Undergraduate Research Long Term Grant Application
Cover Sheet
Amount Requested: $3490.60
Is there more than one person involved in this grant? YES
Student Information
___ Garner, David, Cooney, Mitchell, Fonnesbeck, J. Scott
Project Information
__The Effect of Probiotics in Yogurt on oral Streptococcus mutans
Project Title (10 words or less)
__Wright, Scott
______________________
Faculty Mentor Name (last, first)
_________
3905 _______________________
Mail Code
___ Weber State is the University_______________________Clinical Laboratory Science _________
College (Weber State is the University, NOT college)
Department
This project _X DOES/___ DOES NOT require review by the WSU Institutional Review Board for
Human Subjects or the WSU Animal Care and Use Committee.
__________________________________________
Student Signature
___________________
Date
___________________________________________
Project Mentor Signature
___________________
Date
__3905_______
Campus Mail
___6716 ____
Phone Ext.
___________________________________________
Undergraduate Research Committee Representative
___________________
Date
___________________________________________
Faculty Mentor Department Chair
___________________
Date
OUR Long Term Grant Application
Revised October 28, 2010
Page 1 of 10
UNDERGRADUATE RESEARCH LONG TERM GRANT APPLICATION
Budget Worksheet
BUDGET ITEM
Department or College
Funds
Ray Garner DDS
donated
$195.50
Materials
Equipment
Outside Agency
Funds
Personal
Funds
Undergrad.
Research Funds
GRAND
TOTAL
$3225.60
$3421.10
$265 for
incentive prize
$265.00
$3490.60
$3686.10
Sodexo Fridge
CLS dept incubator
CLS dept autoclave
Benefits
Mileage to gather
Data (.36 per mile)
GRAND TOTAL
OUR Long Term Grant Application
$195.50
Revised October 28, 2010
Page 2 of 10
UNDERGRADUATE RESEARCH LONG TERM GRANT APPLICATION
Body of Proposal
Project Description
The aim of the research project is to determine if probiotics contained in commercial
yogurt can decrease the count of Streptococcus mutans in the oral cavity. Bacteria such as
Streptococcus mutans cause common and extensive oral infections in 95% of the general
population.1 Surgical means have been the most common form of treatment for dental caries
(also known as cavities) along with fluoride and other public preventative measures which have
resulted in a reduction of dental caries.2, 3 Unfortunately, those measures do not address the
bacterial infections that are the root cause of oral health problems.1
According to the World Health Organization and United Nations Food and Agricultural
Organization probiotics are “living microorganisms, principally bacteria, that … when ingested
in sufficient quantities, have beneficial effects on human health.”4 Studies showing the effect of
probiotics on oral health are becoming more prominent as additional information is learned about
probiotics. 5, 6 Previous studies in Europe and Asia showed that the probiotics found in yogurt
affect the population of cavity causing bacteria within the mouth.7-10 One such study showed
there was “a statistically significant reduction of salivary mutans streptococci after probiotic
yogurt consumption.”10
There have not been any studies done in the U.S. that have examined the effect of
probiotics on S. mutans. The previous studies conducted in Europe and Asia each studied
populations with distinctive environments, differences of lifestyles and genetic makeup, and
various mixtures of probiotics used in the yogurt. A study within the U.S. would show the effect
that probiotics have in a U.S. population, specifically a set of WSU students. This information
could help prevent the occurance of dental carries and S. mutans infections in the U.S.
OUR Long Term Grant Application
Revised October 28, 2010
Page 3 of 10
The project will consist of forty WSU college students, ages 18-40. As a double-blinded
study, participants will be divided into a group that eats yogurt with live probiotic organisms
(commercial product) and a group that eats the same yogurt without live probiotic organisms
(commercial product that has been autoclaved to kill all bacteria.) The participants will be
evaluated for the amount of S. mutans in the oral cavity nine times over the course of nine
weeks: three weeks pre-study with no yogurt, three weeks of eating yogurt daily, and finally for
three weeks without yogurt. The concentration of S.mutans will be quantitated by a tooth
scraping using Dentocult® SM Strip Mutans kits manufactured by Orion Diagnostica.11 The
results will then be evaluated using statistical analysis.
Student information:
We are currently undergraduate students in the Clinical Laboratory Science (CLS)
Department at WSU. Scott Wright is the faculty mentor and will serve as an advisor for the
project when needed. Our project falls on the research continuum as shown below:
Dependent_____________________X_ Independent
Our research team consists of three members, all with varied clinical experience. One
has been working for two years in a dental laboratory performing model work. He also has a
brother currently enrolled in dental school. Another has been employed by Ogden Clinic for one
year as a laboratory assistant. His father is a dentist and he also a brother enrolled in dental
school. Another is the present media chef for the CLS Department at WSU. All three have
completed their AAS degree in CLS and are currently enrolled in the baccalaureate program. All
three have also completed over 100 semester hours at Weber State and have learned the basic
skills needed to work in a clinical laboratory and conduct scientific research.
OUR Long Term Grant Application
Revised October 28, 2010
Page 4 of 10
At the end of the research project the results will be submitted for potential publication in
the WSU undergraduate research journal ERGO. We will also present the project at the WSU
Undergraduate Research Symposium in the spring of 2011 and apply to present at the National
Society of Clinical Laboratory Sciences conference in the summer of 2011. In addition we will
apply to present at NCUR and UCUR in the fall of 2011. We also plan to submit the final result
to the Journal of Dental Research for publication.
Project Methods & Timeline
To recruit participants for this project, the research team will announce the project to each
CLS class during the first week of the spring 2011 semester. Individuals who want to participate
in the study will be contacted one or two days prior to commencement of the project. Each
potential participant will be evaluated to determine if they are eligible and willing to participate
(see Addenda 1 and 2). Those selected to participate will also be given a demonstration of
proper teeth brushing and flossing by Stephanie Bossenberger, the Dental Hygiene Department
Chair at WSU.12
During the study which will begin January 12, the forty participants will be required to
brush their teeth twice daily and floss once daily for three weeks. At the end of the three weeks
the amount of S. mutans on their teeth will be measured three times in one day (morning,
afternoon, and evening) using Dentocult® SM Strip Mutans kits. This will serve as the baseline
count for the amount of S. mutans. For the next three week course of the study, as a doubleblinded study, the participants will be randomly divided into two groups. One group will be
instructed to consume yogurt once a day that contains live probiotic organisms (unaltered
commercial product) and one group will be instructed to eat yogurt that does not contain live
organisms (yogurt that has been autoclaved to kill all bacteria.) At the end of the three weeks the
OUR Long Term Grant Application
Revised October 28, 2010
Page 5 of 10
amount of S. mutans will be measured three times (morning, afternoon, and evening.) During the
final three weeks, the participants will not consume any yogurt, but will continue to brush and
floss as instructed. At the conclusion of the three weeks, S. mutans will again be measured three
times as before. The project will be completed by the end of March. The data will be statistically
analyzed using a T-test with an alpha level of 0.01 to determine if results are significant or not.
Budget Explanation
The total budget is close to $3500 because we are trying to maximize the amount of patients we
can test with the cost of the test and the cost of the yogurt, while still staying within the budget.
Materials
Dentocult®SM Strip mutans
Kits from Orion Diagnostica/North Bay Bioscience
LLC (US distributors)
60 bottles Toothpaste (donated by Ray Garner DDS)
60 Toothbrushes (donated by Ray Garner DDS)
60 Packages floss (donated by Ray Garner DDS)
Yogurt (Purchased through Sodexo for WSU
discount.)
S. mutans KPSK 2
S. faecium 9790
49.95 per kit (47 kits = $2347.65 = 470
tests)
Shipping $31.95
$66.60 (donated)
$79.80 (donated)
$49.10 (donated)
$756 (6 oz. cups, $0.75 per cup, 1008
cups)
$45 (positive control)
$45 (negative control)
Equipment
Fridge to store yogurt in
Incubator, Autoclave
Provided use by Sodexo
Provided use by the CLS department
Benefits
Incentive for completing the study
$265 for gift cards to the WSU
bookstore. We are attempting to contact
the WSU bookstore to match the amount
we contribute to increase the incentive.
References
1.
2.
3.
4.
Caglar, E., B. Kargul, and I. Tanboga, Bacteriotherapy and probiotics' role on oral
health. Oral Diseases, 2005. 11(3): p. 131-137.
Adair, S.M. and Q. Xie, Antibacterial and probiotic approaches to caries management.
Advances In Dental Research, 2009. 21(1): p. 87-89.
Anderson, M.H. and W. Shi, A probiotic approach to caries management. Pediatric
Dentistry, 2006. 28(2): p. 151-153.
Bonifait, L., F. Chandad, and D. Grenier, Probiotics for oral health: myth or reality?
Journal (Canadian Dental Association), 2009. 75(8): p. 585-590.
OUR Long Term Grant Application
Revised October 28, 2010
Page 6 of 10
5.
6.
7.
8.
9.
10.
11.
12.
Burton, J.P., et al., A preliminary study of the effect of probiotic Streptococcus salivarius
K12 on oral malodour parameters. Journal Of Applied Microbiology, 2006. 100(4): p.
754-764.
ÇAglar, E., et al., A probiotic lozenge administered medical device and its effect on
salivary mutans streptococci and lactobacilli. International Journal of Paediatric
Dentistry, 2008. 18(1): p. 35-39.
Caglar, E., et al., Effect of yogurt with Bifidobacterium DN-173 010 on salivary mutans
streptococci and lactobacilli in young adults. Acta Odontologica Scandinavica, 2005.
63(6): p. 317-320.
Busscher, H.J., A.F. Mulder, and H.C. van der Mei, In vitro adhesion to enamel and in
vivo colonization of tooth surfaces by Lactobacilli from a bio-yoghurt. Caries Research,
1999. 33(5): p. 403-404.
Nikawa, H., et al., Lactobacillus reuteri in bovine milk fermented decreases the oral
carriage of mutans streptococci. International Journal Of Food Microbiology, 2004.
95(2): p. 219-223.
Caglar, E., et al., Salivary mutans streptococci and lactobacilli levels after ingestion of
the probiotic bacterium Lactobacillus reuteri ATCC 55730 by straws or tablets. Acta
Odontologica Scandinavica, 2006. 64(5): p. 314-318.
Jensen, B. and D. Bratthall, A new method for the estimation of mutans streptococci in
human saliva. Journal Of Dental Research, 1989. 68(3): p. 468-471.
Bossenberger, S., MS, RDH Personal Interview, D. Garner and S. Wright, Editors. 27
Oct 2010: Ogden.
OUR Long Term Grant Application
Revised October 28, 2010
Page 7 of 10
Addendum 1
The Effect of Probiotics in Yogurt on oral Streptococcus mutans
Informed Participant Consent Document
Study Overview:
This research study will address the ability of certain ingredients (probiotics) in common
yogurt to lower the levels of specific cavity causing bacteria found in the mouth. The study will
involve about forty WSU college students and last approximately nine weeks. You will be
expected exhibit good oral health by brushing your teeth twice a day and flossing once a day
during the entire nine weeks of the study. You will be required to attend a ten minute
instructional meeting conducted by a dental hygienist faculty member on the proper technique
for brushing and flossing your teeth. For the first three week section, you will only brush and
floss. At the onset of the second three week period, you will meet with researchers to collect a
tooth scraping sample (using a toothpick), obtain a three week supply of yogurt to be consumed
daily, and receive instructions about yogurt consumption. The final three week period, you will
to stop yogurt consumption, but continue to maintain your tooth brushing and flossing regimen.
You will be required to meet with the research team at the end of each three week time period to
collect teeth scraping samples.
By signing this Informed Consent Document you agree to the following terms and
conditions during the nine week research study:
•
•
•
•
•
Brush teeth twice daily and floss once daily,
Meet with the research team a minimum of four times throughout the course of the study,
Follow instructions for yogurt consumption provided by the researchers,
Inform research team if any health complications arise (such as a cold, the flu, need for
antibiotics etc.),
Inform research team of any desire to terminate participation early.
Due to the nature of the study you shall be excluded from the project if you:
•
•
•
•
•
•
•
Fail to honestly report all actions pertinent to the study including failure to comply with study
requirements (fill out the form that will be handed out with the supplies),
Are currently pregnant or become pregnant during the course of the study,
Smoke or chew tobacco,
Have any allergies to dairy products,
Have any chronic illness (i.e. asthma, diabetes, physical disabilities that would prevent
brushing of teeth, etc),
Currently taking or need to take antibiotics during the course of the study,
Consume products that contain xylitol (i.e. Orbit gum, Stride gum, Trident gum, to name a
few.)
Risks:
While participating in this study, you may develop pain and sensitivity in the mouth due
to daily brushing and flossing of teeth. If this occurs consult a qualified dentist before continuing
the study. Additionally live probiotic cultures have been shown to occasionally affect the
OUR Long Term Grant Application
Revised October 28, 2010
Page 8 of 10
intestinal tract. If any discomfort or issue occurs contact your primary care provider to determine
if continuing with the study is recommended. Weber State University and research team
members cannot be held responsible for any accidental harm or injury which may occur during
the course or in connection to the study.
Benefits:
You will receive a complimentary toothbrush, tube of toothpaste, and package of dental
floss. In addition, you will receive 6 oz cup of yogurt for every day of the three week testing
period. You will also receive a 10-15 minute demonstration by the Department Chair of the
Dental Hygiene Department on how to properly brush and floss. If you fulfill all requirements
and complete the study, you will be entered in to a grand prize drawing to include one 200 dollar
gift card, one 50 dollar gift card, and one 15 dollar gift card from WSU bookstore. Finally, the
information garnered from this study will benefit the dental community by providing information
about oral health care and the possible ways of combating dental cavities.
Participation in this study is voluntary, and refusal to participate will involve no penalty
or loss of benefits to which you are otherwise entitled. You may discontinue participation at any
time without penalty or loss of benefits to which you are otherwise entitled. Dismissal from the
study may occur at anytime and for any reason which the researchers deem appropriate. The
study results and personal information of participants will be kept confidential and not be given
to anyone outside of the research team. The Food and Drug Administration may inspect study
records if necessary. Results of the overall study will be available upon request at the termination
of the project. For answers to any pertinent questions about the research and your rights as a
participant, feel free to call Scott Wright in the WSU Clinical Laboratory Science Department
(801-626-6716).
-----------------------------------------------------------------------------------------------------------
By signing this form I consent to the information previously presented.
______________________________
Name (last, first)
_______________________________
Signature
___________
Date
_____________________________
Phone number
*Star the best way to contact you
________________________
Email address
OUR Long Term Grant Application
Revised October 28, 2010
Page 9 of 10
Addendum 2
Please keep track of the items listed below during the course of the study. Please highlight
those things that are missed. If you do not turn this paper in at the end of the study, your
name will not be entered into the drawing for the gift cards.
Brush
Teeth
Floss
Yogurt
Consumption
Brush
Teeth
Floss
Yogurt
Consumption
Jan 12
13
13
14
14
Feb 15
15
16
16
17
17
18
Jan 18
19
19
20
20
21
21
Feb 22
22
23
23
24
24
25
Jan 25
26
26
27
27
28
28
Mar 1
29
2
30
3
31
4
Feb 1
5
2
6
3
7
4
Mar 8
5
9
6
10
7
11
Feb 8
12
9
13
10
14
11
Mar 15
12
Mar 16
Please list anything you feel could have affected your results of this study:
OUR Long Term Grant Application
Revised October 28, 2010
Page 10 of 10