Kevin Cunningham, Donald Fisher, Jack Spreen

Professor Teh

Biology 3b

November 25, 2015

Oral Bacteria Inhibition With the Use of Listerine®

Title should be in the following format:

Title

Your NamesDepartment of Biological Sciences

Saddleback CollegeMission Viejo, California 92692

Abstract goes hereMake sure your abstract contains:

● Less than 1 page, 200 words● Adequate background coverage● Clearly stated hypothesis● Brief, sufficient methods● Appropriately stated data, discuss data● A conclusion related to hypothesis● No spelling or grammar errors● Single spaced, bold

For Materials & Methods, Results, and Discussion:● Use Columns: Leave a space after your Abstract and then insert

columns. • Inserting columns: Format tab → Columns → Click on the “two columns” box → Adjust the width to 3.1″ and the spacing to 0.3″ → Apply it to “this point forward” → Click “OK” • Cut & paste Introduction, Materials & Methods, Results, and Discussion on to page.

● Font size: 12 (for title) and 10 (for text). ● Make sure that it this single spaced.

Introduction (in bold, no underline, 12 font, centered)

(Space) Humans and various types of bacteria form symbiotic relationships virtually

everywhere throughout our bodies. A notable symbiotic relationship occurs in the mouths

of humans. Traditionally, humans brush their teeth to rid themselves of harmful bacteria

that can cause bad breath and gingivitis. However, most dentists recommend also adding

an oral antiseptic mouthwash to individuals’ daily mouth care regimen to reduce plaque

and biofilms (Arweiler, (no comma before et al) et al., 2001). Also, the consistent use of

oral antiseptics may greatly increase the health of gums and teeth as well as reduce the

possibility of contracting oral disease. (Gunsolley, 2010) Period here the clinical benefits

of anti-plaque mouth rinse are similar to the benefits of oral hygiene instructions at six

month recall appointments (citation?). One of the more prominent brands is Listerine®.

Listerine® claims that brushing only rids you of twenty-five percent (25%) numbers only

is OK of the bacteria that attributes to tooth decay, gingivitis and bad breath. While

adding Listerine® to an individual’s daily regimen, will aid in the removal of the

remaining seventy-five percent (75%) of harmful bacteria. The goal of this experiment

was to attain if Listerine® would in-fact remove significantly more bacteria in the mouth

over only brushing shorten the sentence by putting a period here, only brushing means no

Listerine without the use of Listerine®. The intellectual merit that can be obtained testing

Listerine’s effectiveness as an antiseptic mouthwash can provide helpful data to those

who suffer from oral diseases caused by bacterial growth period here, oral=mouth within

the mouth. (Yang, (no comma before et al)et al., 2015) if this citation belongs to the

previous sentence, period must come after the parenthesis, alcohol rinse free solutions

were shown to be effective in reducing plaque and bacteria levels immediately, and the

same was shown in long term patients as well (Gaffer, (no comma before et al)et al.,

1997). While using an oral antiseptic changes levels of bacteria in the mouth, there was

not found to be a significant difference in product effectiveness (Strydonck,(no comma

before et al) et al. 2005). The overall goal of this research is to aid in clarifying if

Listerine® will infact reduce the amount of bacterial colonies found within the human

mouth. The specific hypotheses and experimental design should be described.

Methods (in bold, no underline, 12 font, centered)

( Space) Ten (10) Since it is the beginning of the sentence use words for numbers;

numbers only is OK in the text subjects were swabbed a total of four (4) times to gather a

total of forty (40) swabs. Subjects were swabbed on the inside of the mouth swabbing

(avoid repeating the same word in a sentence) over gums, teeth, and inside of cheek.

Subjects were swabbed upon waking up in the morning, the first swab was taken without

brushing or drinking water, the second swab was taken after subject had brushed for sixty

(60) seconds. The third and fourth swabs were taken a day later. The third swab was

taken before rising in the morning before the subject had brushed, the fourth and final

swabs was were taken after subject had both brushed for sixty (60) seconds and swish

with Listerine® for sixty (60) seconds (how much Listerine was used?). Each Swab was

labeled in correspondence to their test number and subject initials,  then isolated into bags

, and kept chilled (in what degree?) until brought into the lab. Test numbers were

chronologically assigned in the same sequence as test were conducted. Swabs were

placed in test tubes with sterile water. Dilutions (how much solvent was added? what was

the concentration?) were performed on each sample tube a total of two (2) times. One (1)

ml of the final dilution was then mixed vigorously and added to petri dishes with nutrient

agar using the pour plate method, aseptic technique and incubated (temperature?)for

forty-eight (48) hours. Visual bacterial counts were then taken of developed bacterial

colonies and entered into Microsoft Excel and statistical analysis was performed using

descriptive statistics and a paired one-tailed student's T-test. (Include the statistical

analysis that was used including the p-values for statistical significance)

Check your grammar (make sure past tense is used in all sentences), avoid repeating

words, some sentences could be shortened, be more specific about where and when the

swabs were taken, did the subjects brush before bed? did they brush between 2nd and 3rd

swab?

Results (in bold, no underline, 12 font, centered)

( Space)This experiment produced a lot of raw data that was collected and analyzed. The

recorded amount of bacterial growth in colonies per each agar was multiplied by 121 to

account for the solution dilutions for a more accurate representation of the actual amount

of bacteria recorded per each subject. Pouring each agar plate directly after each solution

was diluted would have happened in an ideal world. Due to lab and time limitations, each

dilute solution waited approximately 72 hours until the actual pour which could have

affected bacterial growth (this should be discussed in the discussion).  After examining

each agar plate, the mean was taken of all the subjects bacterial count per agar plate. The

control pre-brush (Test 1) instead of test1, test2 or test3 use more clear labels, for

example with brushing/without brushing resulted in a higher mean bacterial count

compared following post bush with no rinse (Test 2) . As expected, our results showed a

similar trend between our second pre-brush control (Test 3)  compared to the post brush

with the antiseptic rinse (Test 4).  Recording from each individual agar plates indicated a

decrease in bacterial count was observed between test 1 and 2 (observation should be in

the discussion section). A similar trend observed from test 1 and 2 was also recorded

from test 3 and 4. The test recordings between test 1 -2 and 3-4 were very consistent in

term of results. The two control groups for pre brush test contained higher bacterial count

because brushing alone will decrease bacterial count. When oral antiseptic is added to

brushing, the gap between bacterial amounts in each subject increases significantly

compared to brushing alone. A visual representation of bacterial amounts observed is

( check your grammar, some sentences can be shortened, no discussion of results in the

results section) represented in Figure1.  No box around figures

Figure 1. Graph shows mean of bacterial colonies observed on agar nutrient plates. (P-

value of 0.006 was found using a one-tailed student T-test. Error bars are ±SEM).

(Make sure to label your graph by placing the figure and number in bold and the text in

italics, put a title on the graph, isn’t the y-axis supposed to be the mean number of

colonies as supposed to only numbers of colonies? Statistical significance?)

Note the decrease in bacterial amount between test 2 and 4 (clarification on what test 2

and 4 are is needed). Another visual will demonstrate the change in bacteria count in

terms of percentage to demonstrate the effect of oral cleaning technique practiced in this

experiment.

No titles on top (figure caption only) and no box around figure

Figure 2. Shows the reduction in bacterial colonies found using bacterial colony counts

on Agar nutrient plates. (Make sure to label your graph by placing the figure and number

in bold and the text in italics, where is the x-axis? label the axis, more description is

needed on the graph)

Discussion (in bold, no underline, 12 font, centered)

( Space)The data found in the experiment resulted in showing that supplementing oral

healthcare with a product such as Listerine® will decrease the amount of bacteria found

within the mouth, shown in Figure 2, the resulting findings state that there was found to

be a mean bacterial count of 181.5 colonies or a reduction of 42.3% of bacteria compared

to the baseline of 100%, using the group without brushing or swishing (sentences too

long, check your grammar, rephrase). The control group, which consisted of only

brushing, yielded 205.7 colonies or only a reduction of 34.6%. Johnson and Johnson

argues on their campaign ad that brushing removes 25% of bacteria in the mouth, and that

adding Listerine® will remove 99.9% of bacteria (citation), this statement was found to

be untrue.  Continual use of the product is necessary to inhibit bacterial buildup and

healthy gums. A factor during this test was the lack of a quick interchange between

dilution and placing the samples into the agar nutrient plates to culture an accurate

amount of bacteria. There was a delay of 48 hours between dilution and proper placement

in agar plates which may have affected the accuracy of the results. However, the results

did quantify the hypothesis that there would be a significant decrease in bacteria found in

samples taken after swishing with Listerine®. Another pattern arose that certain

individuals in the test groups were on different schedules. Subjects would brush their

teeth prior to going to bed and swabs were taken when subjects first arose in the morning

before ingestion of food or water, usually around 8AM. However, certain subjects in the

group would not retire to bed until 4AM. Meaning bacteria only had roughly 4 hours to

re-cultivate in the mouth from the previous days brushing. These individuals’ counts

looked to be lower in both pre and post brushing samples than those that had 8 or more

hours from their last brushing. A necessary adjustment to testing parameters would be to

swab subjects only after an allotted time from the previous days brushing. A recent

publication by Johnson and Johnson, owners of the product Listerine®, have recently

begun a campaign stating that it takes 21 days of supplementation to remove all of the

harmful bacteria (citation). This would be found to be true seeing as it may take more

time for the complete removal of the biofilms found in the mouth (you don’t know if this

would be true, you haven’t tested for 21 days). The company states that during the first

initial swishing of the product 99.9% of harmful bacteria are killed, but 0.1% remaining

in the mouth can reproduce and come back again stronger than previously (citation). This

would have to do with the remaining bacteria’s genetic coding that gave it resistance to

the antiseptic found in Listerine®. However, prolonged use of Listerine® will (or

could?)eventually lead to the removal and destruction of those bacterial cells. Another

factor that affects bacterial counts in the inhibition of biofilms found on the teeth created

by bacteria (grammar mistake). Once the initial treatment with Listerine® occurs,

individuals must continue to use the antiseptic to treat and control the bacteria,

discontinuing usage of the antiseptic could lead to a greater bacterial response to

treatment at a later time (Gordon, et al. 1985). However, different types of plaque and

oral bacteria can be found within individuals based on dietary and various oral activities,

involving smoking, drinking, or other activities involving the mouth, The individual

response by oral bacteria to these stimuli could affect the strength of the antibacterial

properties of some mouthwashes (Burt, B. A.. 1983).

This (what is?) is beneficial to individuals over the long term. However, a secondary

thought would be the removal of symbiotic bacteria found within the mouth that is

beneficial to the host. Several strains of beneficial bacteria contain antimicrobial

functions that remove unhealthy (missing the rest of the sentence)

Work on the grammar, this section needs to be heavily cited, use other journal articles

besides Johnson and Johnson, is there other articles that support your hypothesis? more

info should be given discussing the results

Conclusion (no need for a separate title, conclusion should be stated within the

discussion)

This experiment yielded beneficial results showing that Listerine® will infact remove a

greater percentage of the bacteria from the mouth. Though it does not remove the amount

Johnson and Johnson has advertised and stated through their own scientifical research.

Further testing should be conducted using long term effect analysis to validate the

statement made by the company. However, the results quantify the hypothesis that there

would be a drastic reduction of bacterial colonies found within the mouth.

Acknowledgment section?

Citations should be Literature Cited ( follow CBE Style Guide, alphabetize by authors’

last names)

Articles from Journals: Print

Format: Author(s). year. Article title. Journal title volume

number(issue number):inclusive pages.

Articles from Journals: Electronic

Format: Author(s). Date of publication. Title of article. Abbreviated

journal title [type of medium]; volume number(issue

number):pagination. Availability statement. [Date of accession if

needed].

Holt, R., Roberts, G., & Scully, C.. (2000). ABC Of Oral Health: Dental Damage,

Sequelae, And Prevention. BMJ: British Medical Journal, 320(7251), 1717–1719.

Retrieved from http://www.jstor.org/stable/25224907

        Mouth-Washes. (1909). Mouth-Washes. The British Medical Journal, 2(2548),

1300–1301. Retrieved from http://www.jstor.org/stable/25288365

       Johannes van Houte. (1983). Bacterial Adherence in the Mouth. Reviews of

Infectious Diseases, 5, S659–S669. Retrieved from http://www.jstor.org/stable/4453200

Eveillard, M., Schmit, J., & Eb, F.. (2002). Antimicrobial Use Prior to the

Acquisition of Multiresistant Bacteria. Infection Control and Hospital Epidemiology,

23(3), 155–158. http://doi.org/10.1086/502029

[Review of The Mycology Of The Mouth: A Textbook Of Oral Bacteria]. (1903).

[Review of The Mycology Of The Mouth: A Textbook Of Oral Bacteria]. The British

Medical Journal, 2(2240), 1470–1471. Retrieved from

http://www.jstor.org/stable/20278627

Robert S. Hirsch, & Nigel G. Clarke. (1989). Infection and Periodontal Diseases.

Reviews of Infectious Diseases, 11(5), 707–715. Retrieved from

http://www.jstor.org/stable/4455332

Volker, J. F.. (1950). Dental Caries Can Be Prevented. The American Journal of

Nursing, 50(2), 97–99. http://doi.org/10.2307/3467477

Genco, R. J., Plaut, A. G., & Moellering, R. C.. (1975). Evaluation of Human

Oral Organisms and Pathogenic Streptococcus for Production of IgA Protease. The

Journal of Infectious Diseases, 131, s17–s21. Retrieved from

http://www.jstor.org/stable/30106307

Gordon, J. M., Lamster, I. B. and Seiger, M. C. (1985), Efficacy of Listerine

antiseptic in inhibiting the development of plaque and gingivitis. Journal of Clinical

Periodontology, 12: 697–704. doi:10.1111/j.1600-051X.1985.tb00941.x

Filoche, S. K., Soma, D., Van Bekkum, M. and Sissons, C. H. (2008), Plaques

from different individuals yield different microbiota responses to oral-antiseptic

treatment. FEMS Immunology & Medical Microbiology, 54: 27–36. Retrieved from

http://onlinelibrary.wiley.com/doi/10.1111/j.1574-695X.2008.00443.x/full

Mandel, I. D.. (1979). Dental Caries: Although we seem to be paying for the

sweet life through the teeth, resistance to caries is possible, both as a natural and an

induced phenomenon. American Scientist,67(6), 680–688. Retrieved from

http://www.jstor.org/stable/27849533

Sognnaes, R. F.. (1965). Fluoride Protection of Bones and Teeth. Science,

150(3699), 989–993. Retrieved from http://www.jstor.org/stable/1717893

14. Burt, B. A.. (1983). The Scientific Basis for Plaque Control in the Prevention of Oral

Disease. Journal of Public Health Policy, 4(3), 298–312. http://doi.org/10.2307/3342111

Arweiler, N. B., L. Netuschil, and E. Reich. 2001. "Alcohol-free Mouthrinse Solutions to

Reduce Supragingival Plaque Regrowth and Vitality”. A Controlled Clinical

Study." Journal of Clinical Periodontology. 168-174.

http://europepmc.org/abstract/MED/11168742

Gaffar, A., J. Afflitto, and N. Nabi. 1997.  "Chemical Agents for the Control of

Plaque and Plaque Microflora: An Overview." European Journal of Oral Sciences.

<http://europepmc.org/abstract/MED/9395116>.

Gunsolley, John C. 2010. "Clinical Efficacy of Antimicrobial

Mouthrinses."Journal of Dentistry.  http://europepmc.org/abstract/MED/20621242

Strydonck, D. A. C. Van, M. F. Timmerman, U. Van Der Velden, and G. A. Van

Der Weijden.   2005. "Plaque Inhibition of Two Commercially Available Chlorhexidine  

Mouthrinses." Journal of Clinical Periodontology. 305-09.

http://europepmc.org/abstract/MED/15766375

Yang, Su-Jeong, Sang-Ha Han, Ah-Ra Lee, Joon-Ho Jun, Mi-Won Son, Se-Hwan

Oh, Jaehong Kim, and Soon-Young Paik. 2015. "Evaluation of Antimicrobial Effects of

Commercial Mouthwashes Utilized in South Korea." BMB Reports. 42-47

http://europepmc.org/abstract/MED/11168742

Review FormDepartment of Biological Sciences

Saddleback College, Mission Viejo, CA 92692

Author (s): Cunningham, Fisher and Spreen

Title: Oral Bacteria Inhibition With the Use of Listerine®

SummarySummarize the paper succinctly and dispassionately. Do not criticize here, just show that you understood the paper.

In this experiment the efficiency of Listerine® in significantly inhibiting bacterial growth over brushing only was tested. Ten subjects were swabbed in the mouth upon waking up before brushing and after brushing for two days. The samples were diluted, added to petri dishes and incubated. Mean number of bacterial growth was taken, a paired one-tailed t-test was performed. The results showed that using Listerine® decreased the amount of bacterial growth in the mouth but were not as drastic as the claim made by Johnson and Johnson. Further research was suggested, an increased percentage was speculated if Listerine® was used consistently for 21 days. This study is helpful for everyone especially those with oral diseases.

General Comments Generally explain the paper’s strengths and weaknesses and whether they are serious, or important to our current state of knowledge.

Weaknesses: A lot of grammar errors were observed, many sentences need to be rephrased. There were missing citations. Hypothesis wasn’t clearly stated in the introduction. Methods weren’t described specifically. Results were discussed in the results section. Figures weren’t labeled appropriately. Not enough citation was used in the discussion section. Not enough journal articles were used in the paper.

Strengths: The topic was interesting. The findings are helpful and important for anyone. Appropriate methods were used to make accurate results. Results were discussed well. Conclusions were clearly made. A claim by Johnson and Johnson was tested and dismissed.

Technical Criticism Review technical issues, organization and clarity. Provide a table of typographical errors, grammatical errors, and minor textual problems. It's not the reviewer's job to copy Edit the paper, mark the manuscript.

This paper was a final version This paper was a rough draft

Grammar errors should be addressed, sentences need to be rephrased. Some work needs to be done on clarification and organization ( specific comments regarding clarification and organization are made within the text). Literature cited needs to be alphabetized. Figures need to be labeled. Research format paper needs to be followed. (Refer to the paper for specific comments)

Recommendation

□ This paper should be published as is This paper should be published with revision□ This paper should not be published