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Title of Journal: Curr Microbiol

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Abbravation: Current Microbiology

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Springer-Verlag

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DOI

10.1016/0956-7151(91)90250-5

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1432-0991

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RealTime PCR Assays for Monitoring Anaerobic Fung

Authors: Khin Ohnmar Lwin Mika Hayakawa Tomomi BanTokuda Hiroki Matsui
Publish Date: 2010/12/14
Volume: 62, Issue: 4, Pages: 1147-1151
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Abstract

The relationship between copy numbers of internal transcribed spacer 1 ITS1 and biomass or zoospore count of anaerobic fungi was studied to develop a quantitative realtime PCRbased monitoring method for fungal biomass or population in the rumen Nine fungal strains were used to determine the relationship between ITS1 copy number and fungal biomass Rumen fluid from three sheep and a cow were used to determine the relationship between ITS1 copy number and fungal population ITS1 copy number was determined by realtime PCR with a specific primer set for anaerobic fungi Freezedried fungal cells were weighed for fungal biomass Zoospore counts were determined by the rolltube method A positive correlation was observed between both ITS1 copy number and dry weight and ITS1 copy number and zoospore counts suggesting that the use of ITS1 copy numbers is effective for estimating fungal biomass and population density On the basis of ITS1 copy numbers fluctuations in the fungal population in sheep rumen showed that although the values varied among individual animals the fungal population tended to decrease after feeding In the present study a cultureindependent method was established that will provide a powerful tool for understanding the ecology of anaerobic fungi in the rumenThe authors would like to thank Dr Yasunari Yamamoto Mie Prefecture Livestock Research Institute Mie Prefecture Japan for providing rumen fluid and Professor Kazuo Sakka Graduate School of Mie University Japan for his instruction on realtime PCR This study was supported by the Integrated Research for Developing Japanesestyle Forage Feeding System to Increase Forage SelfSupport Ratio of the Ministry of Agriculture Forestry and Fisheries of Japan

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