207297 Application of tangential flow ultrafiltration and molecular detection for the microbial evaluation of 100L surface and ground water samples impacted by industrial food animal production

Monday, November 9, 2009

Kristen E. Gibson, BS , Department of Environmental Health Sciences/Division of Environmental Health Engineering, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD
Kellogg J. Schwab, PhD , Department of Environmental Health Sciences/Division of Environmental Health Engineering, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD
One of the leading causes of death worldwide is infectious diseases, thus emerging human and animal microbial pathogens associated with waterborne transmission are of particular concern to the public health community. A major limiting factor in evaluating the microbial quality of various water sources is the lack of an effective and efficient method for simultaneous collection and recovery of low levels of bacteria, viruses, and protozoa and the subsequent inability of these isolated microbes to be rapidly identified and quantified. Current methods for the assessment of microorganisms in water are targeted towards specific pathogen groups. Our research applied tangential flow ultrafiltration (TFU) for the simultaneous concentration and recovery of microbes from large volumes (100L) of surface (SW) and ground water (GW) potentially impacted by animal wastes—a product of industrial food animal production practices. A TFU system designed to concentrate 100L water samples was optimized using a commercially available capsule filter containing hollow fiber membranes of 70,000-dalton pore size. The final concentrated sample (100-300mL) is analyzed using culture-based methods and/or molecular-based real time polymerase chain reaction (qPCR) and loop-mediated isothermal amplification (LAMP) for the identification of select, endogenous microorganisms including pathogenic bacteria, viruses, and protozoa of human health concern. Lower Yakima Valley, WA—a rural region with the densest concentration of dairy operations and milk cows in WA state—was selected for the application of TFU to 100L water samples (n=21). Bacterial analysis has been completed and viral and protozoa analysis is ongoing.

Learning Objectives:
Demonstrate the usefulness of the TFU method for microbial evaluation of 100L environmental water samples. Develop molecular-based methods, including qPCR and LAMP, for the detection of select microorganisms of public health concern in TFU concentrates. Describe how TFU combined with molecular techniques can help better address the role water plays as a vehicle for infectious diseases in the environment.

Presenting author's disclosure statement:

Qualified on the content I am responsible for because: I am a PhD student and the work that is being presented is part of my dissertation research. I have been involved in similar research in the past, and here are some publications I am an author on: Sapkota AR, Curriero FC, Gibson KE, Schwab KJ. “Antibiotic-resistant Enterococci and Fecal Indicators in Surface Water and Groundwater Impacted by a Concentrated Swine Feeding Operation.” Environmental Health Perspectives. 115:1040-1045. 2007. Schwab KJ, Gibson KE, Williams DL, Kublicki KM, Lo CP, Mihalic JN, Breysse PN, Curriero FC, Geyh, AS. “Microbial and Chemical Assessment of Regions within New Orleans, LA Impacted by Hurricane Katrina.” Environmental Science and Technology. 41:2401-2406. 2007.
Any relevant financial relationships? No

I agree to comply with the American Public Health Association Conflict of Interest and Commercial Support Guidelines, and to disclose to the participants any off-label or experimental uses of a commercial product or service discussed in my presentation.