A Comparative Study: Food (apple, breadand potato chips) that produces the Most

Bacteria in the Mouth Ma. Antonett F. Galdones1, Jia Raschelynn M. Mercado 1

1Biology student, Department of Biology, College of Science2Polytechnic University of the Philippines, Mabini Campus, Sta.

Mesa, Manila

INTRODUCTION

The human body is home tomany indigenousmicroorganisms, with distinctcommunities at differentanatomical sites (Dethlefsenet al., 2007). Recent studieshave shown the importance ofthe gut microbiota indigestion, fat storage,angiogenesis, immune system

development and response,colonization resistance andepithelial architecture(reviewed in Flint et al.,2007; Tappenden and Deutsch,2007; Cogen et al., 2008). Theoral cavity is also home tomicrobial communities, withimportant implications forhuman health and disease.

The dark, wet, and warmenvironment of the mouth, withthe occasional meal

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ABSTRACT

The oral cavity is also home to microbial communities, with important implications for human health and disease. The dark, wet, and warm environment of the mouth, with the occasional meal running through it, makes it an excellent niche for microbes to live. This study determines the food that produces the most bacteria in a person’s mouth and comparesthe reproduction of bacteria among the chosen foods (apple, bread, potatochips). Multiple plated nutrient agars were designated for each individual (sample A, B and C). Plates were divided into four (4) quadrants; Q1 (Apple), Q2 (Commercial bread), Q3 (Potato chips) and Q4 asthe control. The three (3) participants were asked to brush their teeth and gargle with water. They were asked to swab the inside of their mouth by gently rubbing the cotton swab on the buccal mucosa which then streaks

running through it, makes itan excellent niche formicrobes to live. Over thepast 40 years, scientists havebeen arduously working todiscover the over 500different species of bacteriain and around the mouth knowntoday. The mouth is comprisedof an oral cavity, whichincludes the teeth and gums,surrounded by the lips,cheeks, tongue, palate, andthroat. Each of these habitatsoffers differing environmentalconditions, and as such, iscolonized by a differentmicrobial flora. The oralenvironment is constantly influx. From birth to around age12, when the permanentdentition is complete, thelocal oral conditions arecontinuously changing as teethare shed and new ones erupt.[Skinner F.A. and Carr J.G.1974]. In addition,environmental factors such as,nutrition, diet, hygiene,smoking, dehydration, and evenstress, alter the ecologicalconditions of mouth. Salivacovers all surfaces and servesvarious important functions,mechanical and nutritional,(digestive, swallowing,cleansing, lubricant,bactericidal, and excretory)

in the oral cavity. [Wilson M.2007]

This study aimed to knowwhich food among the chosenfoods (apple, bread, potatochips) produces the mostbacteria in a person’s mouth.The foods mentioned above wereopted for the reason thanthey’re one of the frequentlyconsumed foods by thepopulace.

METHODOLOGY

I. Subject Participants

Mouth bacteria werecollected from three (3)individuals with healthy oraltissues and gingival (threewomen; age range 18, 19 and 20years). All participants werein good general health.

II. Methodology

A total of three (3)Multiple plated Nutrient Agarwere used in this studylabelled sample A, B and Cwith a corresponding food foreach quadrant. A fresh apple(Q1), commercial bread (Q2),potato chips (Q3) and cheeksas control (Q4).

Each plated nutrient agarwas designated for everyindividual (sample A, B and

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C). Plates were divided intofour (4) quadrants; Q1(Apple), Q2 (Commercialbread), Q3 (Potato chips) andQ4 as the control (cheeks).

To measure the accuracy ofthese study three (3) trialswere done:

Table 1: Shows three (3)trials in relation to sets offoods per samples A, B and C.

A B C Control

1st

trial

Apple(Q1)

Chips(Q3)

Bread(Q2)

Cheeks(Q4)

2nd

trial

Bread(Q2)

Apple(Q1)

Chips(Q3)

Cheeks(Q4)

3rd

trial

Chips(Q3)

Bread(Q2)

Apple(Q1)

Cheeks(Q4)

Testing procedures: The three(3) participants were asked tobrush their teeth and garglewith water. After a fewminutes they were asked toswab the inside of their mouthby gently rubbing the cottonswab on the inside of theircheek which was streak in Q4and labelled as the controland then the respondents wereallowed to eat the first setof food for the 1st trial.Participants were given 15

minutes only to eat the foodand to let the bacteriamultiply inside their mouthfor about 5 minutes. Sameprocedures were applied forthe 2nd and 3rd trial alongwith the brushing of theparticipants’ teeth before andafter.

Gram staining procedure:Prepare and heat-fix bacterialsmears and place it on astaining tray. Gently floodsmear with crystal violet andlet stand for 1 minute. Tiltthe slide slightly and gentlyrinse with tap water ordistilled water using a washbottle. Gently flood the smearwith Gram’s iodine and letstand for 1 minute and rinsewith tap water or distilledwater using a wash bottle. Thesmear will appear as a purplecircle on the slide.Decolorize using 95% ethylalcohol or acetone. Tilt theslide slightly and apply thealcohol drop by drop for 5 to10 seconds until the alcoholruns almost clear. Immediatelyrinse with water. Gently floodwith safranin to counter-stainand let stand for 45 seconds.Tilt the slide slightly andgently rinse with tap water ordistilled water using a washbottle. Blot dries the slidewith bibulous paper. View the

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smear using a light-microscope.

RESULTS AND DISCUSSIONS

This study demonstrated thatthe food that produces mostbacteria in a human’s mouthwas the apple. Apples havemany types of carbohydratesand almost all of the caloriesin apples come from thismacronutrient. Oral bacteriaconvert sugar into variousacids that degrade enamel,thus facilitating tooth decay.Since oral bacteria live in ina wet environment, sugar’sdehydration effects aresubdued and the bacteriametabolize the substanceinstead. Excessive consumptionof sugar can compromise immunefunction and promote bacterialgrowth in the body even theamount of fluorine in mottledenamel and dentine is notsufficient to inhibitbacterial growth, but mayprevent the formation of acidsfrom carbohydratefermentation.

Dr. Loesche showed that onecould calculate the growthrate of bacteria indirectlyfrom two other known facts. Anaverage person swallows about1 litre (1,000 millilitres) ofsaliva daily and 1 ml. of

saliva contains about 100million microbes. Multiplying1,000 ml of saliva by 100million (microbes per ml.)gives us 100 billion. That’sthe number of microbes in thesaliva we swallow every 24hours. Adopting thiscalculation the researchesassume for about 15 minutes ahuman mouth can produce 250bacteria in the mouth (seefig.2).

The comparison of bacterialproduction was based upon thephysical appearances of thegrowth in the plated culture.

Table 2: Shows the legendthat is to be used for thecomparison of the results.

+ Normal

++ Many bacteria

+++ More bacteria

++++ Most bateria

Table 3: Shows the comparisonbetween the quadrants of PlateA.

Q1(Apple

)

Q2(Bread

)

Q3(Chips

)

Control

++++ ++ +++ +

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Table 4: Shows the comparisonbetween the quadrants of PlateB.

Q1(Apple

)

Q2(Bread

)

Q3(Chips

)

Control

++++ ++ +++ +

Table 5: Shows the comparisonbetween the quadrants of Platec.

Q1(Apple

)

Q2(Bread

)

Q3(Chips

)

Control

++++ +++ ++ +

Upon the observation of thedifferent Plates, there is asignificant difference uponthe size of the bacterialstreak of the differentquadrants, with the Quadrant 1

(Apple), being bigger andwider than the rest.

Fig 1 Food materials that wereused

Fig 2 Bacterial colonystreaked in Q4 (Control).

Fig 3 Bacterial colonystreaked in Q1 (Apple)

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Q1

Bacterial

Q3

Q4

Fig. 4 Bacterial colonystreaked in Q2

Fig 5 Bacterial colonystreaked in Q3

Fig 6 A-B Gram stain of mouthbacteria under LPO with itsgrape-like clusters.

Macroscopic observation ofbacterial smears results togram positive bacteria. UnderLow Power Objectives smearsappears a violet color. Bothare cocci bacteria, one ingrape-like clusters (a) andthe other in chain like (b).

Staphylococcus andStreptococcus are bothpositive bacteria that arepredominant in human mouth(Mac et al., 2011).

CONCLUSION

It was in the Quadrant ofthe apple that has the mostbacteria.

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A

Sugar is the main cause oftooth decay. Apples have alarge amount of sugar in them.Even though apples are healthyfor the body, they can causecavities just as quickly ascandy can because of all thenatural sugars they have. Thebacteria in your mouth thriveon sugar, and after you eatsomething sugary, it producesan acid that damages yourteeth.

The worst time to not brushyour teeth is before bed. Yourmouth is driest when you sleepand, without saliva breakingit down, bacteria willmultiply rapidly.

RECOMMENDATIONS

Eat a well-balanced dietcharacterized by moderationand variety. Choose foods fromthe five major food groups:fruits, vegetables, breads andcereals, milk and dairyproducts and meat, chicken,fish or beans. Avoid fad dietsthat limit or eliminate entirefood groups, which usuallyresult in vitamin or mineraldeficiencies.

Always keep your mouthmoist by drinking lots ofwater. Saliva protects bothhard and soft oral tissues. Ifyou have a dry mouth,supplement your diet withsugarless candy or gum tostimulate saliva. Always brushyour teeth at least twice aday—before bed and after youwake up.

Foods that cling to yourteeth promote tooth decay. Sowhen you snack, avoid soft,sweet, sticky foods such ascakes, candy and dried fruits.Instead, choose dentallyhealthy foods such as nuts,raw vegetables, plain yogurt,cheese and sugarless gum orcandy.

When you eat fermentablecarbohydrates, such ascrackers, cookies and chips,eat them as part of your meal,instead of by themselves.Combinations of foodsneutralize acids in the mouthand inhibit tooth decay. Forexample, enjoy cheese withyour crackers. Your snack willbe just as satisfying andbetter for your dental health.One caution: malnutrition (badnutrition) can result from toomuch nourishment as easily astoo little. Each time you eat,you create an environment for

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B

oral bacteria to develop.Additionally, studies areshowing that dental disease isjust as related to overeatingas heart disease, obesity,diabetes and hypertension. Somaking a habit of eating toomuch of just about anything,too frequently, should beavoided.

REFERENCES

Books

1. Wilson, M. Bacteriology ofHumans: an ecologicalperspective. Massachusetts:Blackwell Publishing Ltd, 2008

2. Avila, M., Ojicus, D. M.And Yilmaz, O. “The OralMicrobiota: Living with aPermanent Guest.” DNA & CellBiology, 2009, DOI:10.1089/dna.2009.0874

Journals

1. doi:10.1128/JB.183.12.37703783.2001J.

Bacteriol. June 2001 vol.183 no. 12 3770-3783

2. Koshy Philip et al.Identification of MajorCultivable AerobicBacteria in the OralCavity of MalaysianSubjects. AmericanJournal of Biochemistryand Biotechnology 4 (4):367-370, 2008

3. van der Mei, H. C.,Rustema-Abbing, M., deVries, J. & Busscher, H.J. (2008). Bondstrengthening in oralbacterial adhesion tosalivary conditioningfilms. Appl EnvironMicrobiol 74, 5511–5515.

4. Skinner F.A. and CarrJ.G. The Normal MicrobialFlora of Man. London:Academic Press Inc, 1974

Websites

1. http://www.scientificamerican.com/article/beat-bad-breath-keep-mouth-bacteria-happy/

2. http://www.education.com/science-fair/article/mouth-bacteria/?page=2

3. http://www.knowyourteeth.com/infobites/abc/article/?abc=N&iid=315&aid=1274

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4. http://www.colgateprofessional.com/patient-education/articles/mouth-healthy-eating

5. http://www.ehow.com/facts_5811712_effect-

sugars-bacterial-growth.html

6. http://www.livestrong.com/article/161300-what-are-the-different-types-of-bacteria-found-in-the-human-mouth

APPENDIX A

Shows the Aseptic Technique and Streaking of specimens

Fig. 1 Aseptic Techniquebefore streaking Fig.2 Preparing a bacterial smear

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Fig. 3 Heating the inoculating loop afterstreaking

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