Journal
WATER AIR AND SOIL POLLUTION
Volume 226, Issue 9, Pages -Publisher
SPRINGER INT PUBL AG
DOI: 10.1007/s11270-015-2560-x
Keywords
MPN-qPCR; Nonspecific enrichment; Pathogen; Detection; Vegetable; Aerosol; Soil; Wastewater
Funding
- US-Israel Binational Agricultural Research and Development Fund [CP-9033-09]
- Israeli Water Authority [87227611, 874130]
- Kreitman School for Graduate Studies
- Zuck Maccabi Fund
- Kraft food group
- MIT International Science and Technology Initiatives (MIT-MISTI)
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A sensitive, high-throughput, and cost-effective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were concomitantly spiked with Salmonella enterica and/or Pseudomonas aeruginosa to determine recovery rates and limits of detection. The various matrices were first enriched with a general pre-enrichment broth in a dilution series and then enumerated by most probable number (MPN) estimation using quantitative PCR for rapid screening of amplicon presence. Soil and aerosols were then tested in non-spiked environmental samples, as these matrices are prone to large experimental variation. Limit of detection in the various soil types was 1-3 colony-forming units (CFU) g(-1); on vegetable surface, 5 CFU per tomato; in treated wastewater, 5 CFU L-1; and in aerosols, >300 CFU mL(-1). Our method accurately identified S. enterica in non-spiked environmental soil samples within a day, while traditional methods took 4 to 5 days and required sorting through biochemically and morphologically similar species. Likewise, our method successfully identified P. aeruginosa in non-spiked aerosols generated by a domestic wastewater treatment system. The obtained results suggest that the developed method presents a broad approach for the rapid, efficient, and reliable detection of relatively low densities of pathogenic organisms in challenging environmental samples.
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