| Project/Area Number |
09670256
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
寄生虫学(含医用動物学)
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| Research Institution | Kobe University School of medicine |
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Principal Investigator |
UGA Shoji Kobe University, Associate Professor, 医学部, 助教授 (90071399)
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| Co-Investigator(Kenkyū-buntansha) |
ONO Kazuo Hyogo Prefectural Institute of Public Health, 微生物部, 次長(研究職)
KATAOKA Nobumasa Kobe University, Associate Professor, 医学部, 助教授 (80071398)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Toxocara canis / Toxocara cati / Toxocara spp. / Escherichi coli / Sandpit / Zoonoses / Cryptosporidium / 犬蛔虫 / トキソカラ / トキソプラズマ / 猫蛔虫 |
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| Research Abstract |
We established a new detection method of Cryptosporidium oocysts from the soil by a centrifugal floatation method using sucrose solution. Because the oocysts in the soil samples were surrounded by many of the particulate sand components, usual centrifugal floatation techniques could not be applied to effective collection of the oocysts. This problem was overcome by adding gelatin to the sucrose solution at a concentration of 0.1%. Although the recovery efficiency of the method varied from 10 to 20%, this technique was used for our survey. Soil samples around the farm, which had been confirmed to be enzootic for cryptosporidiasis, were examined and we found that all of 16 samples were positive. Studies on the contamination with Cryptosporidium in sandpits in public parks are being carried out at present.
Different from Cryptosporidium oocyst, Toxoplasma oocyst could not be identified only by morphological approach. Then, we attempted a new detection method using a polymerase chain reaction. Purification method of Toxoca oocysts from soil sample was the same as that used for Cryptosporidium. Purified samples suspended in 400 μl of TNE buffer (containing 1% PVP and 1% SDS) were used for DNA extraction according to the standard procedure and were subjected to the PCR.
Escherichia coli was detected from 120 out of 144 (80%) sand samples examined. The contamination rate of Toxocara egg examined at the same sandpits was 30% (43/144). which was different from the results obtained through E.coli study. Average number of E.coli found in 100 g of positive sand was 3400, and the numbers of E.coli in 10 sandpits were over the maximum detection limit of E.coli of this test (160,000). In a case of E.coli examination, some of the unused sea sand samples showed positive for E.coli showing that a E.coli examination of the sand will not be a good indicator for the fecal contamination.
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