Biodegrading Acid: A Novel

Approach Using Bacteria

Proponents:
Aljamal, Raneen (School: Austin Peace Academy)

Abstract:
During bitumen extraction, a mixture of fine silts, hydrocarbons, salts,
and soluble organic compounds called oil sands tailings is produced. In Oil
Sand Tailing Ponds, crude oil is present. Crude oil is made up of various
chemicals that make it difficult to clean up. In this project, I wanted to test
and determine which method is most effective at degrading the harmful acids
that are spilled into the environment, such as naphthenic acid. I tested which
method of biodegradation is more effective in breaking down naphthenic
acid: slow sand filters or planktonic batch cultures. I used four types of
bacteria that are present in tailing ponds; E.Coli, Pseudomonas, Acidovorax,
or all three combined. I hypothesized that the slow sand filter biodegrades
the acids more efficiently with the combined bacteria, compared to The
Plankton Batch Culture, because of their tendency to form biofilms, which
will provide a better environment to break down the harmful acids. I also
hypothesized that the bacteria will biodegrade the harmful acids found in
crude oil, as it has biodegradable qualities. In conclusion, both hypothesis
were correct. The slow sand filter biodegraded the acid more effectively with
the average number of colonies after 4 weeks being 152, and the average
pH level after 4 weeks being 6. I also proved that bacteria has biodegradable
qualities and is able to break down acids located in crude oil. In the future, I
can use the studies found in this experiment for future upkeep of oil Tailing
Ponds.
 Optimizing the Ratios of Manuka
Honey to Adhesive in an Antibacterial
Surgical Adhesive
Proponents:
Leyendecker, Peyton (School: Skyview Academy)

Abstract:
The purpose of this science project was to optimize the ratios of
Manuka honey solution to adhesive in an antibacterial surgical adhesive.
Specifically, the different ratios were investigated to determine how they
affected the strength of the adhesive and the bacterial growth on stimulated
wounds sealed with the adhesive. To determine how the differing ratios
affected the amount of bacteria on the simulated wounds, slits were cut in
pieces of meat, on which the different formulations of the adhesives were
applied, and swabs were taken a day after the adhesive was applied. There
was a linear decline of bacteria on the wound as the Manuka honey solution
in the ratio increased, while the adhesive strength differed depending on the
percentage of honey solution added.
Screening of Yeasts in Azalea Nectar
for Bioethanol Production
Proponents:
Ota, Chihiro (School: Notre Dame Seishin Gakuen Seishin Girl’s High
School)
Shimoyama, Natsuno (School: Notre Dame Seishin Gakuen Seishin Girl’s
High School)

Abstract:
Yeast species in floral nectar, especially Saccharomyces cerevisiae,
show high alcohol fermentation properties, with nectar-dwelling non-
Saccharomyces yeast Metschnikowia pulcherrima additionally capable of
assimilating xylose and cellobiose, and fermenting glucose and galactose.
To produce ethanol using woody biomass by yeast, we isolated yeasts from
azalea nectar using an accumulative culturing method. The isolates were
examined for three properties: xylose assimilation, cellulose decomposition
and alcohol fermentation. A total of 44 yeast strains were isolated
successfully and seven of them possessed all three properties. An 18S rDNA
gene sequence analysis revealed that these seven strains were from species
closely related to the genera Metschnikowia and Candida. Finally, we
attempted alcohol fermentation in the lignin-removed woody biomass extract
using the selected strains. Four of seven strains were confirmed for CO2 gas
generation, suggesting that fermentation had occurred. These results
indicate that conversion of woody biomass into ethanol by nectar yeast
independent of non-woody biomass should be possible in the future.
 Identifying the Components in Basil
that Protect Lung Cells from
Pneumonia
Proponents:
Suresh, Arundhathy (School: Pioneer High School)

Abstract:
Basil is well known for its various medicinal properties. For the last 5
years, I have been working on the antimicrobial effect of basil against various
strains of bacteria such as E.coli, Streptococcus mutans, S. pneumoniae and
K. pneumoniae in my science fair projects. The major breakthrough came
when I discovered that basil could protect human lung cells from cell death
following infection by a bacteria that causes a life threatening disease such
as pneumonia. This year, I tried to identify the components in basil that
contribute to this protective effect. I separated proteins from the metabolites
in basil using acetone precipitation. Briefly, one volume of basil was
incubated with 6 volumes of acetone and kept overnight and centrifuged at
high speed (12,000 rpm, 30 minutes). The supernatant was dried and
dissolved in medium. The separated components were then used for
analysis of cell death due to K. pneumoniae. For this, lung epithelial cells
(A549) were cultured in RPMI medium in the presence and absence of basil
components. After 24 hours, 10^8 CFU/ml of K. pneumoniae was added to
it followed by incubation in a 37°C in a CO2 incubator. The viability of cells
was measured using trypan blue dye after 24 hours. I found that in the
presence of K. pneumoniae, lung cell viability was reduced to less than 20%,
but the presence of protein components prevented this significantly.
However, metabolites were unable to prevent cell death due to K.
pneumoniae. Thus, it can be concluded that proteins in basil is responsible
for its protective action against pneumonia infection in lung cells.
 Exploring the Use of Bacteria in
Mitigating Fungal Disease
Proponents:
Skocypec, Hannah (School: Basha High School)

Abstract:
Pseudogymnoascus Destructans, the fungus that causes White nose
syndrome, has killed more than 6.7 million bats in the US since 2006. Local
extinction of multiple bat species in the Northern US and Southern Canada
has been hypothesized. Although the long-term effects of the epidemic are
unknown, it has been estimated that it can cost the agricultural industry $50
billion from reduced insect control and pollination. There is no effective
treatment for this infection. Herein, novel probiotic bacteria, Micrococcus
luteus, antibiotic bacteria, Bacillus megaterium, and antifungal bacteria,
Lactobacillus plantarum, and combinations thereof, are explored to
investigate their potential to combat White nose syndrome. Both preventative
and responsive approaches were investigated. In the preventative approach
the treatment bacteria were inoculated with the fungal infection. In the
responsive approach the fungal infection was exposed to the treatment
bacteria. Since Pseudogymnoascus Destructans is limited to the northeast
and far northwest parts of the United States, it is not directly investigated
herein, to avoid any risk of it escaping in Arizona. Instead, Rhizopus
stolonifer, commonly known as black bread mold, is used as a model system
for Pseudogymnoascus Destructans. Fungal growth rates were qualitatively
observed to substantially decline for both the preventative and responsive
approaches. Spectroscopy was used to quantify the growth results, with
fungal growth rates decreasing by up to 90% for the preventative treatment
and up to 50% for the responsive treatment.