Hepatic disposition of parathion: Uptake by isolated hepatocytes and chromatographic translobular migration

Isolated rat hepatocytes suspended in Waymouth's medium absorbed parathion rapidly and reversibly until the intracellular parathion concentration reached more than 300 times the ambient concentration. The distribution quotient q, defined as the ratio of intra- and extracellular concentrations at equilibrium, decreased when horse serum was added to the medium. The high hepatocyte affinity and rapid uptake of parathion suggested that this compound might be almost completely absorbed by periportal hepatocytes in the perfused liver and migrate downstream chromatographically through the lobule. Parathion infusion experiments verified this prediction and showed that the migration rate is dependent on the q value. The chromatographic feature may be useful as a basis for nonhistological investigation of intralobular hepatocyte heterogeneity. The lobule may function as a reverse-phase chromatograph also for many other unionized xenobiotics. Implications of the findings in the hepatic disposition of xenobiotics in vivo are discussed.     

Genetic diversity of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan based on phylogenetic analyses of rDNA-IGS and <Emphasis Type="Italic">MAT1</Emphasis> sequences

Twenty-eight isolates of Fusarium oxysporum f. sp. spinaciae (FOS; the causal agent of spinach wilt) collected from Japan were assessed for mating type and subjected to phylogenetic analysis. Mating type analysis revealed all isolates to be MAT1-2, suggesting that there is no sexual recombination within the population. Phylogenetic analyses based on nucleotide sequences of the ribosomal DNA intergenic spacer (IGS) and the mating type locus (MAT1) suggested that FOS is polyphyletic. The cluster analysis based on IGS showed four phylogenetic groups (S1–S4) among the isolates. Two distinct lineages, S1 and S3, included FOS isolates both of the vegetative compatibility group (VCG) types, 0330 and 0331, demonstrating that VCG differentiation in FOS may not necessarily reflect the phylogenetic relationships based on IGS and MAT1-2-1.     

A simple method for a mini-preparation of fungal DNA

A simple method was established to prepare DNA from fungal mycelia cultured on an agar plate. The fungi tested successfully with this method contained Zygomycetes, Ascomycetes, Basidiomycetes, and Oomycetes. This method did not require any time-consuming steps to crush or digest mycelia or fractionation in a phenol–chloroform mixture. The DNA was easily extracted by immersing and dispersing the mycelial plugs in a specific buffer (200 mM Tris-HCl, 50 mM ethylenediaminetetraacetic acid, 200 mM NaCl, 1% n-lauroylsarcosine, pH 8.0), then concentrated by ethanol precipitation. The total time to complete the whole procedure was less than 1 h. The quality and quantity were sufficient for polymerase chain reaction amplification and Southern blot analysis.     

Comparison of voluntary feed intake, rumen passage and degradation kinetics between crossbred Brahmam cattle (Bos indicus) and swamp buffaloes (Bubalus bubalis) fed a fattening diet based on corn silage

Three crossbred tropical cattle and three crossbred buffaloes, fitted with rumen cannulas, were used to investigate the differences in feed intake, digestibility and parameters with rumen degradation kinetics between cattle and buffalo fed a fattening diet in the Philippines. The animals were fed a diet consisting of 50% corn silage (CS), 30% brewer's grain (BG) and 20% concentrate mixture (CM) on a dry matter (DM) basis, at a level of 3% bodyweight (BW) as DM. Total DM intake (DMI) and rumen fill were greater (P < 0.05) for buffaloes than for cattle, but significant differences were not detected between the animal species when those were expressed as percentage of BW. The DMI per metabolic body size for BG and CM were greater for buffaloes than for cattle (P < 0.05), whereas that of CS did not differ between the species (P > 0.05). The weight proportion of consumed feedstuffs differed between cattle and buffaloes (P < 0.05), although nutrient intake per total DMI did not differ significantly (P > 0.05) between the animal species. Nutrients digestibility was higher (P < 0.05) for buffaloes than for cattle. Ruminal passage rate constant of feed particles and rumen fluid did not differ (P > 0.05) between the animal species. Higher DM degradation rate constant and effective degradability with CS were prominent for buffaloes than for cattle (P < 0.05). Degradation parameters did not differ (P > 0.05) between cattle and buffaloes, neither with BG nor CM. The results indicate that the greater digestibility with DM and energy for buffaloes are ascribable to greater effective degradability of CS fed ad libitum level.     

Bacterial Wilt of China Aster Caused by Erwinia chrysanthemi

In July 2000, a bacterial wilt was found on China aster (Callistephus chinensis Nees) on Hokkaido, Japan. Bacteriological characteristics of the isolated bacteria were identical to Erwinia chrysanthemi. This is the first report on bacterial wilt of China aster caused by E. chrysanthemi. Received 4 June 2001/ Accepted in revised form 6 September 2001     

Gene Arrangement and Sequence of str Operon of Phytoplasma Resemble Those of Bacillus More Than Those of Mycoplasma

The complete region of a putative streptomycin operon (str operon) of onion yellows (OY) phytoplasma, a phytopathogenic mollicute, was isolated and sequenced. This operon contains four genes, rps12, rps7, fus, and tuf, encoding ribosomal proteins S12 and s7, elongation factor (EF) -G, and EF-Tu, respectively. These four genes constitute the str operon in non-mollicute bacteria, such as Escherichia coli and Bacillus subtilis. In two species of mollicute Mycoplasma, the tuf gene was reported not to be included in this operon, but was located apart, indicating that the gene arrangement of this operon in phytoplasmas resembles that of B. subtilis more than that of Mycoplasma spp. In addition, the deduced amino acid sequence of EF-G of phytoplasmas also resembles that of B. subtilis more than that of Mycoplasma spp. These results suggest that analyses of the gene organization and sequence of the phytoplasma genome will provide valuable insights into evolutionary relationships among the culturable mollicutes, phytoplasmas and other Gram-positive bacteria. Received 25 April 2001/ Accepted in revised form 21 August 2001     

日本北海道东部高位泥沼远东红皮云杉、库页冷杉和日本桤木树干茎流化学及其对沼泽孔隙水化学的影响(英文)

在日本北海道高位泥沼池,调查了远东红皮云杉(Piceaglehnii)、库页冷杉(Abies sachalinensis)和日本桤木(Alnusjaponica)树干茎流液的化学性质和泥炭孔隙水的化学性质,包括了横切和纵切面孔隙水化学性质。沼泽森林树干基部茎流对泥炭孔隙水化学性质有明显的作用,而且树基部的泥炭孔隙水化学性质具有物种特异性。远东红皮云杉茎流液和泥炭孔隙水中盐分含量最高,日本桤木则最低;盐分从茎基部向树冠形成浓度梯度。树冠下泥炭孔隙水化学受化学过程控制,孔隙水充沛条件下,控制作用减弱,因地势平缓(1°)茎流移动缓慢。事实上,泥炭孔隙水充足会使茎流化学物质浓度降低。微生物活动、优势树冠的营养再生介导的表面水和茎流对森林沼泽地微生境化学环境的空间异质性有贡献。图2表2参44。     

Nested PCR‐RFLP is a high‐speed method to detect fungicide‐resistant Botrytis cinerea at an early growth stage of grapes

BACKGROUND: Grey mould caused by the fungus Botrytis cinerea Pers. ex Fr. is one of the major diseases in grapes. The use of fungicides is a simple strategy to protect grapes against B. cinerea disease. However, phenotypes exhibiting resistance to fungicides have been detected in B. cinerea populations. The variation of fungicide‐resistant B. cinerea isolates renders B. cinerea disease control difficult in grapevine fields. RESULTS: The authors have developed a nested polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) method to detect fungicide‐resistant B. cinerea isolates at an early growth stage of grapes in grapevine fields. The nested PCR‐RFLP method was carried out to detect benzimidazole‐, phenylcarbamate‐ and/or dicarboximide‐resistant B. cinerea isolates from grape berries and leaves at Eichorn–Lorenz growth stage 25 to 29. This method successfully detected fungicide‐resistant B. cinerea isolates at an early growth stage of grapes. In addition, only 8 h was required from tissue sampling to phenotyping of fungicide resistance of the isolates. CONCLUSION: It is proposed that the early diagnosis of fungicide‐resistant B. cinerea isolates would contribute to further improvement of integrated pest management against B. cinerea in grapevine fields, and that the nested PCR‐RFLP method is a high‐speed, sensitive and reliable tool for this purpose. Copyright © 2008 Society of Chemical Industry     

Ecological studies on subtropical evergreen broad-leaved forest in Okinawa, Japan: Litter production and nutrient input

Patterns of litterfall and nutrient input in a subtropical evergreen broad-leaved forest in northern Okinawa, Japan, were studied during May, 1996–April, 1999. The mean annual rate of litterfall in the five sampling plots ranged from 6.84 to 8.93 Mg ha⁻¹ yr⁻¹, of which 63.3–68.5% were leaves; 22.4–29.1% woody parts (including branches < 5.0 cm in diameter and bark); 2.8–5.0% sexual organs and 4.6–6.3% miscellaneous material. Significant differences were found among plots and among years. Significantly monthly differences pronounced seasonal patterns in litterfall were observed. Total litterfall and leaf litter showed negative correlations with relative basal area of the dominant species,Castanopsis sieboldii; and showed positive correlations with mean height of the stands. The dominant species,C. sieboldii produced an average of 2.36 Mg ha⁻¹ yr⁻¹ of leaf litter, which covered 30.5% of the annual litter production, and the nutrient input from those litterfall contributed 32.3, 28.3, 30.2, 22.2, 32.5, and 30.5% of total N, P, K, Ca, Mg, and Na, respectively. Nutrient use efficiency in litter production was high, especially for P and K compared with other broad-leaved forests in Japan indicating that P and K may be limiting in Okinawan evergreen broad-leaved forest.