PREVOTELLA

INTERMEDIA
and
FUSOBACTERIUM
NUCLEATUM
Presented by : Dr.D.Surya
PG I
Streptococcus sp.. a.a,p.g,p.i.f.n
 INTRODUCTION
Of more than 1014 cells of which make the human body, only about 10% are

mammalian. The remaining cells are the microorganisms that make the

commensal microbiota of the host.

Periodontitis is a chronic inflammation of the periodontium with multi-

factorial etiology. The periodontal pocket of humans harbors more than 700

bacterial species.
 INTRODUCTION
Socransky et al in 1998 analysed 13261 subgingival plaque samples from 185 adult subjects

Presence of 40 sub gingival species

Checkerboard DNA hybridization

Defined color coded complexes of periodontal micro organisms that tend to be found

together in health and in disease.

Composition of the different complex was based on the frequency with which different

clusters were recovered

CONTENTS
MORPHOLOGY and TAXONOMY
GROWTH
VIRULENCE FACTORS
DIAGNOSIS
ASSOCIATION WITH ORAL INFECTIONS
ASSOCIATION WITH SYSTEMIC INFECTONS
ANTIMICROBIAL THERAPY
RECENT ADVANCEMENTS
PREVOTELLA
INTERMEDIA
MORPHOLOGY
Gram negative bacilli
Obligate anaerobic
Black-pigmented

Rod-shaped bacterium
Non motile
They are pleomorphic, taking on different forms
during life cycle
TAXONOMY
Kingdom: Bacteria
Phylum:
Bacteroidetes
Class: Bacteroidia

Order: Bacteroidales
Family:
Prevotellaceae
Genus: Prevotella
Species:
intermedia
 GROWTH
Habitat is the strictly anaerobic environments in the upper respiratory tract,
female genital tract, gastrointestinal tract and gingival crevice.

Supra and subgingival plaques

Periodontal pockets

Endodontic lesions

Bacteria of this group are frequently found in the oral cavity of both healthy
individuals and patients with periodontal diseases.
 VIRULENCE FACTORS
Virulence is the relative ability of an organism to cause disease or to

interfere with a metabolic or physiological function of its host.

LATIN - VIRULENTUS MEANS FULL OF POISON

Virulence factors refer to the properties that enable a microorganism to establish itself on

or within a host of a particular species and enhance its potential to cause disease.
 VIRULENCE FACTORS OF P.i

All factors that attribute to the colonization,

proliferation, and pathogenicity of microbes will be
considered virulence factors.

fimbriae
Bacterial lipopolysaccharide
protease
s
exopolysaccharide
Bacterial
nucleases
hemolysins

Interpain A, a Cysteine Proteinase from Prevotella intermedia, Inhibits Complement by Degrading Complement Factor C3 Michal Potempa,
Jan Potempa, Tomasz Kantyka, KyAnh Nguyen, Katarzyna Wawrzonek, Surya P. Manandhar, Katarzyna Popadiak, Kristian Riesbeck, Sigrun Eick, Anna M. Blom.
FIMBRIAE
FIM A GENE ENCODE

Four
morphologic
small
ally distinct Antibodies
filamentous
types of fimbriae against P.
cell surface
fimbriae had a fairly intermedia
appendages
using uniform type C
associated
electron ultrastructu fimbriae
with
microscopy, ral and
bacterial
designated compositio inhibited
colonization
as types A, n. bacterial
of host
B, C and D Invasion.
tissues.
based on
their size.
 The interaction is mediated
The type C fimbriae could
by fimbriae and the
promote invasion by
integrin oral epithelial cells
providing a means for the
are specific for fimbriae,
bacteria to attach to the
whereby the binding may
cell surface.
induce internalization.

a separate ligand-receptor
and a cytoskeletal interaction is necessary for
rearrangement are the internalization during
required for invasion of the association between P.
oral epithelial cells by P. intermedia and the oral
intermedia. epithelial cell as promoted
by the type C fimbriae.

Bacterial adhesins to host components in Periodontitis. ATSUO AMANO. Periodontology 2000, Vol. 52, 2010, 1237
LIPOPOLYSACCHARIDE
Lipopolysaccharide is a
major constituent of the
outer membrane of
gram-negative bacteria,
including P. intermedia.

demonstrated the
structure of lipid A

the lipid A activates

immune cells through a
TLR4-mediated
signaling pathway.
Prevotella intermedia lipopolysaccharide stimulates release of tumor necrosis factor-a through mitogen-activated protein kinase signaling
pathways inmonocyte-derivedmacrophages. Sung-Jo Kim1,2, Eun-Young Choi3, Eun Gyung Kim1, Su-Hwa Shin3, Ju-Youn Lee1,2, Jeom-Il Choi1,
2 & In-Soon Choi. FEMS Immunol Med Microbiol 51 (2007) 407413.
 Lipid A
+
TLR4

trigger a number of host cells, especially

mononuclear phagocytes, to produce IL-1b,
IL-6, IL-8, and, most importantly, tumor
necrosis factor a (TNF-).

induce the production of MMPS

Role in inflammation and Pathogenesis of

periodontitis
 EXOPOLYSACCHARIDE
Recently identified virulence factor is an
exopolysaccharide that appears to confer
resistance to phagocytosis by
polymorphonuclear leukocytes.

P. intermedia strain 17 was rarely internalized

by human polymorphonuclear leukocytes, but
other strains were readily engulfed.
production of EPS by P. intermedia strain 17
could contribute to the pathogenicity of this
organism by conferring their ability to evade the
host's innate defence response.
PROTEASES
Bacteria-derived proteases may contribute to tissue destruction,
and are likely to impair host defense by degrading
immunoglobulin's and complement.

P. intermedia causes extensive degradation of IgG-IgA, and

substantial breakdown of transferrin, haptoglobin and
macroglobulin.

The presence of IgG strongly stimulated the bacterial growth,

indicating that these strains used the protein for growth.
 INTERPAIN

A strain of
P.
intermedia
that
produces
interpain A InpA
was found showed a
to be more preference
resistant to for the
Cysteine complement chain of C3
proteases. . and C4.

38 KDa is through Its action is

proteases is the efficient on C3, C4
capable of degradation & C5.
degrading of C3.
igG.
Interpain A, a Cysteine Proteinase from Prevotella intermedia, Inhibits Complement by Degrading Complement Factor C3 Michal Potempa,
Jan Potempa, Tomasz Kantyka, KyAnh Nguyen, Katarzyna Wawrzonek, Surya P. Manandhar, Katarzyna Popadiak, Kristian Riesbeck,
Sigrun Eick, Anna M. Blom.
 AT LOW CONCENTRATION

At low
periodontal
concentrations,
bacteria at low
they were able to
concentrations
activate This may lead to a
appear to cause
complement low grade
nonspecific
factors C3, C4 and inflammation that
activation of C1
C5 as they provides access to
and to generate
preferentially nutrients for
C5a and C3a
target the chains bacterial growth
fragments
of these proteins and colonization.
chemotactic
to cause the
factors for
release of
neutrophils.
anaphylatoxins.
AT HIGH CONCENTRATION

At higher concentrations, they simply

degrade these three complement factors
particularly C3, into smaller fragments
so that they can no longer propagate the
complement cascade.
At higher concentrations of bacteria and
proteinases, the complement system
becomes incapacitated by multiple
cleavages of critical proteins within the
cascade.
 creating a favorable
condition for the
InpA will aid establishment of a
survival of common ecosystem
bystander bacterial for a beneficial
species, habitat

Promotes secondary
colonization of pathogenic
organisms such as P. gingivalis
by providing attachment sites,
growth substrates and reduced
oxygen concentration locally.

Interpain A, a Cysteine Proteinase from Prevotella intermedia, Inhibits Complement by Degrading Complement Factor C3 Michal Potempa,
Jan Potempa, Tomasz Kantyka, KyAnh Nguyen, Katarzyna Wawrzonek, Surya P. Manandhar, Katarzyna Popadiak, Kristian Riesbeck,
Sigrun Eick, Anna M. Blom.
 NUCLEASES of P.i
when compared the DNA degradation activity of
major Gram-negative periodontopathogenic
bacteria, Porphyromonas gingivalis, Prevotella
intermedia, Fusobacterium nucleatum, and
Aggregatibacter actinomycetemcomitans

P. intermedia showed the

highest degradation activity.
nuclease degrades
the NETs to escape
NET killing.

Nucleases from Prevotella intermedia can degrade neutrophil extracellular traps. M. Doke, H. Fukamachi, H. Morisaki, T. Arimoto,
H. Kataoka and H. Kuwata.
major molecular size of Pr.
intermedia nuclease was Nucleases of these bacteria
approximately 3539 kDa, with a are secretary enzymes
minor molecular size of
approximately 42 kDa.

Biochemical analysis revealed that divalent cations, such as

Mg2+ and Ca2+, were required for optimal enzymatic activity
 Hemolytic activity of P.i

For the bacteria to successfully colonize the

host tissue and emerge as a potential
pathogen
after the initial attachment, one of the
prerequisites is that they must be able to
obtain the essential nutrients for growth.
P.INTERMEDIA requires iron for growth.
Human hemoglobin (hHgb) found in
crevicular fluid could be a source of iron
for P. intermedia.
Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens. Masaaki Okamoto a;*, Nobuko Maeda a,
Kenichiro Kondo a, Kai-P. Leung. FEMS Microbiology Letters 178 (1999) 299^304.
the globin moiety of the hemoglobin molecule binds to P. intermedia hemoglobin
binding receptors.

the release of hemin or iron from the surface-bound hemoglobin

internalization of hg by the cell.

P. intermedia possess ferrochelatase,

extracts the ferrous iron out of uptaken hemoglobin enabling the cells to obtain
iron from the host.
MICROBIOLOGICAL TESTS
 A selective medium for P.
intermedia is non-selective
blood agar.

Columbia Agar Base is

supplemented with sheep
blood, bacitracin, colistin and
nalidixic acid as selective
agentsfor the isolation
ofP.intermedia.

This medium is prepared, stored

and dispensed under oxygen-
free conditions to prevent the
formation of oxidized products
prior to use.
Prevotella intermedia lipopolysaccharide stimulates release of tumor necrosis factor-a through mitogen-activated protein kinase
signaling pathways inmonocyte-derivedmacrophages. Sung-Jo Kim1,2, Eun-Young Choi3, Eun Gyung Kim1, Su-Hwa Shin3, Ju-Youn Lee1,
2, Jeom-Il Choi1,2 & In-Soon Choi. FEMS Immunol Med Microbiol 51 (2007) 407413.
OTHER TESTS
IMMUNODETECTION METHODS USING FLUORECENT
MICROSCOPES
SANDWICH ELISA
NUCLEIC ACID PROBE
Polymerase chain reaction (PCR)
ASSOCIATION WITH ORAL INFECTIONS

NUG AND NUP

REFRACTORY PERIODONTITIS

PUBERTY ASSOCIATED GINGIVITIS.

ASSOCIATED WITH A.A COMITANTS IN AGGRESSIVE PERIODONTITIS

Necrotizing ulcerative gingivitis
Caused by specific bacteria Fusiform bacillus and
spirochetes.

Loesche et al described P.intermedia as a constant flora in

NUG.

Rx with metronidazole resulted in significant reduction of P.i

Serum IgG and IgM Levels to Bacterial Antigens in Necrotizing Ulcerative Gingivitis Randal W. Rowland, * Jiri Mestecky,f John C. Gunsoley* and
Ronald . Cogen
 Necrotizing ulcerative periodontitis

They also is the low or The flora found

observed that the variable level of in NUP in AIDS
In HIV positive patients demonstrated destructive spirochetes, patients is more
significantly greater prevalence of P.i. periodontal which is in line with that
lesions seen in inconsistent with of chronic
AIDS patients the NUG flora. Periodontitis
differ
substantially
from NUG
lesions;

Murray and co-workers have reported that cases of

NUP in AIDS patients demonstrate greater numbers
of the opportunistic fungus Candida albicans and
higher prevalence of A.comitans, Prevotella
intermedia, Porphyromonas gingivalis,
Fusobacterium nucleatum, and Campylobacter spp.

Carranza 11th edition

REFRACTORY PERIODONTITIS
Those patients who are unresponsive to any treatment provided,
whatever the thoroughness or frequency.

Refractory periodontitis may be due to an abnormal host response, an

unusually pathogenic and virulent microflora, failure to eliminate
plaque-retentive factors, or a combination of any of these factors.
 High proportion of A. Comitans and P. Intermedia has been reported in
refractory periodontitis cases.

Rodenburg et al.Have noted the following microorganisms and their incidence

in refractory chronic periodontitis:

F. Nucleatum (75% of patients/sites), P. Intermedia (40%), A.

Actinomycetemcomitans (30%), peptostreptococcus micros (30%),
staphylococcus sp. (30%), bacteroides forsythus (25%), campylobacter rectus
(25%), P. Gingivalis (15%), candida sp. (15%), and enterobacteriaceae or
pseudomonadacea spp. (10%).

Carranza 11th edition

Walker et all'

two patterns or rates of attachment loss, associated with refractory periodontitis; each pattern may
be indicative of a different flora

The pattern associated with a relatively rapid loss of attachment was characterized by a gram
negative flora that contained spirochetes, P. intermedia, and Fusobacterium species.

A slow, continuous rate was associated with a predominantly gram-negative flora containing a high
proportion of S.intermedius or S. intermedius- like organisms.
 PUBERTY ASSOCIATED GINGIVITIS.
Puberty occurs between the average ages of 11 to 14 in most women.

An increase in the production of sex hormones (estrogen and progesterone) occurs,

which then remains relatively constant during the remainder of the reproductive phase.

the prevalence of gingivitis increases without an increase in the amount of plaque.

Gram-negative anaerobes, especially P.i, have been implicated in association with

puberty gingivitis.

Kornman and Loesche postulated that this anaerobic organism may use ovarian
hormone as a substitute for vitamin K growth factor .

Delaney and Kornman suggest levels of black pigmented Bacteroides, especially P.i
increase with increased levels of gonadotrophic hormones in puberty.
 Microbiologic factors of LAP
A. actinomycetemcomitans has been implicated as the primary pathogen.

several specific microorganisms are detected in patients with localized aggressive periodontitis

A. actinomycetemcomitans, Capnocytophaga sp., Eikenella corrodens, Prevotella intermedia, and

Campylobacter rectus.

not all reports support the association of A. actinomycetemcomitans and localized aggressive
periodontitis.

In some studies, A. actinomycetemcomitans could not be detected in patients.

studys found elevated levels of P. gingivalis, P. intermedia, Fusobacterium nucleatum, C. rectus, and
Treponema denticola in patients with either Aggressive Periodontitis localized or generalized
aggressive disease.
ASSOCIATION WITH SYSTEMIC INFECTIONS

Bacterial vaginosis (BV) is a disease of the vagina caused by excessive

growth of bacteria.

there may be no symptoms.

Having BV approximately doubles the risk of infection by a number of

other sexually transmitted infections, including HIV/AIDS

It also increases the risk of early delivery among pregnant women.

Bacterial Vaginosis and Anaerobic Bacteria Are Associated with Endometritis Catherine L. Haggerty,1 Sharon L.
Hillier,1,2 Debra C. Bass,1 and Roberta B. Ness1 for the PID Evaluation and Clinical Health (PEACH) Study
Investigatorsa
 CHRONIC SINUSITIS: Sinusitis, also known as asinus infection

isinflammationof thesinusesresulting in symptoms.P.i constitutes a

significant percentage, about 20% of all cases of maxillary sinusitis.

G.I TRACT INFECTION: Gastroenteritis,inflammation of thegastrointestinal

tractthat involves thestomachandsmall intestine.Signs and symptoms

include some combination ofdiarrhea,vomiting, andabdominalpain.

Gastroenteritis can be due to infections byviruses,bacteria,parasites,

andfungus.

Campylobacterspecies , Escherichia coli,Salmonella,Shigella, and P.i.

 ANTIBIOTIC SENSITIVITY
Amoxicillin/ampicillin

sensitive
Highly

Poorly
Fairly sensitive

sensitive
with clavulanate
(100%)
Spiramycin (68%)
Clindamycin (93-100) Penicillin
Tetracycline
Metronidazole (90%)
Ciprofloxacin
Erythromycin (87%)

Antibiotic toxicity, interactions and resistance development. MARILYN C. ROBERTS. Periodontology 2000, Vol. 28, 2002, 280297.
ANTIBIOTIC RESISTANCE

B- LACTAMASE

tet(Q), a gene encoding the ribosome protection protein.

codes for tetracycline resistance

erm(F) which codes for erythromycin resistance.

Antibiotic toxicity, interactions and resistance development. MARILYN C. ROBERTS. Periodontology 2000, Vol. 28, 2002, 280297.
 Recent advances
A recent study found that Pr.intermediashowed the
highest DNA degradation activity. Also Recombinant NucA
and NucD from Pr.intermedia digested DNA and RNA,
which required both Mg2+and Ca2+for optimal activity. In
addition, NucA and NucD were able to degrade the DNA
matrix comprising NETs(neutrophil extracellular traps)
therby escaping noutrophil mediated bacterial killing. (ref
25).
A recent study suggest thatP. intermediaandT.
forsythiainfected hDFSCs(Human dental follicle stem
cells) maintain their stem cell functionality, reduce PMN-
induced tissue and bone degradation via suppression of
PMN-activity, and at the same time allow for the survival
of the oral pathogens.(ref 26).
 CONCLUSION:

Despite the growing evidence implicating this bacterium in the pathogenesis of periodontitis, it is up to

now a rather poorly studied organism, as well as to the lack of molecular tools for genetic manipulation.

A better insight into the mechanisms of these factors in pathogenesis will allow the development of

future strategies to control periodontitis.

 REFERENCES
Bacterial adhesins to host components in Periodontitis. ATSUO AMANO. Periodontology 2000, Vol. 52, 2010, 1237

Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens. Masaaki Okamoto a;*, Nobuko Maeda a, Kenichiro Kondo a, Kai-

P. Leung. FEMS Microbiology Letters 178 (1999) 299^304.

Microbiology and treatment of dental abscesses and periodontal-endodontic lesions. GUNNAR DAHLEN. Periodontology 2000, Vol. 28, 2002, 206239

Prevotella intermedia inducesmatrixmetalloproteinase-9 expression in human periodontal ligament cells. Su-Min Guan1, Lei Shu2, Shan-Min Fu1, Bin Liu1, Xiu-

Li Xu3 & Jun-Zheng Wu. FEMS Microbiol Lett 283 (2008) 4753.

Prevotella intermedia upregulatesMMP-1andMMP-8 expression in human periodontal ligament cells. Su-Min Guan1,2, Lei Shu3, Shan-Min Fu1, Bin Liu1, Xiu-

Li Xu4 & Jun-Zheng Wu. FEMS Microbiol Lett 299 (2009) 214222.

Invasion of Human Oral Epithelial Cells by Prevotella intermedia. BRIAN R. DORN, K.-P. LEUNG, AND ANN PROGULSKE-FOX. INFECTION AND

IMMUNITY,Dec. 1998, p. 60546057.

Prevotella intermedia lipopolysaccharide stimulates release of tumor necrosis factor-a through mitogen-activated protein kinase signaling pathways inmonocyte-

derivedmacrophages. Sung-Jo Kim1,2, Eun-Young Choi3, Eun Gyung Kim1, Su-Hwa Shin3, Ju-Youn Lee1,2, Jeom-Il Choi1,2 & In-Soon Choi. FEMS Immunol

Med Microbiol 51 (2007) 407413.

 Sex and the subgingival microbiome: Do female sex steroids affect periodontal bacteria? PURNIMA S. KUMAR. Periodontology 2000, Vol. 61, 2013, 103
124.

The binding and utilization of hemoglobin by Prevotella intermedia. K.-P. Leung a;b;*, P.S. Subramaniam c, M. Okamoto a, H. Fukushima d, C.-H. Lai.
FEMS Microbiology Letters 162 (1998) 227-233

Ejects of porphyrins and inorganic iron on the growth of Prevotella intermedia. K.-P. Leung _, Shawn P. Folk FEMS Microbiology Letters 209 (2002) 15-21.

Hormonal influences: effects of diabetes mellitus and endogenous female sex steroid hormones on the periodontium. Brian L. Mealey & Alan J. Moritz.
Periodontology 2000, Vol. 32, 2003, 5981

Interpain A, a Cysteine Proteinase from Prevotella intermedia, Inhibits Complement by Degrading Complement Factor C3 Michal Potempa, Jan Potempa,
Tomasz Kantyka, KyAnh Nguyen, Katarzyna Wawrzonek, Surya P. Manandhar, Katarzyna Popadiak, Kristian Riesbeck, Sigrun Eick, Anna M. Blom.

Antibiotic toxicity, interactions and resistance development. MARILYN C. ROBERTS. Periodontology 2000, Vol. 28, 2002, 280297.

Nucleases from Prevotella intermedia can degrade neutrophil extracellular traps. M. Doke, H. Fukamachi, H. Morisaki, T. Arimoto, H. Kataoka and H.
Kuwata.

Text book of Carranza..revised 11th edition

Fusobacterium
nucleatum
MORPHOLOGY
G ve bacilli
Obligate anaerobe
Non-motile rods
Non-spore forming
Latin Fusus means Spindle

Knorr in 1923 Proposed the genus Fusobacterium for

pointed, non-sporing, g-ve bacilli found in the mouth
TAXONOMY
Kingdom: Bacteria

Phylum: Fusobacteria

Class: Fusobacteria
Order:
Fusobacteriales
Family:
Fusobacteriaceae
Genus: Fusobacterium

Species: nucleatum
 Subspecies
Divided into 5 species based on electrophoretic patterns of
Whole cell proteins
21 alloenzymes
Mobility of 2 enzymes namely
Confirmed by analysis of
DNA Homology of 16S-23S internal transcribed spacer region sequences(ITS)
5 subspecies
Nucleatum
Polymorphum
Vincentii
Fusiforme
Animalis
Recently a new subspecies Canifelium has been isolated from the dog & cat bite
wound is humans.
Growth
Fusobacterium need rich media for growth.

It grows well in media containing Trypticase, peptone or yeast extract.

F.nucleatum mainly depends upon aminoacid catabolism & peptides for

their growth unlike other bacteria which depend upon carbohydrates.

All strains used mainly used aminoacids like glutamate, histidine, and
aspartate initially which is then foll0wed by peptide utilisation once
aminoacids become less.
Growth
F.N. utilise glucose only for biosynthesis of intracellular molecules and not
for energy metabolism.
The energy is derived from aminoacid and peptide catabolism.
They have a symbiotic growth relationship with P.gingivalis.
The ability of F.N. to utilize carbohydrates for biosynthesis when AA level
is high and ferment sugar when AA level is low contributes to its survival
in oral cavity & its persistence in periodontal disease.
 Symbiotic Growth With P.gingivalis
In gingival crevices, the sacharolytic bacteria like P.G utilise
carbohydrates.
Peptides are formed by the hydrolytic activity of P.G.
These peptides and AA are in turn utilized by F.N for their
growth and metabolism.
FN PG
Subgingival Supragingival
Utilise AA & peptides Utilise carbohydrates
Asacharolytic Sacharolytic
Increases pH Decreases pH
Utilise carbohydrates for Utilise carbohydrates for energy
intacellular synthesis metabolism
IMMUNOLOGIC ACTIVITY
Produces factors capable of suppressing lymphocyte responses in vitro.
Induces suppressive monocytes resulting in inhibition of host-protective
immune reactions.
Causes polyclonal B-cell activation.
Stimulates IgG, IgA, IgM and T-cell responses.
Activates complement.
IMMUNOLOGIC ACTIVITY
Stimulates cells to produce IL-1, IL-6, TNF a , TGF b, MMP
leading to periodontal destruction.

Enhances PMN adherence and causes release of toxic oxygen radicals

and lysosomal enzymes resulting in periodontal damage.

Also induces apoptosis of PMN & PBMC (peripheral blood

mononuclear cells.
COAGGREGATION
Coaggregation is a phenomenon prevalent among oral bacteria
isolated from the human oral cavity.

Coaggregation is a direct bacterium- bacterium interaction and is

highly specific in that only certain cell types are partners.

The recognition may be intrageneric, intergeneric, or multigeneric in

nature and in all three kinds of coaggregations the cells appear to
interact independently of other cells in the population
 The fusobacteria, which coaggregate with the widest range of genera tested
so far, do not coaggregate with other fusobacteria, (No intrageneric).

Intergeneric coaggregation is defined as cell-to-cell recognition and

adherence between bacterial pairs from different genera.

F.nucleatum coaggregates with P.gingivalis, S.sanguis, Candidia albicans

and Eubacterium.
 F. nucleatum also participates in multigeneric coaggregation, i.e.,
interacting bacterial networks composed of coaggregating cells of three or
more genera.
By multigeneric aggregation, the noncoaggregating cell types are bridged
together by a common partner.
Fusobacteria act as a bridge between early and late colonizers.
The early colonizers adhere to the tooth pellicle and coaggregate with
other early colonizers and also with F. nucleatum.
Late colonizers, such as Selenomonas flueggi, P. gingivalis, and species of
Eubacterium, Actinobacillus, Capnocytophaga, and Treponema,
coaggregate almost exclusively with F. nucleatum.
 Virulence
The virulence mechanisms of F.n can be broadly classified into 2 types.

Colonization and dissemination

Induction of host responses.
COLONISATION AND DISSEMINATION

F.n is a very adhesive bacterium.

The ability to adhere to and invade host cells is believed to play a key role
in F. nucleatums oral and systemic virulence.
F. nucleatum encodes several adhesins for interspecies interactions,
including FadA,Fap2, RadD, and aid1.
F. nucleatum exhibits different types of adhesions and harbors several
adhesins, including
an arginine-inhibitable adhesin,
a mannose-sensitive lectin involved in fusobacterial coaggregation with Candida
species,
Fad, which is required for cell attachment and invasion
Fap2 a galactose sensitive adhesin.
 F. nucleatum also binds to a variety of mammalian cells, i.e.
epithelial and endothelial cells,
PMNs, monocytes, erythrocytes,
fibroblasts, HeLa cells and NK cells,
as well as host molecules such as salivary macromolecules,
extracellular matrix proteins, human IgG, and cadherins.
F. nucleatum invades epithelial and endothelial cells
Adherence and invasion are essential mechanisms for
colonization, dissemination, evasion of host defense, and
induction of host response.
 FadA - best-characterized virulence factor identified from
F.nucleatum.
FadA exists in two forms,
the intact pre-FadA consisting of 129 amino-acid (aa) residues
and
the secreted mature FadA (mFadA) consisting of 111 aa residues.
a predominantly alpha-helical hairpin structure,
Pre-FadA and mFadA form an active complex, FadAc, for
host-cell binding and invasion.
FadA is uniquely encoded by two closely related species, F.
nucleatum and F. periodonticum, absent in most other
species of Fusobacterium making it a potential diagnostic
marker.
 FadA is not only an adhesin but also an invasin.
FadA binds to cell-junction molecules, the cadherins.
FadA binds to VE- cadherin on the endothelial cells and to
E-cadherin on epithelial and colorectal cancer cells.
FadA binding to VE-cadherin on endothelial cells causes
the latter to migrate from cell-cell junction to intracellular
compartments, increasing the permeability of the
endothelial layer
FadA allows both direct invasion into the host cells and
pericellular invasion via loosened cell-cell junctions.
 It is postulated that this is the mechanism employed for systemic
dissemination.
the increased endothelial permeability allows other bacteria in the
vicinity to penetrate through, a likely reason why F. nucleatum is often
found in mixed infections at extra-oral sites.
 Fap2 a galactose sensitive(inhibitable) adhesin.
F. nucleatums attachment to mammalian cells can be
reversed by adding D-galactose, which also inhibits F.
nucleatums ability to coaggregate with the major
periodontopathogen Porphyromonas gingivalis and with at
least nine other oral bacterial species
INDUCTION OF HOST RESPONSES.
It induces human b-defensin 2 from oral epithelial cells via
FAD-I
stimulates factors predisposing to atherosclerosis by GroEL
activates lymphocyte apoptosis by Fap2 and RadD
F. nucleatum is a potent stimulator of inflammatory
cytokines, IL-6, IL-8, and TNF
Binding of F. nucleatum to NK cells activates inflammatory
responses involved in periodontal disease
DISEASES
PERIODONTAL DISEASES
Fusobacterium nucleatum is the Gram-negative species
isolated most frequently from both healthy and
diseased sites in the oral cavity
It is implicated in various forms of periodontal diseases
including the
mild reversible form of gingivitis and
the advanced irreversible forms of periodontitis including
chronic periodontitis, localized aggressive periodontitis and
generalized aggressive periodontitis.
The prevalence of F. nucleatum increases with the
severity of disease, progression of inflammation and
pocket depth
 F.n produces substances like butyrate, propionate,
ammonium and sulfides.
These in turn may result in periodontal pathology by the
following mechanisms.
Inhibits fibroblast proliferation.
Sulfides interfere with opsonisation.
Penetrates gingival epithelium.
Adheres and degrades basement membrane by binding to
type-4 collegen.
Coaggregeates with multiple bacteria and produce
synergistic/additive damage to periodontium.
Adverse pregnancy outcomes
Adverse pregnancy outcome (APO) is a broad term
including preterm labor, chorioamnionitis, preterm
premature rupture of membranes, preeclampsia,
miscarriage, intrauterine growth retardation, low birth
weight, stillbirth, neonatal sepsis, etc.
F. nucleatum is one of the most prevalent species and by
far the most prevalent oral species implicated in APO.
It has been detected in a wide variety of placental and
fetal tissues including amniotic fluid, fetal membranes,
cord blood, neonatal gastric aspirates, fetal lung and
stomach.
 It has been postulated that F. nucleatum translocates
from the maternal oral cavity to the intrauterine cavity
via hematogenous transmission
Hematogenous injection of F. nucleatum resulted in
specific colonization and proliferation of the bacteria in
the fetoplacental unit without causing systemic
infections.
The bacteria colonized initially in the decidua by
crossing the endothelium, followed by spread to
amniotic fluid, fetus and fetal membrane, mimicking
chorioamnionitis, eventually leading to preterm and
term fetal death.
 Only two subspecies have been detected in intrauterine
infections, with the overwhelming majority belonging to
ss animalis, and an occasional few belonging to ss
polymorphum.
Studies show that FadA adhesin in involved in placental
colonisation.
Also studies show that F.n with Fap2 mutation had
impaired ability to invade and colonise placenta.
GI disorders
The main GI disorders with which F.n is implicated are
Colorectal carcinoma
IBD Inflammatory bowel disease
Appendicitis.
F. nucleatum has been detected in colonic biospsies of
patients with IBD. F. nucleatum strains isolated from
inflamed tissues of the IBD patients are more invasive
than those from the normal tissues .
studies have reported association of F. nucleatum in
appendicitis.
 F. nucleatum was first discovered to be enriched in CRC
carcinomas and the rectal swabs of CRC patients.
Studies report that levels of F. nucleatum are also
elevated in adenomas, in stools of patients with
adenoma and carcinoma, and associated with stages of
colorectal neoplasia development.
Rubinstein et al. demonstrate that F. nucleatum
stimulates CRC cancer growth by modulating the E-
cadherin/ catenin signaling via its unique FadA
adhesion
Other infections
F. nucleatum has been implicated in cardiovascular
diseases (CVD). It is frequently detected in the
atherosclerotic plaques, and is also one of the most
common periodontal pathogens detected in ruptured
cerebral aneurysm.
F. nucleatum, is a major cause of the well- known
Lemierres Syndrome, a rare form of upper airways
infection with a life threatening secondary septic
thrombophlebitis of internal or external jugular veins,
usually developed in previously healthy young adults.
 Fusobacterium nucleatum is associated with a wide
spectrum of infections and abscesses including
infections of the head and neck (Lemierres syndrome, acute
and chronic mastoiditis, chronic otitis and sinusitis, tonsillitis,
peritonsillar and retropharyngeal abscesses, postanginal
cervical lymphadenitis, periodontitis),
Infections of brain, lungs, abdomen, pelvis, bones, joints, and
blood.
F. nucleatum has been involved in include rheumatoid
arthritis and Alzheimers disease. Periodontal treatment
has been shown to improve clinical outcomes of
rheumatoid arthritis.
Diagnosis
Gram stain
Culture
Biochemical tests
Produces indole
Produces propionate from threonine
Doesnot hydrolyse esculin.
Gas Liquid chromatography
Phylogenetic grouping by 16s-rRna PCR
Treatment & Antimicrobial therapy
Penicillins
Beta lactams with beta lactam inibitors like
amoxy/clavulanic acid
Metronidazole
Clindamycin
Mechanism of antibiotic resisitance
Beta lactams Beta lactamases
Tetracyclines Efflux & ribosomal protection
Macrolides, lincosamides and streptogramins rRNA
methylases.
 Recent advances
FadA adhesin is unique to Fn (Han et al., 2005),hence, is an ideal
potential diagnostic marker to identify individuals at risk for
developing adenomas and/or adenocarcinomas. (Ref 20).
The development of antibiotic resistance by this bacterium demands
novel therapeutic intervention.
A recent study has reported UDP-N-acetylglucosamine 1-
carboxyvinyltransferase (MurA) as one of the potential target
proteins inF. nucleatum. In this study, they proposed two novel MurA
inhibitors namely orientin and quercetin-3-O-D-glucuronide (Q3G)
throughin silicoscreening and evaluated their mode of inhibition
byin vitroexperiments. They found that that Q3G may interrupt the
PEP binding, thereby inhibiting the MurA activity. (ref 21).
Rubinstein MR, Wang X, Liu W, Hao Y, Cai G, Han YW. Fusobacterium nucleatum promotes colorectal carcinogenesis by modulating E-cadherin/-catenin signaling
via its FadA adhesin. Cell Host Microbe. 2013 Aug 14;14(2):195-206.
Kumar A, Saranathan R, Prashanth K, Tiwary BK, Krishna R. Inhibition of the MurA enzyme in Fusobacterium nucleatum by potential inhibitors
identified through computational and in vitro approaches. Mol Biosyst. 2017 Mar 30.
 Recent advances
A recent study investigated the effects of green and black
tea extracts as well as two of their bioactive components,
EGCG and theaflavins, on the growth and virulence
properties ofF. nucleatum. The green and black tea
extracts, EGCG, and theaflavins decreased the adherence
ofF. nucleatumto oral epithelial cells and matrix proteins,
attenuatedF. nucleatum-mediated hemolysis and hydrogen
sulfide production. this study showed that tea polyphenols
may be of interest for treatingF. nucleatum-associated
disorders. (Ref 22)

Ben Lagha A, Haas B, Grenier D. Tea polyphenols inhibit the growth and virulence properties of
Fusobacterium nucleatum. Sci Rep. 2017 Mar 21;7:44815
Conclusion
F.nucleatum is an oral commensal associated with
periodontal pathogenesis.
This bacterium is also associated with many other
systemic infections due to its systemic spread.
Recent evidences pointing to its role in adverse
pregnancy outcomes and colorectal carcinogenesis
have garnered great interest.
Future studies are needed to study whether the control
of F.nucleatum is associated with decreased systemic
infections.
References
Bolstad AI, Jensen HB, Bakken V. Taxonomy, biology, and
periodontal aspects of Fusobacterium nucleatum. Clin
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Han YW. Fusobacterium nucleatum: a commensal-
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Thank you