Grantee Research Project Results

Development of Rapid Somatic and F+ Coliphages Detection Methods and Comparisons With Existing EPA Methods for Microbial Water Quality Monitoring and Contamination Source Tracking

EPA Grant Number: FP917278
Title: Development of Rapid Somatic and F+ Coliphages Detection Methods and Comparisons With Existing EPA Methods for Microbial Water Quality Monitoring and Contamination Source Tracking
Investigators: Yuen, Yvonne Jia Jia
Institution: North Carolina State University
EPA Project Officer: Packard, Benjamin H
Project Period: August 1, 2011 through July 31, 2014
Project Amount: $126,000
RFA: STAR Graduate Fellowships (2011) RFA Text |  Recipients Lists
Research Category: Academic Fellowships , Fellowship - Water Quality: Coastal and Estuarine Processes

Objective:

The objectives of this research are to develop rapid coliphage detection methods and improve current methods for water quality monitoring and contaminate source tracking. Rapid coliphage detection methods can assist recreational site operators in identifying possible fecal contamination early enough for timely responses of corrective actions or public health warnings. The validity of the developed rapid detection methods will be compared to the current standard EPA coliphage detection methods.

Approach:

Two different approaches will be used to develop new and improved rapid coliphage detection methods: (1) the use of a liquid enrichment culture coupled with coliphage latex-agglutination test (CLAT) or real-time polymerase chain reaction (RT-PCR) assays, and (2) the direct plaque detection approach by improving the single agar layer (SAL) and direct membrane filtration methods to detect color changing or fluorescent signals from the developing plaques. The liquid enrichment and detection procedure involves the amplification of low concentrations of coliphages (as low as one PFU/L) during enrichment with E. coli hosts to levels that can be effectively detected by confirmation steps such as CLAT or RT-PCR. The improvement of the current SAL and direct membrane filter methods involve using fluorogenic or chromogenic additives in non agar-based media and identifying the combinations of optimal water sample volume, incubation conditions, E. coli host strain and membrane filter.

Expected Results:

This research is expected to develop rapid, reliable, cost-effective and simple coliphage detection methods for water quality monitoring or contaminate source tracking. The new rapid methods should have a detection limit that would recognize possible viral contamination early enough for action without compromising the safety of the consumers. These methods should provide simple and direct ways to monitor water quality and are cost effective enough to be used routinely by recreational site operators. The validity of the developed methods will be compared with the current EPA 1601 and 1602 methods using both laboratory spiked samples and field samples. The results from this research also will produce a toolbox of analytical methods that enable users to choose the combinations of procedures that best match their purposes, training and resources by evaluating the critical components of coliphage detection methods.

Potential to Further Environmental / Human Health Protection

Consuming fecally contaminated water can cause diarrheal diseases, which is the fifth leading cause of death in the world. The development of a reliable, simple and cost-effective rapid coliphage detection method can enable communities in low-income countries to test and monitor their drinking, irrigation and recreational water quality before use. The products from this research can also be used by many households in low-income communities or rural areas of the United States that are served by private ground water wells that are not regulated by the states and are not monitored for contamination.

Supplemental Keywords:

water quality, coliphage, fecal contamination

Progress and Final Reports:

2012

2013

Final