Abstract
The ALU gene, whose name comes from Alu 1 restriction enzyme
recognition site, occurs in the human genome about 500,000 times.
Its origin and function is not yet known. It is hypothesized that this
gene passed down from a common ancestor, which could possibly be
used to determine familial relations. The gene may occur as
homozygous positive (+,+), homozygous negative (-,-), or
heterozygous (+,-) in the genome. The objective of this study was to
determine the heritability of the ALU gene among related individuals
as compared to a control of unrelated individuals. In this study,
there were two families of six, and 12 unrelated individuals
(control), who’s DNA was collected from their hair follicles.
Polymerase chain reaction (PCR) was used to amplify the ALU gene
sequence. Agarose gel electrophoresis was used to determine which
individuals have the ALU gene. The genes were measured and
analyzed. The Chi-square test was used to compare the groups of
individuals (family members, control, and the entire group) with the
US population Alu gene frequency. The results indicated that family
members do exhibit heritability of the ALU gene. Although analyzing
the ALU gene is not enough to pinpoint familial relationships
between people, it is one indicator that can be examined. Although
not completely reliable, since there are only three ways the Alu gene
can be analyzed, and unrelated individuals may show a
commonality, the inheritance of the ALU gene in families may be
observed and used as one indicator of relationship.
Experiment Outline
Alexandra Kokovich
Marietta Wright, M.S.
Waynesburg University,
Waynesburg, PA
•
•
•
•
•
Picture 1: This
picture shows the
electrophoresis
results for
Family 1
Figure 1:This picture shows the
pedigree of the inheritance of
the Alu gene for Family 1. This
was produced using the
electrophoresis results.
Main Results
•
•
Introduction
•
The Alu gene name is derived from the Alu 1
Restriction enzyme recognition site
•
It is about 300 base pairs long
•
The origin and function is not known, but
may be involved with expression of
other genes
•
Most abundant SINE
•
Short Interspersed Elements
•
Small dispersed DNA sequence
found throughout the genome
Picture 2: This
picture shows the
electrophoresis
results for
Family 2
•
Alu alleles are identical by descent, and individuals
that share the Alu gene inherited it from a common
ancestor
There is no evidence that there is a process that
removes the Alu element
Purpose and Objective of Experiment
•
•
•
To determine the allelic frequencies within a family
and among non-related individuals
To determine the heritability of the Alu gene
among family members
To determine whether the population has a similar
frequency in the Alu gene as the US population
•
Picture 4: This
picture shows the
electrophoresis
results for
Individuals 8-13
Figure 2: This figure shows the
pedigree of the inheritance of the Alu
gene for Family 2. This was
produced using the electrophoresis
results
Family 1 showed a significant difference compared to the US
totals
Family 2 showed inheritance of the gene throughout several
generations
Only Family 2 showed a non-significant difference, whereas
Family 1, Control, and Total Individuals Sampled were
significantly different
Conclusions
•
Family 2
Positive
Negative
Why the Alu Gene?
•
•
•
Picture 3: This
picture shows the
electrophoresis
results for
Individuals 1, 3-7.
12 Related Individuals
•
2 separate families (6 each)
12 Unrelated Individuals
Hair follicle with bulb was collected from each
individual
Instagene Matrix (Biorad) with Protease added to
each sample
PCR Reaction
•
Denaturing of DNA Strands at 94°C
•
Anneal Primers at 60°C
•
Extends DNA Strands at 72°C
•
Repeats 40 times
1% agarose gel electrophoresis
Chi-Square Test to determine whether population
frequency was similar to U.S. population frequency
sample
Observed (O)
Expected (E)
O–E=d
d2
d2/E
4
2
0.50<p<0.30
6(.79) = 4.74
6(.21) = 2.74
0.74
2.74
0.55
7.51
0.12
5.96
6.08
•
Discussion & Future Studies
•
Table 1: This table shows the chi-square results for Family 2. 0.50<p<0.30
•
All
Individuals
Observed (O)
Expected (E)
O–E=d
d2
d2/E
Positive
10
24(0.79) = 18.96
8.96
80.28
4.23
Negative
14
24(0.21) = 5.04
8.96
80.28
15.93
p<0.01
There is a high allelic commonality between family members of
the Alu gene as compared to non-related individuals
This gene could be used as an indicator or relatedness by
following the inheritance of the gene between family members.
•
The ALU gene may be one indicator that can be examined to
determine relationships in families.
More studies need to be conducted in order to determine the
function of this gene and its purpose in the human genome
Current studies indicate that the Alu gene may play a role in
Hemophilia A, regulation of developmental processes, and
human gene regulation
20.16
Table 2: This table shows the chi-square results for All Individuals involved in this study.
p<0.01
Acknowledgements: We would like to thank Waynesburg University Biology Department for the funds
to perform this research. We would also like to thank the Center for Research and Educational
Development for the printing of this poster.
The Effects of Natural Repellents On Mice Using a Series of Simple Mazes
Andrea Strohecker
Waynesburg University Department of Biology: Waynesburg, PA
Abstract
Mice rely primarily on their sense of smell to make decisions
about their environment. Consequently, it has been reported
that peppermint and other natural oils may be useful and safe
rodent repellents. To determine the validity of these claims,
white mice were placed in simple plastic mazes to determine
whether they would actually select pathways leading away from
a variety of natural “repellents.” The results of this study may
be useful to homeowners searching for safe and simple ways
to keep rodents away; they also provide insights into the
sensory systems of mice.
Results
Cayenne Pepper
and Mouse Food
17%
Lemon and
Mouse Food
33%
Methods
Mouse Food
Only
67%
Mouse Food Only
83%
Figure 1: Cayenne Pepper across all 4 mice
Background
Odor particles (odorants) bind to special receptors on the cilia
of the olfactory neurons
This triggers a neural response that travels to the brain
There are 500-1000 types of olfactory receptors that code for
thousands of genes
1% of a rodent’s DNA is utilized just for smell
 A message from the olfactory neurons speeds along a
pathway to the olfactory bulbs (stem-like projections from the
forebrain)
A different pattern of glomeruli is activated when a mouse
smells different odors
These activation maps also change when odor concentration
increases.
Lemon vs. Food Only
Cayenne Pepper vs. Food Only
Figure 2: Lemon across all 4 mice
Mint vs. Food Only
Discussion
Cinnamon vs. Food Only
Mint and
Mouse
Food
31%
**** Please note that this research is still being conducted
and is not yet finished
Cinnamon and
Mouse Food
44%
Mouse Food Only
56%
Mouse Food
Only
69%
Figure 3: Mint across all 4 mice
Figure 4: Cinnamon across all 4 mice
Mouse 1
Mouse 2
25
45
40
20
35
30
15
% of ov erall 25
choice
20
% of ov erall
choice
10
15
10
5
5
Four lab mice were used in this experiment
PVC pipes were used to construct simple mazes
A plastic box was placed on each exit of the maze
One box contained food and the other contained both food and
a repellent
For each mouse 15 trials will be run for each repellent totaling
240 runs
The four repellents used were lemon extract, mint extract,
cinnamon, and cayenne pepper
The effects of the repellent on chamber
choice were
determined
0
0
Mint
Cinnamon
Lemon
Repellents
Cayenne Pepper
Figure 5: Comparison of all four repellents for mouse 1
Mint
Cinnamon
Lemon
Repellents
Cayenne Pepper
Figure 6: Comparison of all four repellents for mouse 2
Mouse 3
25
20
20
15
15
% of ov erall
choice
% of ov erall
choice
10
10
5
5
0
0
Mint
Cinnamon
Lemon
Repellents
Cayenne Pepper
Figure 7: Comparison of all four repellents for mouse 3
Mint
Cinnamon
Lemon
Repellents
Acknowledgements
Special Thanks To:
The Waynesburg University Biology Department, Dr.
Christopher Cink, Renee Wiltrout, and The Center for
Research and Economic Development
Mouse 4
25
According to the results thus far, cayenne pepper seems to
be the most effective repellent on the mice
The expectations were that the mint and lemon extracts
would serve as the best repellent due to the acid content
Other studies suggest that natural foods that are highly
acidic such as lemon and mint, agitate the vomeronasal
organ of rodents and has appeared to be unpleasant in high
concentrations
Figures 2 & 3 suggest that mint and lemon have the same
affect on all four mice
The activation maps seem to be similar for the mint and
lemon, but are drastically different for cinnamon and cayenne
pepper
Cinnamon seems to have little to no effect and cayenne
pepper seems to have the most effectiveness
Further research must be conducted in order to determine
which natural repellent has the most effectiveness on
household mice
Cayenne Pepper
Figure 8: Comparison of all four repellents for mouse 4
Occurrence of Escherichia Coli and Staphylococcus aureus on Shopping Cart Handles from
an Anonymous Grocery Store in Washington, Pennsylvania
Ashley Thomas and Dr. Chad Sethman
Department of Biology, Waynesburg University, Waynesburg, Pennsylvania 15370
Abstract / Introduction
Conclusions
According to the Centers for Disease Control (CDC), each year
in the United States 76 million people suffer from foodborne
disease. In previous studies it has been shown that grocery
store shopping cart handles are among the top ten common
surfaces that are contaminated with bacteria. Escherichia coli
(E. coli) and Staphylococcus aureus (S. aureus) are two
common foodborne pathogens which have been detected on
grocery store shopping cart handles. In this study, shopping
cart handles were swabbed prior to being disinfected, after
being disinfected, and after every one hour period for four
consecutive hours. The samples collected were then tested
for the presence of E. coli and S. aureus on desoxycholate
agar (DOC) and mannitol salt agar (MSA), respectively. The
hypothesized results affirmed that the S. aureus accrued more
readily than the E. coli. This resulted due to the higher
resistance of S. aureus and its ability to remain viable longer
on the surfaces of the shopping cart, which are unable to
absorb and retain moisture. This assessment of the bacterial
accumulation demonstrated that there is a need for periodic
disinfection of grocery store shopping carts to reduce and
prevent the spread of these foodborne pathogens.
• A large accumulation of bacteria on
surfaces that were not disinfected indicates
that carts need to be disinfected at a
MINIMUM of once a day.
• Every individual shopper should use
disinfectant wipes on their cart to prevent
the transfer of bacteria.
• A cart that contained a baby was found to
have fecal coliforms present, indicating that
babies could possibly be a means for
transferring fecal coliforms.
• The number of shoppers may have an
effect on the amount of bacteria present.
• The species and amount of bacteria present
are influenced by the individual shopper
and what they are transferring to the cart.
Aim
To
assess
the
bacterial
accumulation
of
Staphylococcus
aureus
and
Escherichia coli on
the
surfaces
of
grocery
store
shopping carts.
Figure 1. The blue bar represents the average of the 8 carts from the
initial swabbing of the handles. Fecal coliforms were not found on the
initial handles of the carts. The green bar represents the total number
of bacteria found on the handles.
Figure 2. The blue and red bars each represent the average of 8 carts
from the initial swabbing of the seats. The green bar represents the
total of the two averages.
Figure 3. The blue bar represents the average of the 8 carts from the
initial swabbing of the control (small advertisement sign on the front
of the cart). Fecal coliforms were not found on the initial controls of
the carts. The green bar represents the total number of bacteria found
on the controls.
Acknowledgments
We thank the Waynesburg Univ. Biology Dept. for their
resources and equipment, the grocery store that allowed me to
utilize their facility to conduct my project, the CRED building
for the printing of this poster, and Candace Swiantek and
Andrea Strohecker for their swabbing and laboratory
assistance.
Method
•
Desoxycholate agar (DOC): is
selective, differential, and used
to differentiate various Gram
negative enteric pathogens.
•
Mannitol Salt Agar (MSA): is
selective, differential, and used
to isolate and identify the
pathogenic
bacterium,
S.
aureus.
Figure 4. Chart of the number of bacteria present on cart handles based
on the number of shoppers. Each line is representative of the 4 carts
used for each run. Fecal coliforms were not found for run 2 and are not
represented in this chart.
Figure 5. Chart of the number of bacteria present on cart seats
based on the number of shoppers. Each line is representative of the
4 carts used for that run. S. aureus and fecal coliforms were not
found for run 2 and are not represented in this chart.
Figure 6. Chart of the number of bacteria present on the cart
controls based on the number of shoppers. Each line is
representative of the 4 carts used for each run. Fecal coliforms were
not found for run 2 and are not represented in this chart.
Figure 7. Fecal coliforms on DOC agar
Figure 8. S. aureus on MSA
Analysis of Marcellus Shale in Greene County Pennsylvania
Jessica Husman & Dr. Evonne Baldauff
2007 Energy Consumption in America
Abstract.
The use of hydro‐fracturing by oil‐drilling companies has
caused quite an uproar by the local residents. The hydro‐fracturing fluid
contains numerous chemicals and many seem to believe that these
chemicals are contaminating their water supply. Not only is this water
used for consumption and other daily house‐hold tasks, it is also what
hydrates our wildlife. Another issue is the shale itself. After the drilling
process, elements originally found in the rock far beneath the Earth’s
surface, rise to the top with the drill shavings and fracking fluid. The
objective of this experiment is to determine what elements are found in
the shale that could potentially leach out. The scanning electron
microscope was used for the analysis. The SEM is a type of microscope
that uses electrons instead of light rays to view an image. A tungsten
filament above the specimen shoots a ray of electrons. Two lenses‐ the
condensing and the objective lens aligns the ray of electrons so that they
shoot directly toward the sample. As this energy is introduced, it causes
electrons from the sample to be released. A positively charged detector
then attracts the second order electrons. A photomultiplier amplifies the
results and a visual image is created. From this, the elemental composition
can be determined.
Figure 2.
The figure above shows the fuel distribution
consumed by Americans in 2007. As you can see natural gas
constitutes 22% of our fuel consumption. Only liquids
surpasses this portion of our energy.
This map indicates the primary locations of all the
shale basins in the United States. The shale analyzed in this
study is highlighted.
Figure 4.
This gives the general case of how fracking
fluid works. It keeps the fissures open so that the natural
gas can flow into the well more efficiently.
Not all of the chemical components of fracking fluid
are known. It has been established, although that
many of the chemicals are detrimental to human
health. They can cause:
• damage to the liver
• skin and eye irritation
• respiratory infections
• damage to the cardiovascular and blood system
• damage to the reproductive system
• problems with embryonic development
• malfunction of the immune system
• brain and nervous tissue damage
• and more (Lebron 6).
Violations
Shale Basins Spanning the United States
Figure 1.
The Drilling Process
The SEM
Figure 3.
The gun shoots a beam of electrons that is then
condensed and directed toward the specimen by the lenses.
The deflecting coil allows for the movement of the beam in the
x and y directions. When the beam hits the sample, energy is
emitted in the form of electrons and x‐rays. This is allows for
the production of an image.
Work Cited
Figure 5.
The violations shown in the figure above only
represent 1056 of the total 1614 violations. These 1056 violations
were considered likely to directly endanger the environment
and/or the safety of the community. These violations occurred
between 1/1/2008 and 8/20/2010 and do not include traffic
violations recorded during a 3‐day enforcement blitz. During this
period an additional 1487 violations were accumulated.
"Cumulative Environmental Effects of Oil and Gas Activities On Alaska's North Slope." Cumulative Environmental Effects of Oil and Gas Activities On Alaska's North Slope (n.d.): netLibrary. EBSCO. Web. 22 Jan. 2011.
Kell, Scott. Modern Shale Gas Development in the United States a Primer. Rep. Oklahoma City, 2009.
Lebron, Michael, and Joel Kupferman. "The Marcellus Shale: a paradigm moment in the struggle for environmental justice?." Guild Notes 34.1 (2009): 6‐7. Academic Search Premier. EBSCO. Web. 1 Dec. 2010.
Pennsylvania Land Trust Association. "1056 Identified as Most Likely to Harm the Environment." Marcellus Shale Drillers in Pennsylvania Amass 1614 Violations Since 2008
(2010): 1‐3.
Resnikoff, Marvin, Jackie Travers, and Ekaterina Alexandrova. "Radioactivity in Marcellus Shale." Radioactive Waste Management Associates, 19 May 2010. Web. 30 Nov. 2010. <http://www.rwma.com/Marcellus%20Shale%20Report%205‐18‐2010.pdf>.
"Swapp, Susan. "Scanning Electron Microscopy (SEM)." SERC. 18 Mar. 2011. Web. 26 Mar. 2011. <http://serc.carleton.edu/research_education/geochemsheets/techniques/SEM.html>.
The Alberta government and oil industry say disposing oil drilling wastes by spreading them on farmland is safe. But others worried about environmental risks warn they could be...Spreading the seeds of soil contamination." The Ottawa Citizen (n.d.): C4. LexisNexis Academic. EBSCO. Web. 22 Jan. 2011.
Acknowledgements
Thanks to:
National Energy Technology Laboratory and Department of Energy for providing funding.
Separation Design Group for permitting the use of their facilities and equipment
RJ Lee Group and Waynesburg University for aiding in the research
The Effects of Muscular Stretching on sEMG Electrical Activity Results of the Hamstring Muscles
Julie M. Smith
Bryan Hamilton, PhD
Waynesburg University, Department of Biology
BACKGROUND
•Electromyographs (EMG) are used for diagnosis and
biofeedback therapy of muscular ailments and for
biofeedback information during therapy. They record
muscle action potentials created in the vicinity of the
electrode.
•A common physical therapeutic regimen also involves
muscle stretching for reasons involving a decrease in muscle
stiffness, decrease post-exercise pain, increase blood flow,
stability, and aid with muscle relaxation.
•During a stretch, the elongation effects the muscle spindles.
These muscle spindles play a role in contracting the muscle.
•The body of the hamstring muscle is located halfway
between the ischial tuberosity and the medial and lateral
epicondyles.
•Understanding the muscular benefits of stretching can aid
in targeting therapy to maximize patient rehabilitation.
Determine whether a significant change occurs in the following after a guided hamstring
stretch:
RESULTS
Figure 1
CONCLUSION
Figure 2
•The p-value in Figure 1 shows no significant difference between the
mean of electrical results of the hamstring muscles after stretching in
comparison to before stretching.
• Increased average maximum height
•The p-value of Figure 2 shows no significant difference between the
average maximum peak heights of the hamstring muscles after
stretching.
• Increased mean of electrical results
• Decrease in downward slope after the maximum peak height.
MATERIALS AND METHODS
•An sEMG test was performed on 20 subjects before and after a guided muscle stretch of
the hamstring muscles.
•The test involved two surface electrodes on the dry skin of the upper posterior left leg,
and one on the ankle.
•Each subject did multiple leg-curl exercises at 30lbs. The weight was held between
concentric and eccentric contractions.
•The 40-second stretch was repeated three times, with ten second breaks between.
•Results were collected using the BIOPAC software and integrated EMG graph.
•Match-paired tests were performed to find statistical significance between pre-stretch and
post-stretch sEMG results.
•For the mean of results data, each individual sEMG graph set was analyzed separately,
from the beginning of the first contraction signal to the end of the final. The base line
signal was eliminated to avoid lowering the mean electrical value. A Matched-pair T-test
was performed to compare post-stretch results to pre-stretch.
•The maximum value of each contraction curve was recorded for a subject during the
trial. The values were averaged for both before and after stretching, and compared
using a Matched-pair T-test.
•Possible error may be attributed to sensor misplacement. Muscles may
have been stretched during the day, which would have limited the effects
during the research. Amount of difference found may have been caused
by the pre-stretch exercises.
Indicates a maximum peak
The range indicated is the area calculated in the mean of results
Figures 1 and 2 show the results obtained from subject number 13. Figure 1 is the contraction graph taken before
stretching, and Figure 2 after stretching. The difference between the mean maximum heights is equal to 0.00085mV. The
difference between the average mean of results is -0.00217385.
•Further research could be found by looking at:
•Larger subject size
Graph 1
0.016
Mean of Results
•Muscles with obvious muscle sensor placement
P = 0.168795
0.014
Mean of Electrical Results, mV
Electromyography (EMG) is a useful tool used in both
diagnosis and biofeedback therapy of muscular ailments,
such as disease or weakness. A surface-electromyograph
(sEMG) is a device that is used to measure the electrical
activity in a skeletal muscle fiber with electrodes placed on
the surface of the skin. This information can then be
compiled to see how the muscle is contracting. It is also
used for its biofeedback possibilities in rehabilitation, in
order to see the progress of the muscles electrical activity as
therapy continues. Another inclusion in physical therapy is
stretching. Stretching is capable of lowering muscle
stiffness, increase range of motion, decrease post-exercise
pain, increase blood flow, stability, and aid with muscle
relaxation. Since relaxation of the muscle is measured with
an electromyograph, it can be hypothesized that these
results may vary after stretching has been introduced to the
muscle. The hamstring muscles: the bicep femoris,
semitendinosus, and the semimembranosus were chosen
because of their availability to be picked up by the EMG
with little background noise. The study will look at EMG
results both before and after a passive hamstring stretch
and compare the results to each other, based on the
individual subject. The comparison results of each subject
will then be compiled together in order to draw conclusions.
SPECIFIC AIMS
•Leg dominance for better coordination
0.012
•Static vs. Passive stretching results
0.01
•Reciprocal inhibition effects after stretching
0.008
Before
After
0.006
REFERENCES
0.004
0.002
1.
Ankrum, Dennis R. “Questions to ask when interpreting surface electromyography (SEMG)
research”. Proceedings of the IEA 2000/HFES 2000 Congress. 5,530-533.
2.
Bang, J.K., Hwang S.J., Ko C.Y., Kim C.H. “Heat Treatment and Rest-Inserted Exercise Enhances
EMG Activity of the Lower Limb”. World Academy of Science, Engineering and Technology.
Volume 11; 466-469. 2005.
3.
Brandy, William D. Physical Therapy. Volume 74, Number 9. September 1994.
4.
Behm, David PhD. “Factors Affecting Electromyographic Signals”. Memorial University of
Newfoundland.
5.
Criswell, Eleanor. Cram’s Introduction to Surface Electromyography. Jones and Bartlett Publishers.
Sudbury, MA. 2nd edition. 2011.
6.
Marek, Sarah M, Cramer j, Fincher A, Massey L, Dangelmaier S, Purkayastha S, Fitz K,
Culbertson J. “Acute Effects of Static and Proprioceptive Neuromuscular Facilitation Stretching on
Muscle Strength and power Output”. Journal of Athletic Training. 40(2): 94-103. Apr-Jun 2005.
7.
Neblett, Rnady. “Theory and Rationale for Surface EMG-Assisted Stretching as an Adjunct to
Chronic Musculoskeletal Pain Rehabilitation”. Applied Psychophysiology and Biofeedback.
Volume 28, Number 2. June 2003.
0
1
2
3
4
5
6
7 8
9 10
Subject number
11
12 13
14
15 16
Graph 1 shows the mean of results for each individual subject, before and after stretching. The error bars show a significant
error. The p-value was found using a one-tail, equal variance t-test and showed no significant difference of post-stretch
results in comparison to the pre-stretch. The bars show that six subjects had decreasing post-stretch mean results, while ten
had increasing results.
Graph 2
0.06
Average Maximum Peak Height (mV)
ABSTRACT
Average Maximum Heights
P=0.306148
0.05
0.04
0.03
Before
After
0.02
0.01
ACKNOWLEDGEMENTS
0
1
2
3
4
5
6
7
8
9 10
Subject number
11
12
13
14
15
16
Graph 2 shows the average maximum values for each individual subject, before and after stretching. The error bars indicate
significant error. The p-value was found using a t-test and showed no significant difference of post-stretch results.
A special thank you is extended to Dr. Bryan Hamilton for the time spent to help formulate and plan my
project. The Biology Department and the Center for Research and Economic Development for the
materials to perform the research and presentation. Also, to Dr. James Bush for help with data analysis.
The Inhibitory Effects of Various Oral Hygiene Products on the Cavity‐Causing (Cariogenic) Bacterium Abstract:
Streptococcus mutans
200
Doubling Times Vs. Max. Densities
†
70
•Gram positive
•Anaerobic bacterium
•Found in oral cavity of human beings
Doubling Times
1.4
****
Max. Densities
1.2
60
50
***
1
***
****
0.8
40
†
††
†††
30
††
0.6
0.4
20
†††
10
0.2
Doubling Times of Scope Mouthwash
180
160
Doubling Time (in minutes)
80
Max Density (Absorbance)
Streptococcus mutans:
Tonya Tanner*, Ryan Schell*, and Dr. Chad R. Sethman
Waynesburg University Department of Biology
Doubling Time (min)
A dental carie or cavity is damage to tooth enamel resulting from bacterial
metabolism. Immediately following brushing, layers of multiple species of bacteria
begin to accumulate on the surface of teeth, ultimately forming a stable community, or
“biofilm”, commonly known as dental plaque. Streptococcus mutans (S. mutans) is one
of the early colonizers of dental biofilms. S. mutans binds to the surface of teeth and
subsists on a diverse group of carbohydrates.
When S. mutans specifically
metabolizes the sugar, sucrose, lactic acid is produced which weakens enamel leading
to the formation of dental caries. Oral hygiene products such as mouthwashes and
toothpastes can be useful for inhibiting the formation of dental bio-films and preventing
dental caries. The specific aim of this research was to determine the efficiency of
various oral hygiene products at inhibiting the growth of S. mutans. We first determined
that Trypticase Soy Broth (TSB) was the most efficient media for the growth of S.
mutans. We then analyzed bacterial growth curves of S. mutans grown in TSB
containing various concentrations of different oral hygiene products in order to
determine which product most effectively inhibited the growth of S. mutans.
140
120
100
80
60
40
20
0
0
0
TS
BHI
TYC
N
Figure 2:
Analysis of the doubling-time and maximum density of S.mutans grown in various
media. Data represent averages of five sample runs. Error bars represent standard deviation.
***p<0.05, ****p<0.01, † p<0.001, †† p<0.05, ††† p<0.01.
160
•Find the most desired media for the Growth of Streptococcus mutans (S. mutans).
•Find the desired concentration of various oral hygiene products that doubles the
original doubling time in TS broth.
•Compare the ability of various oral hygiene products to inhibit the growth of S. mutans.
Methodologies:
•This research study was performed by growth curves using a spectrophotometer.
Doubling times in Listerine® mouthwash
0.10%
1%
3%
5%
V/V% Scope added to 12.5 mL of TS broth/ S. mutans
10%
Figure 4:
Analysis of the growth of S. mutans in TS broth with various concentrations
of Listerine. Data represented by the average of 3 sample runs.
Results:
140
Doubling time (minutes)
Goals/ Specific Aims:
0%
•S. mutans prefer Trypticase Soy broth.
•S.mutans has a doubling time of 42.88 minutes in TS broth.
•Figures 3 and 4 indicate that Scope has a higher doubling time than Listerine.
•The more Listerine added slows down the doubling time of the S. mutans.
120
100
80
Future Studies:
60
•Use toothpastes and other various oral hygiene products to determine the most
desired product that doubles the original doubling time in TS broth.
•Test the oral hygiene products to determine if the products are bacteriastatic or
bacteriacidal.
•Determine the cost efficiency of the most preferred inhibitor of S. mutans.
40
20
0
0%
Figure 3:
0.10%
1%
3%
5%
V/V% Listerine ® added to 12.5ml of TS broth/S.mutans
10%
Analysis of the growth of S.mutans in TS Broth with various concentrations of Listerine.
Data’s represented by the average of 3 sample runs.
Acknowledgements:
I would like to thank the Waynesburg Universities Center for Research and Economic
Development for poster printing.
The Maximum Carrying Capacity of Staphylococcus aureus and Other Bacteria on Gasoline Pumps in Southwestern Pennsylvania
Victoria Danielczuk, Dr. Chad Sethman
Biology Department, Waynesburg University, Waynesburg, PA
ABSTRACT
SPECIFIC AIMS
RESULTS
Staphylococcus aureus is a type of bacteria carried on human skin. It
has the potential to cause severe infections in individuals if it enters
the tissue through an open wound. Staphylococcus and other various
types of bacteria can be transmitted among people when the bacteria
are deposited on surfaces by one individual and picked up by another.
A common surface that many people come in contact with is a pump
at a gasoline station. The following study examined gasoline pumps
and the number of colony forming units (CFUs) of Staphylococcus
aureus and other bacteria that are deposited on them. The purpose of
this research was to determine an appropriate time interval during
which gasoline station attendants should sanitize the pumps in order
to prevent the spread of potentially infectious bacteria. In this study,
eight gasoline pumps at one station in southwestern Pennsylvania
were sanitized and individually sampled by swabbing at 30-minute
intervals. Mannitol salt agar was used to identify and count colonies
of Staphylococci species and trypticase soy agar was used to count
other colonies of bacteria. A hypothesis was postulated that the
number of colonies taken from the pumps will continue to increase
over three hours or every twenty-five patrons and will then level out
when the pump has reached its maximum capacity.
The aim of this research was to determine the time interval at which a gasoline station pump
reaches its maximum carrying capacity for Staphylococcus aureus. Scientific techniques
were used to collect specimens from gasoline pump handles and to plate the specimens to
determine how many bacterial colonies would grow after a specified time interval. Aims for
this research included:
The data that was collected throughout the course of this research is represented in Figures
1-6. Figures 1 and 2 represent the colonies that were observed on TSA plates, and Figures 3
and 4 represent the colonies that were observed on the MSA plates. The number of people
versus bacteria are depicted in Figures 5 and 6. All plates were innoculated with 0.2
milliliters of the sample solution and were incubated for 48 hours before being observed.
• To determine whether or not the maximum carrying capacity for different types of
bacteria is reached within four hours
140
Colonies on Plates
• If the maximum carrying capacity is reached within four hours, the number of colony
forming units (CFUs) of Staphylococcus aureus collected from the pump handle will be
compared to its infective dose to determine whether infection is possible
TSA, Day 1
100
100
80
80
60
60
40
40
TSA Controls
20
20
TSA Pumps
0
0
60
90
Bacteria are present on almost every surface and environment
on Earth. In addition to harsh hydrothermal vents and
uninviting conditions of the world’s highest mountains, bacteria
inhabit surfaces as common as human skin where they thrive as
normal flora. The palms of individuals carry millions of diverse
species of bacteria, the including Staphylococci, Streptococci,
and Proprionibacteria species.2 Bacteria are sensitive to pH,
dryness, hormone levels, use of lotion, and hand washing.
These factors can play a part in the numbers of colony forming
units (CFUs) of bacteria that are present on human hands.
The following research studied the amount of bacteria that was
deposited onto gasoline station pumps over the course of four
hours. Based on the amount of bacteria that commonly inhabits
the human hands, it was hypothesized that as more people used
the gasoline station pumps, the amount of bacteria collected
from the pump handles would increase, and the amount
collected from the control areas would remain constant or
would increase slightly due to environmental contamination.
• Mannitol salt agar (MSA) is a medium that is used to isolate different species of
staphylococci. The medium contains mannitol, a sugar alcohol used in medicine as a
therapeutic agent or in food production as a sweetener. Generally, staphylococcal strains
grow in small, round white colonies on the surface of the red MSA plate; however,
mannitol is fermented by Staphylococcus aureus. MSA is impregnated with a pH
indicator that changes from red to yellow when the mannitol is fermented.
• Methods:
• The spread plate technique was used to spread the bacteria evenly over the surface of
the agar to facilitate enumerating colonies of bacteria. This technique was done by
depositing a liquid sample onto the agar plates using a sterile pipette, and immediately
spreading the liquid with a flame sterilized spreader. The spreader had been immersed in
pure ethanol and removed, passed over a Bunsen burner to catch fire, and allowed to
burn off and cool for a few seconds before it was used to spread the liquid.1 This
technique is useful in identifying individual colonies of bacterial growth
150
180
210
240
30
60
90
120
Figure 2
MSA, Day 1
150
180
210
240
MSA, Day 2
100
• Test bacteria grown on TSA plates
80
60
60
40
40
MSA Controls
20
20
MSA Pumps
0
0
-20
-20
Colonies on Plates
• Attempt to execute the research on days with similar temperatures
and humidity
Future research:
Time (minutes)
80
30
60
90
120
150
180
210
• Gram stain to identify bacteria
• Extend research to other potentially contaminated surfaces:
• ATM keypads, door handles, buttons, light switches, etc.
30
240
60
90
Figure 4
Time (minutes)
• Test for other types of bacteria on different media
120
150
180
210
240
Time (minutes)
The figures above indicate the relationship between the bacteria collected from the pump
handles and control areas when the trypticase soy agar plates (Figures 1 and 2) and mannitol
salt agar plates (Figures 3 and 4) were examined.
Day 1: Patrons v. Bacteria
Bacterial Colonies on Plates
INTRODUCTION
120
Time (minutes)
Figure 3
• Trypticase soy agar (TSA) is selective for aerobic microorganisms, which are found
on surfaces that are frequently touched. TSA media can be mixed with different amounts
of blood to select for various types of bacteria. This media is used in industries such as
sewage treatment and food preparation, and is useful in identifying contaminants in
water, food, and even cosmetics. TSA was used in this research to broadly identify
microorganisms left behind on the gas station pump handles.
• Set a number of people rather than time limit
-20
-20
100
• Media used in this research was selected and prepared:
• Longer time period between swabbing and monitoring pumps
120
Figure 1
• Permission to sample the pumps at a nearby gas station was obtained and documented
before any research was performed.
140
It was hypothesized that the number of bacterial colonies would
increase over time. This hypothesis was not supported because the
numbers of colonies fluctuate over the four-hour time period. Also,
the amount of Staphylococcus aureus collected from the pump
handles was nowhere near that of its infective dose.
Improvements to this research:
TSA, Day 2
120
30
METHODS
CONCLUSION
Day 2: Patrons v. Bacteria
140
130
120
110
100
90
80
70
60
50
40
30
20
10
0
140
130
120
110
100
90
80
70
60
50
40
30
20
10
0
0
Figure 5
2
4
6
8
Number of Patrons
10
12
14
Patrons vs
TSA
Patrons vs
MSA
0
5
Figure 6
10
15
20
REFERENCES
1.
Buck, John D. and Robert C. Cleverdon. “The Spread Plate as a Method for the Enumeration of Marine
Bacteria.” Limnology and Oceanography. 5.1 (1960): 78-80.
2.
Fierer, Noah, Micah Hamady, Christian L. Lauber, and Rob Knight. “The influence of sex, handedness, and
washing on the diversity of hand surface bacteria.” PNAS, 105.46 (2008):17994-17999.
3.
Rutala, William A., Matthew S. White, Maria F. Gergen, and David J. Weber. “Bacterial Contamination of
Keyboards: Efficacy and Functional Impact of Disinfectants.” Infection Control and Hospital Epidemiology.
27.4 (2006): 372-376.
4.
Tiller, Joerg C., Chun-Jen Liao, Kim Lewis, and Alexander M. Klibanov. “Designing surfaces that kill
bacteria on contact.” PNAS. 98.11 (2001): 5981-5985.
25
Number of Patrons
Figures 5 and 6 display the number of patrons that used the pumps versus the bacteria
collected from the pumps. The number of bacterial colonies that grew on the MSA plates
remained fairly constant for the most part. The number of colonies on the TSA plates
sharply increased then decreased between four and seven patrons on day 1 (Figure 5) and
increased gradually on day 2, but then sharply decreased.
ACKNOWLEDGEMENTS
I would like to thank Sarah Rizzi, Cain Stone, Christina Hecker, and Julie Smith for allowing me the
use of their automobiles for transportation to and from the gas station. A special thanks is extended
to the Center for Research and Economic Development for printing this poster, and I would finally
like to thank the Science Department at Waynesburg University for allowing me the use of their
equipment, media, and resources to complete this research.
Analysis of Peroxidase Activity in Various Fruits and Vegetables
Enzymes catalyze reactions in biological systems by speeding up the metabolic processes properly. Vitamins, minerals, and hormones all require enzymatic activities to function. Many enzymes are acquired through intake of certain foods, such as fruits and vegetables. Peroxidases are enzymes that catalyze the oxidation of a substance by peroxide. Enzymatic activity of fruits and vegetables can be determined by measuring the activity of peroxidase. The relationship between enzyme concentration and kinetics of peroxidase can be used to determine which type would be beneficial to a human’s diet. Guaiacol is a chemical compound used to monitor the activity of peroxidase. The optical density of the various extracts were measured at a wavelength of 500 nm. Three vegetables: potatoes, turnip, and radishes were examined. Measuring absorbance over a time period of one hundred twenty seconds, it was determined that potatoes contained a higher optical density, resulting in a greater enzymatic activity. Three fruits: apples, pomegranates, and oranges, were examined. Of the three, apples were experimentally determined to contain a higher optical density and therefore, a higher enzymatic activity. In the final part of the experiment it was determined which of the fruit or vegetable had a higher enzymatic activity. It was expected that potatoes would have a higher peroxidase activity because they contain a higher amount of catalase. The importance of this study was to determine whether certain fruits or vegetables contribute more enzymatic function to a human diet.
RESULTS
INTRODUCTION
•
•
•
•
•
•
•
•
Enzymes serve as catalysts in organisms to increase the metabolic rate
Enzymes can be proteins, ribozymes
Enzymes increase the rate of a reaction by a factor of 1020
Enzymes make life possible and are needed for every chemical reaction that occurs in the human body
Enzymatic activity is affected by temperature, change in pH, and the concentration of the substrate or the enzyme
Rate of a chemical reaction is dependent upon both the substrate and enzyme concentrations
Peroxidases catalyze the oxidation of a substance by peroxide
Changing substrate concentrations has an effect on enzyme activity
Time Vs. Absorbance
1.00
0.90
0.80
0.70
0.60
Absorbance
ABSTRACT
Jeff Fenton & Marietta Wright, M.S.
Waynesburg University, Department of Biology, Waynesburg, PA 15370
Potato
0.50
Turnip
0.40
Radish
0.30
0.20
0.10
0.00
0
20
40
60
80
100
120
Time (sec)
FIGURE 1 represents the data obtained from the vegetable experiment. Potatoes were shown to have higher absorbances than compared to the turnips and radishes. SPECIFIC AIMS
 Determine the relation ship between enzyme concentration and kinetics of the peroxidase reaction in various vegetables and fruits to further understand the importance of various food types in a human’s diet
 To determine which vegetables, and which fruits have higher enzymatic activity
 To determine whether vegetables or fruits have higher enzymatic activity and which contributes more to a diet
Time Vs. Absorbance
0.80
0.70
0.60
MATERIALS & METHODS
Substrate binds to the enzyme which induces a conformation change to form an enzyme‐substrate complex, which is lower in energy and increases the reaction rate. Substrate is transformed into products and released from the enzyme. The enzyme remains unchanged and therefore can catalyze more reactions.
 Fruits (Apples, Pomegranates, Oranges)
 Vegetables (Potatoes, Turnips, Radishes)
 Citrate Buffer
 0.1 M Citric Acid
 0.2 M Sodium Phosphate
 Spectrophotometer (λ= 500nm)
 25 mM Guaiacol
Absorbance
0.50
Apple
0.40
Pomegranate
0.30
Orange
DISCUSSION/CONCLUSION
• In an enzyme‐catalyzed reaction, the enzyme binds to the substrate to form a complex in the active site on the enzyme.
• The substrate, H2O2, induces a conformational change in the enzyme, peroxidase.
• Hydrogen peroxide and peroxidase must bind for to form a complex before the reaction occurs.
• The hydrogen peroxide is transformed into products, hydrogen peroxide and oxygen, when bonds are formed and broken in the complex.
• Peroxidase is a principle enzyme involved in the metabolism of hydrogen peroxide, which could be toxic to the cell.
• Along with catalase and superoxide dismutase, these enzymatic reactions are important in the digestions of fats and in the detoxification of alcohol and other poisons.
• Peroxidase is also important in the diet in that it helps oxidize particles that can damage the DNA, such as free radicals.
• Guaiacol was used to monitor the activity of peroxidase by undergoing a color change.
• The concentration of each extract was then able to be measured with spectrophotometer at a wavelength of 500 nm.
• Potatoes were determined to contain a higher enzymatic activity than turnips and radishes.
• Apples were determined to contain higher enzymatic activity than pomegranates and oranges.
• Potatoes are expected to have higher enzymatic activity than apples because they contain a higher amount of catalase.
ACKNOWLEDGEMENTS
0.20
0.10
0.00
0
20
40
60
80
100
120
Time (sec)
FIGURE 2 represents the data obtained from the fruit experiment. Apples were shown to have higher absorbances than compared to the pomegranates and oranges. I would like to thank the Waynesburg University
Biology Department for funding this project,
the Center for Research and Development building for printing the poster, and Mrs. Marietta Wright
for mentoring this project.