Effect of nitrate fertilization of gametophytes of the kelp Undaria pinnatifida on growth and maturation of the sporophytes cultivated in Matsushima Bay, northern Honshu, Japan

In northern Japan, the early production of high-quality specimens of kelps Undaria pinnatifida prior to the outbreak of pinhole disease is needed. To address this need, we tested the effect of nitrate fertilization of gametophytes on the growth and maturation of sporophytes using experimental and control gametophytes derived from sporophytes cultivated in Matsushima Bay, northern Honshu, Japan. From mid-August to October 2008, experimental gametophytes were cultured on a rope indoors at a nitrate concentration of 1 mg/L at the optimal seawater temperature of 20 °C, whereas control gametophytes were cultured without the addition of nitrate in normal seawater at an average temperature of 23.5 °C during this period in the past 30 years. Next, the seedlings were cultivated in Matsushima Bay until March 2009. For the experimental kelps, the total length, stipe length, stipe width, sporophyll length, sporophyll width, and dry weight of the blade, midrib, and sporophyll increased significantly from December to January compared to those of control kelps. Matured sporophylls of the experimental kelps were more than those of the control kelps from early December to late January. Significant higher photosynthesis occurred at seawater temperatures of 10 and 15 °C, as well as higher uptake rates of NO₃–N of the experimental kelps in December and January. These results show that nitrate fertilization of the gametophytes makes it possible to harvest the high-priced blade, midrib, and sporophyll in January, which is earlier than the traditional harvest date.     

Acoustic pressure sensitivities and effects of particle motion in red sea bream Pagrus major

The auditory pressure thresholds of red sea bream were examined using cardiac response in the field by placing fish subjects far from the sound source to prevent particle motion. Pressure and particle motion thresholds were also obtained using the auditory brainstem response (ABR) technique. The thresholds at 100 and 200 Hz were significantly higher when measured using the cardiac response in the far field than those obtained in previously conducted experiments in experimental tub. However, thresholds obtained using ABR from 200 to 500 Hz were not remarkably lower, although significantly different (0.01 < P < 0.05), compared with those obtained using cardiac response in the far field. Furthermore, calculated particle velocity thresholds indicated that fish probably detected particle motion within the frequency range of 50–200 Hz, even in fish with a deactivated lateral line. Although the ABR method is widely applied in fish auditory study, hearing thresholds are apparently affected by particle motion.     

Development of a microbial community on cellulose buried in waterlogged soil

Summary The development of a microbial community on cellulose (cellophane film and filter paper) buried in waterlogged soil was observed under a microscope. Throughout the decomposition of the cellulose, the biomass, immobilized N, ATP and gas metabolism of the microbial community were examined. As cellulose decomposition progressed, a microbial succession was recognized. This succession was divided into two stages. In the first stage, a few types of cellulolytic microorganisms predominated on the cellulose. Vigorous decomposition of the cellulose was accompanied by a rapid increase in microbial biomass, and H₂ was evolved from the microbial community on the cellulose. In the second stage, the rate of cellulose decomposition was slow. The cellulose remaining was thickly covered with various types of microoganisms. The H₂ produced was consumed by the microorganisms closely adhering to the remaining cellulose. In addition, non-cellular organic N accumulated on the remaining cellulose. A large part of the microorganisms seemed to be dormant in this stage. The trends in this microbial succession were similar to those found in ecosystem successions.     

Spatial variations in nitrous oxide and nitric oxide emission potential on a slope of Japanese cedar (Cryptomeria japonica) forest

To quantify the spatial variation and spatial structure of nitrous oxide (N₂O) and nitric oxide (NO) emission from forest soils, we measured N₂O and NO emission rates from surface soil cores taken at 1 m intervals on a cross-line transect (65 m × 20 m) on a slope of Japanese cedar ( Cryptomeria japonica ) forest in a temperate region of central Japan and analyzed the spatial dependency of N oxide gas emissions using geostatistics. We divided N₂O emission into N₂O from denitrification and N₂O from nitrification using the acetylene inhibition method. According to the geostatistical analysis, N₂O emission rates on the slope had large spatial variation and weak spatial dependency. This weak spatial dependency was caused by the inordinately high N₂O emissions on the slope, which were derived mainly from denitrification. In contrast, NO emission rate on the slope had large spatial variation, but strong spatial dependency and a distinct spatial distribution related to slope position, that is, high in the middle of the slope and low in the shoulder and the foot of the slope. The CN ratio and water-filled pore space were the dominant factors controlling NO emission rate on a slope. Our results suggest that spatial information about topographic factors helps to improve the estimation of both N₂O emission and NO emission from forest soils.     

Quantitative fecal culture for early diagnosis of Corynebacterium (Rhodococcus) equi enteritis in foals.

Quantitative culture of Corynebacterium (Rhodococcus) equi from feces of 17 foals on a farm (A) with an endemic C. equi infection problem and 26 foals on a farm (B) without the disease in the past decade was done with a selective medium at weekly or monthly intervals from April to August of 1984. Corynebacterium equi was observed in the feces of 16 of 17 foals on farm A, and 19 of 26 foals on farm B. The mean viable count of C. equi in one gram of feces was 4.1 +/- 3.7 (log10) on farm A, and 3.9 +/- 3.4 (log10) on farm B. Corynebacterium equi was recovered from feces of foals as young as two weeks old. Almost all foals at an age between two to four weeks shed the bacteria in the feces. During the observation period two foals showed clinical signs: fever, diarrhea, and cough, at four or five weeks old. At the same time the bacterial count per gram of feces increased from 4 to 7 or 8 (log10). They shed large number of bacteria in the feces and continued to show the clinical signs until death at 10 or 11 weeks old. One of the foals was diagnosed as having had C. equi enteritis and pneumonia by the postmortem recognition of lesions with bacteriological confirmation. The quantitative culture of the feces of foals at weekly intervals after birth on farm A was found to be very useful as an aid in early diagnosis of C. equi enteritis in foals.     

Biological roles of monoterpene volatiles derived from rough lemon (<Emphasis Type="Italic">Citrus jambhiri</Emphasis> Lush) in citrus defense

Volatile compounds of plants, including monoterpenes, are a possible source of signal molecules that induce defense systems to protect plants from tissue damage. Volatile compounds from rough lemon leaves were trapped by solid-phase microextraction fibers in sealed vials, and subsequent gas chromatography–mass spectrometry and gas chromatography analyses identified the profile of the major components, mainly various monoterpenes. Among several monoterpenes examined, citral, citronellal, and linalool significantly inhibited the spore germination and hyphal growth of Alternaria alternata. The effect of linalool was fungistatic, while the effects of citral and citronellal were partially fungicidal. Wounding of rough lemon leaves induced a significant increase in release of monoterpenes. The release of linalool was the most abundant and was 14.5 times that of unwounded rough lemon leaves. Unlike the wounding treatment, microbe attack did not significantly change monoterpene releases, and there was statistically no difference found in the peak areas from microbe-treated and untreated leaves. Linalool, limonene, and β-pinene also had insect-repellant effects on wild-type Drosophila melanogaster. Expression patterns of defense-related genes in rough lemon and rice significantly changed after treatment with vapors of monoterpene volatiles. Taking these results together, monoterpene volatiles are likely to play roles in the defense of rough lemon against microbe and insect pathogens.     

Development of a water-soluble preparation of emamectin benzoate and its preventative effect against the wilting of pot-grown pine trees inoculated with the pine wood nematode, Bursaphelenchus xylophilus

Water-soluble preparations have been investigated to develop a trunk injection agent based on the poorly water-soluble anti-nematode emamectin benzoate. Following tests on the phytotoxicity of some solvents and solubilizers and demonstration of the ability of some solubilizers to dissolve emamectin benzoate in water, acetone + methanol was selected as the solvent and Polysorbate 80 as the solubilizer. This water-soluble preparation of emamectin benzoate prevented the wilting of pot-grown 4-year-old trees of the Japanese black pine, Pinus thunbergii, artificially inoculated with the pine wood nematode, Bursaphelenchus xylophilus, at a dose of 20 g emamectin benzoate per cubic metre of pine tree.     

Activation with ethanol improves embryo development of ICSI-derived oocytes by regulation of kinetics of MPF activity

Developmental potential of bovine embryos that are not artificially activated after intracytoplasmic sperm injection (ICSI) is generally very low. In this study, we investigated effects of artificial activation with ethanol on kinetics of maturation promoting factor (MPF) activity (p34(cdc2) kinase activity) and development of bovine oocytes following ICSI. Treatment of oocytes with ethanol at 4 h after ICSI improved their first cleavage and further preimplantation development (51% vs. 13%, 14% vs. 4%: treatment with vs. without ethanol, respectively). MPF activity of oocytes was lowered until at least 2 h after ICSI. In oocytes without activation after ICSI, MPF activity temporarily elevated at 6 h after ICSI, whereas this phenomena was not observed in the oocytes treated with ethanol. Furthermore, MPF activity was elevated 20 h after ICSI in oocytes activated with ethanol, whereas this elevation of MPF activity was not shown in oocytes without activation. These results indicate that the stimulus of sperm was sufficient to lower MPF activity of oocytes following ICSI, and moreover the activation treatment of bovine oocytes with ethanol after ICSI served to maintain the low levels of MPF activity until the next cell cycle started.     

Association of disease with isolation and virulence of Rhodococcus equi from farm soil and foals with pneumonia

OBJECTIVE: To determine whether isolation and virulence of Rhodococcus equi from soil and infected foals are associated with clinical disease. DESIGN: Cross-sectional and case-control study. SAMPLE POPULATION: R equi isolates from 50 foals with pneumonia and soil samples from 33 farms with and 33 farms without a history of R equi infection (affected and control, respectively). PROCEDURE: R equi was selectively isolated from soil samples. Soil and clinical isolates were evaluated for virulence-associated protein antigen plasmids (VapA-P) and resistance to the beta-lactam antibiotics penicillin G and cephalothin. Microbiologic cultures and VapA-P assays were performed at 2 independent laboratories. RESULTS: VapA-P was detected in 49 of 50 (98%) clinical isolates; there was complete agreement between laboratories. Rhodococcus equi was isolated from soil on 28 of 33 (84.8%) affected farms and 24 of 33 (72.7%) control farms, but there was poor agreement between laboratories. Virulence-associated protein antigen plasmids were detected on 14 of 66 (21.2%) farms by either laboratory, but results agreed for only 1 of the 14 VapA-P-positive farms. We did not detect significant associations between disease status and isolation of R equi from soil, detection of VapA-P in soil isolates, or resistance of soil isolates to beta-lactam antibiotics. No association between beta-lactam antibiotic resistance and presence of VapA-P was detected. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of soil microbiologic culture and VapA-P assay results, it is not possible to determine whether foals on a given farm are at increased risk of developing disease caused by R equi.     

Effect of alkali extraction on the lignin monomeric composition inEucalyptus

The effect of alkali extraction on the lignin monomeric composition was examined inEucalyptus camaldulensis andE. globulus by thioacidolysis using extractive-free samples as a control. Results showed that the effect onEucalyptus is different among species and among sample positions in the trunk, although a small amount of lignin is solubilized during the extraction in all samples. In addition, it was proved that lignin extracted by the alkali extraction is not always guaiacyl-rich, probably relating to the original lignin monomeric composition, which depends on the sample species or the sample position in the trunk.