Competitive enzyme-linked immunosorbent assay for the detection of the antibodies specific to akabane virus.

A competitive enzyme-linked immunosorbent assay (C-ELISA) using neutralizing monoclonal antibodies (MAbs) against Akabane virus (AKAV) was developed to detect antibodies to AKAV in cattle sera. The performance of the test using 7 different competitor MAbs was evaluated in sequential serum samples and sera from cattle infected with various bovine arboviruses. The dynamics of the antibody response expressed by percentage of inhibition (PI) in C-ELISA coincided with those of neutralizing antibody titers in sequential serum samples from 2 cattle experimentally infected with AKAV. The value of PI in C-ELISA for convalescent sera from cattle infected with arboviruses correlated with the neutralizing antibody titer to AKAV but was unaffected by the antibodies to other arboviruses. In the validation experiment of C-ELISA using 286 bovine sera previously examined for the AKAV antibody by serum neutralization (SN) test, the relative specificity of C-ELISA was more than 98%, whereas the relative sensitivities of individual MAbs ranged from 49% to 82.2%. Overall agreement between C-ELISA and the SN test varied from 72% to 90% depending on the MAb. These results suggest that the C-ELISA is acceptable as a rapid and specific method for detecting antibodies to AKAV and is a potential alternative to the SN test.     

Genetic analysis for sow stayability at different parities in purebred Landrace and Large White pigs

Genetic parameters for sow stayability were estimated from farrowing records of 10,295 Landrace sows and 8192 Large White sows. The record for sow stayability from parity k to parity k + 1 (k = 1, …, 6) was 0 when a sow had a farrowing record at parity k but not at parity k + 1, and 1 when a sow had both records. Heritability was estimated by using single-trait linear and threshold animal models. Genetic correlations among parities were estimated by using two-trait linear–linear and single-trait random regression linear animal models. Genetic correlations with litter traits at birth were estimated by using a two-trait linear–linear animal model. Heritability estimates by linear model analysis were low (0.065–0.119 in Landrace & 0.061–0.157 in Large White); those by threshold model analysis were higher (0.136–0.200 & 0.110–0.283). Genetic correlations among parities differed between breeds and models. Genetic correlation between sow stayability and number born alive was positive in many cases, implying that selection for number born alive does not reduce sow stayability. The results seem to be affected by decisions on culling made by farmers.     

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions were compared between the immunity‐selected Large White line and the non‐selected Large White line. The selected Large White line showed a higher level of pulmonary MPS lesions compared with the non‐selected Large White line. Subsequent to vaccination, the percentage of natural killer cells and T cells (CD3⁺CD4⁺CD8^‐ and CD3⁺CD4^?CD8⁺ T cells) were significantly increased in the non‐selected line but remained unchanged in the immunity‐selected Large White line. Secretion of Mycoplasma hyopneumoniae vaccine‐specific immunoblogulin G and phagocyte activity in peripheral blood were significantly higher in the immunity‐selected Large White line than in the non‐selected line. Expression of interleukin (IL)‐4 and IL‐6 messenger RNA in hilar lymph nodes was significantly lower in the immunity‐selected Large White line than in the non‐selected line. However, expression of IL‐10 in all immune tissues was significantly higher in the immunity‐selected Large White line. These results suggest that the selection for high immunity was not effective in increasing resistance to MPS lung lesions.     

Strong antiviral activity of heated and hydrated dolomite--preliminary investigation

Heated and hydrated naturally occurring dolomite showed very strong antiviral activity. Infectivity of avian and human influenza, avian infectious bronchitis (coronavirus), Newcastle disease (paramyxovirus) and avian laryngotracheitis (herpesvirus) viruses dropped at least 1,000 fold following contact with the dolomite for five minutes at 4 degrees C. Dolomite is expected to be useful to inhibit the incidence of emerging and re-emerging infectious diseases.     

Effect of ovary lipid of skipjack tuna on rat serum components after stress application

Lipid extracted from the ovary of skipjack tuna by the method that we developed is rich in phospholipid-type docosahexaenoic acid. The ovary lipid of skipjack tuna (OLS) was studied for its anti-stress activity in male Wistar rats, focusing on stress-related blood components: recovery from stress was examined after application of water immersion restraint stress. As a result, serum corticosterone (CORT) secretion was inhibited and decreased rapidly after stress application in rats given OLS compared with control rats. As CORT acts as a glucocorticoid, non-esterified fatty acid (NEFA) is expected to increase by stress application. However, the concentration tended to be lower in rats given OLS than in control rats. With respect to OLS concentration, OLS increased serum dehydroepiandrosterone, secretion concentration-dependently. In addition, as with the recovery study, it tended to inhibit the increase in NEFA. These results indicate that OLS may have an anti-stress activity against acute stress.     

Nutrient leaching from the plow layer by water percolation and accumulation in the subsoil in an irrigated paddy field

To estimate the impact of water percolation on the nutrient status in paddy fields, the seasonal variations of the concentrations of cations, anions, inorganic carbon (IC), and of dissolved organic carbon (DOC) in percolating water that was collected from just below the plow layer (PW-13) and from drainage pipes at the 40 em depth (PW-40), as well as in irrigation water were measured in an irrigated paddy field. Total amounts of Ca, Mg, K, Fe, and Mn leached from PW-13 during the period of rice cultivation were estimated to range from about 390 to 770, 65 to 130, 33 to 66, 340 to 680, and 44 to 87 kg ha^-1, respectively. Amounts of losses that were estimated from the differences between the input by irrigation water and the output by percolation water from the plow layer corresponded to 11 to 26, 22 to 47,5.9 to 12, and 13 to 26% of exchangeable Ca and Mg, amorphous Fe, and easily reducible Mn in the plow layer, respectively. The concentrations of Ca, Mg, K, Fe, and Mn in PW13 were higher than those in PW-40. The amounts of these nutrients that were retained in the subsoil between the 13 em and 40 em soil depth corresponded to 83, 86, 61, 99, and 89% of the amounts that percolated from the plow layer, respectively. Total amounts of IC and DOC that percolated from the plow layer ranged from 750 to 1,500 and 85 to 170 kg-C ha^-1, which corresponded to 5.0 to 10.0% and 0.6 to 1.1% of the total carbon content in the plow layer, respectively. Eighty eight % of IC in the percolating water from the plow layer was also retained in the subsoil.     

Improvement of preservation method for ammonia-oxidizing bacteria by freeze-drying

Long-term preservation of ammonia-oxidizing bacteria had been achieved until now by using the silica gel-freezing method. Recently, the freeze-drying method for the preservation of microorganisms has been widely used, and has been applied to the preservation of ammonia-oxidizing bacteria. The 10% skim milk-l% pyruvate or glutamate ratio was optimum as a dispersion-medium, and the preservation temperature of the freeze-dried cells was effective at -80°C. This method of preservation was found to be considerably more effective than the silica gel-freezing method.     

Identification of Trichoderma SKT-1, a biological control agent against seedborne pathogens of rice

Trichoderma SKT-1 was previously reported as a powerful biological control agent against seedborne pathogens of rice, but the taxonomic disposition of the fungal isolate was not clear. Trichoderma SKT-1 produced irregular pyramidal warts on conidia and had an optimum growth temperature of 30°C. Morphological characteristics and colony growth were identical to those of known species of Trichoderma, including the newly recognized species T. asperellum. The 5.8S rDNA with the internal transcribed spacer (ITS) region (ca. 514 bp) of the fungus was compared with those of known species to determine the phylogenetic placement of the fungus. The length and sequence of the regions from Trichoderma SKT-1 were completely identical to those of an isolate of T. asperellum NRRL 5242 (AJ230669). On the basis of these results, we concluded that Trichoderma SKT-1 was T. asperellum.     

Occurrence of chrysanthemum virescence caused by “<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma aurantifolia” in Okinawa

In 1999, a disease of chrysanthemum [Dendranthema grandiflorum (Ramat.) Kitamura], characterized by virescence of flowers, occurred in Okinawa Prefecture. The causal agent was identified as “Candidatus Phytoplasma aurantifolia” based on 16S rDNA sequencing. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB247462.     

Genetic Variability in Phosphorus Acquisition and Utilization Efficiency from Sparingly Soluble P-Sources by Brassica Cultivars under P-Stress Environment

Plants display an array of classical strategies to maximize phosphate (Pi) acquisition from sparingly soluble P sources. Acclimation to Pi-stress via elegant Pi-starvation induced (PSI) adjustments would reduce our current overreliance on expensive, polluting and non-renewable Pi-fertilizers. Nevertheless, differences in the ability of various species to solubilize sparingly soluble P-sources have been often evidenced; inter-cultivar variations are scarcely documented. Brassica is known as an effective, non-mycorrhizal user of sparingly soluble P-sources. Various growth parameters and biomass accumulation by genetically diverse Brassica cultivars were determined in four experiments using hydroponics and quartz sand culture media. Role of PSI root mediated pH changes, organic anions (OAs) exudation and altered root architecture in mobilization and acquisition of sparingly soluble P-forms [Jordan rock-P (RP) at 2 g l⁻¹ and Ca₃(PO₄)₂ (TCP) at 0.2 g l⁻¹ respectively] was investigated. Cultivars showed considerable genetic variations in biomass accumulation, various growth parameters and root–shoot ratio. Concentration and total uptake of P, specific absorption rate of P, P-transport rate and P-utilization efficiency (PUE) were also significantly (P < 0.001) different for various cultivars and their dry matter was significantly correlated with P-uptake [r = 0.94** (significant at 1% level)]. P-tolerant cultivars showed substantial decrease in solution media pH because of H⁺ efflux and exuded more carboxylates than low P-sensitive cultivars under P-starvation. P-uptake by cultivars increased linearly with decreasing pH. The amount and types of OAs exuded from the roots of P-starved plants differed from those of plants grown under P-sufficient environment. In split pot study, with TCP and RP supplied spatially separated from other nutrients, efficient cultivars were still able to mobilize RP and TCP more efficiently than inefficient cultivars. In rhizobox study, the elongation rates of primary roots decreased but the elongation rates of the branched zones of primary roots and the length of lateral roots increased under P-starvation. Tested cultivars showed genetic diversity in accessing, mobilization, acquisition and utilization of Pi from sparingly soluble P forms. An arrange marriage of plant traits can explain cultivar’s access to different forms of sparingly soluble P, and in addition to altered lateral root topology and enhanced P-uptake and PUE, enhanced H⁺ efflux and OAs exudation are key factors in Pi scavenging from extra cellular sparingly soluble P-forms.