samples in 17 of 36 cases, although the presence of gross lung lesions correlated relatively well with the detection of organisms from the samples. Our results indicated that the diagnosis of mycoplasmal pneumonia by nPCR in individual pigs with nasal swabs is reliable under these experimental conditions. At present, nPCR with nasal swabs should only be used for monitoring the disease status at the herd level under field conditions.     

Vulnerability of feline T-lymphocytes to charged particles

An analysis of ionizing radiation-induced damage in peripheral lymphocytes has been employed to predict the prognosis of radiotherapy in terms of toxicity in normal tissues. Therefore, understanding the sensitivity of lymphocytes to high linear energy transfer (LET)-charged particles would be indispensable for utilizing charged particle therapy in veterinary medicine. However, the availability of such information is very limited. This study aimed to compare the radiosensitivity of feline T lymphocytes to gamma-rays (0.2 keV/microm) and 4 different types of charged particles with LET values ranging from 2.8 to 114 keV/microm. It was observed that the relative biological effectiveness, inactivation cross-section, and isodose-induced apoptosis increased in an LET-dependent manner. On the other hand, no difference in apoptosis frequency was observed in the cells exposed to an isosurvival dose of all the radiation types tested. This is the first study that demonstrates the LET dependence of cell killing and apoptosis induction in feline T lymphocytes. Our results suggest that lymphocytes can be effectively used to predict the prognosis of charged-particle therapy in cat patients.     

耐热优质黄瓜新品种夏盛的选育

利用耐热、抗病的雌性系ＧＥ作母本，优质的绿宝选１作父本配制而成的一代杂种——夏盛，夏季栽培从播种到始收３３天，一般６６７ｍ２产２５００ｋｇ，高产的可达３５００ｋｇ，耐热性强，抗病毒病（ＣＭＶ）和中抗疫病。果实圆筒形，长２１．５ｃｍ，横径４．７ｃｍ，肉厚２．８ｃｍ，质脆，皮色深绿有光泽，适于华南地区夏秋露地栽培     

Movements of dams milked for fermented horse milk production in Mongolia

Airag, (Fermented horse milk) is a traditional milk product in Mongolia. Herders separate foals from their dams and tie them at a milking site during the daytime to produce airag. To evaluate the effects of horse management on the movement of dams, we tracked three dams in a herd in camp 1 during summer and camp 2 during autumn of 2013 and analyzed their movements during the milking (daytime) and non‐milking (nighttime) periods in an area famous for its high‐quality airag. Dams were gathered every 1.7 ± 0.0 h between 07.46 and 15.47 hours at the milking sites and milked 4.6 ± 0.2 times/day during the study period (86 days). Daily cumulative and maximum linear distances from the milking sites were longer (P < 0.01) during the non‐milking period than during the milking period. Daily home ranges were 91 and 26 times greater during the non‐milking period (P < 0.001) in camps 1 and 2, respectively. The greater range during the non‐milking period would reflect the spatial distributions of water, salt and forage. The dams initially used similar areas and gradually shifted their daily home ranges after several days. This shift suggests that the dams grazed farther afield as forage availability declined around the milking site. For better airag production and sustainable pasture use, our results provide insights useful for evaluating the effects of milking management on vegetation and soil in those pastures, for selecting the appropriate milking times and frequency, and for choosing the right timing to shift milking sites.     

HPLC fluorometric determination of natural phytoplankton phycocyanin and its usefulness as cyanobacterial biomass in highly eutrophic shallow lake

A method for HPLC determination of phycocyanin in phytoplankton samples using gelchromatography with a fluorescence detector was developed to examine the use of phycocyanin as an index of the appearance and progress of cyanobacterial blooms in highly eutrophic lake. At least two types of phycocyanin with different molecular weights, each spectroscopically different from phycoerythrin and allophycocyanin, were found in natural phytoplankton samples. Changes in phycocyanin concentrations were clearly coupled to changes in chlorophyll-a concentrations during June to October while cyanobacterial blooms were occurring. The chlorophyll-a to phycocyanin relationship was linear at chlorophyll-a concentrations of less than 250 μg L^?1. The relationship between cyanobacterial carbon and phycocyanin concentration was also linear, suggesting that phycocyanin content may be a useful index of cyanobacterial biomass in highly eutrophic lakes where large cyanobacterial blooms occur.     

Evaluation of soybean germplasm conserved in NIAS genebank and development of mini core collections

Genetic variation and population structure among 1603 soybean accessions, consisted of 832 Japanese landraces, 109 old and 57 recent Japanese varieties, 341 landrace from 16 Asian countries and 264 wild soybean accessions, were characterized using 191 SNP markers. Although gene diversity of Japanese soybean germplasm was slight lower than that of exotic soybean germplasm, population differentiation and clustering analyses indicated clear genetic differentiation among Japanese cultivated soybeans, exotic cultivated soybeans and wild soybeans. Nine hundred ninety eight Japanese accessions were separated to a certain extent into groups corresponding to their agro-morphologic characteristics such as photosensitivity and seed characteristics rather than their geographical origin. Based on the assessment of the SNP markers and several agro-morphologic traits, accessions that retain gene diversity of the whole collection were selected to develop several soybean sets of different sizes using an heuristic approach; a minimum of 12 accessions can represent the observed gene diversity; a mini-core collection of 96 accession can represent a major proportion of both geographic origin and agro-morphologic trait variation. These selected sets of germplasm will provide an effective platform for enhancing soybean diversity studies and assist in finding novel traits for crop improvement.     

Cell-Associated Adsorption of Thorium or Uranium from Aqueous System Using Various Microorganisms

The cell-associated adsorption of thorium or uranium from the solution containing each metal only at pH 3.50 using various microorganisms was examined. Among the species tested, high thorium adsorption abilities were exhibited by strains of the gram-positive bacteria Arthrobacter nicotianae IAM12342, Bacillus megaterium IAM1166, B. Subtilis IAM1026, Micrococcus luteus IAM1056, Rhodococcus erythropolis IAM1399, and Streptomyces levoris HUT6156. And high uranium adsorption abilities were found in some gram-positive bacterial strains S. albus HUT6047, S. levoris HUT6156, and A. nicotianae IAM12342. Among these highly efficient thorium and uranium adsorbing microorganisms, S. levoris, which could adsorb the largest amount of uranium from the aqueous solution at pH 3.50, could adsorb about 383 mol thorium and 390 mol uranium per gram dry weight of microbial cells from the solution containing thorium or uranium at pH 3.50. The amount and time course of thorium adsorbed were almost unaffected by co-existing uranium; however, the adsorption of thorium was faster when carried out after the adsorption of uranium. Thorium adsorption also became faster when uranium was added after thorium adsorption. The effect of pH on thorium adsorption was also discussed.