Detection of viral, bacterial, and protozoan pathogens and microbial source tracking markers in paired large- and small-volume water samples

When sampling for waterborne microbes, researchers may need to diverge from recommended sample volumes due to logistical constraints, novel targets, or challenging matrices, with little guidance about the potential impact on results. In field studies, we measured bacteria, viruses, and protozoa (15 quantitative polymerase chain reaction assays) in paired large- and small-volume samples to evaluate method performance and relevant factors. Concordance between methods was low. Large-volume ultrafiltration yielded more detections than small-volume sampling, especially for pathogens in groundwater. Greater microbial concentrations were associated with more frequent detections in small-volume samples and greater concordance between paired samples. Large-volume samples appeared to be more susceptible to diminished sensitivity from complex sample matrices. In laboratory studies, recovery of microbes was poorer for large- than small-volume methods, although large-volume methods more reliably detected low-concentration targets. Large-volume samples were less stable than small-volume samples during storage. Overall, large-volume sampling was superior for detecting pathogens but may underestimate concentrations; small-volume sampling was more prone to false negatives but was adequate when concentrations were relatively high, like we observed for microbial source tracking in surface waters.