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Applied and Environmental Microbiology, November 2009, p. 7107-7114, Vol. 75, No. 22
0099-2240/09/$08.00+0     doi:10.1128/AEM.00464-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of Bacteriophages for Microbial Source Tracking in Korea

Jung Eun Lee,¹ Mi Young Lim,¹ Sei Yoon Kim,¹ Sunghee Lee,¹ Heetae Lee,¹ Hyun-Myung Oh,² Hor-Gil Hur,³ and GwangPyo Ko^1*

Institute of Health and Environment, Department of Environmental Health, School of Public Health, Seoul National University, 28 Yeongeon-dong, Chongro-gu, Seoul, South Korea 110-799,¹ Department of Marine Sciences, College of Natural Sciences, Inha University, Yong-Hyun-Dong, Nam-Gu, Incheon, South Korea 402-751,² Department of Environmental Science & Engineering, Gwangju Institute of Science and Technology, Gwangju, South Korea 500-712³

Received 25 February 2009/ Accepted 11 September 2009

We investigated coliphages from various fecal sources, including humans and animals, for microbial source tracking in South Korea. Both somatic and F⁺-specific coliphages were isolated from 43 fecal samples from farms, wild animal habitats, and human wastewater plants. Somatic coliphages were more prevalent and abundant than F⁺ coliphages in all of the tested fecal samples. We further characterized 311 F⁺ coliphage isolates using RNase sensitivity assays, PCR and reverse transcription-PCR, and nucleic acid sequencing. Phylogenetic analyses were performed based on the partial nucleic acid sequences of 311 F⁺ coliphages from various sources. F⁺ RNA coliphages were most prevalent among geese (95%) and were least prevalent in cows (5%). Among the genogroups of F⁺ RNA coliphages, most F⁺ coliphages isolated from animal fecal sources belonged to either group I or group IV, and most from human wastewater sources were in group II or III. Some of the group I coliphages were present in both human and animal source samples. F⁺ RNA coliphages isolated from various sources were divided into two main clusters. All F⁺ RNA coliphages isolated from human wastewater were grouped with Qβ-like phages, while phages isolated from most animal sources were grouped with MS2-like phages. UniFrac significance statistical analyses revealed significant differences between human and animal bacteriophages. In the principal coordinate analysis (PCoA), F⁺ RNA coliphages isolated from human waste were distinctively separate from those isolated from other animal sources. However, F⁺ DNA coliphages were not significantly different or separate in the PCoA. These results demonstrate that proper analysis of F⁺ RNA coliphages can effectively distinguish fecal sources.

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* Corresponding author. Mailing address: Department of Environmental Health, School of Public Health, Seoul National University, 28 Yeongeon-dong, Chongro-gu, Seoul, South Korea 110-799. Phone: 82 2 3668 7881. Fax: 82 2 745 9105. E-mail: gko{at}snu.ac.kr

Published ahead of print on 18 September 2009.

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Applied and Environmental Microbiology, November 2009, p. 7107-7114, Vol. 75, No. 22
0099-2240/09/$08.00+0     doi:10.1128/AEM.00464-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.