Genotyping of Malassezia pachydermatis isolates from canine healthy skin and atopic dermatitis by internal spacer 1 (IGS1) region analysis

Isolates of Malassezia pachydermatis from healthy dog skin and from dogs with atopic dermatitis were molecularly characterized using internal spacer 1 (IGS1) region analyses, and their phospholipase A2 activity and pH growth profiles were then characterized in vitro. The percentage of isolates from healthy dogs that had the following IGS1 subtypes (isotype, %) were as follows: 1A, 6%; 1B, 27%; 1C, 11%; 2A, 6%; 2B, 6%; 3A, 11%; 3C, 3%; and 3D, 24%. In contrast, 9% of isolates from dogs with atopic dermatitis were isotype IB and 91% were isotype 3D, indicating that isolates of subtype 3D were the most prevalent in dogs with atopic dermatitis. Production of phospholipase A2 was statistically higher in isolates of subtype 3D than in the other subtypes. The subtype 3D isolates showed enhanced growth on alkaline medium compared with non-3D subtype isolates. The main clinical sign of canine Malassezia dermatitis is waxy exudates on the skin, which predispose the patient to development of a yeast overgrowth of the subtype 3D. Increased phospholipase A2 production may be involved in the inflammatory process associated with Malassezia dermatitis.     

An azole-resistant isolate of Malassezia pachydermatis

Canine Malassezia dermatitis (MD) is frequently treated with systemic ketoconazole (KTZ) and itaconazole (ITZ). However, the antifungal susceptibility of clinical isolates of M. pachydermatis from dogs and cats to the azoles has not been well investigated. In the present study, the in vitro susceptibility of the standard strain (CBS1879: the neotype strain of M. pachydermatis) and 29 clinical isolates of M. pachydermatis to the azoles was measured by a modified CLSI M27-A2 test using modified Dixon medium as well as by the E-test. The minimum inhibitory concentrations (MICs) of the 30 isolates of M. pachydermatis (including the neotype strain) against KTZ and ITZ were <0.03 μg/ml by the two methods. The MICs of 1 clinical isolate (ASC-11) were 1 and 2 μg/ml against KTZ, and 2 and 8 μg/ml against ITZ, by the modified CLSI M27-A2 test and the E-test, respectively. Thus, isolate ASC-11 may be resistant to these azoles, making this the first report of a resistant isolate of M. pachydermatis to KTZ and ITZ.     

Establishment of a novel feline leukemia virus (FeLV)-negative B-cell cell line from a cat with B-cell lymphoma

There is significant lack of basic hematologic and immunological data in adult sows. Therefore, aim of this study was to provide respective reference intervals. 32 clinically healthy multiparous Large White sows aged 33.5 ± 9.6 months and all of them two months postpartum were included in this study. Mean erythrocyte count was 5.5 ± 0.7 × 10(6)/μl and total leukocyte count was 12.1 ± 2.1 × 10(3)/μl. Proportion of lymphocytes was 44.7 ± 10.2% and of neutrophils 41.6 ± 11.0%. The ratio of na?ve T helper (Th) cells to memory Th cells was 1:3.1 and the ratio of Th cells to cytotoxic T cells (CTLs) was 1:4.2. Proportions of regulatory T cells, NK cells, and CD21(+) B cells were lower (3.1, 2.6, and 6.0%) than those of memory Th cells ranging from 8.8 to 27.5% depending on the activation status and CTLs with 37.3%. γδ T cells were found at comparably high numbers (19.1%). Flow cytometric measurement of intracellular cytokines in PBMCs revealed marginal levels for IL-1β, IL-2, IL-4, IL-6, IL-10, and IL-12p35, but remarkable levels for TNF-α and IFN-γ. Highest mRNA levels were found for IL-1, IL-10, and TNF-α, with TNF-α showing the least inter-individual variation.     

Vision-based Navigation System for Autonomous Transportation Vehicle

Autonomous navigation using global positioning system (GPS) or geomagnetic direction sensor (GDS) to detect absolute position is widely used. An alternative system that detects boundary lines and intersections information can be achieved using machine vision based navigation systems. The objectives of this research are the development of the intersection detection method using machine vision and the approaching/turning at the intersection using dead reckoning method. Intersection guidance markers (IGM) were installed on farm road intersections. The method of intersection recognition involved the search to detect the IGM instead of the actual intersection, which would be difficult because the latter changes appearance through the season. Hough Transform and coordinate transformation was used for this purpose. A dead reckoning system for approaching and turning on the intersection was developed. Rotary encoders installed on both sides of the crawler sprocket are used as guidance sensors. RTK-DGPS and fiber optic gyroscope (FOG) were used for evaluating the track of autonomous traveling. The result of field tests (the speed was 0.8 m/s) showed the rms of intersection detection error and the dead reckoning error were 0.12 m at 8.64 Hz processing speed and 0.063 m respectively.     

Effects of Controlled-Release Fertilizers and Their Application Methods on Germination and Seedling Growth of Dent and Sweet Corns

Effects of controlled-release fertilizers (CRFs) (C-AS, polyolefin coated ammonium sulfate, 50-day-type; Dd-LP, polyolefin coated urea with dicyandiamide, 40-day-type; C-ANP, polyolefin coated ammonium nitrate phosphate, 40-day-type; and C-DAP, polyolefin coated diammonium acid phosphate, 40-day-type), ammonium sulphate and no fertilizer control, and their application methods (spot, band, surface and mixed) on germination and seedling development of sweet corn (Zea mays L.var. saccharata Sturt.) and dent corn (Zea mays L.var. indentata Sturt.) were investigated in a greenhouse. Under co-situs application (band and spot) of CRFs, there were no obvious differences in the germination speed and rate for both dent corn and sweet corn relative to control. Mortality rates of sweet corn seedlings under co-situs application were high in experiment 1, but were very low in experiment 2, because the environmental conditions were different in the two experiments. That is, under lower temperature and weaker sunlight, young seedlings easily die due to high soil nutrient concentration and slow growth speed of corn. Shoot weight of both dent and sweet corn did not greatly decrease in experiment 1. In experiment 2, there were no significant differences in shoot and root weight of both corns between cositus and surface or mixed application methods. However, with spot and band application of ammonium sulfate, shoot and root weight were significantly reduced. Soil EC and pH were considerably affected by co-situs application, especially at the fertilizer application site. For both dent and sweet corn, EC in the 0-3 cm soil was significantly higher under co-situs application and surface application than that under mixed application, whereas in the 3-6 cm soil depth the situation was reversed. Compared with control, mixed application of CRFs decreased soil pH slightly (0-3 cm depth) or greatly (3-6 cm depth).     

Effect of seed source elevation on bud opening of Yezo spruce (<Emphasis Type="Italic">Picea jezoensis</Emphasis>)

Spring frost damage is one of the obstacles to be overcome in establishing a Yezo spruce (Picea jezoensis Carr.) plantation. The timing of bud opening of Yezo spruce seedlings cultured from seeds collected at three different elevations (420, 700, and 1200m) in the Tokyo University Forest in Hokkaido was observed at two planting site elevations (610 and 750m) to investigate whether seed source elevation or planting site elevation affects the bud opening date. At both planting sites the bud opening dates were not significantly different among the seedling groups from different seed source elevations. On the other hand, bud opening dates of seedlings planted at the lower elevation site were significantly earlier than those planted in the higher elevation site. It could be deduced from this study that the environment of the planting site rather than seed source elevation affects the bud opening date of Yezo spruce.     

Analysis on residue formation during wood liquefaction with polyhydric alcohol

Liquefactions of cellulose powder, steamed lignin, alkali lignin, and their mixtures were carried out to analyze the reaction process of wood using polyhydric alcohol. The liquefaction of wood proceeded immediately and wood components were converted to N,N-dimethylformamide (DMF)-soluble components. After that, the condensation reaction occurred with increasing reaction time. However, none of cellulose powder, steamed lignin, and alkali lignin condensed by themselves during their liquefaction. The mixture of cellulose and lignin was also liquefied, and condensed after a long reaction time. The results of analysis showed that the behavior of the mixture resembled that of wood with respect to molecular weight distribution and the main functional groups. Lignin was converted to DMF-soluble compounds in the initial stage of wood liquefaction, followed by cellulose gradually being converted into soluble compounds. After that, condensation reactions took place among some parts of depolymerized and degraded compounds from cellulose and lignin, and were converted into DMF-insoluble compounds. It was concluded that the rate-determining step of wood liquefaction was the depolymerization of cellulose. Furthermore, it was suggested that the condensation reaction was due to the mutual reaction among depolymerized cellulose and degraded aromatic derivatives from lignin or due to the nucleophilic displacement reaction of cellulose by phenoxide ion.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002     

Analysis of current forest operations in the Okumikawa Forestry Area by location and topography

We analyzed current forest operations in Toei, in the Okumikawa Forestry Area, Japan, by examining actual achievement, location, and topography. We considered five basic forest operations conducted by the Forest Owners Association between 1991 and 2000 in a subsidized reforestation and silviculture project: planting, weeding, pruning, cleaning, and thinning. In addition, we considered the road network, including public, forest, and strip roads. The area in which forest operations have been conducted over the past 10 years amounted to less than two-thirds of the area targeted in the Toei Forest Improvement Plan. The mean access distance of the work sites was 164m, which was 52m less than the 10-year average for the whole forest (216m). Forest operations tended to ignore remote forest areas where the topography was steep, due to cost, the logging techniques required, and labor requirements. We determined that the limitations on forest operations in Toei were a 300m access distance, an elevation of 800masl, and a 90% slope. In accessing work sites, the utilization ratios of forest and strip roads were higher than their composition ratios, and public roads were used most. Newly constructed strip roads were used more than forest roads. Based on current forest operations, we calculated the road density and length required to be 7.1m/ha and 79.2km, respectively, with an estimated cost exceeding 720 million yen for new construction.