A Novel Screening Tool Using Microarray and PCR to Detect Pathogens in Agriculturally Impacted Waters

R. L. Crawford; E. R. Coats; C. K. Brinkman; T. L. Green; M. M. Ederer; F. S. Alhamlan
Microbial populations are present in high concentrations in dairy wastewater treatment lagoons and thus pose a potential public health risk. A broad and robust detection assay is needed to monitor the presence of human pathogens in such systems. The objective of this study was to develop and corroborate a microarray-based assay for screening and monitoring of agriculturally-impacted waters for immediate detection of > 1,500 specific viruses and bacteria. Water samples from two dairy lagoons (Lagoons I and II) were screened using molecular assays (such as ElectroChemical Detection (ECD) 12K microarray) in conjunction with traditional analytical chemical methods. Bacterial signature DNA sequences and viral signature DNA/RNA sequences that are associated with human diseases were detected in higher numbers in both lagoons. The presence of some of these microbial signatures in the lagoons was confirmed using PCR and qPCR techniques. Based on microscopic analyses, most probable number (MPN) counts, total viable counts (CFU), 16S rRNA gene analysis, microarray signal intensity analyses, and qPCR assays, the microbial community density was found to trend higher in Lagoon I than Lagoon II; however, microbial diversity in Lagoon I trended lower. Results from this research confirm the usefulness of ECD-based microarray technology as a pathogen screening method for agriculturally-impacted waters.
Electrochemical Detection Microarray, Pathogen Detection, Monitoring, Dairy Lagoons
Download | Back to Issue| Archive