Department of Oral Pathology And

THESIS SYNOPSIS

Dr.SUPRIYA.H

DEPARTMENT OF ORAL PATHOLOGY AND

MICROBIOLOGY

KVG DENTAL COLLEGE AND HOSPITAL

KURUNJIBAGH, SULLIA,DK – 574327

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

4th BLOCK, JAYANAGAR,BANGALORE,KARNATAKA

ANNEXURE

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

NAME OF THE CANDIDATE AND ADDRESS / DR.SUPRIYA.H
POST GRADUATE STUDENT,
DEPT OF ORAL PATHOLOGY AND
MICROBIOLOGY,
K.V.G DENTAL COLLEGE AND HOSPITAL
SULLIA,DK-574327
NAME OF THE INSTITUTION / K.V.G DENTAL COLLEGE AND HOSPITAL
SULLIA,DK-574327
COURSE OF THE STUDY AND SUBJECT / MASTER OF DENTAL SURGERY
DEPARTMENT OF ORAL PATHOLOGY AND MICROBIOLOGY.
DATE OF ADMISSION TO COURSE / 24-05-2013
TITLE OF THE TOPIC: AN EVALUATION OF ORAL CANDIDA ALBICANS IN SMOKERS WITH AND WITHOUT LESIONS–A MICROBIOLOGICAL STUDY.
BRIEF RESUME OF THE INTENDED STUDY:
6.1) Need for the study:
Candida albicans isan ubiquitous, yeast like fungus and an opportunistic pathogenwith potential to cause disease under specific conditions and circumstances. Oral candidiasis serves as a useful clinical marker for the presence of significant predisposing conditions.1
Smoking is clearly recognized as a predisposing factor for oral candidosis.The smoking habit may provoke increased oral epithelial keratinization and subsequent enhancement ofhydrophobicity which predisposes the smoker to higher oral yeast colonization.2Studies have reportedincreased oral candidal load to 30-70% in smokers.3
The presence ofCandida in mouth with epithelial changes may predispose to candidal infection which,along with other cofactors may induce epithelial dysplasia.4 Certain strains of Candida have been shown to produce nitrosamines, a chemical carcinogen which either act directly on oral mucosa or interact with other chemical carcinogens to activate specific proto-oncogenes and thereby initiating oral neoplasia.5
Recently, an interest in the study of Candida albicans has markedly increased mainly because of its association with human immune-deficiency viral infection, but also because of its relation with potentially malignant lesions of oral mucosa.6 In view of these changes, there is a need to establish risk,specifically associated withoral candidosis in order to implement appropriatepreventive program. This study is undertaken to isolate,identifyand evaluate the roleofCandidaalbicans insmokers with and without oral lesions.
6.2Review of Literature:
A study ofsmoking effects on the prevalence and intraoral distribution of Candida albicans was done,swabs and saliva samples from hundred healthy persons, smokers and non-smokers, were cultured. Among carriers, the mean concentration of Candida albicans colonyforming units in saliva of smokers was twice that of non-smokers. This study suggested that the effect is twice that of the non-smokers, and the isolation frequency of Candida albicans at each of five mucosal sites was also higher in smokers than in non-smokers. Men were mucosal carriers more often than women (p < 0.025), and the mucosal site from which Candida albicans was recovered most often was the posterior dorsum of the tongue.7
A study to assess and compare the quantitative and qualitative oral colonization of Candida species between a group of healthy tobacco smokers and a comparable group of non-smokers, and to investigate a possible correlation between oral candidal colonization and the quantity or duration of the smoking habit was done. Fifty smokers and fifty non-smokers were included in the study. Candida species were isolated using the concentrated oral rinse technique and identified using the germ tube test. Overall candidal transmission was 84 %. Candida species were isolated from forty two (84 percent) of the smokers and thirty seven (74 percent) of the non-smokers (p>0.05). The mean colony forming unit/ml were 333 (SD=358) and 268 (SD=332), respectively (p>0.05)concluding a variation of colonisation of candida in smokers.8

In another studytwo hundred and twenty-three patients who were undergoing an incisional biopsy for the diagnosis of an oral mucosal lesion were enrolled. Mouth swills were obtained from each patient for the presence and amount of oral yeast present. Some of the patients (44.6%) had a histopathological diagnosis of either oral epithelial dysplasia or oral squamous cell carcinomaand the frequency of oral yeast carriage was significantly greater (P<0.001) in these patients than those without histopathologically detected dysplastic or neoplastic oral lesions. Furthermore, significantly (P<0.001) more patients with oral epithelial dysplasia or oral squamous cell carcinoma had a higher number of yeast (over 1000 colony forming unit/ml) in their oral cavity than patients without any evidence of epithelial dysplasia or neoplasia histopathologically. It was concluded that the degree of epithelial dysplasia present in these patients correlated with higher amounts of yeast in the oral cavity (P=0.017). The results of the study revealed that there is an interaction between oral carriage of yeast and oral epithelial dysplasia.9

The effect of cigarette smoke condensate (CSC) on ten clinical isolates of Candida albicans obtained from non-smokingvolunteers werestudied in vitro. Cigarette smoke condensate was generated by complete burn of five commercial cigarettes in an in-house smoking machine and used to prepare the culture broth in which the strains were grown. In 24-h intervals (T(24), T(48), and T(72)), the cells were harvested, washed, sub cultured, and the resultant growth were evaluated for possible variations .The results indicated a temporal increase in the secretion rates of enzymes, particularly when yeast cells were exposed tocigarette smoke condensate for 48-72 h (P < 0.05). It was concluded that cigarette smoke condensate may significantly enhance the secretion of candidal histolytic enzymes and adherence to surfaces, thereby promotingoralyeast carriage and possible infection in smokers.2

6.3 Aim of the study
To evaluate the prevalence of oral Candida albicans insmokers and theirassociationwith and without oral lesions.
6.4Objectives of the study

1. To isolate and identify Candida albicans from saliva.
2. To compare thecolonization oforal Candida albicans in non-smokersand smokers withoutlesions.
3. To study the prevalence of oralCandida albicans in smokers with lesions.
4. To compare the prevalence of Candida albicans among non-smokers, smokers without lesions and smokers with premalignantor malignant lesions.

7. / MATERIALS AND METHODS
7.1 Source of the Data
The study will include total 100 subjects (20non-smokers without any lesions as healthy controls, 40 smokers without lesions, 40smokers with lesions) from the outpatient Department of Oral Medicine and Radiology, K.V.G Dental College and Hospital, Sullia, after obtaining an informed consent from the patients.
The patients will be grouped as:
Group A: Healthy controls without smoking habits and without any lesion.
Group B:Smokers without oral lesions.
Group C: Smokerswith oral lesions.
7.2 Method Of Collection Of Data:
All subjects enrolled will be screened by using the concentrated oral rinse technique. Oral rinse samples will be obtained by asking patients to rinse their mouths with 10 mL of phosphate-buffered saline (PBS; pH 7.2, 0.1 M) for 60 seconds and to expectorate the rinse into a sterile container.
Saliva culture technique:
Saliva sample are centrifuged and sediment will be inoculated ontwo Sabouraud’s dextrose agar slants incubated atroom temperature and37°C for 3-5 days.
Identification of Candida:
Growth on Sabouraud’s dextrose agar will be stained with PAS and examined under light microscope. Buddingyeasts are subjected to germ tube test. A small inoculum from an isolated Candidal colonyis picked up with a sterile inoculating loop and suspended in glass tube containing normal human serum (0.5 ml). The mixture is incubated at 37°C for 2-3 hrs. A drop of mixture will be placed in a clean glass slide and covered with a cover slip. This is first examined under a low-power objective to locate the group of cells and later, the presence of germ tube will be confirmed under high-power objective of the microscope.
Identification of Candida albicans by chlamydospore production:
Germ tube-positive samples were further taken for chlamydospore productionincorn meal- Tween 80agar media at room temperature .After24-48 hourschlamydospores will be observed under wet mounts for isolation of candida albicans.
Histopathological diagnosis:
Histological sections of oral lesion processed according to standardized laboratory procedures and stained with Haematoxylin and Eosin for histopathological diagnosis.
Inclusion criteria:
1. Patients in the age group of above20 years.
2.Patients with smokinghabits.
3. Biopsy of oral lesions associated with smoking.
4. Control group will comprise of healthy individuals without any smoking habit and oral lesions.
Exclusion criteria:
1. No ongoing treatment with antifungals or antimicrobial mouth rinses.
2.Individuals with a history of prolonged antibiotic or steroid therapy, iron deficiency anemia, diabetes, xerostomia, HIV, salivary gland diseases.
Statistical analysis
Chi- square test
7.3 Does the study require any investigations (or) interventions to be conducted in patients (or) other humans (or) animals?
Yes.
7.4 Has ethical clearance been obtained from your institution?
Yes. Institutional Ethics Committee clearance enclosed.
REFERENCES:

1. Lynch DP. Oral candidiasis: History, classification, and clinical presentation. Oral surg oral med oral pathol 1994;78(2):189-93.

1. Baboni FB, Barp D, Izidoro AC. Enhancement of Candida albicans virulence after exposition to cigarette mainstream smoke. Mycopathologia. 2009;168(5):227-35.

1. Bernhard D.Cigarette Smoke Toxicity: Linking Individual Chemicals to Human Diseases. Wiley-VCH;2011.

1. Scully C,El Kabir M. Samaranayake LP. Candida and Oral Candidosis: A Review.CROBM 1994;5(2):125-157.

1. Saigal S, Bhargava A, Mehra SK, Dakwala F. Identification of Candida albicans by using different culture medias and its association in potentially malignant and malignant lesions. Contemp Clin Dent2011;2(3):188–193.

1. Vuckovic N, Bokor-Bratic M, Vuckovic D,Picuric I. Presence of Candida albicans in potentially malignant oral mucosal lesions. Arch Oncol 2004;12(1):51-54.

1. Oliver DE, Shillitoe EJ.Effects of smoking on the prevalence and intraoral distribution of Candida albicans.J Oral Pathol1984;13(3):265-70.

1. Darwazeh AMG, Al-Dwairi ZN, Al-Zwairi AAW. The relationship between tobacco smoking and oral colonization with Candida species.J Contemp Dent Pract 2010 1;11(3):017-24.

1. McCullough M, Jaber M, Barrett AW. Oral yeast carriage correlates with presence of oral epithelial dysplasia. Oral Oncol 2002;38(4):391-3.

1. Fisher F, LookMB. Fundamentals of diagnostic mycology. 2nded. WB Saunder;1998.p.200-202.vol (1)..

INFORMED CONSENT FOR PARTICIPATION IN RESEARCH
I, Dr. SUPRIYA.H Post Graduate Student in Department of Oral Pathology and Microbiology, am conducting a dissertation work for award of M.D.S degree in OralPathology and Microbiology. The topic for the study is-AN EVALUATION OF ORAL CANDIDA ALBICANS IN SMOKERS WITH AND WITHOUT LESIONS –A MICROBIOLOGICAL STUDY.
AIMS AND OBJECTIVES OF THE STUDY:
AIM:
To evaluate the prevalence of oral Candida albicans in smokers and their association with and without oral lesions.
OBJECTIVES
1.To isolate and identify Candida albicans from saliva.
2.To compare the colonization of oral Candida albicans in nonsmokerand smokers without oral lesions.
3.To study the prevalence of oral Candida albicans in smokers with oral lesions.
4.To compare the prevalence of Candida albicans among normal controls, smokers without lesions and smokers with premalignant or malignant lesions.
MR/MS______, we are requesting you to enroll yourself in the study conducted by Dr. Supriya. H, Post Graduate Student in Department of OralPathology and Microbiology under the guidance ofProf. (Dr.) Harishchandra Rai. K.V. G. Dental College and Hospital, Sullia D.K.
You are requested to participate. During the study you will be asked some questions and you are supposed to answer to the best of your knowledge.
Your participation in this research is voluntary. Your decision whether to participate will not affect your relationship with K. V. G. Dental College and Hospital, Sullia. If you decide not to participate you are free to withdraw at any time. The purpose of research is toevaluateoral Candida albicans in smokers and their association in smokers with and without oral lesions.
PROCEDURE INVOLVED:
If you agree to participate in this research study we would need to collect saliva from your mouth.
RISK AND BENEFITS:
This procedure is done in routine and do not cause any harm to you.
ALTERNATIVES:
Even if you decline in participation, you will get the routine line of management.
PRIVACY AND CONFIDENTIALITY:
The only people to know that you are a research subject are members of the research team.
No information about you or provided by you during the research will be disclosed to others without your written permission except:
1. In emergency to protect your rights and welfare.
2. If required by law.
AUTHORIZATION TO PUBLISH RESULTS:
When the results of the research are published or discussed, in a conference, no information will be displayed that would disclose your identity. Any information that is obtained in a connection with this study and that can be identified with you will remain confidential.
FINANCIAL INCENTIVES FOR PARTICIPATION:
You will not be paid/offered any free gifts for participating in the research. You will not be reimbursed for expenses.
Signature of investigator
Name
Address
Phone no.
I have been told in a language that I understand about the study. I have been told that this is for a research procedure, that my participation is voluntary and I/he/she reserve the full right to withdraw from the study at my own initiative at any time, without having to give any reason, and that right to participate or withdraw from study at any stage will not prejudice my/his/her, rights and welfare. I have been assured by the investigator that confidentiality will be maintained and only be shared for academic purposes. Also I have been told that no incentive either in the form of cash or gift for participating in the study will be given.
I hereby give consent to participate in the above study. I am also aware that I can withdraw this consent at any later date, if I wish to. This consent form being signed voluntarily indicating my agreement to participate in the study, until I decide otherwise. I understood that I will receive a signed and dated copy of this form.
If I have any doubts/questions pertaining to the above study, I have been asked to contact
Dr. Supriya. H, Mobile no: 9481754528.
Signature of the research subject:
Date :
Place:
Signature of the witness:
Date:
Place:
SIGNATURE OF THE CANDIDATE

REMARKS OF THE GUIDE

NAME & DESIGNATION OF (In block letters)
11.1 Guide / PROF.(DR.) HARISHCHANDRA RAI
HEAD OF THE DEPARTMENT
DEPARTMENT OF ORAL PATHOLOGY & MICROBIOLOGY
K.V.G DENTAL COLLEGE AND HOSPITAL
SULLIA, DK-574327
11.2 Signature of guide
11.3 Co-Guide ( if any ) / PROF.(DR). SUBBANNAYYA KOTIGADDE
PROFESSOR
DEPARTMENT OF MICROBIOLOGY
KVG MEDICAL COLLEGE ,SULLIA
11.4 Signature
11.5 Head of the Department / PROF.(DR.) HARISH CHANDRA RAI
HEAD OF THE DEPARTMENT
DEPARTMENT OF ORAL PATHOLOGY & MICROBIOLOGY,
K.V.G DENTAL COLLEGE AND HOSPITAL
SULLIA,DK-574327
11.6 Signature
12.1 Remarks of the Chairman andThe
Principal
12.2 Signature / PROF. (DR.) MOKSHA NAYAK