Seedling blight of Glycyrrhiza uralensis caused by Pythium myriotylum,P. aphanidermatum and P. spinosum and identifying primary inoculum sources using multiplex PCR detection

Pythium species, isolated from seedlings of Glycyrrhiza uralensis with blight, were identified as P. myriotylum, P. aphanidermatum, and P. spinosum on the basis of morphological characteristics and sequences of the internal transcribed spacer regions of rDNA. In pathogenicity tests, the isolates of the three Pythium species caused blight, producing the original disease symptoms. The primary inoculum source was determined using a multiplex PCR to detect the pathogen. All the Pythium species were detected in the soils of fields with the diseased plants and in soils of adjacent field soils.     

Demographic and height growth response of native broad-leaved deciduous tree saplings to overhead canopy release in a coastal Pinus thunbergii forest in Hokkaido, northern Japan

Prevention efforts to combat pine wilt disease must be instituted to avoid destruction of coastal forests in Hokkaido, northern Japan. We examine an alternative method involving the conversion of Pinus thunbergii forests to native broadleaf species through advancing their growth, but we have little knowledge about the response of advanced growth to overhead canopy release. To demonstrate the demographic and height growth response of advanced growth to the light change in a coastal Pinus thunbergii forest, canopy trees were removed and the sapling populations were monitored for 6?years. Inhibition in height growth caused by light increase was observed in Cerasus sargentii and Kalopanax septemlobus saplings. Especially in Kalopanax septemlobus, a remarkable decline was observed in the saplings growing in the darker positions which experienced light change. Therefore, large light changes should be carefully avoided for this species especially in the darker positions. In contrast, relative height growth rates of Quercus crispula and, especially, Quercus dentata increased with increasing light, and saplings tended to die in the darker positions. No dead trees of Sorbus commixta were observed during the study period. The saplings also showed good height growth even when under closed canopy. This species seems to be adaptive to a dark environment. Since Quercus dentata and Kalopanax septemlobus are the major components of the natural coastal forests in Hokkaido, gradual canopy release is available to foster advanced growth in coastal Pinus thunbergii forest, in accordance with the concept of density control in coastal forests.     

Augmented Activity of the Pelvic Nerve Afferent Mediated by TRP Channels in Dextran Sulfate Sodium (DSS)-Induced Colitis of Rats

Enteritis has been recognized as a major symptom in domestic animals and human patients suffering from feed and food poisonings. The aim of the present study was to clarify the excitatory mechanism of the pelvic nerve afferent which may influence the occurrence of enteritis in response to nociceptive chemical stimuli of the colon in normal and abnormal rats with colitis induced by dextran sulfate sodium (DSS). The pelvic nerve afferent activity was markedly increased by colonic instillation of solution (0.5 ml) of acetic acid (5-25%) and capsaicin (100μg/ml). The nerve activity was augmented by colonic instillation of capsaicin to a greater extent in rats with DSS-induced colitis than in normal control rats. This augmented activity by capsaicin was more prominent at one day (DSS-1) than at 8 day (DSS-8) after the administration of DSS. The increased nerve activity caused by capsaicin in DSS-1 and DSS-8 was significantly inhibited by pretreatment with ruthenium red, which is a nonselective inhibitor of TRP channels of unmyelinated C-fibers (nociceptors). In conclusion, it was elucidated that the nociceptive function of the pelvic nerve was largely elevated at one day after DSS-induced colitis and such increased function was mostly mediated by TRP channels.     

Changes in heart rate variability in horses during immersion in warm springwater

OBJECTIVE: To determine the effects of immersion in warm springwater (38 degrees to 40 degrees C) on autonomic nervous activity in horses. ANIMALS: 10 male Thoroughbreds. PROCEDURE: Electrocardiograms were recorded from horses for 15 minutes during a warm springwater bath after being recorded for 15 minutes during stall rest. Variations in heart rate (HR) were evaluated from the power spectrum in terms of low frequency (LF, 0.01 to 0.07 Hz) power and high frequency (HF, 0.07 to 0.6 Hz) power as indices of autonomic nervous activity. RESULTS: Mean (+/- SE) HR during stall rest and immersion in warm springwater was 31.1 +/- 1.7 and 30.3 +/- 1.0 beat/min, respectively. No significant difference was found between the HR recorded during stall rest and that recorded during immersion in warm springwater. The HF power significantly increased from 1,361 +/- 466 milliseconds2 during stall rest to 2,344 +/- 720 milliseconds2 during immersion in warm springwater. The LF power during stall rest and immersion in warm springwater was 3,847 +/- 663 and 5,120 +/- 1,094 milliseconds2, respectively, and were not significantly different from each other. Similarly, the LF:HF ratio did not change during immersion in warm springwater. The frequency of second-degree atrioventricular block, which was observed in 2 horses, increased during immersion in warm springwater, compared with during stall rest. CONCLUSIONS AND CLINICAL RELEVANCE: Increases in HF power indicates that the parasympathetic nervous activity in horses increases during immersion in warm springwater. Thus, immersion in warm springwater may provide a means of relaxation for horses.     

PCR primers for identification of opine types of <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> in Japan

The polymerase chain reaction (PCR) is a rapid, precise method for detecting and identifying pathogenic bacteria. In addition to the published primers for identification of Agrobacterium tumefaciens up to species level, two sets of primers were designed to identify the nopaline and octopine types of Agrobacterium tumefaciens. The RBF-RBR primer set designed based on the nopaline type T-DNA right border detected the nopaline type A208 and R225f strains, and the ocsF-ocsR primer set derived from the ocs gene of the octopine type A. tumefaciens detected the octopine type A348 strain. After polymerase Chain reaction (PCR) amplification by the RBF-RBR primers, the A208 and R225f strains could be differentiated from each other by restriction fragment length polymorphism digestion using the restriction enzymes DraI and XbaI. Multiple colonies can be screened at one time in a single PCR tube with satisfactory efficiency, thereby allowing rapid detection of pathogenic A. tumefaciens. Following a rough screening by classical biovar medium and -methyl-d-glucoside medium, the developed PCR system was introduced to identify isolates collected from soil and crown gall samples. Of 42 isolates determined to be A. tumefaciens, 7 were found to be octopine type; all the rest were R225f type.     

Adsorption of OP<Subscript>1</Subscript>-related bacteriophages requires the gene encoding a TonB-dependent receptor-like protein in <Emphasis Type="Italic">Xanthomonas</Emphasis><Emphasis Type="Italic">oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis>

Xanthomonas oryzae pv. oryzae strain T7174R is lysed by bacteriophage OP_1h and OP_1h2. Three mutants tolerant to both OP_1h and OP_1h2 were isolated by transposon mutagenesis. The mutants had an insertion of the transposon in XOO1687, which is predicted to encode a TonB-dependent receptor gene. Plasmid pHMIroNB that contained XOO1687 of T7174R was constructed, and the mutant was transformed with the plasmid. The transformant recovered sensitivity to OP_1h and OP_1h2. Electron microscopic analysis demonstrated that OP_1h and OP_1h2 can adsorb to the wild type and the transformant, but they could not adsorb to the phage-tolerant mutant. These results suggest that the TonB-dependent receptor gene relates to adsorption and infection of T7174R by OP_1h2 and OP_1h. Y. Inoue and S. Tsuge have contributed equally to this work.     

Relationship of prepartum plasma concentrations of estrone sulfate and estradiol-17β with the weight of the calf and placental parameters in Holstein–Friesian cows

The aim was to evaluate the relationship of prepartum plasma estrone sulfate (E₁S) and estradiol‐17β (E₂β) concentrations with the weight of the calf and the placental parameters. Holstein–Friesian cows (n = 33) inseminated artificially with Japanese Black beef bull semen at Hiroshima University Farm in Japan were used for the experiment. Blood samples were taken every day from day 270 of gestation until the day after calving. The plasma samples were analyzed for E₁S and E₂β by enzyme immunoassay. The calf birth weight was taken immediately after calving. Complete fetal membranes were collected from 19 cattle and the weights of the placental components and the number of cotyledons were recorded. All 33 cattle delivered singleton normal calves. The prepartum plasma E₁S concentration was found to correlate significantly (P < 0.01) with the calf birth weight (r = 0.83), total fetal membrane weight (r = 0.81), cotyledonary weight (r = 0.79) and inter‐cotyledonary membrane weight (r = 0.64), but it did not correlate significantly with the number of cotyledons, whereas prepartum plasma E₂β was not found to correlate significantly with the weight of either the calf or any of the placental components except the number of cotyledons. In conclusion, prepartum plasma E₁S, not plasma E₂β, was found to correlate significantly with the weight of the calf and the placental components.     

Analysis of the equine ovarian structure during the first twelve months of life by three-dimensional internal structure microscopy

A three-dimensional internal structure microscopy (3D-ISM) can clarify the anatomical arrangement of internal structures of equine ovaries. In this study, morphological changes of the equine ovary over the first 12 months of life were investigated by 3D-ISM in 59 fillies and by histological analysis in 2 fillies. The weight and volume of the paired ovaries initially decreased from 0 to 1 months to 2 to 3 months of age and then significantly increased at 8 to 12 months of age. The ovulation fossa was first observed around the 3rd month and became evident after the 6th month. The number of follicles with a diameter of ≥10 mm and the diameter of the largest follicle increased gradually after 6 months of age. On a volume basis, the medulla accounted for nearly 90% of the whole ovary at 0 to 1 months of age, but significantly decreased from 2 to 3 months of age. The volume of the cortex increased progressively after birth and reached approximately 60% of the total volume at 8 to 12 months of age. This significant development of the cortex coincided with the increased number and size of large follicles observed from 6 months of age. These results suggest that the development of the cortex plays a role in the maturation of the follicles and the equine ovary undergoes substantial morphological changes postnatally until puberty.