Spread from the Sink to the Patient: In Situ Study Using Green Fluorescent Protein (GFP)-Expressing Escherichia coli To Model Bacterial Dispersion from Hand-Washing Sink-Trap Reservoirs

ABSTRACT There have been an increasing number of reports implicating Gammaproteobacteria as often carrying genes of drug resistance from colonized sink traps to vulnerable hospitalized patients. However, the mechanism of transmission from the wastewater of the sink P-trap to patients remains poorly understood. Herein we report the use of a designated hand-washing sink lab gallery to model dispersion of green fluorescent protein (GFP)-expressing Escherichia coli from sink wastewater to the surrounding environment. We found no dispersion of GFP-expressing E. coli directly from the P-trap to the sink basin or surrounding countertop with coincident water flow from a faucet. However, when the GFP-expressing E. coli cells were allowed to mature in the P-trap under conditions similar to those in a hospital environment, a GFP-expressing E. coli-containing putative biofilm extended upward over 7 days to reach the strainer. This subsequently resulted in droplet dispersion to the surrounding areas (<30 in.) during faucet operation. We also demonstrated that P-trap colonization could occur by retrograde transmission along a common pipe. We postulate that the organisms mobilize up to the strainer from the P-trap, resulting in droplet dispersion rather than dispersion directly from the P-trap. This work helps to further define the mode of transmission of bacteria from a P-trap reservoir to a vulnerable hospitalized patient. IMPORTANCE Many recent reports demonstrate that sink drain pipes become colonized with highly consequential multidrug-resistant bacteria, which then results in hospital-acquired infections. However, the mechanism of dispersal of bacteria from the sink to patients has not been fully elucidated. Through establishment of a unique sink gallery, this work found that a staged mode of transmission involving biofilm growth from the lower pipe to the sink strainer and subsequent splatter to the bowl and surrounding area occurs rather than splatter directly from the water in the lower pipe. We have also demonstrated that bacterial transmission can occur via connections in wastewater plumbing to neighboring sinks. This work helps to more clearly define the mechanism and risk of transmission from a wastewater source to hospitalized patients in a world with increasingly antibiotic-resistant bacteria that can thrive in wastewater environments and cause infections in vulnerable patients.

[1]  P. Borella,et al.  Aetiology, source and prevention of waterborne healthcare-associated infections: a review. , 2014, Journal of medical microbiology.

[2]  A. Pantel,et al.  Persisting transmission of carbapenemase-producing Klebsiella pneumoniae due to an environmental reservoir in a university hospital, France, 2012 to 2014. , 2016, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[3]  A. Melhus,et al.  Minor outbreak of extended-spectrum β-lactamase-producing Klebsiella pneumoniae in an intensive care unit due to a contaminated sink. , 2012, The Journal of hospital infection.

[4]  J. Rodríguez-Baño,et al.  Wastewater drainage system as an occult reservoir in a protracted clonal outbreak due to metallo-β-lactamase-producing Klebsiella oxytoca. , 2013, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[5]  S. Dancer Dos and don’ts for hospital cleaning , 2016, Current opinion in infectious diseases.

[6]  T. Palmore,et al.  The role of water in healthcare-associated infections , 2013, Current opinion in infectious diseases.

[7]  P. Savelkoul,et al.  The sink as a correctable source of extended-spectrum β-lactamase contamination for patients in the intensive care unit. , 2014, The Journal of hospital infection.

[8]  Camille Lemieux,et al.  Outbreak of Multidrug-Resistant Pseudomonas aeruginosa Colonization and Infection Secondary to Imperfect Intensive Care Unit Room Design , 2009, Infection Control &#x0026; Hospital Epidemiology.

[9]  C. Fusch,et al.  Self‐disinfecting sink drains reduce the Pseudomonas aeruginosa bioburden in a neonatal intensive care unit , 2015, Acta paediatrica.

[10]  Kaisen Lin,et al.  Aerosolization of Ebola Virus Surrogates in Wastewater Systems. , 2017, Environmental science & technology.

[11]  A. Rickard,et al.  Microbial Characterization of Biofilms in Domestic Drains and the Establishment of Stable Biofilm Microcosms , 2003, Applied and Environmental Microbiology.

[12]  S. Dancer,et al.  Controlling Hospital-Acquired Infection: Focus on the Role of the Environment and New Technologies for Decontamination , 2014, Clinical Microbiology Reviews.

[13]  E. de Jonge,et al.  An integrated approach to control a prolonged outbreak of multidrug-resistant Pseudomonas aeruginosa in an intensive care unit. , 2014, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[14]  Y. Li,et al.  Multi-zone modeling of probable SARS virus transmission by airflow between flats in Block E, Amoy Gardens. , 2005, Indoor air.

[15]  K. Vickery,et al.  Presence of biofilm containing viable multiresistant organisms despite terminal cleaning on clinical surfaces in an intensive care unit. , 2012, The Journal of hospital infection.

[16]  E. Leitner,et al.  Contaminated Handwashing Sinks as the Source of a Clonal Outbreak of KPC-2-Producing Klebsiella oxytoca on a Hematology Ward , 2014, Antimicrobial Agents and Chemotherapy.

[17]  William A Rutala,et al.  Role of hospital surfaces in the transmission of emerging health care-associated pathogens: norovirus, Clostridium difficile, and Acinetobacter species. , 2010, American journal of infection control.

[18]  T. Planche,et al.  Multidrug-resistant Pseudomonas aeruginosa outbreaks in two hospitals: association with contaminated hospital waste-water systems. , 2012, The Journal of hospital infection.

[19]  N. Kamar,et al.  Transmission of metallo-β-lactamase-producing Pseudomonas aeruginosa in a nephrology-transplant intensive care unit with potential link to the environment. , 2016, The Journal of hospital infection.

[20]  S. Solomon,et al.  Antibiotic resistance threats in the United States: stepping back from the brink. , 2014, American family physician.

[21]  J. Soothill Carbapenemase-bearing Klebsiella spp. in sink drains: investigation into the potential advantage of copper pipes. , 2016, The Journal of hospital infection.

[22]  P. Edmonds,et al.  Epidemiology of Pseudomonas aeruginosa in a Burns Hospital: Surveillance by a Combined Typing System , 1972, Applied microbiology.

[23]  G L French,et al.  Hospital outbreak of Klebsiella pneumoniae resistant to broad-spectrum cephalosporins and beta-lactam-beta-lactamase inhibitor combinations by hyperproduction of SHV-5 beta-lactamase , 1996, Journal of clinical microbiology.

[24]  D. Flournoy,et al.  Prevalence of gentamicin- and amikacin-resistant bacteria in sink drains , 1980, Journal of clinical microbiology.

[25]  R. Kumar,et al.  A survey of hand-washing facilities in the outpatient department of a tertiary care teaching hospital in India. , 2011, Journal of infection in developing countries.

[26]  K. Wigginton,et al.  Survivability, Partitioning, and Recovery of Enveloped Viruses in Untreated Municipal Wastewater , 2016, Environmental science & technology.

[27]  J. Bartley,et al.  Evaluating hygienic cleaning in health care settings: what you do not know can harm your patients. , 2010, American journal of infection control.

[28]  M. Mörgelin,et al.  Acetic acid as a decontamination method for sink drains in a nosocomial outbreak of metallo-β-lactamase-producing Pseudomonas aeruginosa. , 2016, The Journal of hospital infection.

[29]  Ki Bae Hong,et al.  Investigation and Control of an Outbreak of Imipenem-resistant Acinetobacter baumannii Infection in a Pediatric Intensive Care Unit , 2012, The Pediatric infectious disease journal.

[30]  L. Wright,et al.  “Down the drain”: carbapenem‐resistant bacteria in intensive care unit patients and handwashing sinks , 2013, The Medical journal of Australia.

[31]  A. Sundsfjord,et al.  A Long-Term Low-Frequency Hospital Outbreak of KPC-Producing Klebsiella pneumoniae Involving Intergenus Plasmid Diffusion and a Persisting Environmental Reservoir , 2013, PloS one.

[32]  A. Robicsek,et al.  Management of a multidrug-resistant Acinetobacter baumannii outbreak in an intensive care unit using novel environmental disinfection: a 38-month report. , 2010, American journal of infection control.

[33]  W. Rutala,et al.  Healthcare Outbreaks Associated With a Water Reservoir and Infection Prevention Strategies. , 2016, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[34]  R. Weinstein,et al.  Pseudomonas in the sinks in an intensive care unit: relation to patients. , 1984, Journal of clinical pathology.

[35]  A. Chapuis,et al.  Outbreak of Extended-Spectrum Beta-Lactamase Producing Enterobacter cloacae with High MICs of Quaternary Ammonium Compounds in a Hematology Ward Associated with Contaminated Sinks , 2016, Front. Microbiol..

[36]  B. Willey,et al.  Outbreak of Extended-Spectrum β-Lactamase–producing Klebsiella oxytoca Infections Associated with Contaminated Handwashing Sinks , 2012, Emerging infectious diseases.

[37]  D. Roux,et al.  Contaminated sinks in intensive care units: an underestimated source of extended-spectrum beta-lactamase-producing Enterobacteriaceae in the patient environment. , 2013, The Journal of hospital infection.

[38]  W. Rutala,et al.  Water as a Reservoir of Nosocomial Pathogens , 1997, Infection Control &#x0026; Hospital Epidemiology.