Molecular identification of fecal pollution sources in water supplies by host-specific fecal DNA markers and Terminal Restriction Fragment Length Polymorphism profiles of 16S rRNA gene.
Specific fecal DNA markers were investigated for major pollution sources, cow, human, and pig, and occurrence of the identified markers was analyzed in river waters using Terminal Restriction Fragment Length Polymorphism (T-RFLP) techniques and sequencing of 16S rDNA of Bacteroides-Prevotella. The unique and specific DNA markers for cow and human were identified as a 222 bp and 60 bp peak in HaeIII T-RFLP profiles, respectively, and the pig-specific marker was not identified but the unique T-RFLP profile of pig could be used as a substitution. Human-specific marker was detected in most of the river waters tested (92.1%) and T-RFLP profiles of river waters were shown to be similar to those of human feces. Cluster analysis of T-RFLP data showed that the fecal sources were multiple (human plus cow and human plus dairy cow) in most of the river waters. The phylogenetic analysis for the clones recovered from the fecal and water samples showed that the clones from cow formed a discreet cluster from those of other sources. The other clones from human, pig, and river water formed two groups all together. The results of this study could be used to identify and control the fecal pollution source in the bodies of water in Korea.
Jeong
Ju-Yong
JY
Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-742, Republic of Korea.
Gil
Kyung-Ik
KI
Lee
Kyong-Hee
KH
Ka
Jong-Ok
JO
D000328:Adult; D000818:Animals; D001439:Bacteroides / Q000235:genetics / Q000302:isolation & purification; D002417:Cattle; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific; D005243:Feces / Q000382:microbiology*; D020459:Genes, rRNA; D005819:Genetic Markers / Q000235:genetics*; D006801:Humans; D007723:Korea; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D018720:Prevotella / Q000235:genetics / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D045483:Rivers / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013552:Swine; D014876:Water Pollution / Q000032:analysis*
D004269:DNA, Bacterial; D005819:Genetic Markers; D012336:RNA, Ribosomal, 16S; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases