Towards the standardization of Enterococcus culture methods for waterborne antibiotic resistance monitoring: A critical review of trends across studies

Davis, Benjamin C.; Keenum, Ishi; Calarco, Jeanette; Liguori, Krista; Milligan, Erin; Pruden, Amy; Harwood, Valerie J.

doi:10.1016/j.wroa.2022.100161

Cited by 7 publications

(8 citation statements)

References 131 publications

(171 reference statements)

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“…Overall, the SBA demonstrated a selectivity for Enterococcus spec. of 97.9%, which is higher than that found in other studies [29,30]. The high proportion of E. faecium in the present report is in contrast to the summarized data of a recent review calculating a rate of this species of around 42% in municipal wastewaters [29] but is in approximate agreement to the results of other investigations [31,32].…”

Section: Characterization Of Strainssupporting

confidence: 76%

“…However, in all VREs of the present study, van operon types were confirmed, which are the most common in clinical isolates [ 43 ]. VanA as well as vanB genes are located on specific transposons, and for vanA -carrying strains, an additional resistance to teicoplanin has been confirmed [ 29 ]. With the PCR protocol used here, amplification products cover 95.3% of vanA and 57.3% of vanB genes in comparison to sequences deposited in GenBank.…”

Section: Resultsmentioning

confidence: 99%

“…The results of the sequencing suggest a relative uniformity of van genes in the isolated E. faecium strains from the sampled wastewater treatment plant. Further molecular characterization of strains, like multilocus sequence typing (MLST), would allow a comparison of enterococci of clinical, veterinary, and environmental origin [ 29 , 41 , 42 ]. Unfortunately, typing data on isolated strains from patients in the catchment are not available.…”

Section: Resultsmentioning

confidence: 99%

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Enterococci, Van Gene-Carrying Enterococci, and Vancomycin Concentrations in the Influent of a Wastewater Treatment Plant in Southeast Germany

Geissler,

Schröttner,

Oertel

et al. 2024

Microorganisms

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Vancomycin-resistant (VR) Enterococcus spp. can be detected in high concentrations in wastewaters and pose a risk to public health. During a one-year study (September 2022–August 2023), 24 h composite raw wastewater samples (n = 192) of a municipal wastewater treatment plant were investigated for cultivable enterococci. After growth on Slanetz–Bartley agar (SBA), a mean concentration of 29,736 ± 9919 cfu/mL was calculated. Using MALDI-TOF MS to characterize randomly picked colonies (n = 576), the most common species were found to be Enterococcus faecium (72.6%), E. hirae (13.7%), and E. faecalis (8.0%). Parallel incubation of wastewater samples on SBA and VRESelect agar resulted in a mean rate of VR enterococci of 2.0 ± 1.5%. All the tested strains grown on the VRESelect agar (n = 172) were E. faecium and carried the vanA (54.6%) or vanB gene (45.4%) with limited sequence differences. In susceptibility experiments, these isolates showed a high-level resistance to vancomycin (>256 µg/mL). Concentration of vancomycin was determined in 93.7% of 112 wastewater samples (mean: 123.1 ± 64.0 ng/L) and varied between below 100 ng/L (the detection limit) and 246.6 ng/L. A correlation between the concentration of vancomycin and the rate of VR strains among the total enterococci could not be found. The combination of incubation of samples on SBA and a commercial vancomycin-containing agar applied in clinical microbiology with a multiplex PCR for detection of van genes is an easy-to-use tool to quantify and characterize VR Enterococcus spp. in water samples.

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Section: Characterization Of Strainssupporting

confidence: 76%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Enterococci, Van Gene-Carrying Enterococci, and Vancomycin Concentrations in the Influent of a Wastewater Treatment Plant in Southeast Germany

Geissler,

Schröttner,

Oertel

et al. 2024

Microorganisms

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“…Among the different target bacteria, the following have commonly been utilised and/or proposed for antimicrobial resistance (AMR) monitoring: Escherichia coli, Enterococci, Enterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter baumannii and Aeromonas spp. (Berendonk et al, 2015;Davis et al, 2022;Huijbers et al, 2020;Liguori et al, 2022).…”

Section: Microbial Cecsmentioning

confidence: 99%

Selection of indicator contaminants of emerging concern when reusing reclaimed water for irrigation — A proposed methodology

Verlicchi

Grillini

Lacasa

et al. 2023

Science of The Total Environment

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“…While the general advantage of culture-based methods is that they indicate a viable endpoint, a limitation of the WHO Tricycle protocol is its singular focus on E. coli . Other targets have been suggested for inclusion, such as ESBL Enterobacterales, Enterococcus spp., Pseudomonas , Aeromonas , or Acinetobacter [ 16 , 17 , 29 , 30 , 31 ]. Still, no single or combined set of culture-based targets will be able to fully capture all resistant bacteria in a given water sample.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Cefotaxime-Resistant Escherichia coli and sul1 and intI1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water

Liguori,

Calarco,

Maldonado Rivera

et al. 2023

Antibiotics

Self Cite

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Awareness of the need for surveillance of antimicrobial resistance (AMR) in water environments is growing, but there is uncertainty regarding appropriate monitoring targets. Adapting culture-based fecal indicator monitoring to include antibiotics in the media provides a potentially low-tech and accessible option, while quantitative polymerase chain reaction (qPCR) targeting key genes of interest provides a broad, quantitative measure across the microbial community. The purpose of this study was to compare findings obtained from the culture of cefotaxime-resistant (cefR) Escherichia coli with two qPCR methods for quantification of antibiotic resistance genes across wastewater, recycled water, and surface waters. The culture method was a modification of US EPA Method 1603 for E. coli, in which cefotaxime is included in the medium to capture cefR strains, while qPCR methods quantified sul1 and intI1. A common standard operating procedure for each target was applied to samples collected by six water utilities across the United States and processed by two laboratories. The methods performed consistently, and all three measures reflected the same overarching trends across water types. The qPCR detection of sul1 yielded the widest dynamic range of measurement as an AMR indicator (7-log versus 3.5-log for cefR E. coli), while intI1 was the most frequently detected target (99% versus 96.5% and 50.8% for sul1 and cefR E. coli, respectively). All methods produced comparable measurements between labs (p < 0.05, Kruskal–Wallis). Further study is needed to consider how relevant each measure is to capturing hot spots for the evolution and dissemination of AMR in the environment and as indicators of AMR-associated human health risk.

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Towards the standardization of Enterococcus culture methods for waterborne antibiotic resistance monitoring: A critical review of trends across studies

Cited by 7 publications

References 131 publications

Enterococci, Van Gene-Carrying Enterococci, and Vancomycin Concentrations in the Influent of a Wastewater Treatment Plant in Southeast Germany

Enterococci, Van Gene-Carrying Enterococci, and Vancomycin Concentrations in the Influent of a Wastewater Treatment Plant in Southeast Germany

Selection of indicator contaminants of emerging concern when reusing reclaimed water for irrigation — A proposed methodology

Comparison of Cefotaxime-Resistant Escherichia coli and sul1 and intI1 by qPCR for Monitoring of Antibiotic Resistance of Wastewater, Surface Water, and Recycled Water

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