Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Analysis of intracellular distribution of Borna disease virus glycoprotein fused with fluorescent markers in living cells

Borna disease virus (BDV) is a non-segmented, negative-strand RNA virus that is characterized by nuclear replication and persistent infection. A unique feature of BDV is that it releases only a small number of infectious particles from infected cells. Although these characteristics might make it difficult to obtain a large amount of recombinant viruses in a reverse genetics system, the mechanism underlying the budding or assembly of BDV particle has remained largely unknown. In this study, as a first step toward understanding the virion formation of BDV, we investigated the intracellular distribution and mobility of the fluorescent marker fusion envelope glycoprotein (G) of BDV in living cells. Expression analysis revealed that fusion proteins seem to cleave into functional subunits and localize in the endoplasmic reticulum (ER)/Golgi apparatus, as well as the authentic BDV G. Furthermore, we demonstrated using fluorescence recovery after photobleaching analysis that BDV G fluorescence shows rapid recovery in both the ER/Golgi and plasma membrane regions, indicating that BDV G fusion protein may be a useful tool to investigate not only the maturation of BDV G but also the budding and assembly of BDV particles in living cells.     

Inhibitory action of dissolved humic substances on the growth of soil bacteria degrading DDT

Abstract

Dissolved humic acid (HA) and fulvic acid (FA) prepared from a Dando brown forest soil (Typic Dystrochrept) inhibited the growth of soil bacteria degrading DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) in the culture. The population of DDT-degrading Gram-variable rod Bll6 decreased by the application of both HA and FA, suggesting the presence of bactericidal effect. Such inhibitory effect was stronger for HA and resulted in a lower degrading activity of DDT in the culture of Bll6. No inhibitory effect was observed on the growth of DDT-degrading Bacillus sp. B75. The electron spin resonance spectra showed the presence of organic free radicals in both HA and FA. The relative concentration of the radicals was higher in HA. Storage of HA solution for 3 months at 4°C decreased the concentration of the radicals as well as the inhibitory action. The addition of catalase decreased the inhibitory effect of humic acid. It is suggested that a hydroxy radical, which is derived from free radicals of humic substances, is involved in the inhibition of bacterial growth and degradation of DDT.     

Impacts of acute imipramine treatment on plasma and brain amino acid metabolism in mice given graded levels of dietary chicken protein

Several studies have shown a relationship between depression and animal protein intake. To evaluate whether the difference of dietary chicken protein levels induces an antidepressant‐like effect and potentiates acute antidepressant effects, three levels of dietary chicken protein were used as the representative animal protein with imipramine used as the antidepressant. In addition, the effects of dietary chicken protein on brain metabolism were evaluated. Open field test (OFT) and forced swimming test (FST) were conducted on the 27th and 28th days, respectively. OFT and FST were not influenced by both imipramine and dietary protein levels. However, characteristic effects of imipramine treatment on brain monoamine metabolism were observed in the cerebral cortex and hypothalamus. In addition, dietary protein significantly increased taurine and L‐ornithine levels even though these amino acids were not contained in the diets. In conclusion, the metabolism of several amino acids in the plasma and brain were altered by dietary chicken protein.     

Differences in catecholamine metabolism and behaviour in neonatal broiler and layer chicks

1. To clarify the difference in behavioural activities and catecholamine metabolism between layer and broiler-type chicks two experiments were conducted. 2. In experiment 1, 1-d-old male layer and broiler chicks were placed in an open-field area and their responses were investigated for 10 min. The responses of the two strains were remarkably different, with broilers being less active than layers. Vocalisations rapidly decreased in broilers whereas those of layers remained elevated during the 10 min. 3. In experiment 2, 1-d-old chicks of both strains were killed and brain catecholamine concentrations were determined in three parts of the brain: telencephalon, optic lobe and brain stem. 4. In the whole brain, dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) were significantly higher in broilers. However, the values for norepinephrine (NE), epinephrine (E) and 4-hydroxy-3-methoxyphenylacetic acid (HVA) were similar between strains. The ratios of metabolite/precursor were also calculated: HVA/DOPAC was higher in layers, while NE/DA, E/NE and DOPAC/DA were not significantly different between strains. 5. These results suggest that behavioural activities differ greatly, while there are some differences in catecholamine metabolism between the two strains.     

Dietary β-alanine enhances brain, but not muscle, carnosine and anserine concentrations in broilers

Carnosine (β‐alanyl‐L‐histidine) and its derivative anserine (β‐alanyl‐1‐methyl‐L‐histidine) are present in high concentrations in the muscle and brain of chickens. They are known as antioxidants and putative neurotransmitters in the brain. If administration of β‐alanine (β‐Ala), one of the constituents of carnosine, could increase the concentrations of these dipeptides in the brain and muscles, it could improve brain function and increase the commercial value of the meat in chickens. As an early step in investigating this hypothesis, in the present study, the effect of dietary β‐Ala on these dipeptide concentrations in the brain, Musculus pectoralis superficialis, Musculus pectoralis profundus and Musculus biceps femoris was investigated in broilers. Four‐week‐old broilers were given a commercial diet or diet containing 0.5, 1 or 2%β‐Ala for 4 weeks. At the end of the experiment, concentrations of both dipeptides were increased in the brain, while taurine concentration was decreased. In the muscles, concentrations of these dipeptides were unchanged. These results indicate that dietary β‐Ala might influence brain function, but is ineffective in increasing the concentrations of carnosine and anserine in the muscles of broilers.     

Efficacy of atovaquone against Babesia gibsoni in vivo and in vitro

The therapeutic efficacy of atovaquone against Babesia gibsoni was examined in three dogs experimentally infected with B. gibsoni isolated from naturally infected dogs in Aomori Prefecture, Japan. Once parasitemia reached 10%, atovaquone was administered orally (30 mg/kg twice daily for 7 days). Within 2 days of atovaquone treatment, the parasite disappeared from blood smears without any clinical side effects. Anemia and thrombocytopenia were significantly improved in all the dogs. However, a polymerase chain reaction assay revealed that a B. gibsoni marker gene was intermittently present in peripheral blood after atovaquone therapy, indicating that the organism had not been eliminated, and parasites reappeared in blood smears 33 days after the last treatment. To investigate the change in sensitivity against atovaquone, an in vitro sensitivity test was performed using peripheral blood obtained from an untreated dog that was infected with the original parasite isolate, and from two of the experimentally infected and atovaquone-treated animals (blood was collected at the time of the post-treatment recurrence of the B. gibsoni infection). Atovaquone was added to the culture medium to final concentrations of 0.1, 1, 10, 100, and 1000 nM. For the untreated parasites, complete growth inhibition occurred at 1000 nM of atovaquone, whereas the recurrent parasites were inhibited by only 39.52 +/- 8.34% and 31.31 +/- 8.14% at this concentration after 48 h of incubation. Thus, the recurring parasites were less sensitive to atovaquone than the untreated originally isolated parasites.     

Chemical studies on soil humic acids

The levels of alcoholic and phenolic hydroxyl, methoxyl, carboxyl, and carbonyl groups of 33 to 38 humic acids obtained from various types of soils were determined to analyze the relationships between the amounts of these functional groups and the degree of humification or the types of soils. The amounts of various oxygen-containing functional groups examined were all proven to be significantly different among the various types of humic acids by analyses of variance. During humification. generally, the carboxyl and carbonyl groups increased while alcoholic and phenolic hydroxyl and methoxyl groups decreased. Linear and logarithmic regression analyses of carboxyl group contents on RF values (optical density of the alkaline solution of humic acids at 600 nm) gave very significant positive correlations. The carboxyl group contents of Rp type humic acids and humic acids from calcareous soils were largely distributed in the upper side of the regression curve. The carbonyl group contents showed a very significant linear positive correlation with carboxyl group contents. and both of them showed high linear positive correlation with RF values. Phenolic hydroxyl group contents decreased with humification in Rp(l). B. and A type humic acids. Alcoholic hydroxyl group contents showed a significant negative linear association with _RF values. Methoxyl group contents decreased rapidly with increasing humification in low humified humic acids, and their negative correlation with _RF values were proven to be very significant by logarithmic regression analysis. The complicated relationship between oxygen content and RF value which was reported previously (7) has been accounted for by the results obtained in the experiments conducted here.     

Behavior of phenolic substances in the decaying process of plants

The changes in amount of ether-extractable phenolic substances, individual phenolic adds, organic matter content, and pH value in the decaying process of rice straw were compared at 10, 30, and 50°C under moist and flooded conditions for 70 days.

1) The changes in pH value with time varied markedly according to temperature and water conditions.

2) Phenolic substances in the ether-extractable fraction obtained from the acidified solution of alkaline methanol extract contained undetected phenolic compounds in amounts larger than the phenolic adds detected by gas chromatography. pH value during incubation was hardly attributable to these phenolic substances because of their small quantity.

3) The changes in amount of the ether-extractable phenolic substances with time was mainly due to phenolic compounds other than phenolic acids, and influenced largely by the temperature and water conditions. Their formation and degradation took place rapidly at high temperatures under flooded conditions.

4) From the plots of individual phenolic acids vs. time, it was tentatively concluded that phenolic acids contalned in non-decayed straw were rapidly degraded in the earlier stage, and some of them were newly produced from ether-unextractable precursors in the later stage. These reactions occurred rapidly at high temperatures, especially under flooded conditions.

5) The content of p-hydroxybenzolc acid increased simultaneously with a rapid decrease in p-coumaric acid during the incubation ot straw. The relationship between vanillic acid and ferulic acid was the same.