Progress 09/15/06 to 03/14/09
Outputs
OUTPUTS: The research team evaluated different formulations of pectin-based medium using a texture analyzer to determine gel strength and compared these formulations with 0.9% agar. Different bacterial stains and chromgenic substrates have been evaluated and an optimal formulation has been determined and used to produce prototype kits. The prototype of EasyPhage kit has been designed and produced. The kit was first introduced in the General Meeting of American Society for Microbiology, May 2007 in Toronto, Canada. More than 50 kits were sent out for ASM attendants to try out. Beta-testing with 3 independent laboratories has been executed and the results are analyzed. A national wide survey has been completed and more than 200 samples were tested for E. coli, male-specific and somatic coliphages, and human enteroviruses. All field samples were analyzed human enteroviruses by RT-PCR and all RT-PCR positive samples were analyzed by the cell culture infectivity assay. Selected negative samples from each type of vegetables were also analyzed by cell culture infectivity assay. All results are analyzed and final report is preparing. PARTICIPANTS: Dr. Fu-Chih Hsu, PI Dr. Choi-Iok, Co-PI Ms. Alicia Jones, Technician Ms. Elizabeth Angerman, Technician Ms. Susan Shepard, Technician Mr. Everett Chu, Technician Dr. Jonathon Roth, partner, Micorology Laboratories LLC Dr. Daniel Fung, Consultant, Kansas State University TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: The research team has underestimated the time and labor involved in the field study. It was not possible to analyze 800 samples proposed in the research plan; therefore, we had to scale back to 200 samples and request 6 month no-cost extension. During the field study, the research team found out that somatic coliphages (not proposed in the original research plan) were more prevailing than male-specific coliphages. We decided to analyze somatic coliphages as well that had an impact to be able analyze more field samples. However, the research team have made a right decision because the results from somatic coliphages provide additional information.
Impacts
The Easyphage kit (pectin-based medium) developed in this research has shown several great advantages in detection of fecal contamination in fresh produce as well as water samples. It is easy and more sensitive than current methods available. Two different kits have been developed for different market purposes and these kits have been used in education, research, and monitoring. Several states have shown interest to use Easyphage for monitoring ground water. In the field study, the research team has generated many useful findings in ready-to-east fresh produce. Some of fresh produce poses a significant high risk for human consumption due to extremely high concentrations of E. coli, coliphages, or positive for human enteroviruses. The method developed in this research is effectively to recover and detect human enteroviruses from fresh produce. It can be a standard method for recovery and detection of human enteric viruses from fresh produce. Human enteroviruses were detected in some samples either by RT-PCR or cell culture infectivity assay. These findings should help the regulatory agents to implement a better regulation to monitor biological safety for fresh produce.
Publications
- Choi-Iok Wong, Dave Battigelli, Fu-Chih Hsu. Development of a simple, quantitative kit to test vegetables and fruits for evidence of fecal contamination. Poster in the General Meeting of American Society for Microbiology 2007, Toronto, Candida.
- C.-I. R. Wong, J. Roth, E. Angerman, J. Larkin , and F.-C. Hsu. Detection of Coliphages from Water Using Pectin-Based Medium. Poster in the General Meeting of American Society for Microbiology 2008, Boston, MA.
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Progress 09/15/06 to 09/14/07
Outputs
SMI has demonstrated EasyPhage kits during May 2007 at the General Meeting of American Society for Microbiology and also present a poster. Many microbiologists from education and research universities as well as other research laboratories visited the SMI booth and requested a sample kit. SMI has sent out more than 50 kits. Largely as a result of significant interest at the 2007 ASM General Meeting, researchers at the following companies and universities have received one or more EasyPhage kit: Ivy Tech College, Tufts University, Codexis (CA), Metropolitan Water Reclamation District, University of Washington -Seattle, Texas A & M, North Carolina State Univ -Food Science Dept., Utica College, Los Angeles County Sanitation Districts, Dalynn Biologicals, Texas A & M University, Auburn University, Ohio Wesleyan University, Virginia Tech, University of Guelph, University of South Florida, American Water Works SVC Co., Inc, Computer Science Corp, Food & Drug Administration,
BCS Laboratories, University of Washington, University of Wisconsin/State Lab of Hygiene, USGS, Southeast Missouri State University, US Geological Survey, 3M Center Canadian Research Institute for Food Safety, Cook College, Rutgers University, North Shore Community College, Mary Mount University, Public Health Agency of Canada, Natl. Center for Toxicological Research, University of Mississippi State, Kirkegard & Perry Laboratories. Inc, Sul Ross State University We are actively following up to receive customer feedback and determine their interest in purchasing additional EsayPhage products. About 30 EasyPhage kits have been sold for educational and research purposes. It is worth noting that a pharmaceutical company has recently purchased 7 kits to detect somatic coliphages from genetically modified E. coli fermentation.
Impacts
The research continues to closely follow the Phase II project's work plan. Currently, samples of the EasyPhage educational kit have been offered to science teachers and college professors. A soft launch of the product has taken place and a dozen 10 test kits have been purchased. The EasyPhage 10 test kit can be purchased at www.scientificmethods.com and a permanent website with an automated shopping cart is nearly complete at www.micrologymethods.com. A second product configuration, having 100 ml of medium rather than 10 ml, is under development for food testing labs and for water testing labs. This new format will be allow a more economical approach to testing food for the presence of these indicator viruses that are documented to be associated with the presence of fecal contamination and pathogenic viruses. These viruses are also more resistant to desiccation than E. coli bacteria and should indicate fecal contamination even if the E.coli has perished en route to the
consumer. After SMI conducts next summer's national food basket survey, meaningful range values will be generated that will permit users to compare their coliphage results to historical food samples results. This tool will allow food distributors and processors to better protect their customers from food that may be contaminated with pathogenic viruses and fecal matter.
Publications
- EVALUATION OF FUNG DOUBLE TUBE TEST FOR CLOSTRIDIUM PERFRINGENS AND EASYPHAGE TEST FOR F-SPECIFIC RNA COLIPHAGES AS RAPID SCREENING TESTS FOR FECAL CONTAMINATION IN RECREATIONAL WATERS OF HAWAII, Journal of Rapid Methods and Automation in Microbiology Vol 15, pp 217, 2007
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