Relationship Between Gutkha Chewing People and Their Saliva Composition Including Bacterial Microflora

INDIAN DENTAL JOURNAL
Offical Publication of Society of Medical Dental & Public Health
RELATIONSHIP BETWEEN GUTKHA CHEWING PEOPLE AND THEIR

SALIVA COMPOSITION INCLUDING BACTERIAL MICROFLORA
Vandana Maurya1 Bhaskar Agarwal 2

1
M Sc (Biochemistry) 2Senior Resident, Department of Prosthodontics, King George’s Medical University, UP, Lucknow, India.
Address for Correspondence: Dr Bhaskar Agarwal, 1060 Sadar Bazar, Cantt., Lucknow, UP, India.
E-mail: [email protected]
ABSTRACT
Background: Chewing tobacco has been a tradition in India for centuries. But oral diseases are still a neglected
epidemic. Of the total amount of tobacco produced in the country, around 48% is in the form of chewing tobacco,
38% as bidis, and 14% as cigarettes. Chewing mixes without tobacco are termed as ‘Pan Masala’ and with tobacco
as ‘Gutkha’. In the present study an attempt was made to find a relationship between gutkha chewing people and
their saliva composition including bacterial microflora.
Materials and Methods: 50 subjects were chosen randomly in a college which also included gutkha chewing people.
Saliva was collected, inoculated on media and observed.
Results: Decreased salivation and mucous formation was observed in 20 gutkha chewing subjects. Hence reduced
number of normal symbiotic saliva microflora. Many bacterial species were commonly present both in the healthy
and gutkha chewing subjects.
Conclusion: We expect the exploration of composition and microbial diversity in the saliva will direct the
development of efficient and noble prevention methods of oral diseases due to eating gutkha.
KEYWORDS Gutkha, Oral Cavity, Microflora
I NTRODUCTION - Tobacco- a plant native to

America, introduced by Portuguese in India in
1605, was used initially for ceremonial and
medical purposes.2 It is now used in various forms all
over the world. Chewing tobacco has been a tradition
directly as a result of cardiovascular diseases, chronic
obstructive lung diseases and tobacco related
cancers.4 So tobacco causes almost one-third of all
deaths. During the recent years, in India, both in
urban and rural, consumption of tobacco in the form
in India for centuries. It is the third largest tobacco- of direct chewing of gutkha is alarmingly increasing
producing country (13%) after China (40%) and USA especially in the young adults as which is major
(15%).3 The Indian National Sample Survey revealed reason for subsistence of oral cancer. Saliva is a
that 35% men and 12 % women use tobacco in one complex biological fluid composed of a wide variety
form or another.6 Several types such as smoking of organic and inorganic constituents. The major
practices (e.g. cigarette, bidi, hukka and pipe), organic constituents of saliva are proteins and
tobacco chewing (e.g. tobacco-beetle i.e. Pan, gutkha glycoproteins while main inorganic constituents are
and khaini), rubbing tobacco powder/paste on gums phosphate and calcium ions. Interest in saliva as a
and teeth, and snuffing tobacco are prevalent with diagnostic fluid has grown exponentially in recent
varying frequency in different communities.9 Thus, years. The oral cavity provides two types of surfaces
bidis, snuff and chewing tobacco (such as gutka, for microorganisms’ colonization – soft tissue and
khaini and zarda) form the bulk (86%) of India’s total hard tooth enamel/exposed root surfaces. It comprises
tobacco production. Chewing mixes without tobacco of different environments such as the tongue, cheek,
are termed as ‘Pan Masala’ and with tobacco as palate, tooth surfaces and gingival crevice, which are
‘Gutkha’. Oral diseases are still a neglected epidemic. colonized by different microfloras. These microfloras
About 8 lakhs deaths occur every year in India are influenced by host factors such as tooth eruption
3
found that gutkha chewing subjects had decreased

or loss of teeth, oral hygiene, and hormonal changes
salivation and mucous formation. So overall there
of the host. Because the microorganisms present in
was reduced normal symbiotic microflora in gutkha
saliva are removed from different surfaces of the
chewing subjects (Figure 1)
mouth, saliva can provide an overview of the
microflora in the mouth of an individual. Here, we
present an attempt to find a relationship between
DISCUSSION
gutkha chewing people and their saliva composition
including bacterial microflora. During the recent years, in India, both in urban and
rural, consumption of tobacco in the form of direct
chewing of gutkha is alarmingly increasing especially
MATERIALS AND METHODS in the young adults as which is major reason for
subsistence of oral cancer. Sangita Sushree Nayak
The study was performed at PL Mishra Public Inter
found that more males use gutkha than females.10 We
College, Gooja, Kanpur. Subjects were chosen
too found the same during our study. Out of 20
randomly which included students, lecturers and
subjects, 16 were males and 4 females. Our main
other office staff. The control subjects were selected
focus, however, was to find relationship between
from healthy subjects. Data was collected by
gutkha chewing people and their saliva and bacteria
distributing questionnaire to all the subjects. Dental
microflora and how it is different from that of non-
check up was conducted. Saliva was collected from a
gutkha chewing people. It was found that even
group of 50 subjects by mouth rinse with 10 mL
teenagers too had gutkha as time pass. Avasn et al.
sterile saline for 30 sec. The saline solution had been
conducted this sort of study for gutkha chewing
UV-irradiated to avoid DNA contamination. Samples
including smoking and oral micro flora in some slum
collected in sterilized eppendorf tubes were placed on
dwellers of Visakhapatnam. The results revealed that
ice immediately. They were plated as soon as
decrease in salivation and mucous formation in
possible otherwise stored at -80°C until further use.
gutkha chewers, which further resulted in reduction
Salivation and mucous formation was observed under
in number of oral micro flora. 1 Our findings were the
microscope. Ten fold serial dilutions of samples were
same. Genotoxcity of Pan masala and gutkha among
prepared in saline solution. Microorganisms were
chewers was examined by Smita Jyoti et al.11. In the
isolated by spreading diluted samples (100μl) in
young, changes in the oral microflora that have been
small petri dishes containing Nutrient Agar (NA). All
related directly to age are reasonably well established
plates were incubated at 37°C for 3-5 days.
and are usually associated with major perturbations to
Following incubation, microbial colonies were
the habitat (e.g., tooth eruption, hormonal changes,
purified by sub-culturing on the same media used for
etc.) while in adults, the resident microflora is
isolation. In total, 70 bacterial microflora strains were
believed to remain relatively stable unless the
isolated.
environment is disturbed, as occurs during disease
was examined by Percival et al.8. We used nutrient
agar media to isolate bacteria from saliva samples. A
RESULTS total of 70 bacterial strains were isolated. Three
A total of 50 questionnaires were administered. 25 genera-Prevotella, Streptococcus and Veillonella
were students, 15 lecturers and 10 office staff. Total constituted about 50% of the salivary microflora.
of 27 males and 23 females, out of which 20 were Similar findings were by Jung-Gyu Kang et al.5
gutkha chewing subjects. 70 bacterial strains were Streptococcus species were more abundant in gutkha
isolated. Many bacterial species were commonly chewing subjects. Prevotella and Veillonella species
present both in the healthy and gutkha chewing were abundant in non gutkha chewing subjects. It
subjects. Such as Peptococcus niger, Fusobacterium, was also found that gutkha chewing subjects had
Streptococcus, Campylobacter, Pseudomonas, decreased salivation and mucous formation. It was
Veillonella and Prevotella species. Three genera- also observed that gutkha chewing subjects had one
Prevotella, Streptococcus and Veillonella constituted or the other dental issues. Therefore, a saliva-based
about 50% of the salivary microflora. Streptococcus test could prove very useful for early detecting and
species were more abundant in gutkha chewing monitoring of health effects of these habits since
subjects. Prevotella and Veillonella species were saliva is a readily available specimen7 so we can say
abundant in non gutkha chewing subjects. It was also that an understanding of the composition of the oral
4
microbial ecosystem in relation to oral health is lead to an increase in the oral pathogens by reducing
essential for the prevention and treatment of oral the normal symbiotic microflora. Our results provide
diseases. a framework for further studies of saliva microflora
and for investigations of various factors that
CONCLUSION influence salivary microbiome. These in turn will
direct the development of efficient and noble
Drug addiction in general and gutkha use in prevention methods of oral diseases due to eating
particular represent a complex phenomenon, which gutkha. The alarming scenario demands that federal
has various social, cultural, biological, geographical, regulatory and health agencies and NGOs should
historical and economic aspects. The use of gutkha is launch awareness programmes to inform and educate
growing day by day and teenagers are the main the public regarding the adverse health consequences
consumers. The main users are males and people and possible cancer risk associated with gutkha.
belonging to nuclear families. Gutkha chewing may
0.3
0.25
0.2
0.15 Gutkha chewing subjects
0.1
Non gutkha chweing subjects
0.05
0
Prevotella Veillonella Streptococcus
species species species
Figure 1: Mean CFUs of various microflora in gutkha and non-gutkha chewing subjects
REFERENCES
7) Ministry of Planning, Government of India, New
1) Avasn Maruthit Y, Rao RS, Palivela H, Thakre S. 8) Delhi. National Sample Survey Organization.
Impact of gutkha chewing & smoking on microbial Sarvekshan 1991; 15-48.
environment of oral cavity: a case study on slum
dwellers of selected areas in Visakhapatnam. J Environ 9) Naciye Kurtul and Engin Gökpınar. Salivary Lipid
Sci Eng. Oct 2004; 46(4):268-73. Peroxidation and Total Sialic Acid Levels in Smokers
and Smokeless Tobacco Users as Maraş Powder.
2) Council of Scientific and Industrial Research Mediators of Inflammation, 2012; Article ID 619293: 8.
(CSIR). Drug Addiction with Special Reference to
India, 1965; New Delhi: Publication and Information 10) R.S Percival, S.J Challacombe and P.D Marsh.
Divison (PID). Age-related microbiological changes in the salivary and
plaque microflora of healthy adults. Jr Med. Microbiol.
3) Department of Agriculture and Co-operation, 1991; 35: 5-11.
Government of India. Tobacco in India- A Handbook of
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Development, Ministry of Agriculture. India-Part 1: The detailed habit. Jr of Oral Biology and
Craniofacial Research, Dec 2011; 1: 24-30.
4) Director General of Health Services, Ministry of
Health and Family Welfare, Government of India. 12) Sangita Sushree Nayak. Risk Factors Associated
Health Information of India, 1993; New Delhi: Central with Gutkha Addiction: An Empirical Study in Cuttack
Bureau of Health Intelligence. City of Odisha. Jr Sociology Soc Anth. 2011; 2(2): 89-
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5) Jung-Gyu Kang, Seong Hwan Kim, and Tae-Young
Ahn. Bacterial Diversity in the Human Saliva 13) Smita Jyoti, Mohammad Afzal and Yasir Hasan
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INDIAN DENTAL JOURNAL

Offical Publication of Society of Medical Dental & Public Health

RELATIONSHIP BETWEEN GUTKHA CHEWING PEOPLE AND THEIR

Vandana Maurya1 Bhaskar Agarwal 2

KEYWORDS Gutkha, Oral Cavity, Microflora

I NTRODUCTION - Tobacco- a plant native to

found that gutkha chewing subjects had decreased

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