Development and an application of a PCR-baaed method for the identification of cosumed Japanese flounder, Paralichthys olivaceus, juveniles form the gut contents of crustacean predators
| Project/Area Number |
14560160
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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 |
General fisheries
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| Research Institution | Kitasato University |
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Principal Investigator |
ASAHIDA Takashi Kitasato University, School of Fisheries Sciences, Associate professor, 水産学部, 助教授 (00296427)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASHITA Yoh Kyoto University, Field Sciences Education and Research Center, Professor, フィールド科学教育研究センター, 教授 (60346038)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Predation / Crustacean / mitochondrial DNA / Species-specific PCR / Southern blot hybridization / gut contents / Japanese flounder / Identification of species / ヒラメ / 甲殻類捕食者 / 密度依存 / サイズ選択的捕食 / イシガレイ / PCR / 仔稚魚 |
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| Research Abstract |
It is recognized that predation by benthic crustaceans is one of the major causes of mortality of flatfishes. However, it is difficult to identify ingested larval fish species from the stomach contents of crustacean predators, because crustacean predators finely macerate their prey. We have developed a method for mass screening to detect consumed Japanese flounder-specific DNA from the stomach contents of sand shrimps. The PCR primers used in the present study are designed to amplify a 191 bp region of the mitochondrial control region and are Japanese flounder specific. We applied Southern blot hybridization and an immunological detection method for mass screening. The PCR products were blotted onto a nylon membrane and hybridize with appropriate probes labeled with digoxigenin. The predation experiment was performed in the laboratory under natural light conditions and the temperature was maintained at about 20℃ through the rearing and experiment period. In a laboratory experiment, successful amplification and detection of Japanese flounder-specific DNA were possible up to 15 hrs, with 100% detection within 8 hrs, after the termination of predation. Microscopic examination of the samples revealed the presence of debris, vertebrae and other bones. However, as digestion progressed, we could not observe any debris and bones from the stomach contents sampled after 13 hrs after the termination of predation, and the percentage of successful amplification decreased with time. Since the predator is nocturnal, shrimps should be sampled within 12 hours after sunset for practical use of the method on field-collected shrimp samples. Also, we have got some new knowledge about predator-prey relationship between flounder juveniles and crustaceans in the field. We suggest that this method could be applied to a range of flatfish and crustacean predators.
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Report
(4 results)
Research Products
(4 results)
