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and landfills. It was later found to migrate through the subsurface, contaminate groundwater systems, and persist for decades after initial disposal. Today, there
are approximately 202 hazardous waste sites in the U.S. containing 20 million tons of this dangerous compound. Biotic transformation of CT has been
demonstrated with cell exudates (biomolecules) of Methanosarcina thermophila, a methanogenic microorganism. These biomolecules have been found to
contain iron and zinc. In an effort to optimize and determine the most efficient and cost-effective method of CT degradation, the current research hypothesizes
that supplementing increased concentrations of iron and zinc in the growth medium of this methanogen will result in increased production of the biomolecules,
thereby resulting in higher rates of CT degradation. Pure cultures of the microorganisms were grown in the presence of 4 variables (high & low concentrations
of both iron and zinc), and the growth medium was filtered to obtain the biomolecules once the organisms were at stationary phase. CT was then added to this
filtrate, and the degradation of CT was measured using gas chromatography. Comparison (in triplicates) of the concentrations of CT in the liquid cultures, with
and without the metals over 7 time periods, indicated that the presence of high iron, low iron, high zinc, and low zinc increased the rate of degradation of CT by
49.4%, 14.1%, 231.0% and 52.2% respectively. ANOVA and t-tests indicated that the data was significant for P< 0.01. The hypothesis was, therefore,
supported. It is estimated that this process will reduce the cleanup costs of CT by almost $3 million.<br><br>
Awards won at the 2003 ISEF
Second Award of $1,500 - Microbiology - Presented by Intel Foundation

2003 - MI008
CHARACTERIZATION OF THE SECRETED ASPARTIC PROTEINASES OF CANDIDA ALBICANS USING A NOVEL COMBINATORIAL APPROACH
Jamie Elyce Rubin
Canterbury School, Fort Myers, FL, USA
Candida is a dimorphic fungus that is responsible for a wide variety of mycotic infections. The secreted aspartic proteinases (Saps) are among the organisms
putative virulence factors, and inhibition of these enzymes has been shown to have a curative effect upon candidiasis. In this study chromogenic combinatorial
libraries of octapeptides were used to investigate the extended substrate interactions of Sap1 and Sap2 of Candida albicans, the most common cause of
candidiasis in humans. This novel technique revealed a key difference between the preference of these enzymes and the preferences of human aspartic
proteinases, a characteristic that could be exploited to design effective inhibitors of Sap activity. Additionally, existing aspartic proteinase inhibitors were tested
against these enzymes to demonstrate the feasibility of targeted inhibition of Sap activity. These data were used to construct peptidomimetic inhibitors that
demonstrated marked inhibition of Sap activity and no interaction with human enzymes.
Awards won at the 2003 ISEF
Award of $500 U.S. Savings Bond - Ashtavadhani Vidwan Ambati Subbaraya Chetty (AVASC) Foundation
Second Award of $1,500 - Microbiology - Presented by Intel Foundation
Second Awards of $1,500 - U.S. Air Force

2003 - MI009
THE EFFECT OF X-RADIATION (X-RAYS) ON BACILLUS SUBTILIS
Anthony Michael Barnett
McDonaugh High School, LaPlata, Maryland USA
Research was performed to determine the effect of X-Radiation (X-Ray) exposure on observable and testable characteristics of Bacillus subtilis. With a
sufficient exposure, X-Rays are known to have a lethal affect on bacteria, and this has a potential application as a method to control pathogenic organisms
(such as Bacillus anthracis). This research was conducted to better understand how X-Rays affect Bacillus subtilis, which was selected due to its close
relationship to the anthrax bacillus. This research observed cellular functions at varying levels of low-dose, 70 kilovolts pressure (kVP), X-Rays using a dental
X-Ray machine. Easily observable (testable) characteristics were observed, before and after X-Ray exposure, including sugar metabolism (dextrose and
mannitol), cellular motility, catalase production, spore production, and colony morphology after X-Ray exposures ranging from 120 seconds up to 360 seconds.
This experiment showed that exposure to a non-lethal X-Ray dose has a significant affect on individual bacterial cell motility by direct observation (wet mount)
immediately after X-Ray exposure. It is also noted that X-Rays affect motility in subsequent generations based on testing in motility media. In addition, this
experiment noted that spores were produced by both the control (no X-Ray exposure) and by all of the cultures exposed to the various doses of X-Rays.
However, the spore count data suggests that the higher X-Ray doses appear to stimulate or accelerate spore production. Further research is needed to
determine the mechanism of the X-Ray effect on motility and to verify and explore the effect X-Rays on the spore generation process.

2003 - MI010
BACTERIAL BREAKDOWN
Hattie Bluestone Smith, Charlottesville High School
Charlottesville, Virginia (United States)
I sought to determine the effectiveness of different microorganisms (psychrophiles, mesophiles, and thermophiles) in decomposing a carbon-based compost
heap. I researched the conditions of a compost heap, the way the decomposition process works, and the three main categories of bacteria that help this
process. I found that in a compost heap the type of bacteria that is decomposing the organic matter is determined by the temperature of the heap.
Thermophiles are activated to decompose at 95 to 160 degrees F, Mesophiles are activated to decompose at 68 to 90 degrees F, and Psychrophiles are
activated to decompose at 55 to 62 degrees F. Therefore, I could isolate the work of a single type of bacteria by controlling the temperature of its compost
heap. I put a mixture of paper, wood shavings, and microbes into three compost bins. To control the temperatures of the bins, I used different amounts of
insulation. Then, to accommodate the climactic demands of a compost heap, I continuously pumped in water vapor. For three weeks I took temperature

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readings and made observations. After three weeks, I found that the bin with Psychrophiles at work looked similar to the original mixture, the bin with
Thermophiles at work had many parts of rich brown compost, and the bin with Mesophiles at work had some of each. I concluded that Thermophiles are the
most effective at composting a carbon-based compost heap, while Psychrophiles are the least effective.

2003 - MI011
DEVELOPMENT OF AN IMPROVED ENTEROVIRUS DETECTION PROTOCOL FOR TOMATOES: A SECOND YEAR STUDY
Hannah Mary Kaufman
Newberry High School, Newberry, Florida, United States of America
The purpose of this project was to test the effectiveness of Polyethyleneimine (PEI) on enumeration and recovery of viruses from produce and the effectiveness
of chlorine dioxide on decontamination of seeded tomatoes.<br><br>MA104 (ATCC #CRL-2378) monolayer cells were tested. Pre-filtered viruses were
procured from sewage wastewater. Rotavirus was also used. Ten tomato stem scars were inoculated with 0.1 mL of viral solution. After drying for one hour,
select tomatoes were subjected to 10 ppm chlorine dioxide treatment. An activated chlorine dioxide pouch and three tomatoes were covered with a container
and incubated for 15, 30, 60, or 120 minutes. For the aqueous treatments the pouch was activated and immersed in distilled water for one hour; tomatoes were
submerged into the solution for one minute, then briefly into a solution of 0.001% sodium thiosulfate in distilled water. The stem scars were placed in 50 mL
tubes with 20 mL Phosphate Buffered Saline and shaken for 15 minutes. Pre-Treatment with PEI and a Standard Method of cell culturing was then tested.
Three mL of .004% PEI was added to flasks for Pre-Treatment. After ten minutes, three mL of the produce/viral solution was placed into all flasks. One hour
later 10 mL of 2% medium was added. They were incubated at 370 Celsius and checked every 72 hours for cytopathic effect development. <br><br>The
hypothesis that Polyethyleneimine would enhance the susceptibility of tissue cell culture to enterovirus infection and that chlorine dioxide would effectively
decontaminate seeded tomatoes, proved to be correct.<br><br>
Awards won at the 2003 ISEF
First Award of $1,000 - Institute of Food Technologists

2003 - MI012
XYLELLA FASTIDIOSA GENETIC ANALYSES PROVIDE HOST-RANGE INSIGHTS
Mary Melissa Gardner
Spalding High School, Griffin, Georgia, United States
Preliminary data from the 2002 science fair project showed that AFLP molecular markers could be used to differentiate among host-specific Xylella fastidiosa
strains. But, without population genetic analyses of the raw molecular dataset, there was no way to determine gene flow patterns, genetic partitioning or to gain
insight into infection patterns that could be vector-specific and environment-specific. To determine if Xylella fastidiosa is geographically biased based on hostspecificity, a new research project was created to accomplish the overall goal, which is to master population genetics computer software analyses that can be
applied to many types of molecular datasets and which will facilitate continued research on Pierces disease. Data read from the ABI 377 automated sequencer
were reconstructed using a computer program that turned the dye-specific bands into a gel image that was straightened and sized. Three population genetics
programs were then used to analyze these data based on population genetics concepts and assumptions. The purpose of this study was to test the hypothesis
that population genetic analyses of host-specific Xylella fastidiosa strains will not demonstrate similarities across geography or host range to include systemic
(causing symptoms) and nonsystemic (symptomless) hosts. After evaluating the phylogeny, genetic identity and genetic distance resulting from analyses of the
molecular dataset, the original hypothesis was rejected in favor of an alternative hypothesis that population genetic analyses of host-specific Xylella fastidiosa
demonstrates similarities across geography as well as host range. The results also provided insight into nonsystematic hosts for vector acquisition of the
bacterium.<br><br>
Awards won at the 2003 ISEF
Fourth Award of $500 - Microbiology - Presented by Intel Foundation

2003 - MI013
A STUDY OF QUORUM SENSING ACITIVITY IN E. COLI (JM101) AFTER LACZ TRANSFORMATION
Caitlin Hannah Walls
South Pemiscot High School, Steele, Missouri,
Bacteria utilize a low molecular weight chemical belonging to the N-oxo-acylhomoserine Lactone (HSL) family for communication by quorum sensing. In this
project HSL production was observed in E. coli with the aid of a reporter strain modified with a LacZ reporter gene. The reporter strain produces bgalactosidase in response to exogenous HSL generated by the test organism. This production was measured quantitatively by using the lactose analog ONPG
(o-nitrophenyl-galactopyranoside) in a spectrophotometric assay. E. coli JM101 lacks both the amp and lacZ genes. A plasmid, pBLU, containing the genes for
lacZ and ampicillin was used to transform the E. coli JM101. Transformants were isolated on selective media, which contained LB/Amp/X-gal. The transformed
bacteria grew in blue colonies illustrating the uptake of the pBLU plasmid. Transformed bacteria with the reporter gene were used as the test organism.
Cultures of transformed bacteria, (-) lacZ bacteria and + lacZ bacteria were grown for 24 hours. Cell free extracts were taken every 4 hours for 4, 8, 12,16,20,
and 24-hour samples. Cell free extracts were combined with a fresh culture of transformed bacteria for 30 minutes. B-galactosidase enzyme activity was
assayed with ONPG substrate in a spectrophotometer at 420nm for 5 minutes. Optical Density of the culture was measured at 600nm. Specific activity of the
enzyme was calculated and compared to a control sample of transformer bacteria for each cell free extract sample collected over the 24-hour period. Results
indicate an increase in specific enzyme activity through the 8-hour extracts for each group tested. This corresponds to the maximum growth of the E. coli and
maximum cell density reached at 8 hours of growth.

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