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
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