Strategies of newcastle disease vaccination for commercial ostrich farms in Japan

Strategies of Newcastle disease (ND) vaccination were demonstrated in a commercial ostrich farm in Japan. Three of 13 seven-month-old ostriches kept in a pen were vaccinated with a live ND vaccine by eye dropping for the 1st and 2nd vaccinations and spraying for the 3rd to 5th vaccinations. Antibodies against ND virus (NDV) were detected in all of the unvaccinated ostriches by virus neutralization test. At 2.5 months post final vaccination, 2 ostriches introduced into the pen raised antibodies against NDV. These data indicate that NDV may be transmitted from vaccinated to unvaccinated ostriches in the flock and that the virus may be sustained for a certain period in the flock. These data may be helpful for ND vaccination management in ostrich farms.     

Situation of serum antibodies against Newcastle disease virus in slaughter-age ostriches after vaccination campaign in Japan

A total of 516 slaughter-age ostrich sera were collected in Japan during 2006-2009. Sixty-one of five hundred and sixteen were positive by virus neutralization (VN) test and the titer of most positive samples was low level. Within the 61 positive sera, 35 sera were collected from unvaccinated ostriches. This result implies that these ostriches might have been infected naturally with low-virulent Newcastle disease virus (NDV). Within the 455 negative samples, 125 samples were from vaccinated ostriches. Since ostrich farmers use live attenuated vaccines, it is reasonable that the titer decreased to below detection level by 1 or 1.5 year-old. The above data indicate that NDV has infiltrated into ostrich farms in Japan, and that the efficacy of ostrich ND vaccination is often time-limited.     

Surveillance and characterization of Newcastle disease viruses isolated from northern pintail (Anas acuta) in Japan during 2006-09

A total of 38 Newcastle disease virus (NDV) isolates were obtained from 6060 fecal samples from northern pintail (Anas acuta) ducks collected in the Tohoku district in Japan during 2006-09. One isolate from each sampling location and date was selected for a total of 38 isolates, then 15 of these were characterized for their pathogenicity by mean death time of minimum lethal dose (MDT/MLD) using chicken embryos and by plaque formation on chicken embryo fibroblasts. Furthermore, nine isolates were randomly selected from these 15 isolates, and the fusion protein genes were sequenced to characterize amino acid sequences around the cleavage site. All 15 were confirmed to be nonvirulent by MDT/MLD test, and nine isolates were also confirmed as nonvirulent by the cleavage site of the fusion protein 112G/E-K/R-Q-G/E-R*L117 that was specific for nonvirulent NDVs. The characteristics of nine isolates identified by phylogenic analysis of the fusion protein gene indicated that the isolates belong to genotype I or II. In addition, we also isolated 68 avian influenza viruses and 28 other hemagglutinating viruses. Our data indicate that northern pintails are subclinically infected by, perpetuate, and distribute NDV along with different subtypes of avian influenza viruses and other hemagglutinating viruses during their migrations across vast areas over the Northern Hemisphere to Japan.     

Deficiency of N,P, K,Ca, Mg,or Fe Mineral Nutrients in Narcissus cv. “Garden Giant”

Narcissus cv. “Garden Giant” bulbs were grown in N, P, K, Ca, Mg, or Fe-deficient solutions and compared with bulbs grown in the control solution containing all these nutrients. Plants were sampled at 4 stages: (I) at planting, (II) at sprouting, (III) after flower senescence and/or after visible deficiency symptoms developed, and (IV) at lifting. Observation of visible deficiency symptoms showed that leaves of narcissus displayed chlorosis in the —N, —Mg, and —Fe treatments, while roots were most susceptible to Ca-deficient conditions. Root tips in the —Ca treatment showed brown in followed by root rot. In the —N treatment, shoot growth was markedly retarded and leaves were small and yellow. On the other hand, visible deficiency symptoms were not evident in the —P treatment except for early senescence. K deficiency symptoms were also not evident. Narcissus flowers were not affected by the mineral deficiencies and mineral contents in full-bloom flowers were not different between samples.

In the control plants, there was a large accumulation of N in the roots at sprouting and the content decreased thereafter. Large amounts of K accumulated in roots more than in any other organs. On the other hand the Ca content was high in the tunic and Ca in the scales was hardly mobilized thoughout the growth period. A large proportion of each element eventually accumulated in new inner scales whereas only a small fraction in the old outer scales. The -N, -Ca, and -Mg treatments severely depressed dry matter accumulation, unlike the -P, -K, and -Fe treatments. The -N treatment did not affect the concentration of other minerals, but the -P treatment tended to decrease N, K, Ca, and Mg concentrations. On the other hand, the -K treatment increased Ca and Mg concentrations. -Ca caused an increase in Mg concentration and -Mg raised Ca concentration. These results may be due to compensation effects of cation absorption for the maintenance of the cation/anion balance.     

Characterisation of proteins in the storage organs of Curcuma alismatifolia Gagnep.

Summary

Curcuma alismatifolia rhizomes are exported to world markets for ornamental use. Rhizome size and the number of storage roots affected shoot sprouting, plant growth, and inflorescence yield and quality, indicating an important role for storage food reserves such as the carbohydrates and proteins in these organs. An understanding of the proteins used for growth and development in rhizomes and in storage roots is lacking, so changes in the storage proteins in these organs were investigated at the planting stage, the sprouting stage, the second-leaf stage, the flowering stage, and the dormancy stage. The protein concentrations in both these organs decreased rapidly until the flowering stage in order to support the initial growth of roots and shoots. The intensity of Coomassie Brilliant Blue staining of tissues was stronger in rhizomes than in storage roots. Protein staining was present in the cytosol and in the cell walls in rhizomes, and was intense in small particle-like protein bodies in the cytosol. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles showed that peptides of 10.6 kDa and 12.0 kDa were the two major soluble proteins in rhizomes and were different from the proteins in storage roots.The 10.6 kDa and 12.0 kDa bands contained five peptides and one peptide, respectively, when separated by 2D-PAGE. The amino acid sequences of these six peptides were not homologous to any known proteins using BLASTP or FASTA analysis. The N-terminal amino acid sequences of these two polypeptides did not start with methionine, indicating that they had undergone posttranslational modification. Knowledge of the major storage proteins in the rhizomes and storage roots of C. alismatifolia may provide information to improve N-fertilisation.     

IDENTIFICATION OF ENDOPHYTIC BACTERIA ASSOCIATED WITH N2 FIXATION AND INDOLE ACETIC ACID SYNTHESIS AS GROWTH PROMOTERS IN CURCUMA ALISMATIFOLIA GAGNEP.

Endophytic bacteria carrying out dinitrogen (N₂) fixation and indole acetic acid (IAA) synthesis were firstly identified in C. alismatifolia, a globally important flower crop. Their potential as growth promoters to stimulate the rapid growth of host plant was also examined. It will be beneficial to reduce the propagation period of tissue culture plantlets, and also utilize as a biofertilizer for rhizome production in the field. Seven endophytic bacteria were isolated from the leaf, four isolates from the leaf base, and two from the rhizome. ECS203, a gram-negative bacterium with a round shape, showed the highest N₂ fixation at 4.2 nmol C₂H₄/10⁶ cells/hr, and ECS202 showed the highest IAA synthesis at 296 μL μg ^{? 1} protein. Three selected isolates of N₂-fixing and IAA synthesizing endophytic bacteria, i.e., ECS202, ECS203, and ECS204, isolated from the leaf base, were used to reinoculate Curcuma plantlets derived from tissue culture. Then, plants were grown in sterilized sand for 2 months and weekly supplied with N-free nutrient solution. Plant growth, colonization, nitrogen fixation, and IAA synthesis were measured at two months after planting. The inoculated plants clearly showed a better performance of plant growth and yield in terms of the plant height, plant weight, leaf area, and diameter of new rhizomes compared with uninoculated plants. The chlorophyll content and N concentration of leaves and roots also increased in inoculated plants. Endophytic bacteria from inoculated plants colonized the roots, rhizome, and leaf base. Partial sequence analysis using 16S rDNA indicated that the isolate ECS202 corresponded to Sphingomonas pseudosanguinis (99.2% similarity over 1,371 bp), ECS203 to Bacillus drentensis (99.4% similarity over 1,450 bp) and ECS204 to Bacillus methylotrophicus (99.9% similarity over 13,06 bp).     

Site of nitrogen accumulation in tulip (Tulipa gesneriana L.) roots during winter

Abstract

Plants (60 species in 37 genera, 27 families) grown on granite weathered soils of temperate natural forest in central Japan were sampled and analyzed for Fe, Mn and Cu. Soil samples coIlceted from the site of plant stands were also analyzed. Results showed that considerable difference existed amons plant species with less variations amons plant samples of the same plant species. Similar responses were frequently found among plant species in the same genus and sometimes in the same family. Variations due to soU also occurred but to a lesser degree. Pe, Mn and Cu were generaIly low in coniferous trees. On the other hand Acantiropanax sciadophylloides accumulated higher amount of Mn in the leaves (4.6 × 10³ ppm, dry matter basis) which is about 180 times more than that of low content species. Other Mn accumulating species were found in Anacardiaceae and Aceraceae. Cryptomeria japonica was the lowest in Mn content (26 ppm). Mean concentration ratio for Mn was 113. Cu was found to be slightly rich in scattering species including Lastrea japonica, Magnolia salleifolia, Acer mono var. connivens and Callicarpa japonica. Mean concentration ratio for Cu was 17.     

Nutrients,Free Amino Acids,and Sugar Contents in Narcissus Roots Affected by N,P, K Deficiency during Winter

Non-rooted bulbs of Narcissus L. cv. “Garden Giant” were planted on October 25, 1993 and cultivated under N, P, or K deficiency or in complete medium (control). The roots emerged on November 5 and were sampled on March 7, 1994. The X-ray microanalyzer images showed the distribution of N, P, K, Ca, Mg, S, and Cl in a control root. The distribution of Cl was similar to that of K and also a similar distribution was found between Ca and S. There were significant differences in the concentration of N, P, K, Ca, Mg among treatments. Interaction among nutrient levels was observed. Control roots accumulated a large amount of nitrogen (84 mg g DW^-1) which was partitioned almost equally between the 80% ethanol soluble and insoluble fractions. N-deficient roots in the -N treatment, on the other hand, accumulated only about 12 mg g DW^-1 nitrogen, mostly in the insoluble fraction (83% of total N). Total amino acid contents responded to N, P, or K deficiency. Glutamine was a major free amino acid in control roots (17 mg N g DW^-1) and K-deficient roots (29 mg N g DW^-1), while arginine seemed to be the predominant amide in P-deficient roots. Content of total soluble sugars in the control, -P and -K treatments that was 49, 46, and 49 mg g DW^-1, respectively, was lower than in the -N treatment (137 mg g DW^-1). Mono- and disaccharides; fructose, glucose, and sucrose, were found in narcissus roots.     

Identification of long needle crystals and distribution of oxalate salts in Narcissus cv. ‘Garden Giant’

Long needle crystals abundantly present in the bulb scales and the shoot sap of Narcissus cv. ‘Garden Giant’ were identified as Ca-oxalate by ¹³C-nuclear magnetic resonance and capillary electrophoresis. The Ca-oxalate crystals were distributed in every part of the plant, including tunics, shoot, flowers, basalplate, and roots. A large proportion of water-soluble oxalate was present in the roots, but not in the scales and shoot. Oxalate accumulation occurred mainly in the young new scales originating from the basal part of shoot. Most of all the oxalate in the inner new scales and the outer old scales was in the form of Ca-oxalate. The fact that Ca and oxalate contents in old scales remained constant after planting to the flowering stage indicated that Ca and oxalate in bulb scales are not reserve nutrients supporting plant growth. Alternatively Ca-oxalate accumulated in scales appeared to be a stable end-product.