Project Proposal for Water Resources Research Institute Program under

Project Proposal
for
Water Resources Research Institute Program
under
Section 104, Water Resources Act of 1984
to the
Alabama Water Resources Research Institute
In support of the
Research Proposal
Identification of pollution sources on agricultural farms and evaluation
of new fecal indicators for surface water quality monitoring
by
Luxin Wang, Ph.D.
Principal Investigator
Assistant Professor
Department of Animal Sciences
Auburn University, Alabama
Telephone (334) 844-8146
Eric Reutebuch
Co-Principal Investigator
Director, Alabama Water Watch
Auburn University, Alabama
Telephone (334) 844-4785
December 2014
A. Project Category: 2 ($20K max.)
B. Project Number
C. Title
Identification of pollution sources on agricultural farms and evaluation of new fecal
indicators for surface water quality monitoring
D. Focus Category
REC; MET; WQL; NPP
E. Descriptors
Agricultural land use, pollution loads, water quality, indicator microorganisms, Enterococcus
F. Duration From March 1, 2015 to February 29, 2016
G. Fiscal Year Federal Funds
Total $19,963
Direct
$19,963
Indirect
H. Non-Federal Funds Allocated
Total $39,290
Direct
$20,073
Indirect
$0
$19,217
I. Name, University, and City of Principal Investigator
Luxin Wang, Auburn University, Auburn, Alabama
J. Congressional District of University Performing the Research
Congressional District 3
K. Water Problem and Need for Research:
As indicated by the “State Water Resources Competitive Grant Program” RFP, one of the
major water resources problems in Alabama is surface water point-source and non-point
source contamination. Although the Good Agricultural Practices have been implemented for
years, studies are still needed in order to develop efficient strategies to minimize the
pollution from agricultural, industrial and urban non-point sources. The research team led by
the two PIs of this proposal conducted a survey in 2014 (funded by USDA) and found that E.
coli concentrations downstream of cattle farms were significantly higher than those upstream
of the farms. The results indicate that, 1) a more efficient surface water quality monitoring
protocol is still needed to better understand the sources and transport of fecal pollution, and,
2) improved methods that can identify the contamination sources are of great importance for
future development and implementation of BMPs.
L. Expected Results, Benefits, Information, etc.:
Upon completion of the proposed research, two outcomes will be generated. The first
outcome will be a better understanding of contamination routes and pollution sources of
surface water will be generated and provide the basis for development of efficient
intervention strategies and on- farm practices. The second will be development of an easy-touse, sensitive and accurate protocol for the Alabama Water Watch and citizen volunteer
2
water monitors to use in addition to the current Coliscan Easygel (E. coli) protocol. The
outcomes of this research will, without a doubt, benefit the sustainability and quality of the
Alabama water resources.
M. Nature, Scope, and Objectives of the Research
The determination of the sources of fecal pollution is a critical issue in complying with the
Clean Water Act (Federal Water Pollution Control Act amendments of 1973 and 1977). It is
necessary to have the ability to differentiate fecal microbial contamination of water runoff
from animal operations versus that from human sources. Escherichia coli (E. coli) has been
used as an indicator microorganism for fecal source tracking because it is easily cultured and
is used as the primary regulatory indicator for pathogen contamination in recreational waters
(Simposon et al., 2002; U.S. EPA, 2002). However, there are problems associated with using
E. coli as a source identifier. These problems include a high degree of genetic diversity not
attributable to a specific host animal source, the potential for E. coli to replicate outside the
host, and geographic and temporal visibilities (U.S. EPA, 2005).
A recent study conducted by Wang and Reutebuch (unpublished) found that the E. coli
concentrations downstream of cattle farms were significantly higher than the E. coli
concentrations upstream of the farms (Figure 1). These results indicated that on-farm
management and good agricultural practices need to be improved in order to lower the fecal
contamination of surface water. The genus Bacteroides has been suggested as an alternative
fecal indicator to replace E. coli or fecal coliforms because they make up a significant portion
of the fecal bacterial population (Fiksdal et al., 1985; Kreader, 1998). Most importantly,
Bacteroides are host specific and can be used as to track the contamination sources (Layton
et al., 2006). Objective 1 of this research is to identify water contamination sources by
collecting and analyzing surface water samples (upstream and downstream) from
different farms and detecting host specific Bacteroides groups via real-time PCR assays.
Farm 1
Farm 2
Farm 3
Farm 4
Figure 1. The concentrations of E. coli present in the upstream (blue bar) and downstream
(green bar) of four cattle farms in Alabama and Georgia (Wang and Reutebuch,
unpublished).
3
Due to the concerns with using E. coli as the indicator organisms, several other genera have
been proposed for use as an alternative indicator for fecal contamination of surface waters.
One of them is Enterococcus. An epidemiological study performed by U.S. EPA
demonstrated a direct relationship between the density of E. coli and Enterococci in surface
waters and an increase in swimmer-associated gastroenteritis (U.S. EPA, 1986). For
freshwater, the current single-sample advisory limits are 235 CFU/100 ml for E. coli and 61
CFU/100 ml for Enterococci (U.S. EPA, 2000). Another recent study conducted by Wang
and Reutebuch (unpublished) found that the concentrations of Enterococci present in
recreational waters were higher than E. coli, which indicates that Enterococcus may serve as
a better indicator microorganism for monitoring fecal contamination in fresh waters, and a
better indicator for AWW volunteer monitors to use. Because the higher the concentration of
the indicator microorganisms, the better the chance of recovering them, we feel that
Enterococcus may make AWW enumeration more reliable. Therefore, Objective 2 of this
research is to develop a water monitoring protocol that is credible and easy-to-use for
AWW water quality monitoring groups using Enterococci as the indicator
microorganism. We will also investigate the feasibility and reliability of volunteer monitors
using Bacteriodes as an indicator of fecal contamination.
N. Methods, Procedures, and Facilities
1. Conducting Bacteriological Sampling on farm
Water samples from upstream and downstream of four farms will be collected in the spring,
summer, fall, and winter of 2015. These water samples will be kept on ice and delivered to
Wang’s microbiology lab located on the Auburn University campus.
2. Enumeration of E. coli
The AWW Coliscan Easygel method will be used to enumerate the number of E. coli present
in the water samples. Concentrations of E. coli measured in the water column will be
interpreted relative to human health by comparison with USEPA and Alabama Department of
Environmental Management water quality criteria (USEPA 2012; ADEM 2012).
3. Enumeration of Enterococci and Comparison of different enterococci enumeration methods
Three Enterococci enumeration methods will be evaluated:
Method 1: Enterolert® Method
One pack of Enterolert® reagent will be added to each 100-ml water sample. The mixture
will be poured into a Quanti-Tray and the tray will be sealed and incubated at 41°C for 24
hours. Enterococci present in the water will react with the Enterolert® reagent and release 4methylumbelliferone that can exhibit fluorescence. The results will be read under a UV lamp
at 365nm and the data will be interpreted following the 51-well Quanti-Tray MPN table.
Method 2: Membrane Filtration Agar Method
A100-ml water sample will be filtered through a 0.45 µm membrane. Following filtration,
the membrane will be placed on mEI agar (membrane- Enterococcus Indoxyl-β-D-Glucoside
Agar) and incubated at 41°C for 24 h. All colonies with blue haloes (regardless of color) will
be recorded as suspect Enterococci colonies. To confirm the suspect colonies, 10 wellisolated colonies will be taken and transferred onto Bile Esculin Agar (BEA). Gram staining
4
will be conducted as well. Colonies will be confirmed as Enterococci by being gram-positive
cocci, and growth and hydrolysis of esculin on BEA.
Method 3: Quantitative Polymerase Chain Reaction (qPCR) Assay
Every 100-ml water sample will be filtered through a 0.45 µm membrane. After that, the
membrane containing the bacterial cells will be used for DNA extraction following the
protocol described by Zhao et al., 2013. Forward primer: 5'GAGAAATTCCAAACGAACTTG-3’ and Reverse primer: 5'CAGTGCTCTACCTCCATCATT-3’ will be synthesized by Integrated DNA Technologies,
Inc. (Coralville, USA). SYBR® Green Master Mix (Life technologies, USA) will be used
and the reaction conditions will follow the Wang lab protocol.
4. Water contamination source tracking
A TaqMan®-based real-time PCR assay will be modified and adopted for the contamination
source tracking. The DNA extraction step will follow the protocol by Zhao et al. 2013.
Primers and probes used for the TaqMan® assay are listed in Table 1. Reaction conditions
will follow Layton et al. 2006.
Table 1. Primers and probes used for the TaqMan® real-time PCR.
5. Facilities
Molecular work will be conducted in Dr. Luxin Wang’s lab. The main 1500-square-foot
research lab is located in Upchurch Hall on the Auburn University campus. It is a certified
BSL II lab equipped with a biosafety hood, one fume hood, -80 and -20 °C freezers, one
double-door refrigerator, one shaking incubator, several regular incubators, Nanodrop, pH
meters, microscopes, a stomacher homogenizer, two centrifuges (high speed, benchtop, with
temperature control), waterbaths, FastPrep®-24 Instrument (a bead beating system for DNA
extraction from environmental samples), ABI thermocycler Veriti Fast, regular gel
electrophoresis units, a denaturing gradient gel electrophoresis unit (DGGE), and a pulse
field gel electrophoresis unit (PFGE). The PI also has easy access to an ABI 7500 real-time
PCR system, a Pathatrix® cell recover system, an Agilent 2100 bioanalyzer and an
autoclave. The lab has two computers installed with different DGGE/PFGE gel analysis
software to facilitate data analysis.
Project bacteriological analyses will be conducted in the Alabama Water Watch Watershed
Stewardship Laboratory in the Center for Advanced Science, Innovation and Commerce
located in the AU Research Park. The building was completed in mid-2013, and the AWW
laboratory is a state-of-the-art facility and is ideal for bacteriological research.
5
O. Related Research
The most closely related research was done by Wang and Reutebuch, the two PIs of this
proposal. In a recent study conducted by Wang and Reutebuch (unpublished), they found that
the concentrations of Enterococci present in recreational waters were higher than E. coli,
which indicated that Enterococci may serve as a better indicator microorganism for fecal
contamination. Another study conducted by Wang and Reutebuch found that the E. coli
concentrations downstream of cattle farms were significantly higher than the E. coli
concentrations upstream of the farms. This finding indicates that more research needs to be
conducted in order to investigate the pollution sources for Alabama surface waters.
Other related research:
Heaney et al. (2014) evaluated the associations between water quality, weather and
environmental factors. The strongest correlation between fecal indicator density in sand and
water was fecal Bacterioides, followed by Enterococci, Clostridium, and Bacteroidales.
Layton A. et al. (2006) designed a real-time PCR assay that targeted Bacteroides species
present in all animals, in humans, and in cattle (AllBac, HuBac, BoBac), which enabled the
quantitative detection of each specific Bacteriodes group. By using this Bacteroides real-time
PCR, one can track the major contamination sources of the water.
References
1. Fiksdal, L., Maki, J. S., LaCroix, J., and Staley, J. T. 1985. Survival and detection of
Bacteroides spp., prospective indicator bacteria. Appl. Environ. Microbiol. 49: 148-150.
2. Heaney, C. D., Exum, N. G., Dugour, A. P., Brenner, K. P., Haugland, R. A., Chern, E.,
Schwab, K., Love, D. C., Serre, M. L., NOvle, R., and Wade, T. J. Water quality, weather
and environmental factors associated with fecal indicator organism density in beach sand at
two recreational marine beaches. Sci. Total Environ. 1: 497-498.
3. Kinzelman, J., Ng, C., Jackson, E., Gradus, S., and Bagley, R. 2003. Enterococci as
indicators of lake Michigan recreational water quality: comparison of two methodologies and
their impacts on public health regulatory events. Appl. Environ. Microbiol. 69(1): 92-96.
4. Kreader, C. A. 1995. Design and evaluation of Bacteriodes DNA probes for the specific
detection of human fecal pollution. Appl. Environ Microbiol. 67: 1171-1179.
5. Layton, A., McKay, L., Williams, D., Garrett, V., Gentry, R., and Sayler, G. 2006.
Development of Bacteroides 16S rRNA gene TaqMan-based real-time PCR assays for
estimation of total, human, and bovine fecal pollution in water. Appl. Environ. Mcirobiol.
72(6): 4214-4224.
6. Simpon, J. M., Santo Domingo, J. W., and Reasoner, D. J. 2002. Microbial source tracking:
state of the science. Environ. Sci. Technol. 36:5279-5288.
7. U.S. Environmental Protection Agency. 2002. Implementation quidance or ambient water
quality criteria for bacteria. EPA-823-B-02-003. http://www.
Epa.gov/ost/standards/bacteria/bacteria.pdf.
6
8. U.S. Environmental Protection Agency. 2005. Microbial source tracking guide document.
EPA/600-R-05-064. www.epa.gov/ORD/NRMRL/pubs/600r05064/600r05064.pdf.
9. U.S. Environmental Protection Agency. 2000. Bacterial water quality standards for
rectreational waters (fresh and marine). http://www.epa.gov/OST/beaches/.
10. U.S. Environmental Protection Agency. 1986. Ambient water quality criteria for bacteria.
EPA/440/5-84/002. Office of Water, U.S. U.S. Environmental Protection Agency,
Washignton, DC.
11. Zhao, L., Tyler, P. J., Starnes, J., Bratcher, C. L., Rankins, D., McCaskey, T. A., and Wang,
L. 2013. Correlation analysis of shiga toxin producing Escherichia coli shedding and fecal
bacterial composition in beef cattle. J. Appl. Microbiol. 115(2): 591-603.
P. Progress Review
N/A
Q. Investigator’s Qualifications (See attached vitae)
R. Training Potential
Wang’s graduate students will be active in field and laboratory activities of this research
project and will be trained in both field and laboratory bacteriological monitoring and
molecular analyses techniques.
Once an Enterococcus, and possibly Bacteriodes, protocol is established, a training workshop
for Alabama Water Watch volunteer monitors will be developed and implemented for
statewide training of citizen monitors.
7
LUXIN WANG,
Ph.D.
Assistant Professor
Department of Animal Sciences
Auburn University
Auburn, AL 36849
EDUCATION
Ph.D. Food Science (Food safety emphasis)
M.S. Biological Engineering
M.S. Food Science (Microbiology emphasis)
B.S. Microbiology
Email: [email protected]
Phone: 334-844-8146 (office)
Fax: 334-844-1519
University of Missouri, USA
University of Missouri, USA
University of Missouri, USA
Shandong Agricultural University, China
EXPERIENCE
Assistant Professor 1/2012-present
Department of Animal Sciences, Auburn University, Auburn, Alabama
Postdoctoral Research Associate 8/2010-12/2011
Mentor: Dr. Linda J. Harris
Department of Food Science and Technology, University of California-Davis, Davis,
California
FDA/CFSAN Fellow 9/2009-8/2010
Mentors: Dr. Samir Assar (U.S. Food and Drug Administration) and Dr. Linda J. Harris (UCDavis) Western Center for Food Safety, Davis, California
Graduate Research Assistant 8/2004-8/2009
Advisor: Dr. Azlin Mustapha
Division of Food Systems and Bioengineering, University of Missouri, Columbia, Missouri
Graduate Research Assistant 8/2007-5/2009
Advisor: Dr. Sherman X. Fan
Department of Biological Engineering, University of Missouri, Columbia, Missouri
AWARDED RESEARCH GRANTS
• Title: Examination of bacterial levels in water and sediment for the development of
refined monitoring protocols for inland recreational waters. (With E. Reutebuch)
Budget: $41,832.00 (2014-2015)
Grant agency: US Geological Survey
• Title: AAES Equipment grant (with Bratcher, C. L., Price, S., and Feng, Y.)
Budget: $14,408.00 (2013)
Grant agency: Alabama Agricultural Experimental Station, USA
• Title: Pathogenicity of Listeria monocytogenes biofilm on meat (with Schwartz, E.)
Budget: $7,500.00 (2013)
8
2009
2009
2006
2004
•
•
•
•
Grant agency: Office of the vice president for research, Auburn University, USA
Title: A systems approach to identifying and filling gaps in and between knowledge
and practice in production and distribution of local and regional foods for a more
secure food supply chain (Bratcher, C. L. (Leading PI), Deutsch, W., Mulvaney, D.,
Singh, M., Weese, S. J., Worosz, M. R., Hanna, J., Tackie, N. O., Bartlett, J. R., and
Halpin, R.)
Budget: $4,818,915.00 (2012-2017)
Grant agency: United State Department of Agriculture (USDA), USA
Title: Investigation of the prevalence and antimicrobial resistant profiles of O157 and
non-O157 shiga toxin producing Escherichia coli from young calves before and after
weaning stresses (with Bratcher, C. L., and Rankins, D.)
Budget: $49,970.00 (2012-2014)
Grant agency: Alabama Agricultural Experimental Station, USA
Title: Animal management as a strategy to limit carriage of E. coli O157:H7 and
Salmonella in beef cattle (with McCaskey, T.)
Budget: $21,000.00 (2012)
Grant agency: Alabama Beef Forage Initiative, USA
Title: Influence of the pre-harvest environment on the physiological state of
Salmonella and its impact on increased survival capability (with Harris, L. J.)
Budget: $97,078.00 (2010-2012)
Grant agency: The Center for Produce Safety, USA
PUBLICATIONS
1. Guo, M., Jin, T. Z., Wang, Luxin, Scullen, C. J., and Sommers, C. H. Antimicrobial
films and coatings for inactivation of Listeria innocua and Salmonella Typhimurium on
Ready-to-eat meat. Accepted by Food Control in November 2013.
2. Guo, M., Jin, T. Z., Geveke, D., Fan, X., Site, J., and Wang, Luxin. 2013. Evaluation of
microbial stability, bioactive compounds, physicochemical properties, and consumer
acceptance of pomegranate juice processed in a commercial scale pulse electric field
system. Food and Bioprocess Technologies. Published online Oct. 1st.
3. Zhao, L., Tyler, P. J., Starnes, J., Rankins, D., McCaskey, T. A., and Wang, Luxin.
2013. Evaluation of weaning stress on Escherichia coli O157 shedding, body weight and
fecal bacterial communities in beef calves. Foodborne Pathogens and Disease. Epub
ahead of print, Oct. 18th.
4. Zhao, L., Tyler, P. J., Starnes, J., Bratcher, C. L., Rankins, D., McCaskey, T. A., and
Wang, Luxin. 2013. Correlation analysis of shiga toxin producing Escherichia coli
shedding and fecal bacterial composition in beef cattle. Journal of Applied Microbiology.
115(2): 591-603.
5. Harris, L. J., Berry, E. D., Blessington, T., Erickson, M., Jay-Russell, M., Jiang, X.,
Killinger, K., Michel, F.C., Millner, P., Schneider, K., Sharma, M., Suslow, T.V.,
WANG, Luxin, and Worobo, R. W. 2013. A framework for developing research
protocols for evaluation of microbial hazards and controls during production that pertain
to the application of untreated soil amendments of animal origin on land used to grow
produce that may be consumed raw. Journal of Food Protection. 76(6): 1062-1084.
9
6. Kimber, M. A., Kaur, H., WANG, Luxin, Danyluk, M. D., and Harris, L. J. 2012.
Survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on
inoculated almonds and pistachios stored at -19, 4, and 24 °C. Journal of Food
Protection. 75(8): 1394-1403.
7. WANG, Luxin, Wu, C., Fan, X., and Mustapha, A. 2012. Detection of Escherichia coli
O157:H7 and Salmonella in ground beef by a bead-free quantum dot-facilitated method.
International Journal of Food Microbiology. 156: 83-87.
8. WANG, Luxin, and Mustapha, A. 2010. EMA real-time PCR as a reliable method for
detection of viable Salmonella in chicken and eggs. Journal of Food Science. 75(3):
134 - 139.
9. WANG, Luxin, Li, Y., and Mustapha, A. 2009. Detection of viable Escherichia coli
O157:H7 by ethidium monoazide real-time PCR. Journal of Applied Microbiology.
107(5): 1719 - 1728.
10. WANG, Luxin, Li, Y., and Mustapha, A. 2007. Rapid and simultaneous quantitation of
Escherichia coli O157:H7, Salmonella and Shigella in ground beef by multiplex real-time
PCR and immunomagnetic separation. Journal of Food Protection. 70(6): 1366 - 1372.
PROFESSIONAL ACTIVITIES
Reviewer, the “Food Control”
9/2013-present
Reviewer, the “Journal of Food Measurement & Characterization”
2013-present
Reviewer, the “Journal of Animal Science”
3/2013-present
Reviewer, the “Sensing and Instrumentation for Food Quality and Safety”
1/2011-present
Reviewer, the “Journal of Food Science”
4/2011-present
Reviewer, the “International Journal of Food Microbiology”
8/2011-present
Reviewer, the “Journal of Food Processing and Preservation”
6/2011-present
PROFESSIONAL MEMBERSHIPS
Member, the American Meat Science Association
Member, the Alabama Cattlemen’s Association
Member, the International Association for Food Protection
Member, the Institute of Food Technologists
10
6/2012-present
12/2012-present
7/2010-present
12/2004-present
Eric Martin Reutebuch
Research Associate IV
Auburn University Water Resources Center
Auburn University, Auburn, AL 36849
E-mail: [email protected]; Phone: (334) 844-1163
Education:
B.S., 1981, Purdue University, Indiana (Biology)
M.S., 1988, Auburn University (Fisheries)
Professional Experience:
2014-present Director, Alabama Water Watch Program
2013-present Associate Director, Alabama Water Watch Program
1989-2013
Research Associate, Department of Fisheries and Allied Aquacultures, AU, AL.
1988-1989
Aquaculture Specialist, U of Georgia Coastal Plain Experiment Station, Tifton, GA.
1986-1987
Graduate Researcher in Fisheries, Auburn University, AL.
1982-1984
Fisheries Extensionist, U.S. Peace Corps/Nepal.
1980
Research Technician, Botany Department Purdue University, IN.
Significant Honors and Awards:
• President, Save Our Saugahatchee, Inc., 2014-present
•
Board of Directors, Lake Watch of Lake Martin, Inc., 2010-present
•
Woodfin Martin Lake Stewardship Award from Lake Watch of Lake Martin, 2009
•
Board of Directors, Save Our Saugahatchee, Inc., 2003-2014.
•
Board of Directors, Alabama Water Watch Association, 1997-1998.
•
H. S. Swingle Award for Highest Academic Achievement at the Master of Science Level 1987, from the Department of Fisheries and Allied Aquacultures, Auburn University, AL.
•
National Dean’s List, Phi Eta Sigma, Purdue 500.
•
Graduated Summa Cum Laude, Purdue University, 1981
Professional Development Activities (Post-Master’s Degree):
• STEPL and Region 5 Modeling and Load Reduction Training, ADEM, 2005.
•
MS Access 2000: Understanding Relational Database Design, AU at Montgomery, 2002.
•
Understanding GIS, the Arc/Info Method, Auburn University, 1996.
11
•
Interpretation of Aerial Photography and Remote Sensing Imagery, Auburn University,
1995.
•
Geographic Information Systems, Auburn University, 1995.
•
Problem Solving with the Instream Flow Incremental Methodology, U.S. Fish and Wildlife
Service, 1992.
•
Hydrology and Climatology, Auburn University, 1991.
•
Management of Aquatic Flora, Auburn University, 1991.
•
Management of Fish Populations in Small Impoundments, Auburn University, 1991.
•
SAS Programming, Auburn University, 1991.
•
Stream Ecology, Auburn University, 1990.
Synergistic Activities:
1. Reutebuch leads the Alabama Water Watch Program, a citizen volunteer water quality
monitoring program that promotes improved water quality and water policy through training
and certifying volunteer monitors throughout the state (see www.alabamawaterwatch.org).
2. Reutebuch facilitates the National Integrated Drought Information System’s Drought Early
Warning for the Apalachicola-Chattahoochee-Flint River Basin Webinar on a monthly basis.
3. Reutebuch serves as President of Save Our Saugahatchee, a volunteer water monitor
organization that conducts water monitoring throughout the Saugahatchee Creek Watershed
in the Auburn/Opelika area. Other watershed stewardship activities include stream cleanups,
outreach presentations, and working on improvements in local and state water policy.
4. Reutebuch serves on the Board of Directors of Lake Watch of Lake Martin, a volunteer water
monitor organization that conducts water monitoring throughout the Lake Martin Watershed.
Other watershed stewardship activities include outreach presentations, environmental
education at local schools, and working on improvements in local and state water policy.
Products (select):
(i) Peer-Reviewed Publications:
1. Saafeld, D. T., E. Reutebuch, R. J. Dickey, W. C. Seesock, C. E. Webber, and D. R.
Bayne. 2012. Effects of Landscape Characteristics on Water Quality and Fish
Assemblages in the Tallapoosa River Basin, Alabama. Southeastern Naturalist,
11(2):239-252.
2. Dougherty, M., D. R. Bayne, L. Curtis, E. Reutebuch, W. C. Seesock. 2007. Water
quality in a non-traditional off-stream polyethylene-lined reservoir. Journal of
Environmental Management, 85:1015–1023.
3. Deutsch, W., E. Reutebuch, and S. Ruiz-Córdova. 2007. Validity and Applications of
Citizen Volunteer Water Quality Data: A Case from Alabama. Water Resources Impact,
9(5) 16-20.
12
4. Bayne, D. R., E. Reutebuch and W. C. Seesock. 2002. Relative motility of fishes in a
Southeastern Reservoir based on tissue polychlorinated biphenyl residues. North
American Journal of Fisheries Management, 22:122-131.
5. McGregor, M. A., D. R. Bayne, J. G. Steeger, E. C. Webber and E. Reutebuch. 1996.
The potential for biological control of water primrose (Ludwigia grandiflora) by the
water primrose flea beetle (Lysathia ludoviciana) in the southeastern United States.
Journal Aquatic Plant Management, 34:74-76.
(ii) Other significant products:
1. Dyson, D., T. Muenz and E. Reutebuch. 2014. Forest Management and Stream Water
Quality. In Alabama’s Treasured Forests - A Publication of the Alabama Forestry
Commission, pp. 21-23.
2. Reutebuch, E. M. 2012. Citizen Volunteer Watershed Stewardship of Alabama’s
Reservoirs – Lake Martin Watershed. Alabama Water Watch, Auburn, AL. 32 pp.
3. Reutebuch, EM, W Deutsch, W Seesock, G Lockaby, C Anderson, R Governo, C Nagy,
and J D’Angelo. 2010. Saugahatchee Watershed Management Plan - Final Report.
Alabama Department of Environmental Management, Montgomery, AL. 183 pp.
4. Reutebuch, EM, WG Deutsch and S Ruiz-Córdova. 2008. Community-Based Water
Quality Monitoring- Data Credibility and Applications. Alabama Water Watch, Auburn,
AL. 24 pp.
5. Deutsch, W, EM Reutebuch, and S Ruiz-Córdova. 2007. Validity and Applications of
Citizen Volunteer Water Quality Data: A Case from Alabama. Water Resources Impact,
9(5): 16-20.
6. Reutebuch, EM, WG Deutsch and S Ruiz-Córdova. 2007. Citizen Volunteer Water
Monitoring on Wolf Bay. Alabama Water Watch, Auburn, AL. 8 pp.
7. Reutebuch, EM and WG Deutsch. 2007. A Transferable Model of Stakeholder
Partnerships for Addressing Nutrient Dynamics in Southeastern Watersheds – Final
Report – 2006-2007. U. S. Department of Agriculture Cooperative Research, Education
and Extension Service, Washington D. C. 96 pp.
8. Reutebuch, E., R. Estridge and W. Deutsch. 2005. Saugahatchee Creek Watershed –
Past, Present, & Future. Sponsored by the ADEM, Montgomery, AL, 20 pp.
13
Budget Form
BUDGET
REGIONAL WATER RESOURCES RESEARCH PROJECT
FISCAL YEAR
Proposed Starting Date: March 1, 2015
Proposed Completion Date: February 29, 2016
Project Title: Identification of pollution sources on agricultural farms and evaluation of new
fecal indicators for surface water quality monitoring
Principal Investigator: Luxin Wang
Budget
Federal
1. Salaries & Wages
Principal Investigator (L. Wang, 1 month)
Other Professional Staff (E.Reutebuch, 0.5 months)
Hourly worker (100 HOURS @ 10$/hr)
AWW volunteer time (training and monitoring $20.50/h*)
Total Salaries & Wages
Non-Federal
$
$
$
7,891
4,017
1,000
$
$
$
$
$
16,571
9,909
1,000
984
28,464
$
12,908
$
3,691
$
8,209
Coliscan bacteria testing supplies for AU researchers (3-4
farms*4 dates*2 samples/site)
Standard Methods bacteria testing supplies
Rea-time PCR reagents
Total Supplies
$
$
$
$
800
1,260
560
2,620
$
$
$
$
-
$
$
$
$
800
1,260
560
2,620
4. Equipment
$
-
$
-
$
-
5. Subcontracts/Consultants
$
-
$
-
$
-
6. Travel
4 trips to 4 farms (200 milesx4 @ $0.82/mile)
Per diem ($11/day*4 trips*2 researchers)
Total Travel
$
$
$
656
88
744
$
$
$
-
$
$
$
656
88
744
7. Other Direct Costs
$
-
$
-
$
-
8. Total Direct Costs
$
19,963
$
20,073
$
40,036
9. Forfeited IDC on WRRI FEDERAL FUNDING
$
-
$
9,582
$
9,582
$
9,635
$
9,635
$
39,290
$
59,253
2. Fringe Benefits (31% for faculty & staff)
$
$
$
$
$
8,680
5,892
984
15,556
Total
4,517
3. Supplies
10. IDC on AU COST SHARE
11. TOTAL ESTIMATED COST
$
14
19,963
Budget Justification
Item #3: Supplies:
1. AU Researcher Coliscan Easygel sampling supplies:
(4 farms*4 dates*2 samples/site) = 32 samples done in triplicate (32*3=96)
Order enough for 100 samples @ $8 per triplicate sample = $800
2. Enterococcus monitoring, three methods
For 32 samples done in duplicate (32*2=64). Order enough supplies for 70 samples
$18 per sample for disposables/agar/petrifilms = $18*70= $1,260
3. Real-time PCR supplies
70 Samples*$8= $560
_______________________________________________________________________________________________________________________________________
Total = $2,620
Item #6: Travel:
1. 4 trips to 4 farms (200 miles*4 @$0.82/mile)=$656
2. Per diem @ $11/day * 4 trips * 2 researchers = $88
_____________________________________________________________________________________________________________________________________________
Total = $744
15
Monday, December 1, 2014
Dr. Wang and Eric Reutebuch,
The AU Water Resources Center will match 1.1 months at $5,356/month plus 31% fringe of
Eric Reutebuch’s salary and fringe for your 2015 WRRI proposal.
Sincerely,
Samuel R. Fowler, Director
Auburn University Water Resources Center
And Associate Professor, Ag. Econ & Rural Soc.