InSites is a quarterly newsletter that highlights the personalities and projects of the Waste Management Research and Education Institute (WMREI) of The University of Tennessee. WMREI is an affiliate of the EERC.
WMREI was created in 1985 as a state-funded Center of Excellence. Research areas include solid-, hazardous-, and nuclear-waste management; waste minimization; and pollution prevention.
Biotechnology is the focal point of the institute's technical research, while issues involving public attitudes and federal/state policies related to waste-management issues are the primary concerns of the institute's policy research.
For additional information about InSites, or to be added to our mailing list, please write InSites, WMREI, The University of Tennessee, 311 Conference Building, Knoxville, TN 37996-4134, call 865-974-1156, or fax 865-974-1838. Or, if you prefer, email us.
Table of Contents
Drinking-Water
Dragnet ,
By Kris Christen
Nothin but Blue Skies
,
Chris Wohlwend Probing
Outer Space from Deep Underground
Staff Citings 
Do waterborne pathogens leave fingerprints? UT investigators insist they do and are determined to help resource managers identify—and eradicate—the microbes that pollute Tennessee’s rivers and lakes.
By Kris Christen
All too often, stream banks are dotted with signs warning against physical contact with the water because of bacteriological contamination. Such warnings underscore the fact that pathogenic (disease-causing) microorganisms, together with sediments and nutrients, make up the top three pollutants stressing Tennessee’s waterways.
And once testers have documented the presence of such microorganisms, state waterquality managers have no choice but to close a lake or stream to recreation until the risk of human infection passes. Meanwhile, drinkingwater utilities that draw water from these sources must often apply an extra dose of disinfecting chlorine before distributing the water to customers.
Determining the presence of these bacterial contaminants is relatively easy. Identifying their sources represents a stiffer challenge. Indeed, current methods for detecting pathogens can indicate only their presence, not where the organisms might be coming from, says Alice Layton, a microbiologist at the University of Tennessee’s (UT’s) Center for Environmental Biotechnology (CEB).
And without that key information, water-quality managers are often hampered in their efforts to quickly and efficiently clean up polluted waterways.
“If you can identify that the pathogens are coming from human sources, then you can look at where sewer lines or septic fields might be leaking and whether you need to do something to repair them,” says Larry McKay, a CEB hydrogeologist.
“If, on the other hand, it turns out that the microbes are actually coming from cattle in the upper reaches of a watershed or just dogs and cats, then fixing the problem would cost millions of dollars less,” he says, because repairing sewer lines can be particularly expensive.
FEDERAL LAW,
STATE RESPONSIBILITY
Under the federal Clean Water Act, the Tennessee Department of Environment and
Conservation (TDEC) is required to assess the quality of all the state’s
waterways. Those found to be impaired wind up on a cleanup list.
“Once we put a stream on that list, we have to do a study on it called a total maximum daily load (TMDL),” says Jonathon Burr, one of TDEC’s environmental specialists. TMDLs are essentially a pollution budget designed to help states determine how much various pollutants must be reduced to meet water-quality standards in a given waterway.
Burr’s agency is hoping that CEB researchers will be able to develop a testing tool that will help distinguish whether bacterial pollution loads are coming from human or animal sources, and, in the latter case, what kind of animals.
If successful, the work will not only help Tennessee better manage its contaminated watersheds, it will also assist other states with their water-quality goals, says Gary Sayler, CEB’s director.
“This is pretty significant in the sense that virtually
all communities across the United States are dealing with the issue of trying
to characterize sources of different wastes, because they have to meet these
TMDL limits,” Sayler explains.
CURRENT TECHNIQUES
The U.S. Environmental Protection Agency has approved a number of assays for
determining pathogenic presence in water samples, Layton notes. Often, these
assays work indirectly, identifying classes of organisms rather than specific
pathogens. For instance, the presence of E. coli, bacteria commonly found in
feces, indicates that favorable conditions are present for other enteric pathogens
as well. Other assays test for total coliforms, fecal coliforms, and Enterococcus.
Such indicator tests are used because direct monitoring of individual pathogens is expensive, and so many pathogens exist that it would be impossible or impractical to monitor for all of them. But, Layton cautions, the presence of indicator bacteria does not mean that other pathogens are present in a stream; it means only that conditions are favorable for supporting them.
“Certain strains of E. coli are pathogenic, but the vast majority aren’t,” she explains, which raises the risk of false-positive tests, using current detection methods.
“The assay also doesn’t indicate the quantity
of feces present in a waterway or where the feces are coming from,” says
Layton. “But it’s currently the best measure we have.”
REFINING THE TOOLS
The CEB researchers aren’t looking to replace existing indicator tests
but merely to take them a step further using other fecal bacteria, such as Bacteroides,
to quantify the amount of fecal contamination present, as well as its source.
These bacteria typically exist in high concentrations, composing as much as 10–20 percent of fecal mass, and are host specific, Layton says. Moreover, they don’t survive long outside of their hosts, which means that they don’t accumulate over time, “giving you a better indicator of how long ago a contaminating event occurred.”
Using a technique called the real-time polymerase chain reaction (PCR) method, researchers are able to actually locate an organism’s genes. “So, we’re going from just seeing if an organism is there to being able to quantify it and identify its source,” Layton says. The advantage of real-time PCR is that it offers a quantitative, specific, sensitive, and fast measurement, with analysis times of less than two hours, compared with 18 to 24 hours for traditional culture- based methods.
Using PCR, the CEB researchers have been developing and testing
a series of different microbial assay techniques specific to cattle, horses,
humans, dogs, fish, and birds. So far, their initial studies indicate that the
different sources can be readily distinguished, and they’ve been testing
these techniques in a study of fecal contamination in a small, rural Tennessee
watershed.
FIELD TESTING
Rural areas are especially at risk, says McKay, because so many people drink
untreated water from wells and springs in these places.
“We want to improve the source water quality so those people aren’t getting exposed to dangerous contaminants,” he says.
With help from TDEC, the researchers have chosen Stock Creek, part of a typical watershed in East Tennessee, as the site for testing their method.
“It’s rural and very diverse, with a few houses, some small farms, and people who own horses,” Layton notes. It’s also an area likely to grow rapidly over the next few years.
Over the past year, the group has sampled 16 sites on a monthly
basis, with six sites being on the main creek and 10 on tributaries and an impounded
area of the creek. While Layton focuses on the occurrence of pathogen indicators,
McKay and others are focusing on the hydrology of the system and trying to understand
the mechanisms responsible for transporting these pathogens.
UNIQUE HYDROLOGY
“We’ve found that the hydrology of East Tennessee is particularly
susceptible to microbial contamination,” McKay says. “We have a
lot of very fast runoff, with a lot of stuff getting washed off the soils in
fields and into streams quickly because of the steep slopes and clay rich soils.”
Additionally, the karst bedrock that underlies the region is pocked with large fractures, caves, and conduits where flow can travel extremely fast–as much as a mile a day in some cases, McKay notes. “So there’s very little time for filtration or natural die-off of pathogens to occur.” As a result, another part of the project involves collecting flow measurements at each of the sampling sites, as well as data about basic water chemistry, including pH, dissolved oxygen, and conductivity measures.
“These chemical signatures can tell us what the source of the water is and what other stresses might be present in the environment besides the pathogens we’re worried about,” says John McCarthy, a CEB research professor with UT’s Department of Earth and Planetary Sciences.
For example, he says, “if you think water may be entering the creek from some of the cracks and fissures in the karst drain, there would be telltale signatures in the chemistry.”
These signatures could point to where the pathogens are coming from, what controls their transport, and how waterquality managers might mitigate them. Most of the sampling to date has involved base stream flows, but “we’re going to be doing more storm chasing this year,” McCarthy notes. As water moves across the landscape during large rain events, “things are mobilized or fall down into the karst terrain and come back up in the creek,” he says, “and we want to evaluate how storms versus base flows contribute to the pathogen problem.”
A nice feature of the project lies in the fact that although some of the findings will be specific to the Stock Creek watershed, others will be transferable.
“Overall, I think this approach is going to provide
a good model for doing source identification and evaluating waste loading into
streams from different types of landuse areas,” says Randall Gentry, an
assistant professor with UT’s Department of Civil and Environmental Engineering.
QUICK, CONVENIENT TESTING
One of the next items on the research agenda is to develop the Bacteroides assays
so they are easily deployable in the field.
“We’d like to get these tools in the hands of people who are more routinely going out and sampling,” Layton says. “If you can get results out in the field, you can decide right there if you need to collect more samples or do something else.”
This would also solve a practical problem in that water samples have a limited holding time of six hours, which puts TDEC in a tight spot.
“It can be difficult to get samples taken from some rural parts of Tennessee back to a laboratory within the required timeframe,” McKay explains. “If we had portable field tests, we’d be able to sample over a much wider range.”
Additionally, over the long-term, “we’d like to link this research more directly to health-related issues,” McKay notes. Currently, waterborne disease outbreaks are notoriously difficult to diagnose because the source of the disease may have disappeared.
“We’d like to be able to set up monitoring systems, so that when an outbreak does occur, we could test a whole series of wells very rapidly and very specifically,” says McKay, “not just for E. coli, but for specific types of viral pathogens, like hepatitis.”
* * *
Contact Alice Layton, CEB, The University of Tennessee, 676 Dabney Hall, Knoxville, TN 37996-1605, call 865-974- 8080, or e-mail alayton@utk.edu. Or contact Larry McKay, CEB, The University of Tennessee, 306 Earth and Planetary Sciences Bldg., Knoxville, TN 37996-1410, call 865-974-0821, or e-mail lmckay@utk.edu.
The Department of Energy recently awarded Clean Cities designation and potential federal dollars to the University of Tennessee’s East Tennessee Clean Fuels Coalition.
by Chris Wohlwend
EAST TENNESSEE’S AIR IS AMONG THE worst in the nation, but that might start improving now that the region has been designated an official participant in the U.S. Department of Energy’s (DOE) Clean Cities Program.
A plan submitted by the East Tennessee Clean Fuels Coalition (ETCFC) qualified the region to participate. ETCFC counts 32 area organizations among its membership, ranging from the Tennessee Valley Authority (TVA) to Great Smoky Mountains National Park to Knoxville Area Transit system, better known as KAT.
The designation, according to ETCFC Executive Director Jonathan Overly, means that the coalition “will become eligible for a bevy of benefits, including access to federal funding.
“We can now apply for a pocket of funds that is available only to designated coalitions,” Overly explains. “It’s not guaranteed, but at least we’re now eligible to apply. We will also be able to tap into the network of 70-plus active coalitions across the country that have already addressed many of the hurdles we will encounter.”
The Clean Cities Program is an outgrowth of the Energy Policy Act of 1992, which grew out of the Gulf War. The objectives are to reduce U.S. dependence on foreign oil, improve air quality, and enhance local economic activity by supporting public-private partnerships that deploy alternative fuel vehicles (AFVs) and build supporting infrastructure.
To become eligible to participate, organizations must put together a coalition of stakeholders, assess the local or regional alternative fuels markets, ensure a strong market foundation by building stakeholder commitments, and create a plan to expand alternative fuels use. Nationwide, 83 organizations have received the DOE designation since 1993.
A Tiger on the Team
Besides TVA, Great Smoky Mountains National Park, and KAT, the ETCFC membership
includes the Knoxville Utilities Board (KUB), Oak Ridge National Laboratory
(ORNL), the State Energy Program (SEP), two propane distributors, five environmental
organizations, one car dealership, and the cities of Knoxville, Gatlinburg,
Pigeon Forge, and Sevierville.
One of the benefits of Clean Cities designation is that DOE provides “tiger teams” to help member organizations. “When you get into a sticky problem,” Overly explains, “they will send a technical person or team to help at no cost to us; that’s not something we would need often, but when you need it, it’s important.”
East Tennessee’s designation fills a geographic hole in the program.
“The southeast has been behind the rest of the nation, but we’re rapidly catching up,” says David Dunagan, program manager for DOE’s Southeast regional office in Atlanta. “East Tennessee is central to our plan to form clean-fuel corridors along the Interstates. This designation should be a strong catalyst for the rest of the state and for the region, so that we can form a network of cities along the Interstates.”
Cynthia Oliphant, director of the energy division of the Tennessee Department of Economic & Community Development, agrees. The designation, she says, is “a positive step for Tennessee and a strong addition to the national Clean Cities initiative. By having designated Clean Cities groups in Tennessee, it gives alternative fuel use, and subsequently air quality and energy security, a boost statewide.”
Fill ’er up Green, Please
Alternative fuels currently in use are natural gas, biodiesel, propane, ethanol,
and electricity. Ethanol and biodiesel are renewable. Ethanol can be made from
potatoes, scrap wood, and other biomass materials, though it is primarily made
from corn in the United States. Biodiesel is typically made from waste cooking
oils—“yellow grease”—or virgin oils made from vegetables
such as soybeans. Though natural gas, propane, and electricity are not renewable,
they are produced domestically and burn much cleaner than gasoline.
“We hope that hydrogen will be the long-term answer,” says Overly, “but the other five are being used now.”
Each of the five fuels has pluses and minuses. “Natural gas is the cleanest,” Overly says, “but it’s still a non-renewable resource. Biodiesel will work in any diesel engine and it can be made from used vegetable cooking oil, or even from soybeans, so it’s renewable. But the most economical, practical version available now is B20, which is only 20 percent biodiesel, so it’s not replacing diesel fuel gallon for gallon. B100 will happen, but cost reductions are necessary first. Electricity, ethanol, and propane all require either a new vehicle or at least some vehicle conversion.”
Refueling Facilities
East Tennessee organizations already using AFVs include KAT, Chattanooga Area
Regional Transit Authority, the cities of Sevierville and Gatlinburg, ORNL,
KUB, AmeriGas Propane, and Schwan’s Fine Foods. Typically, problems for
such fleets include limited refueling facilities, increased fuel costs, and
limited range. Therefore, centrally fueled fleets such as buses or local delivery
services are logical places to introduce AFVs.
Three government fleets in Blount County have started a test program using biodiesel, with Maryville’s Calloway Oil supplying the fuel. In addition to Blount County, the cities of Alcoa and Maryville are participating. Four hybrid electric/ propane-fired trolleys have started service for KAT in downtown Knoxville, running both current and new routes.
In Chattanooga, the coalition is helping the city set up a station-car program, which would allow transportation around downtown in four-passenger pollutionfree electric vehicles.
The coalition is seeking funds from the Congestion Mitigation and Air Quality program to install a public biodiesel station at Walker Springs Road off I- 40/75 in Knoxville. And there is an ETCFC proposal before the SEP for funds to build additional biodiesel filling points, in Blount, Cocke, Knox, and Loudon counties. Another SEP proposal would convert new police cruisers for the city of Sevierville to run on propane.
Special events are helping area residents learn more about ETCFC’s initiatives and about alternative fuels in general. For example, ETCFC staged the National AFV Day Odyssey and East Tennessee Fair on the World’s Fair Park Festival Lawn in early April. The fair showcased more than 20 alternative fuel vehicles and featured games and information about alternative fuels, clean air, and foreign oil dependence.
The next week, the Run for Clean Air, a 5K run/walk in Sequoyah Hills, set out from Cherokee Boulevard. Cohosted by the ETCFC and the Regional Clean Air Coalition, the event focused on what area citizens can do to improve air quality. The event featured live music, cheerleaders, an AFV display, and booths presenting information on steps people can take to improve the air they breathe.
* * *
Contact Jonathan Overly, ETCFC, The University of Tennessee, 311 Conference Center Building, Knoxville, TN 37996-4134, call 865-974-3625, or email jgoverly@utk.edu. Visit the ETCFC at http://eerc.ra.utk.edu/etcfc/index.html.
Probing Outer Space from
Deep Underground
The Earth’s most extreme environments, including ultra-deep South African gold mines, may help guide future research and exploration on Mars. by Kris Christen
Using techniques developed for detecting microorganisms in deep, subsurface environments on Earth, University of Tennessee (UT) and Oak Ridge National Laboratory (ORNL) researchers are now working on tools to aid the National Aeronautics and Space Administration (NASA) in its search for life on Mars and beyond.
The idea is that just as subsurface microbial communities on our own planet have been isolated from the surface environment for tens to hundreds of millions of years, so too have any microbes that may exist deep beneath the Martian surface, says Susan Pfiffner, a microbiologist with UT’s Center for Environmental Biotechnology (CEB) and Center for Biomarker Analysis. Hence, methods developed to identify such microbes and the specific genes critical to their survival in extreme terrestrial environments could be brought to bear on any scientific and technological difficulties that may be encountered during the exploration of life on Mars.
With a 5-year, $5 million renewable NASA grant, the researchers are planning a series of laboratory and field experiments in extreme environments on Earth to help determine what types of lifedetection instruments are needed for unmanned subsurface drilling probes on future missions to Mars.
As part of this effort, the UT and ORNL researchers have teamed up with others from Indiana University and Princeton University to form the Indiana- Princeton-Tennessee Astrobiology Initiative (IPTAI).
Their collaboration stems from previous work sponsored by the National Science Foundation to study life in South Africa’s deep gold mines. Other research institutions involved with the IPTAI project include the Pacific Northwest National Laboratory, Lunar and Planetary Institute, Lawrence Berkeley National Laboratory, and University of Toronto.
“This is a big inter-institutional initiative in which UT has a pretty significant role,” says Gary Sayler, CEB’s director. “IPTAI is just one of a network of astrobiology institutes across the globe, and this really gives UT, and particularly the microbial ecology component of CEB, a place at the table for looking at different kinds of astrobiology-related issues and the potential for life occurring in other systems.”
UT will play a key role in the scientific, educational,
and public outreach ends of the project, says Lisa Pratt, a geoscientist at
Indiana University and IPTAI’s principal investigator.
LIFE IN THE EXTREME
Until it can get at Mars, the assembled IPTAI team of geochemists, chemists,
microbiologists, and hydrologists will continue its investigations of life’s
origins and physical and chemical limitations in the deep and ultra-deep South
African gold mines, Pfiffner says. Here, they’ve recovered microbes from
ancient, highly saline fracture waters at depths of up to 3,200 meters.
“We’re looking at biogeochemical cycling…how are the bacteria going about their business, what are they eating, what wastes do they generate, how is that used, and how are things being recycled?” Pfiffner notes.
The team’s next stop will be an Arctic field site whose environmental conditions are the most analogous to what one could expect to find on Mars. The team is currently considering as possible sites those hosting a thick permafrost cap under which gaseous brines are sequestered.
Here too, according to the group’s research proposal, they’ll be examining the microbial composition of the subsurface ecosystem and performing in situ experiments to see how these communities evolve in response to environmental changes.
“We really want to know more about the dissolved constituents…in
those brines that could support metabolic activity by bacteria, as well as the
diversity of life,” Pratt says. The researchers will then compare the
results from the Arctic site to those obtained from the much warmer South
In addition to the subsurface explorations and experiments, the researchers will also be conducting laboratory studies on the bacterial strains found at these sites to determine the processes that control energy and nutrient cycles in the deep subsurface. These experiments, which will rely on ORNL high-pressure bioreactors, will simulate the environmental conditions likely to be present at the Martian subsurface, such as very cold temperatures, low water activity, high salinity, and high carbon dioxide.
The information gleaned will be crucial for designing and field testing geophysical and chemical sensors for detecting life in the Martian subsurface. “If you’re going to be drilling and looking for life on Mars, you really need to test what you’re doing somewhere on Earth,” Pfiffner says. “That’s why so much is being done in extreme environments.”
Indeed, this instrumentation development poses the biggest
technical challenge for the group, Pratt says. “These instruments have
to be able to perform well in brines that are often quite saline and sometimes
caustic in the sense that they’re either highly alkaline or highly acidic,”
she notes. So, “first you have to design the kinds of equipment you want,
test it on Earth, and then re-engineer it so that it’s suitable for Martian
exploration, all the while hoping that the whole process of launching and landing
doesn’t damage the equipment.”
TRAINING FUTURE SCIENTISTS
A key part of the IPTAI project also involves education and public outreach,
which Pfiffner and Kim Davis, assistant director of UT’s Waste Management
Research and Education Institute (WMREI), are heading up and CEB is helping
to fund.
Activities already underway are designed around three areas of emphasis. They include educational workshops targeting minority and female undergraduate students and high school teachers where participants collect and interpret data from laboratory and field experiments; public outreach through a Web site illustrating how and why scientists conduct subsurface research; and mentoring undergraduate and graduate research at Indiana, Princeton, and Tennessee universities.
The workshops are modeled after a highly successful program in South Africa where minority U.S. and previously disadvantaged South African students have participated in a series of hands-on field exercises and laboratory experiments in 5-day and 7-week workshops with U.S. and South African mentors (see “Subterranean Science,” InSites, Spring 2002). More than 75 percent of the workshop participants enroll in advanced degree programs or internships in research laboratories after they complete their bachelor’s degrees, according to Pfiffner and Davis.
The two plan to include lectures on astrobiology in future workshops to show students how they might take their work to a different realm. “In our work on Earth in these environments, we have the potential to look at life on other planets,” Pfiffner notes. “We’re trying to open their eyes to how diverse science can be.”
A filming team from Indiana University is gathering material for the project’s public outreach. “They’re documenting everything we do in research and outreach, through high resolution digital video and audio,” Davis says. The footage will be used on the Web site, as well as for programming on public and community television stations.
Each of the universities is also developing undergraduate and graduate coursework in astrobiology. “The longterm goal,” Davis notes, “is to encourage undergraduates, especially minorities, to continue on to graduate school, eventually get their Ph.D.s, and go into academia where they can serve as mentors for other minority students.”
Overall though, IPTAI’s goal is to “gain a better understanding of our own world through research collaborations and the training of other scientists,” Pfiffner says. “This covers much more than astrobiology. The idea is as Earth changes, is there something we can learn from Mars to clean up our own systems?”
* * *
Contact Kim Davis, WMREI, The University of Tennessee, 311 Conference Center Bldg., Knoxville, TN 37996-4134, or call 865-974-1847. or contact Susan Pfiffner, Center for Biomarker Analysis, The University of Tennessee, 10515 Research Dr., Ste. 300, Knoxville, TN 37932-2575, or call 865-974-8031.
Staff
Citings
PROJECTS. Jack Barkenbus, executive director of UT’s
Energy, Environment and Resources Center (EERC) and director of policy for the
Waste Management Research and Education Institute (WMREI), recently announced
several research policy grants. Awards to UT faculty included: Gregory Reed
(head, civil and environmental engineering), who is leading a project that focuses
on relationships between emissions and health problems; Michael McKee (economics),
who is surveying citizen preferences in response to air-quality issues in East
Tennessee and Kevin Robinson (civil and environmental engineering), who is evaluating
how exposure and risk perception affect attitudes regarding the land application
of biosolids.
WMREI also awarded a grant to Research Ecologist Jack Ranney, who is leading a project to establish Knoxville as a prototype for community-based invasive- species control. Ranney and his partners are using presentations, along with results from a survey conducted last fall, to encourage Knoxvillians to adopt policy changes and implement control efforts, planning, and volunteer activities that target invasive non-native plants throughout the city.
Ranney is planning a Knoxville workshop for this fall, and his team is already developing posters and brochures. Several groups, including the City of Knoxville (greenways), the Izaac Walton League, the Tennessee Exotic Pest Plant Council, UT faculty, West High School, and others are partnering with Ranney on this project.
APPOINTMENTS. Outgoing Knoxville Mayor Victor Ashe recently
appointed Kimberly Davis to Knoxville’s Board of Environmental Appeals.
Davis, WMREI’s assistant director, will serve as the board’s Licensed
Professional Engineer. The board hears cases from contractors/developers who
have been fined for practicing inadequate erosion and sediment control. For
more information, see <http://www.
ci.knoxville.tn.us/boards/env-appeals.asp>.
Jack Barkenbus, EERC Research Leader Mary English, and Research Associate Jonathan Overly have been appointed to the statewide Tennessee Early Action Compact Advisory Committee. The 25-member Committee is evaluating measures that promise cleaner air in Tennessee The Committee will provide advice to the state and its seven Early Action Compacts, formed to assure attainment of new, eight-hour ozone airquality standards.
NEW FUNDING. EERC’s Center for Clean Products and Clean
Technologies (CCPCT) has received funding from the Tennessee Valley Authority’s
(TVA) Public Power Institute to develop a Green Electronics Consortium in the
Tennessee Valley. The project, managed by Jack Geibig, CCPCT acting director,
and Catherine Wilt, director of policy, will build partnerships among electronics
manufacturers and suppliers and identify opportunities for increasing the competitiveness
of the TVA-region electronics industry sector, with an emphasis on energy and
environmental issues.
