CICEET Progress Report for the period 9/02/07 Through 3/01/08
Project Title: Detection of Methanogen Markers for Microbial Source Tracking
Principal Investigator(s): Jennifer A. Ufnar, Ph.D.
Additional Investigator(s): R.D. Ellender, Ph.D.
Project Start Date: May 1, 2007
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Objectives
Test all samples from SCCWRP (Doheny and Avalon Beach) using assays for host-specific methanogens Ufnar et al., 2006; Ufnar et al., 2007; Ufnar et al., 2007) and Bacteroides human primers (Bernhard and Field, 2000).
Tasks to meet objectives
a. Process all samples sent by SCCWRP within 24 hours of receipt.
b. Perform DNA extractions using QBiogene Fast DNA Spin Kit for Soil or MO BIO Powersoil DNA kit) on all samples.
c. Run PCR and gels for Mnif and Bacteroides assays
d. Prepare data sheet for all data collected
e. Run PCR and gels for Swine methanogen primers and M. ruminantium primers
f. Transfer grant from University of Southern Mississippi to Vanderbilt University Medical Center
Progress on Tasks
a. All Doheny and Avalon Beach samples have been processed to date (through DNA extraction, PCR, and gel electrophoresis).
b. PCR and gels have been run on all Doheny Beach samples and data reported in Sepember 1, 2007 progress report.
c. All sample processing, PCR, and gel electrophoresis were completed for Avalon Beach samples.
d. All data are reported for Avalon Beach samples in Table 1.
e. PCR and gel electrophoresis on Doheny and Avalon beach samples using swine and bovine-specific primers is continuing. Data will be reported as it is collected.
Have the results/data gathered during this reporting period changed the project objectives when compared to your original proposal? Please explain.
a. No.
Dissemination activities during this reporting period (please include the number of participants where applicable).
Outreach Activities
Two high school seniors will be trained to perform all assays.
Student activity (e.g. theses, dissertations, etc.) on the project (please identify students as graduate or undergraduate)
Undergraduate student working on this project will complete research hours for the summer using the project as a learning tool.
Difficulties
None encountered in this last reporting period.
Data Generated to date
See Table.
Project Objectives for Next Reporting Period
Objectives
a. Perform swine and ruminant methanogen assays for all Doheny and Avalon Beach samples
b. Report all findings to CICEET
Work Plan to Meet Objectives
a) The grant has been transferred in the last week and work will commence March 1, 2008 using the swine and bovine primers.
b) Retest of any samples will be performed in the first week of April, 2008.
Dissemination Objectives for next reporting period
a) Dr. Ufnar will work with other investigators to compare Bacteroides and Mnif data to determine sensitivity and specificity of each method.
b) Outreach activities will include mentoring gifted high school students on the project and bringing the students into the laboratory to learn techniques.
Overall Project Timeline Update
The project is proceeding according to the original timeline with the exception of the swine and bovine assays. These were delayed due to transfer of grant funds. The timeline is now updated to include laboratory work through April, 2008.
Expenditures
Expenditures are in the anticipated range.