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The effect of Paenibacillus on IDEXX Enterolert results from freshwater stream environments

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Abstract

Enterolert, a fluorogenic substrate test, is used as a quantitative method for determining freshwater concentrations of Enterococcus for water quality indicators. However, there is some evidence from recent studies suggesting that Enterolert may not suppress false positives due to pollution sources in waterbodies. In this study, we evaluated this method by analyzing field water and sediment samples from four freshwater streams. We also performed a laboratory microcosm study from two of the stream sediments. The Enterolert method was investigated by phenotypic and genomic analyses for accuracy of isolating and quantifying Enterococcus and/or Streptococcus. Additionally, we tested isolates from Enterolert panels for antibiotic resistance. Results from the field and microcosm studies from initial to final time points indicated that false positives were predominantly Paenibacillus spp. and other non-fecal indicator bacteria. Furthermore, the microcosm study indicated shifts from lactic acid to non-lactic acid bacteria between initial to final time points, but Enterococcus concentrations from Enterolert panels remained stable for the duration of the study for both stream sediments. Antibiotic resistance indicated no distinct pattern of resistance or susceptibility to a suite of antibiotics. However, all isolates tested were resistant to bacitracin and nalidixic acid. In conclusion, we found that Enterolert was not exclusively selective for Enterococcus from freshwater environments and that sediment and polluted waterbodies have the potential to skew the presumed concentrations. More research is needed to evaluate the effectiveness and selectivity of the medium used for the fluorogenic substrate test for Enterococcus enumeration.

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Acknowledgements

The authors would like to acknowledge contributions from student research assistants at the Oklahoma Water Survey: Julie Bahr, Alejandra Hernandez-Santana, and Kassidy Long.

Funding

This research was partially funded by a USGS 104(b) project–2020 OK198B and through research funding provided by the Office of the Vice President for Research at the University of Oklahoma to the Oklahoma Water Survey at the University of Oklahoma.

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Authors and Affiliations

  1. Civil Engineering and Environmental Science, The University of Oklahoma, 303 Carson Engineering Center, 202 W. Boyd, Norman, OK, 73019, USA

    Grant M. Graves & Jason R. Vogel

  2. Microbiology and Plant Biology, The University of Oklahoma, 716 George Lynn Cross Hall, 770 Van Vleet Oval, Norman, OK, 73019, USA

    Ralph S. Tanner, Kara B. De León & Alex W. Walls

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  1. Grant M. Graves
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  2. Ralph S. Tanner
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  4. Kara B. De León
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  5. Alex W. Walls
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Contributions

All authors contributed to study conception and design; Grant M. Graves and Ralph S. Tanner collected and analyzed data; Kara B. De León and Alex W. Walls provided sequence analysis; Grant M. Graves and Jason R. Vogel provided project oversight; Grant M. Graves and Ralph S. Tanner wrote the manuscript; all authors edited and reviewed manuscript.

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Correspondence to Grant M. Graves.

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Graves, G.M., Tanner, R.S., Vogel, J.R. et al. The effect of Paenibacillus on IDEXX Enterolert results from freshwater stream environments. Environ Sci Pollut Res 30, 91074–91083 (2023). https://doi.org/10.1007/s11356-023-28797-y

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