Comparison of five rep-PCR genomic fingerprinting methods for differentiation of fecal Escherichia coli from humans, poultry and wild birds.
The development of a methodology to identify the origin of fecal pollution is important both for assessing the degree of risk posed to public health and for developing strategies to mitigate the environmental loading of pathogens associated with waterborne disease transmission. Five rep-PCR genomic fingerprinting methods, such as rep-PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR, ERIC2-PCR, BOX-PCR and (GTG)(5)-PCR, were assessed for their potential in differentiation of 232 fecal Escherichia coli isolates obtained from humans, poultry (chicken, duck and turkey) and wild birds (Canada goose and gull). Based on the results of cluster analysis and discriminant function analysis, (GTG)(5)-PCR was found to be the most suitable method for molecular typing of fecal E. coli, followed by BOX-PCR, REP-PCR, ERIC-PCR and ERIC2-PCR. A discriminant function analysis of (GTG)(5)-PCR fingerprints showed that 94.1%, 79.8%, 80.5%, 74.4%, 86.7% and 88.6% of turkey, chicken, duck, Canada goose, gull and human E. coli isolates were classified into the correct host group, respectively. Subsequently, (GTG)(5)-PCR was tested for its ability to track the origin of 113 environmental E. coli isolated from natural pond water. In conclusion, the (GTG)(5)-PCR genomic fingerprinting method can be considered as a promising genotypic tool for epidemiological surveillance of fecal pollution in aquatic environments.
Mohapatra
Bidyut R
BR
Department of Biology, University of Victoria, Victoria, BC, Canada. bmohapat@uvic.ca
Broersma
Klaas
K
Mazumder
Asit
A
D000818:Animals; D000835:Animals, Wild / Q000382:microbiology; D015373:Bacterial Typing Techniques; D001717:Birds / Q000382:microbiology; D016000:Cluster Analysis; D016172:DNA Fingerprinting / Q000379:methods*; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D005243:Feces; D005618:Fresh Water / Q000382:microbiology; D016680:Genome, Bacterial; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry / Q000382:microbiology; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity; D014876:Water Pollution / Q000032:analysis