Molecular Characterization of Bacteriophages for Microbial Source Tracking in Korea

• Published in: Applied and Environmental Microbiology Vol. 75; no. 22; pp. 7107 - 7114
• Main Authors: Lee, Jung Eun, Lim, Mi Young, Kim, Sei Yoon, Lee, Sunghee, Lee, Heetae, Oh, Hyun-Myung, Hur, Hor-Gil, Ko, GwangPyo
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.11.2009; American Society for Microbiology (ASM)
• Subjects: Animals; Bacteria; Bacteriophage Typing; Biological and medical sciences; Coliphages - classification; Coliphages - genetics; Coliphages - isolation & purification; DNA, Viral - genetics; Environmental Monitoring - methods; Environmental testing; Farms; Feces; Feces - virology; Fundamental and applied biological sciences. Psychology; Genetics; Humans; Korea; Microbiology; Molecular Sequence Data; Phylogeny; Polymerase Chain Reaction; Public Health Microbiology; Republic of Korea; Ribonucleases - metabolism; RNA, Viral - genetics; Sewage - virology; Water Microbiology; Asia; Korea; Republic of Korea; South Korea; Virus; Characterization; Microorganism; Phage
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/AEM.00464-09

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.