Immunohistochemical analysis of pseudorabies virus spread through neurons innervating the eyeball

Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread.     

Bacterial black spot caused by <Emphasis Type="Italic">Burkholderia andropogonis</Emphasis> on <Emphasis Type="Italic">Odontoglossum</Emphasis> and intergeneric hybrid orchids

A new bacterial black spot disease was observed on Odontoglossum, Odontioda, Odontocidium, and Vuylstekeara orchids in Japan. Typical symptoms on the leaves were dark or black spots (or both) with a yellow halo. The causal agent was identified as Burkholderia andropogonis (Smith 1911) Gillis, Van Van, Bardin, Goor, Hebbar, Willems, Segers, Kersters, Heulin and Fernandez 1995. The isolates were pathogenic on four original host orchids, Phalaenopsis orchid, and tulip; they were not pathogenic on white clover or corn after needle stab inoculation. An antibiotic bactericide (oxytetracycline/streptomycin mixture WP) was most effective for controlling the disease.     

Meteorological factors involved in Japanese encephalitis virus infection in cattle

From 1982 to 1987 a total of 4,371 dairy cattle in Saitama Prefecture, were examined for levels of hemagglutination inhibition (HI) antibody to Japanese encephalitis virus (JEV) and the correlation with meteorological factors and an antibody positive rate was studied. Positivity rates of HI antibody from July to October each year (Yi) ranged from 58.8 to 88.0% with a considerable annual variation. Simple regression analysis of Yi with the comparative meteorological value (Xi) was determined from mean temperatures (Ti, j-1), rainfalls (Ri, j-1) for 10-day-periods each, and the number of days showing 25 degrees C or above (ti, j-1) from June to September, which yielded a correlation coefficient of 0.8147 (p less than 0.05) and an equation for estimated HI antibody positivity rate: Yi = -0.04Xi+79.9 (p less than 0.05). Multiple regression analysis with the power values of three meteorological parameters as independent variables and Yi as the dependent variable, showed a multiple correlation coefficient of 0.987 (p less than 0.05) and a multiple regression equation: Yi = -3991T1/13-0.0035R-12+1978t1/9+4187 (p less than 0.05), giving estimated positivity rates almost equal to the values from the observations. Therefore, a predictive equation was formulated reflecting positive and negative correlations of sero-positivity with the temperature and the precipitation, respectively.     

A cross-protection experiment in pigs vaccinated with Haemophilus parasuis serovars 2 and 5 bacterins, and evaluation of a bivalent vaccine under laboratory and field conditions.

Cross-protection between Haemophilus parasuis serovars 2 and 5 was examined in pigs using a bacterin based vaccine, and subsequently the safety and efficacy of a bivalent vaccine were evaluated. Upon intratracheal challenge of a serovar 2 or 5 strain, pigs immunized with a monovalent vaccine were protected against challenge with a homologous serovar strain, but not with a heterologous serovar strain. Immunization with a bivalent vaccine containing both serovars 2 and 5 bacterins conferred protection in pigs against lethal challenge with each of the serovar strains. A total of 86 pigs from two SPF herds were injected with the bivalent vaccine intramuscularly twice at a four-week interval. No adverse reactions following the vaccination were observed. On day 7 after the second vaccination, vaccinated and non-vaccinated control pigs from herd A were transferred to herd B, where Glasser's disease had broken out. Pigs in the control group developed clinical signs of the disease, and 6 of 8 (75%) pigs died until slaughter, in contrast with only 4 of 46 (9%) pigs in the vaccinated group. In herd C, where there was no outbreak of Glasser's disease, complement fixation antibody titer was raised only in the vaccinated group. A challenge experiment on days 20 and 79 after the second vaccination showed that only the vaccinated pigs were protected. From these findings, the safety and efficacy of the bivalent vaccine were confirmed under laboratory and field conditions.     

Plant–Water Relations of Kidney Bean Plants Treated with NaCl and Foliarly Applied Glycinebetaine

Salinity is at present one of the most serious environmental problems influencing crop growth. It has been extensively demonstrated that salinity affects several physiological processes in the plant, including the plant–water relations of most salt-sensitive crops species. In this study, the effects of salinity on the plant–water relations of kidney bean ( Phaseolus vulgaris L.) and the possibility that foliarly applied glycinebetaine improves these water relations are examined. Kidney bean plants were grown in a greenhouse and treated with 0, 30, 50 and 100 m M NaCl, combined with 0, 10 and 30 m M glycinebetaine in foliar applications. Increased salinity levels decreased stomatal conductance, photosynthetic rate, transpiration and leaf relative water content in the 30, 50 and 100 m M treatments relative to the control treatment. Glycinebetaine applications of 10 m M increased stomatal conductance at 50 m M NaCl, ameliorating significantly the effect of salinity on water relations through increases in the leaf relative water content. At 100 m M NaCl, 30 m M glycinebetaine applications in particular contributed to osmotic stress, and had an adverse effect on plants. Our experiment suggests that glycinebetaine can be used as an alternative treatment to reduce the effects of salt stress on the water relations of salt-sensitive plants, but only to limited salinity levels. Furthermore, the improvement in the water status of kidney beans was dose dependent, suggesting that the concentration of glycinebetaine essential for the survival of salt-sensitive plants is species specific and must be determined individually for each plant species.     

Botulism among penned pheasants and protection by vaccination with C1 toxoid

Fifty to 100 per cent of about 8000 penned pheasants raised on a farm in Hiroshima Prefecture died annually for three years. Deaths were ascribed to type C botulism. Vaccination of four groups of pheasants with partially purified C1 toxoid effectively protected them against type C botulism. The protective efficacy of the toxoid was emphasised by the relatively high susceptibility of the pheasant to C1 toxin.     

Mapping genes for deep-seeding tolerance in barley

Deep-seeding tolerance, the emergence of seedlings from deep seeded conditions, is involved in stand establishment in semi-arid regions, where the soil surface is too dry for seed germination. Genes determining deep-seeding tolerance in barley were mapped using two doubled haploid populations derived from the following crosses: Harrington × TR306 (H/T)and Step toe × Morex (S/M). Significant quantitative trait loci (QTLs) for deep-seeding tolerance were found in each population. Two QTL sex plained 40% of the phenotypic variation in the H/T population and one QTL (S/M) 8% of the total phenotypic variance. Multiple QTLs accounting for coleoptile length and first internode length were detected in both populations. In the H/T population, there were coincident QTLs for deep-seeding tolerance, coleoptile length and first internode length on the long arm of chromosome 5H. These QTLs correspond with previously reported QTLs for abscisic acid and gibberellic acid response. QTL coincidence may be due to the pleiotropic effects of alleles at a single locus. This information may be useful for breeding programs manipulating morphological and physiological traits in order to develop varieties for semi-arid regions. This revised version was published online in August 2006 with corrections to the Cover Date.