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Award Abstract # 1334359
Collaborative Research: Sunlight Inactivation Mechanisms of Pathogenic Bacteria In Natural Waters

NSF Org: CBET
Division of Chemical, Bioengineering, Environmental, and Transport Systems
Recipient: THE LELAND STANFORD JUNIOR UNIVERSITY
Initial Amendment Date: July 15, 2013
Latest Amendment Date: April 23, 2018
Award Number: 1334359
Award Instrument: Standard Grant
Program Manager: Karl Rockne
krockne@nsf.gov
 (703)292-7293
CBET
 Division of Chemical, Bioengineering, Environmental, and Transport Systems
ENG
 Directorate for Engineering
Start Date: July 15, 2013
End Date: September 30, 2018 (Estimated)
Total Intended Award Amount: $210,510.00
Total Awarded Amount to Date: $220,043.00
Funds Obligated to Date: FY 2013 = $210,510.00
FY 2017 = $9,533.00
History of Investigator:
  • Alexandria Boehm (Principal Investigator)
    aboehm@stanford.edu
Recipient Sponsored Research Office: Stanford University
450 JANE STANFORD WAY
STANFORD
CA  US  94305-2004
(650)723-2300
Sponsor Congressional District: 16
Primary Place of Performance: Stanford University
CA  US  94305-4020
Primary Place of Performance
Congressional District:
16
Unique Entity Identifier (UEI): HJD6G4D6TJY5
Parent UEI:
NSF Program(s): EnvE-Environmental Engineering
Primary Program Source: 01001314DB NSF RESEARCH & RELATED ACTIVIT
01001718DB NSF RESEARCH & RELATED ACTIVIT
Program Reference Code(s): 018Z, 021E, 9102
Program Element Code(s): 144000
Award Agency Code: 4900
Fund Agency Code: 4900
Assistance Listing Number(s): 47.041

ABSTRACT

CBET 1334359/1335673
Alexandria Boehm/Kara Nelson
Stanford University/University of California-Berkeley

Fecal indicator bacteria (FIB), such as Enterococcus and Escherichia coli, are used to assess beach water quality and serve as proxies for human pathogens. FIB concentrations in natural waters vary diurnally with concentrations and are often below assay detection limits in mid-afternoon and orders of magnitude higher at night, which has several implications. First, the time the sample is collected dramatically impacts the measured concentration, which could make the difference between compliance and noncompliance with water quality standards. Second, it is not known whether the concentrations of actual pathogens, and thus associated health risk, also experience such fluctuations. Therefore, it is critical to obtain information on the processes that control the diurnal fluctuations for FIB and human pathogens of concern. Sunlight is believed to be the major cause of the diurnal fluctuations in FIB. However, the dominant mechanisms through which sunlight damages microorganisms are not well understood. At least three mechanisms have been described: endogenous direct damage to cellular components by ultraviolet wavelengths, and indirect endogenous and exogenous photoinactivation caused by reactive species generated inside and outside the cell, respectively, when photons are absorbed by sensitizer molecules. Research to date has primarily focused on FIB photoinactivation and has generally been highly empirical and site-specific so that it is not possible to generalize to predict sunlight inactivation rates in other environmental contexts or for other organisms. Additionally, there is a striking lack of data on the photoinactivation of bacterial pathogens. The objectives of this project are to characterize the susceptibility of FIB and a suite of pathogenic bacteria to endogenous and exogenous photoinactivation and develop a quantitative model for photoinactivation. Laboratory experiments will be used to develop a mechanistic understanding of processes that control inactivation, and to understand the nature of observed differences between organisms. Field and laboratory data will be incorporated into a model to predict inactivation rates, and the model will be tested using a microcosm study. The model will use environmental parameters as inputs to estimate the inactivation of bacteria by sunlight and will be useful for estimating inactivation rates for a wide range of organisms and waters without the need for site- and organism- specific studies.

The project will advance knowledge in several ways. The research will yield essential insights into the fate of FIB and bacterial pathogens in the environment, a high priority research need to protect human health and improve coastal water quality. The work will have immediate implications for the management of recreational water for the protection of human health. The improved understanding of sunlight-mediated inactivation mechanisms and the new modeling approach will also be directly useful for engineered and natural systems in which sunlight plays a major role in disinfection, including solar disinfection of drinking water (SODIS) and wastewater treatment in ponds and wetlands. The results from the proposed work will be shared with policy makers and beach managers and will result in the improved protection of human health. The investigators will integrate the results into their classroom instruction. Graduate and undergraduate students will participate in the research. The investigators will develop new curriculum on the impact of sunlight on the treatment of stormwater runoff for high school students and a module on water and environmental engineering for elementary school students.

PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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(Showing: 1 - 10 of 12)
J. McClary, A. B. Boehm. "Transcriptional response of Staphylococcus aureus to sunlight in oxic and anoxic conditions" Frontiers in Microbiology , v.9 , 2018 , p.10.3389/f
J. McClary, L. M. Sassoubre, A. B. Boehm "Staphylococcus aureus photoinactivation mechanisms and cellular response to sunlight exposure" Applied and Environmental Microbiology , 2017
Katherine E Graham Aaron J Prussin, II Linsey C Marr Lauren M Sassoubre Alexandria B Boehm "Microbial community structure of sea spray aerosols at three California beaches" FEMS Microbial Ecology , v.94 , 2018 , p.fiy005
K. E. Brown and A. B. Boehm "Transport of fecal indicators from beach sand to the surf zone by recirculating seawater: laboratory experiments and numerical modeling" Environmental Science & Technology , v.50 , 2016 , p.12840
K. I. Brown, K. E. Graham, A. B. Boehm "Risk-based threshold of gull-associated fecal marker concentrations for recreational water" Environmental Science & Technology Letters , v.4 , 2017 , p.44
K. I. Brown, K. Graham, J. Soller, A. B. Boehm. "Estimating probability of illness due to swimming in recreational water with a mixture of human- and gull-associated microbial source tracking markers" Environmental Science: Processes and Impacts , v.19 , 2017 , p.1528 10.1039/C7EM00316A
M.C. Mattioli, L. M. Sassoubre, T. L. Russell, A. B. Boehm "Decay of sewage-sourced microbial source tracking markers and fecal indicator bacteria in marine waters" Water Research , v.108 , 2017 , p.106
P. A. Maraccini, D. Wang, J. McClary, A. B. Boehm "Growth-dependent photoinactivation kinetics of Enterococcus faecalis" Journal of Applied Microbiology , v.118 , 2015 , p.1226
P. A. Maraccini, D. Wang, J. McClary, A. B. Boehm. "Growth-dependent photoinactivation kinetics of Enterococcus faecalis." Journal of Applied Microbiology , v.118 , 2015 , p.1226
P. A. Maraccini, J. H. Wenk, A. B. Boehm. "2016. Photoinactivation of Eight Health-Relevant Bacterial Species: Determining the Importance of the Exogenous Indirect Mechanism." Environmental Science & Technology , v.50 , 2016 , p.5050 10.1021/acs.est.6b00074
P. A. Maraccini, J. H. Wenk, A. B. Boehm. "Exogenous Indirect Photoinactivation of Bacterial Pathogens and Indicators in Water with Natural and Synthetic Photosensitizers in Simulated Sunlight With Reduced UVB" Journal of Applied Microbiology , v.121 , 2016 , p.587
(Showing: 1 - 10 of 12)

PROJECT OUTCOMES REPORT

Disclaimer

This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.

The scientific objective of this project was to characterize the susceptibility of pathogenic bacteria suspended in water to inactivation by sunlight. We conducted laboratory experiments to develop a mechanistic understanding of processes that control photoinactivation, and to understand the nature of observed differences between bacteria. We then tested our understanding of bacterial photoinactivation using field studies in natural waters. This work is essential for understanding the persistence of pathogenic bacteria in all sorts of water, and for designing water treatment technologies that rely on sunlight for disinfection. 

 

We found that bacteria are generally inactivated via sunlight directly rather than by reactive chemicals (like oxygen) generated outside the cell by dissolved organic material reacting with energy from the sun (referred to as exogenous indirect photoinactivation). Anaerobic or micro-aerophilic bacteria are particularly sensitive to photoinactivation and inactivate more readily than aerobic bacteria. We found that pigmented bacteria are less sensitive to photoinactivation than unpigmented bacteria. Our work supports the idea that bacteria cannot be considered inert particles in photoinactivation studies as we found that photoinactivation rates vary with bacterial growth rates, and that bacteria actively respond to sunlight stress by changing the genes they express. Field research corroborated the laboratory results and showed that, across a variety of waters from wetlands to oceans, that photoinactivation rate constants could be estimated based on the UVA and UVB fluence in the water column. Additional work explored the extent to which sunlight affects the microbiome of beach sands, water and sea spray aerosols, and whether sunlight affected concentrations of microbial source tracking DNA markers in water and sand. 

 

This project supported graduate and undergraduate students, including under-represented students, to carry out cutting edge scientific research. The research team created modules for engaging underrepresented engineering students in environmental engineering and implemented them every year of the project. The PI attended a number of local, regional, state, and national stakeholder meetings to present results. This grant provided training to a high school teacher, provided an internship for a graduate student at the Center for Disease Control, and provided outreach to preschool children about bacteria. 

 


Last Modified: 10/03/2018
Modified by: Alexandria Boehm

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