Population Screening Using Sewage Reveals Pan-Resistant Bacteria in Hospital and Community Samples.

Lital Meir-Gruber,Roberto Azar,Ella Mendelson,Nathan Keller,Shiraz Gefen-Halevi,Gill Smollan,Fernando Mileguir,Musa Y Hindiyeh,Galia Rahav,Yossi Manor,Ari Shamiss,Natasha Belausov,Jan Kluytmans

doi:10.1371/journal.pone.0164873

Lital Meir-Gruber, Roberto Azar + Show 11 more

Open Access

https://doi.org/10.1371/journal.pone.0164873

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Abstract

The presence of pan-resistant bacteria worldwide possesses a threat to global health. It is difficult to evaluate the extent of carriage of resistant bacteria in the population. Sewage sampling is a possible way to monitor populations. We evaluated the presence of pan-resistant bacteria in Israeli sewage collected from all over Israel, by modifying the pour plate method for heterotrophic plate count technique using commercial selective agar plates. This method enables convenient and fast sewage sampling and detection. We found that sewage in Israel contains multiple pan-resistant bacteria including carbapenemase resistant Enterobacteriacae carrying blaKPC and blaNDM-1, MRSA and VRE. blaKPC carrying Klebsiella pneumonia and Enterobacter cloacae were the most common Enterobacteriacae drug resistant bacteria found in the sewage locations we sampled. Klebsiella pneumonia, Enterobacter spp., Escherichia coli and Citrobacter spp. were the 4 main CRE isolated from Israeli sewage and also from clinical samples in our clinical microbiology laboratory. Hospitals and Community sewage had similar percentage of positive samplings for blaKPC and blaNDM-1. VRE was found to be more abundant in sewage in Israel than MRSA but there were more locations positive for MRSA and VRE bacteria in Hospital sewage than in the Community. Therefore, our upgrade of the pour plate method for heterotrophic plate count technique using commercial selective agar plates can be a useful tool for routine screening and monitoring of the population for pan-resistant bacteria using sewage.

Highlights

Antibiotic-resistant bacteria are a growing problem worldwide and possess a major threat to global health
In order to determine the limit of detection (LOD) for the overlay method, 1.5X108 CFU/ml blaKPC Klebsiella pneumoniae, blaNDM-1 Escherichia coli, Methicillin-resistant Staphylococcus aureus (MRSA) or vancomycin-resistant Enterococci (VRE) positive bacteria were serially dilluted logarithmically in sewage that was negative for these targets
We found that the overlay did not cause bacteria death as the LOD was similar (5–10 CFU/ml) between the two methods for the different bacteria

Summary

Introduction

Antibiotic-resistant bacteria are a growing problem worldwide and possess a major threat to global health. Extensive use of antimicrobial agents in hospital settings and animal husbandry, PLOS ONE | DOI:10.1371/journal.pone.0164873. Resistant Bacteria in Sewage in Israel has led to the emergence of pan-resistant "superbugs", which creates serious ecological and epidemiological challenges for humans [1,2,3,4]. Among the most common resistant bacteria are the Carbapenem-Resistant Enterobacteriaceae (CRE), that have developed resistance to carbapenems (ertapenem, meropenem, imipenem and doripenem), which are most often the last option for therapy. Klebsiella pneumonia carbapenemase (blaKPC) and New-Delhi Metallo-β-lactamase (blaNDM-1) are among the most important carbapenemases. Methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) represent the biggest therapeutic hurdles among the gram-positive organisms. Methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) represent the biggest therapeutic hurdles among the gram-positive organisms. [1, 9, 10,11,12,13]

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