Acid-induced autoagglutination found in chicken pathogenic Escherichia coli strain.

Chicken pathogenic Escherichia coli strains were found to autoagglutinate in a static culture of trypticase soy broth (TSB). One strain, designated PDI-386, was further studied for its autoagglutinating property. Acidity in the cultured medium caused by glucose degradation induced the autoagglutination. The bacterial cells grown in a glucose-free L-broth could be aggregated by adding acid, which suggests a potentiality of autoagglutination of the strain grown in the L-broth. The autoagglutinating parent (Agg) formed small colonies with irregular edges like rough colonies on the TS agar, whereas its non-autoagglutinating variant (Nag) formed larger smooth colonies with a perfectly round edge. The Nag colony was easily generated from the Agg colony on the TS agar. The autoagglutinating property was very unstable when the bacteria was passed in the TSB, but rather stable in the L-broth. Under electron microscope, the Agg were found to possess pili of more than 20 microns in length. However, the phenotypic expression of autoagglutination did not correlate with that of mannose-sensitive hemagglutination against guinea pig erythrocytes. Incubation of the Nag in the L-broth at room temperature for more than 10 days provoked the reversion of the autoagglutination. There was no difference between the Agg and the Nag in terms of surface hydrophobicity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of membrane proteins and LPS, and plasmid profiles. The virulence of the Agg was higher than that of the Nag. The autoagglutination property is, however, so unstable that the pathogenicity of E. coli isolates from chickens should be carefully evaluated.     

Estrone sulfate and progesterone profiles during late gestation in recipient cows transferred embryos produced by nuclear transfer and in vitro fertilization

The aim of present experiment was to evaluate the plasma concentrations of estrone sulfate (E(1)S) and progesterone (P(4)) during late gestation in recipient cows transferred embryos produced by nuclear transfer (NT) and in vitro fertilization (IVF). Blood samples were collected from recipients transferred embryos produced by NT (n=9) and IVF (n=13) at 160, 220, 240, 260 and 270 d of gestation and then at 5 d intervals until parturition. Plasma samples were analyzed for E(1)S and P(4) by ELISA. One NT and three IVF cows aborted between days 220 and 260 of gestation. Two NT and one IVF cow had prolonged gestation (over 290 d). One IVF cow had an overweight fetus (50 kg) after abortion (257 d). The patterns of changes in the concentrations of E(1)S during late gestation in the NT and IVF cows were almost identical. The NT and IVF cows that aborted had prolonged gestation and much higher E(1)S concentrations than the average. One NT cow aborted after 220 d of gestation and had a sudden high increase in its E(1)S concentration from 160 d to 220 d of gestation. The NT and IVF cows that had prolonged gestation also had significantly higher (P<0.05) P(4) concentrations than the average. These results raise the possibility that the E(1)S and P(4) profiles can be used to monitor some late gestational problems, such as higher birth weight, abortion and prolonged gestation.     

Seasonal Changes in Rigor Development and Flesh Texture of Wild Japanese Sea Bass (Lateolabrax japonicus)

The season for finding the firmest sashimi made from wild Japanese sea bass was investigated. Flesh rigor tension and contraction were measured by a “new” device with isometric (muscle length kept constant) and isotonic (balance beam moves freely) transducers. Both flesh rigor tension and contraction peaks appeared at the same postmortem times, with attainment of ultimate pH and adenosine triphosphate/inosine monophosphate ratio in four seasons. Seasonal seawater temperature correlated negatively to seasonal maximum rigor contraction and positively to seasonal flesh breaking strength at 72 h postmortem. After rigor peak tension attainment, the tension relaxation was much lower than that of other wild fish species, especially in summer. The sea bass muscle resisted structure disruption in rigor isometric tension generation. Wild sea bass sashimi was firmest in breaking strength in summer.     

Prediction of grain protein content in winter wheat through leaf color measurements using a chlorophyll meter

A field trial was conducted over a 3-year period at the Hokkaido Kitami Agricultural Experiment Station to examine whether the grain protein content (GPC) of a winter wheat cultivar (Triticum aestivum L. cv. Chihokukomugi) suitable for Japanese noodle-making could be predicted before harvest. The prediction of the GPC was accurate based on the color of the second leaf (just below the flag leaf) at the end of the emergence of the inflorescence, when nitrogen application was graded. In order to evaluate the reliability of this test, a survey of 95 wheat fields in the eastern part of Hokkaido was also carried out during a 3-year period. The prediction of the GPC for this cultivar based on the color of the second leaf was less accurate across many sites. The results of this survey, however, suggested that the leaf color could be used as an index for ranking the GPC as low or high in relation to processing requirements. When the leaf color value of the second leaf measured with a chlorophyll meter at the end of the emergence of the inflorescence was less than 40, it was predicted that the GPC would be lower than the processing requirement. This index could be applied to the cultivars grown in the eastern part of Hokkaido, except for those grown on peat soils.     

Susceptibility of Commercial Tomato Cultivars to Bacterial Wilt in Hydroponic System

Commercially available tomato cultivars were hydroponically cultured for inoculation, with Ralstonia solanacearum (K-101), which causes bacterial wilt, by pouring an inoculum suspension into the nutrient solution. Cultivar susceptibility to the bacteria was evaluated, based on the highest percentage of wilting. Because the length of time for wilt appearance varied among cultivars, some cultivars appeared to be suppressive to the translocation and/or multiplication of the invading pathogen. Thus, this hydroponic inoculation system is effective for examining levels of susceptibility in tomato cultivars to bacterial wilt. Received 13 December 2000/ Accepted in revised form 27 March 2001     

An epidemiological survey of Chrysanthemum chlorotic mottle viroid in Akita Prefecture as a model region in Japan

In epidemiological surveys on Chrysanthemum chlorotic mottle viroid (CChMVd) in various chrysanthemums cultivated in Akita Prefecture, Japan, in 2002–2005, approximately 20% of cultivars harbored CChMVd, including more symptomatic types than nonsymptomatic. Large-flowered cultivars were less frequently infected than small-flowered and spray types. The number of CChMVd-infected chrysanthemums is increasing, and the disease is found throughout major chrysanthemum-producing districts. Chrysanthemums infected only with CChMVd, in general, had no noticeable symptoms of disease. Most of those dually infected with known viruses and/or viroid also had no symptoms characteristic of chlorotic mottle disease. The lack of noticeable symptoms in major Japanese cultivars may have resulted in the unnoticed spread of the viroid.     

Comparative external morphology of cetacean spermatozoa

SUMMARY: To compare size and morphology of spermatozoa in cetaceans, sperm and epididymis samples were collected from 10 species in four families and spermatozoa were observed with phase-contrast and scanning electron microscopes. According to the average total length of the spermatozoa, the 10 species examined were classified into the following four groups in order of increasing size: (i) Baird's beaked (Ziphiidae) and Bryde's whales (Balaenopteridae); (ii) Dall's and finless porpoises (Phocoenidae); (iii) common, bottlenose, and Pacific white-sided dolphins (Delphinidae); and (iv) killer and short-finned pilot whales, and Risso's dolphin (Delphinidae). Spermatozoa head length of Bryde's whale and the finless porpoise were shorter than those of the other species. Spermatozoa head width was widest in the killer whale and thinnest in the Baird's beaked whale. The lateral aspects of sperm heads from the 10 species were characterized as the 'anterior region of the sperm head is thin and flat while the posterior region is thick.' The dorsal aspects of sperm heads were 'paddle-shaped' in Bryde's whales, 'bowling pin-shaped' in Baird's beaked whales, 'Japanese fan-shaped' in killer whales, an 'elongated ellipsoid shape' in Delphinidae except for killer whales, and 'ellipsoid shaped' in Phocoenidae. Size and morphology of the spermatozoa showed interspecific differences among the 10 species examined, which correspond to cetacean taxonomic classification.     

Evaluation of genetic variation among wild rice populations in Cambodia

A total of 448 samples in five natural populations of wild rice (Oryza rufipogon) were collected in Cambodia. They were examined using 12 SSR and two chloroplast markers to evaluate the degree of variation among populations and the genetic structure within populations. In the two annual populations, the number of plants with homozygous alleles at all 12 SSR loci were high (66.3% and 79.5%), suggesting that these plants propagate mainly through self-pollination. In the three perennial populations, no individuals had all homozygous genotypes, but redundant genotypes resulted from clonal propagation were observed. Percentages of the redundant genotypes were highly varied (3.6%, 29.2% and 86.0%). This may be due to the different stable levels of environmental conditions. As for chloroplast genome, most of the wild plants showed the same chloroplast types as most Indica-type cultivars have. However, plants with different chloroplast types were maintained, even in the same population. In tropical Asian countries, many wild rice populations were observed under similar ecological conditions examined in this study. Therefore, the present results concerning population structure will be important to further elucidate genetic features of wild rice, and will also give strong clues to utilize and conserve wild natural genetic resources.     

Influences of Forest Decline on Various Properties of Soils at Mt. Hirugatake, Tanzawa Mountains, Kanto District, Japan

At Mt. Hirugatake in the Tanzawa Mountains, Kanto district, Japan, the deciduous broadleaved forests have rapidly declined. In our previous studies, we reported that the amount of soil organic matter had significantly decreased at the early and final stages of forest decline, and that the soil microbial biomass also showed a large decrease at these stages, suggesting that the composition of soil organic matter might have also changed with forest decline. To clarify the influences of forest decline on the composition of soil organic matter, the amount of humic substances, optical properties of humic acid, and the amount of soil carbohydrates in surface soils at different stages of forest decline were investigated. The amounts of humic acid and fulvic acid decreased to a lesser extent at the early and middle stages of forest decline, and showed a significant decrease at the final stage. As the amount of humin significantly decreased at the early stage, it was plausible that the distinct decrease in the total carbon content of the soil surface horizons at the early stage of forest decline was induced by the decrease in the amount of humin, and at the final stage, by the decrease in the amounts of humic acid and fulvic acid. The amount of soil carbohydrates did not change appreciably with forest decline although the soil organic matter content markedly decreased. It was suggested that most of the carbohydrates in the soil surface horizons were in a stabilized form consisting of complexes with humic substances, metals, and minerals, and would not be affected by the environmental changes associated with forest decline.     

Distribution of different fractions of cadmium,zinc, lead,and copper in unpolluted and polluted soils

McLaren and Crawford's method for fractionating soil Cu was modified, and used to fractionate soil Cd, Zn, Pb, and Cu in 38 soil samples from 11 soil profiles from industrially polluted and nearby unpolluted areas. Pollutant metals, especially Cd and Zn, were more soluble than the native soil metals. On average, approximately 45% of Cd was present in the CaCl₂ soluble (CA) fraction, whereas corresponding values for the other metals were below 10%. The percentages of each metal in the CA fraction followed the order Cd > Zn > Pb > Cu. The same order was observed for the acetic acid soluble (AAC) fraction. Approximately 30% of total Pb and Cu were present in the pyrophosphate soluble (PYR) fraction, and only 10% of total Cd and Zn. Approximately 20% of total Zn or Pb and 10% of Cd or Cu were present in the free oxide (OX) fraction. Only 20% of Cd and between 40–50% of the other 3 metals were present in the residual (RES) fraction. The results show that Cd is more labile than the other 3 metals.