Monitoring bacterial indicators and pathogens in cattle feedlot waste by real-time PCR

Marcus Klein, Leearna Brown, Ben van den Akker, Gregory M. Peters, Richard M. Stuetz, David J. Roser

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16 Citations (Scopus)

Abstract

Quantitative microbial health risk assessment requires accurate enumeration of pathogens in hazard-containing matrices as part of the risk characterization process. As part of a risk management-oriented study of cattle feedlot waste contaminants, we investigated the utility of quantitative real-time PCR (qPCR) for surveying the microbial constituents of different faecal wastes. The abundance of Escherichia coli and enterococci were first estimated in five cattle feedlot waste types from five localities. Bacteria were quantified using two culture methods and compared to the number of genome copies detected by qPCR targeted at E. coli and Enterococcus faecalis. Bacterial numbers detected in the different wastes (fresh faeces, pen manure, aged manure, composted manure, carcass manure compost) ranged from 10-7 to 102 g-1 (dry weight). Both indicator groups were detected by qPCR with a comparable sensitivity to culture methods across this range. qPCR measurements of E. coli and E. faecalis correlated well with MPN and spread plate data. As a second comparison, we inoculated green fluorescent protein (GFP) labeled reference bacteria into manure samples. GFP labeled E. coli and Listeria monocytogenes were detected by qPCR in concentrations corresponding to between 18% and 71% of the initial bacterial numbers, compared to only 2.5-16% by plating. Our results supported our selection of qPCR as a fast, accurate and reliable system for surveying the presence and abundance of pathogens in cattle waste.

Original languageEnglish
Pages (from-to)1381-1388
Number of pages8
JournalWater Research
Volume44
Issue number5
DOIs
Publication statusPublished - Mar 2010
Externally publishedYes

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    Klein, M., Brown, L., van den Akker, B., Peters, G. M., Stuetz, R. M., & Roser, D. J. (2010). Monitoring bacterial indicators and pathogens in cattle feedlot waste by real-time PCR. Water Research, 44(5), 1381-1388. https://doi.org/10.1016/j.watres.2009.11.016