Evaluation of Oral Microflora in Obese and Non-
Obese Humans from District Faisalabad, Pakistan
Frisha Abkar1, Sajjad ur Rahman1, Ahsan Naveed1*, Hira Rasheed1 and Syed Ashar Mehfooz2
1Institute of Microbiology, University of Agriculture Faisalabad, Pakistan
2Department of Clinical Medicine and Surgery, University of Agriculture Faisalabad, Pakistan
Submission: January 09, 2019; Published: March 15, 2019
*Corresponding author: Ahsan Naveed, Institute of Microbiology, University of Agriculture Faisalabad, Pakistan
How to cite this article: Frisha A, Sajjad R, Ahsan N, Hira R, Syed A M. Evaluation of Oral Microflora in Obese and Non-Obese Humans from
District Faisalabad, Pakistan. Curre Res Diabetes & Obes J. 2019; 10(1): 555779. DOI: 10.19080/CRDOJ.2019.10.555779
Obesity is accumulation of excessive fat in an adipose tissue that leads to deleterious effects on the body. The oral microbiota may have a correlation with the obesity. Present study was performed to correlate oral microflora and obesity. A total of 100 saliva samples were collected, 50 from obese and 50 from non-obese individuals. The samples were processed according to the standard protocol for DNA extraction through GeneJet Genomic Purification Kit. The extracted DNA were proceeded to polymerase chain reaction (PCR) and sequencing. The bacterial species (Selenomonas noxia) was identified in 93% of the obese individuals at the level >1.05% of the total salivary microbiota. Capnocytophage gingivalis was identified in 89% of obese median while Actinomyces
gerenseriae and Treponema socranskii were present in 71% and 79% of the obese individuals respectively. Some of the bacterial species from all these phyla were recovered in both obese and non-obese individuals, however above-mentioned bacteria were only detected in obese. The overall research evidenced the presence of different oral microflora in healthy and obese individuals. These results support the correlation of oral microflora with the obesity.
Digestive tract is place of residence for a lot more microbes. The oral microbes influence the individual’s health, although there are other factors but the change in oral microflora is evidenced to lead the obesity. Present study was performed to develop a correlation between oral microflora and obesity
The condition in which abnormal or excessive amount of fat accumulate in adipose tissues called obesity. Obesity number was increased around the world in children and adults; it is most common in developed countries Bharmal et al. . The normal and obese people differ from each other in two ways. The abnormal accumulation of fat in adipose tissues and second condition is the regional sharing of fat in the body. Due to obesity in the worldwide mortality and morbidity rates has been increased. Obesity is associated with psycho sociological disorders Burdette & Hill . Human microbiota comprises bacteria viruses arches the universal eukaryotes is more than 1014 microbial cells. According to research the Childs born by C section have great risk of obesity as compared to normal delivery. There is a link between early gut and later obesity bacterial colonization Blustein & Liu .
The aim of study was to evaluate the oral microflora of obese and non-obese individual to check the role of microorganisms in
the obesity. Obesity in worldwide can be controlled by evaluation of bacteria that was involved in the obesity.
Oral Bacteria + Food Gastrointestinal track, it may enhance the metabolic activity
Oral Bacteria it may increase the Appetite enhance the food consumption
Obesity is epidemic, excessive amount of fat causing the disturbance in the blood level of insulin access, bold fat, and sugar that may lead to diabetes, high blood pressure, and causing cardiovascular disorders due to fatty liver. Need to control obesity research focus the microorganisms that involved in obesity.
Numerous and different microorganisms are present in a human oral cavity it is revealed that a relationship between obesity and microbiota. On the intraoral surfaces complex microbial communities are made. Oral cavity of obese and lean people is different. A biological fluid is secreted from salivary glands called saliva Yamanaka et al. . The saliva is containing microbiome from the communities of bacteria from many surfaces like intraoral surfaces. In the saliva the bacteria that are present are not blamed as an agent to cause oral disease Kaplan . In the salivary microbiome indigenous bacteria is present. These bacteria varied from person to person Rasiah et al. . The obesity is associated with the microbiota in saliva Goodson et al.
. For the diagnosis of disease and for health monitoring salivary
microbiome was used as a surrogate indicator Yoshizawa et al. .
In the humans and mice Firmicutes and Bacrerioidetes
are most abundant bacteria. The Bacterioidetes 20- 40 % and
Firmicutes 60-80% present in the humans and mice.
Ley et al.  mostly these bacterial representative phyla
will not grow outside of the host. From surrounding ecosystem
babies got their initial microbiota, especially the fecal microflora
and maternal vaginal microflora Reinhardt et al.  among the
family the human gut microbiome is shared Turnbaugh & Elli et
Saliva samples were collected from obese and non-obese
individuals after prior permission of the individuals. The
selected individuals were thoroughly examined for any disease/
abnormality, furthermore, the individuals were selected on the
basis of their age and body mass index. All the selected individuals
were non-smokers with minor to no habits of fast food intake.
A total of 100 saliva samples were collected, 50 from obese
individuals and 50 from non-obese. The individuals were selected
in age limit between 25 to 60 years. The individuals were having a
body mass index (BMI) of 27 to 32.
Unstimulated 3 ml of saliva sample was collected in 100 sterile
graduated cylinders. A 1 ml of saliva sample was transferred into
the Eppendorf tube with an addition of EDTA that was mixed by
vortex and incubated at 52℃ for 10 minutes.
GeneJet Genomic DNA Purification Kit (Thermo scientific, Cat.
# K0721) was used for the extraction of DNA from saliva samples
according to manufacturer’s protocol. The purified DNA sample
from saliva were quantified through Nano drop techniques and
confirmed on 1% Agarose Gel Electrophoresis (in 100ml TAE
buffer 1gm agarose).
The DNA was processed through conventional PCR using two
oligonucleotide primers the forward 27f and reverse 1492R of a
bacterial 16S ribosomal RNA gene.
27F = 5’AGAGTTTGATCCTGGCTCAG 3’
1492R= 5’GGTACCTTGTTACGACTT 3’
The purified PCR product of samples were sent to Eurofins
Genomics Company for sequence studies. Sequences were
processed through NCBI Blast to identify bacteria and comparison
of bacterial population in obese and non-obese population.
The DNA of obese and non-obese population was identified at
1500bp. The results of gel electrophoresis are depicted in Figure 1.
Lane S1-S3 presented obese population while S4-S6 is presenting
non-obese population. The ladder of 1kb is presented in lane L.
The oral cavity of obese individuals showed presence
of Tanerella forsythia, Capnocytophage gingivalis, and
Capnocytophage sputigena from phylum bacteroides. While nonobese
individuals were free of these bacteria. The Table 1 is
presenting % age prevalence of different bacterial species from
phylum bacteroides in the obese individuals. Tanerella forsythia
was present in 83% of the obese median, Capnocytophage
gingivalis was recovered from 89% while Capnocytophage
sputigena 77% of the obese individuals. Some of the other bacterial
species as Nesseria mucosa, Campylobacter rectus, Campylobacter
gracilis, Campylobacter corrodens from phylum Bacteroides were
recovered from both obese and non-obese individuals at different
percentage as given in Table 1.
The bacterial specie Streptococcus intermedius was present
in 87% of the obese individuals under study while Selenomonas
noxia were detected in the 93% of the obese individuals. These
fermicutes were absent in the oral cavity of healthy human
population as presented in the Table 2.
Streptococcus oralis, Streptococcus sanguis, Eubacterium
nodatum, Eubacterium saburreum were found in saliva samples of both obese and non-obese individuals at varied percentage as
shown in Table 2.
Actinomyces israelii and Actinomyces
gerenseriae were also
detected in the oral microflora of the obese individuals under
study. The oral microflora of the obese was containing Actinomyces
israelii and Actinomyces
gerenseriae in 71% and 63% as depicted
in Table 3. Other notable bacterial species recovered from obese
and non-obese individuals were Actinomyces odontolyticus,
Propionibacterium, Actinobacterium acnes as given in the Table 3.
Treponema socranskii was absent in the healthy individuals
from the phylum Spirochaetae but these bacteria were present
in the oral cavity of the 77% obese individuals as directed in the
The change in oral flora is directly related to obesity, the
relationship of weight gains and changes of oral microflora is
calculated in this research. The bacterial species were different
in the saliva sample of overweighed individuals when compared
with a bacterial species in normal individuals. Goodson et al. 
Studied S.noxia was present in the obese individuals 2.56 % and
0.00% from the non-obese individuals. While in resent study the
results indicate the S.noxia was totally absent from the healthy
individuals and present in 20% of obese individuals.
The bacterium S.noxia was capable of identifying 98% of
obese individuals from healthy individuals. Illness competitor is
S. noxia Selenomonads, it is known as gram negative Bactria, and
it is motile, non-spore forming. The S.noxia participates in release
of inflammatory mediators and inflammatory reaction triggering.
S.noxia. It is intriguing to take note of that Selenomonas noxia was
main Firmicutes which was fundamentally hoisted in the saliva.
Different research has demonstrated that this inclination to put on
weight in the mice is transmissible b/w mice by coprophagia. The
creators guessed that the obese people have in the gut microbiota
the more energy from the food is stored by the Fermicutes.
From Firmicutes phyla Streptococcus intermedius was
absent from the non-obese individuals and present in the obese
individuals, while according to the study of Goodson et al. 
the presence of Streptococcus intermedius in the over weighted
individuals was 0.63% and absent in non-obese individuals.
From phyla Actinobacteria the bacteria Actinomyces
gerenseriae was present in the obese individuals, according to
resent study and absent from the non-obese individuals. The
research results of Goodson et al.  indicated the presence of
gerenseriae in over weighed individuals was 2.28%
while it was absent in healthy individuals. The other bacteria from
Actinobacteria phyla the Actinomyces israelii was absent from
the non-obese medium and present in the obese medium, while
according to Goodson et al. , Actinomyces israelii was present
in the obese individuals with 1.11% while absent in non-obese
Presently, the bacteria Treponema socranskii from phylum
Spirochaetae was absent from non-obese individuals and present
in the obese individuals the results are in accordance with
Goodson et al.  where 1.14% obese individuals were having
Treponema socranskii and it was no recovered from non-obese
Investigation of populaces with periodontitis and without
periodontitis showed that 70% periodontitis were stout compared
to 37% normal people Socransky & Haffajee . Furthermore,
obese people have serious periodontal illness, when assessed by
the plaque, seeping on examining, take profundity, connection
misfortune, and level of the sub gingival T. forsythia. TNFα is one
of some expert provocative cytokines created by the unhealthy
periodontal tissues that may be an essential fiery cytokine
Oral microscopic organisms may be added in the improvement
of weight by no less than three components. Theory is that the
oral microscopic organisms can expand weight pickups by
expanding craving. Indeed, without information, teleology of the
recommendation is smart to the point that it ought to be specified
in unique situation. By empowering host hunger, microscopic
organisms motivate to eat more.
Subsequently, any investigation of disease related weight
pickups ought to incorporate an evaluation of exercise & diet, which was not joined into our pilot contemplate. It should be
perceived that an essential restriction of the examination as
directed is the utilization of a comfort populace as a control,
instead of utilizing chosen accomplices. While it isn’t legitimized
to propose that Selenomonas noxia diseases have an etiologic
part in stoutness, in view of these information, it is sensible to
recommend that Selenomonas noxia might be a pointer of progress
in oral microbial environment Mager et al. .
The connection between the oral bacteria and obesity
was certainly complex and diverse. The relationship might be
fortuitous as being connected with food intake. It is also evidenced
that multiple metabolic process and numerous other factors also
have a positive correlation with obesity. Whatever the reason
behind these microbial populations are diverse and may exist in
obese and healthy individuals taking their part in human health
and physiology [15-28].
The 5 bacterial phyla including, Firmicutes, Actinomyces,
Bacterides, Spirochaetae were evaluated in the obese and healthy
individuals. Presently detected bacterial species from the above
mentioned five phyla were increasingly present in the obese
individuals compared to non-obese. A positive correlation was
observed in the oral microflora and obesity since some of the
bacterial species were only detected in the obese median. The
presence of specific bacterial population in the obese individuals
and their absence in the healthy population is an indication of
relationship among oral microflora and obesity. The further
research studies are needed to develop the correlation of microbial
population in the oral cavity and obesity. It is also expected that
obese individual may be treated through exchange of microbial
communities in their oral cavity.