WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Al-Jebouri et al.
World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 2.786
Volume 3, Issue 6, 1720-1733.
Research Article
ISSN 2278 – 4357
ANTIBIOTICS RESISTANCE AMONG ANAEROBIC PATHOGENS
CAUSING HUMAN PERIODONTITIS
*Mohemid M Al-Jebouri1 and Hadeel M Al-Hadeethy2
1
Department of Microbiology,College of Medicine,University of Tikrit,Tikrit,Iraq
2
Department of Microbiology,College of Dentistry,University of Tikrit
Article Received on
11 April 2014,
Revised on 06 May
2014,
Accepted on 24 May 2014
ABSTRACT
Background:Periodontal disease could be defined as a disorder of
supporting structures of teeth, including the gingiva, periodontal
ligament and alveolar bone. Periodontal disease develops from a preexisting gingivitis. However, not every case of gingivitis develops into
*Correspondence for Author
Dr. Mohemid M Al-Jebouri
Department of
Microbiology,College of
a periodontal disease. The inflammation of gingiva alone is termed
gingivitis, and the severe inflammation of the periodontal ligament
with destruction of alveolar bone is called periodontal disease.
Medicine,University of
Methods: One hundred eighteen samples were examined in the present
Tikrit,Tikrit,Iraq
study. Samples were obtained from periodontal pockets A plaque
sample was inoculated onto different agar plates selected for anaerobic
isolations. Periodontal pathogens were identified utilizing conventional
methods,
API
RapID ANA system II , API 20A ,and Vitek2 ANC systems. Fifteen antibiotics were tested
for their effectiveness on the various anaerobic bacteria isolated utilizing brucella blood agar
plates disc diffusion methods. Results: peptostreptococcus prevotii , prevotella intermedia,
prevotella melani , prevotella disiens, Bifidobacterium sp., Fusibacterium mortiferum
peptostreptococcus tetradius ,and Wolinella sp. fusibacterium varium ,vellionella sp.,
campylobacter gracilis, capnocytophaga sp. ,peptostreptococcus magnus ,peptostreptococcus
micros
,peptostreptococcus
niger,peptostreptococcus
anaerobius
,
staphylococcus
saccharolyticus , streptococcus consellatus , and gemella morbillorum were the most
common anaerobic periodontal pathogens isolated from patients in the present study.The
bacteria isolated were generally highly sensitive to Imipeneme ,erythromcin , doxycycline,
amoxycillin ,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin
,tetracycline and augmentin. Conclusion:The present study showed that 19 different species
were responsible for the periodontal infections and the most common pathogens were
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Peptostreptococcus prevotii, Vellionella sp. and Fusibacterium varium.The antibiotic
sensitivity testing showed that these pathogens were generally sensitive to a wide range of
chemotherapeutic antibiotics including Imipeneme ,erythromcin , doxycycline, amoxycillin
,pipracillin, rodgyle ,spiromycin ,clindamycin , cephalexin ,cephalothin ,tetracycline and
augmentin.Periodontal anaerobic pathogens were very difficult to be isolated and timeconsuming work but it worths doing for the seek of oral health and economy.
Key words: Periodontitis,anaerobic pathogens,antibiotics susceptibility.
1.INTRODUCTION
Periodontal disease could be defined as a disorder of supporting structures of teeth, including
the gingiva, periodontal ligament and alveolar bone. Periodontal disease develops from a preexisting gingivitis. However, not every case of gingivitis develops into a periodontal disease.
The inflammation of gingiva alone is termed gingivitis, and the severe inflammation of the
periodontal ligament with destruction of alveolar bone is called periodontal disease[1].
Healthy gingival sulcus has a flora dominated by equal proportions of gram positive cocci,
especially Streptococcus spp, and Actinomuces sp. Later, plaque "matures" resulting in a flora
consisting from facultative anaerobic microorganisms, spirochetes and motile rods. The
proportions of strict anaerobic, Gram negative organisms increase significantly in accordance
with increasing severity of disease. Disease activity in periodontal disease may range from
slow, chronic, progressive destruction to brief and acute episodic bursts with varying
intensity and duration[2]. The composition of the subgingival microbial flora and the level of
pathogenic species differ from subject to subject as well as from site to site. The search for
the pathogens of periodontal diseases has been underway for more than 100 years, and
continues up today[2].More than 300 species of bacteria colonize subgingival area and their
cell wall components can trigger immune activation[3]. The currently recognized key Gram
negative periodontopathogens include: Porphyromonas gingivalis, Prevotella intermedia,
Bacteroides forsythus ,Aggregatibacter actinomycetemcomitans , Fusobacterium spesies
,Capnocytophaga species , Campylobacter sp.,wollenilla sp.,and Bifidobacterium[3,4,5,6,7].
Also, the following bacteria could be isolated: Eubacterium spp, Peptostreptococcus species
Streptococcus consellatus, and Staphylococcus saccharolyticus[2].
The clinical signs of
periodontitis are changes in the morphology of gingival tissues, bleeding upon probing as
well as periodontal pocket formation. This pocket provides an ideal environment for the
growth and proliferation of anaerobic pathogenic bacteria[8]. The choice of the antimicrobial
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agents in periodontal diseases must be based on the bacterial etiology of the infection .
Several antibiotics have been tested for their clinical and microbiological efficacy in
periodontal diseases[9]. Multiple factors contribute to the development of resistance to
antibiotics. Their widespread and unnecessary use is the greatest contributing factors.
2.MATERIALS AND METHODS
2.1. Patients
One hundred eighteen samples were examined in the present study. They were referred to the
Laboratory of Microbiology, Department of Microbiology, College of Medicine ,Tikrit
University. Patients were of both sexes(140 males and 40 females).Their ages ranged from
16-69 years old. The selection was done randomly among patients in Tikrit
University/college of dentistry/dental teaching hospital . The diagnosis of periodontitis was
made by the clinical examination including taking dental, medical and family history from
the subjects involved in this study, so patients fulfilling the criteria to be diagnosed as
periodontitis. Clinical measurements of periodontal parameters used included dental plaque
index, gingival index, bleeding on probing, probing pocket depth and clinical attachment loss
using (graduated William’s periodontal probe). Clinical diagnosis in each case was according
to the dentist . The interviews were performed for each patient. The questionnaire included
general information about the patient e.g.: name, age, sex, smoking, education, residence and
treatment if the patients treated or not treated by antibiotics befo
2.2.Sampling
Samples were obtained from periodontal pockets after supragingival plaque was removed
from the teeth to be collected[10]. The supragingival dental plaque was removed with sterile
cotton, and the tooth surface was dried with compressed air to prevent contamination with
saliva. The exclusion of moisture in the mouth with sterile cotton rolls and subgingival
plaque was collected from the most inflamed sites by inserting a sterile paperpoint into the
periodontal pocket for 10 seconds , when the pocket depth was from3-7mm.The sample was
mixed with
contamination
1 ml thioglycollate broth (transport medium) , sealed tightly to avoid
and
kept at 4 C0. Samples were processed within 2 days of
collection[11,12,13,14].
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2.3.Cultural Technique
A plaque sample(loopful from thioglycollate broth containing subgingival paperpoint which
inserted into periodontal pocket was inoculated onto Brucella blood agar kanamycin 100
mg/l,vancomycin 7.5 mg/l Brucella blood agar, Enriched blood agar ,brain heart infusion
agar,and trypticas soy agar with haemin and vitamin K1.The inoculated media were
immediately incubated in an anaerobic environment generated with the anaerogens gas pack
CO2 system Chemical, for 3 -7 days[10].
2.4. Isolation and Identification of Bacteria
Isolates were identified using cultural characteristics , gram -stain and biochemichal tests
(convenentional methods) which includes : indole spot test, catalase test , oxidase test ,
arginine hydrolysis, lipase test, nitrate reduction , motility test ,growth in bile , urease test
and growth on kanamycin(1mg),vancomycin(5µg), colistin(10µg) Brucella blood agar
[15,16] .
2.5. Rapid identification methods
2.5.1.API RapID ANA II system [remel,USA].
This system was used as
a qualitative micromethod employing conventional and
chromogenic substrates for identification of anaerobic bacteria.The tests used in RapID ANA
II system were based on microbial degradation of specific substrates detected by various
indicator system[12].
2.5.2.API 20A [Biomereiux,France].
The API 20 A system was utilized for the biochemical identification of anaerobes according
to manufacturer instructions[10].
2.5.3..VITEK 2 ANC System [Biomereiux,France]
The VITEK 2 as an automated microbiological system was utilized for the identification of
anaerobes according to manufacturer instructions[17].
5.4.Antibiotic Sensitivity Testing
A loopfull from colonies of anaerobic periodontal pathogens was inoculated into BHI broth
and incubated in an anaerobic environment generated with the anaerogas pack (CO2)system
chemical, for 2-3 days at 37Co . The bacterial suspension poured on the surface of the
brucella blood agar plates, and left for 10 minutes to settle the bacteria. The excess of the
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bacterial suspension was discarded using Pasteur pipette. The plates were left for one hour at
room temperature to dry [16]. The antibiotic discs were placed by sterile forceps sterilized
with alcohol. The antibiotics used in sensitivity test were shown in Table 6.The diameter of
inhibition zone were measured. Interpretation of the zone of inhibition was done according to
standard measurements[18,19,20] shown in Table 7.
Table 6 : Antibiotics used in sensitivity testing for anaerobic periodontal pathogens
isolates.
Antibiotic
Erythromycin
Ampicillin
Cephalothin
Amoxycillin- clavulanate
potassium[augmentin]
Clindamycin
Vancomycin
Tetracycline
Amoxycillin
Metronidazole
Doxycyclline
Kanamycin
Imipenem
Pipracillin
Cephalexin
Spiromycin
Spiromycin/metronidazol(Ro
dogyl)
Code
E
AM
CEP
Disc potency (mg/disc)
15
10
30
Manufacturer
Oxoid(England)
Himedia(India)
Oxoid(England)
AmC
DA
VAN
TE
AX
MET
DO
K
IPM
PI
CL
SP
30
30
30
30
25
30
30
30
10
100
30
100
Himedia(India)
Oxoid(England)
Oxoid(England)
Oxoid(England)
Himedia(India)
Oxoid (England)
Himedia(India)
Himedia(India)
Himedia(India)
Oxoid (England)
Oxoid (England)
Oxoid (England)
ROD
30
Oxoid (England)
3.RESULTS
3.1. Identification of anaerobic periodontal pathogens
Periodontal pathogens were identified by conventional methods, API RapID ANA system
II , API 20A ,and Vitek2 ANC systems for identification of anaerobic bacteria.
Table 11 showed the types of anaerobic periodontal pathogens isolated. In the present study
the most common anaerobic periodontal bacteria isolated were peptostreptococcus prevotii
which represented 15(8.3%) of all isolates, while prevotella intermedia, prevotella melani ,
prevotella disiens, Bifidobacterium sp., Fusibacterium mortiferum peptostreptococcus
tetradius ,and Wolinella sp. represented only 1 (0.5%) of all isolates.
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Table 11: Types of anaerobic bacteria isolated from periodontal patients(No. of patients
=180)
Types of isolates
prevotella intermedia
prevotella melani
prevotella disiens
Bifidobacterium sp
Fusibacterium nucleatum
Fusibacterium varium
Vellionella sp.
Campylobacter gracilis
Wolinella sp.
Capnocytophaga sp
Peptostreptococcus prevotii
Peptostreptococcus magnus
Peptostreptococcus tetradius
Peptostreptococcus micros
Peptostreptococcus niger
Peptostreptococcus
anaerobius
Staphylococcus
saccharolyticus
Streptococcus conselatus
Gemella morbillorum
Total
3.2. Antibiotic Sensitivity Testing
No. (%) of isolates
1 (0.5%)
1 (0.5%)
1 (0.5%)
1 (0.5%)
1 (0.5%)
10 (5.5%)
14 (7.7%)
3 (1.6%)
1 (0.5%)
2 (1.1%)
15 (8.3%)
2 (1.1%)
1 (0.5%)
3 (1.6%)
4 (2.2%)
2 (1.6%)
5 (2.7%)
4 (2.2%)
2 (1.6%)
73 (40.7%)
Table 18 shows the results of antibiotic sensitivity of periodontal pathogens iso lates. It was
found that all isolates( 100%) were resistant to kanamycin and metronidazole. The
imipeneme ,erythromcin , doxycycline, amoxycillin ,pipracillin, rodgyle ,spiromycin
,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin were the most effective
drugs used in the present study . In the present study ,Prevotella intermedia was highly
sensitive (100%) to doxycycline,imipeneme,amoxycillin,and augmentine .While prevotella
melani was sensitive (100%) to all antimicrobial agents used . Prevotella disiense and
bifidobacterium sp. were 100% sensitive to doxycycline , imipeneme, amoxycillin,
augmentine, cephalthin , and pipracillin. wheras fusibacterium nucleatum only sensitive
(100%) to doxycycline, imipenem, and pipracillin . Fusibacterium varium were 100%
sensitive to amoxycillin, doxycycline, imipenem,ampicillin,cephalothin,and pipracillin.
Vellionella sp.,were 100% sensitive to amoxycillin doxycycline, imipenem, augmentin,
cephalothin,and pipracillin while 50% were sensitive to rodogyle and spyromycin.
Campylobacter gracilis was sensitive to erythromycin (100% ) , but 50% of the isolates were
sensitive to clindamycin, cephalothin and pipracillin. Wollinella sp. was 100% sensitive to
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doxycycline ,clindamycin , cephalothin,and pipracillin. 50% of Capnocytophaga sp. isolates
were sensitive to cephalexin , rodogyle ,spiromycin, erythromycin,clindamycin ,and
cephalothin, but 100% of them were sensitive to doxycycline , tetracycline and pipracillin.
Peptostreptococcs spp. were more sensitive to doxycycline, amoxycillin, cephalothin and
pipracillin .Staphylococcus saccharolyticus was 100% sensitive to doxycycline , imipeneme,
cephalothin and pipracillin but 50% were sensitive to amoxycillin, augmentin and
clindamycin . Also, streptococcus conselatus and gemella morbillorum were 100% sensitive
to doxycycline, imipenem, cephalothin and pipracillin .
Table 18: Antibiotics sensitivity testing for isolated periodontal pathogens using disc
diffusion method
Types of
periodontal
pathogens
isolated(No.)
prevotella
intermedia(1)
Prevotella melani(1)
prevotella disiens(1)
Bifidobacterium
sp(1)
Fusibacterium
nucleatum(1)
Fusibacterium
varium(10)
Vellionella sp.(14)
Sensitivity of isolates to antibiotics( %)
A
M
IP
A
D
E DO K
M
ET M
M
A
C
AX
C
L
R
O
D
SP
0
0
0
0
0
100
0
0
100
100
100
10
0
10
0
10
0
10
0
10
0
10
0
10
0
100
0
0
100
100
100
0
0
0
0
100
0
0
100
100
100
0
0
0
0
100
0
0
100
100
100
0
0
0
0
0
100
0
0
100
0
0
0
0
0
100
0
0
100
0
50
50
0
100
0
0
33
0
0
10
0
10
0
TE
PI
0
0
0
100
10
0
100
100
0
100
0
100
0
100
0
0
0
0
100
100
100
0
100
0
100
100
0
100
0
100
50
100
0
0
0
33
50
50
0
50
0
0
0
0
0
10
0
100
0
100
0
10
0
10
0
Campylobacter
gracilis(3)
0
0
33
33
Wolinella sp.(1)
0
0
0
0
0
50
50
50
50
100
0
0
100
0
0
50
50
50
0
50
50
0
100
0
0
50
0
33.
3
33.
3
0
0
0
0
0
100
0
0
100
100
50
50
100
0
100
0
0
0
0
100
0
0
100
100
100
10
0
100
0
100
0
25
25
0
100
0
0
100
100
100
25
100
0
100
Capnocytophaga
sp.(2)
peptostreptococcus
prevotii(15)
Peptostreptococcus
magnus(2)
Peptostreptococcus
Tetradius(1)
Peptostreptococcus
micros(3)
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10
0
10
0
10
0
10
0
10
0
CE
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33.
3
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100
33.3
Al-Jebouri et al.
Peptostreptococcus
niger(4)
Peptostreptococcus
anaerobius(2)
Staphylococcus
saccharolyticus(5)
Streptococcus
conselatus(4)
10
0
10
0
World Journal of Pharmacy and Pharmaceutical Sciences
0
0
0
0
100
0
0
100
100
100
50
50
0
100
0
10
0
10
0
10
0
0
0
0
0
100
100
10
0
100
10
0
100
50
0
25
25
0
100
0
0
100
50
50
0
100
25
100
50
0
0
0
0
100
0
0
100
50
50
0
100
25
100
Gemella
50
0
0
0
0 100 0
0
100 50 100 0 100
0
100
morbillorum(2)
AX: amoxycillin ; CL: cephalexin; ROD: rodogyle ; SP: spiromycin ; E:erythromycin ; DO
: doxycycline ; K: kanamycin ;MET: metronidazole ;IPM: imipenem; AM: ampicillin , MC:
augmentin ; DA: clindamycin; CEP: cephalothin; TE: tetracycline;PI: pipracillin.
4.DISCUSSION
4.1.Distribution of periodontal pathogens
The oral cavity was characterized by harbouring indigenous microbiota. The ability of
microorganisms to colonize the different oral surfaces depends mainly on their binding
potential[21,22].Various environmental and host factors are involved in the harbouring of
microorganisms and microbial composition[23]. Many indigenous microbiota are anaerobes
and these microorganisms can be associated with oral infections and be the origin of distant
infection[24].The most frequent oral anaerobic infections include gingivoperiodontal
diseases,
pulpal
and
periapical
infections,
peri-implantitis
and
pericoronarities
[25].Gingivoperiodontal diseases, including gingivitis and periodontitis were caused by
dental plaque, which was a biofilm[26]. It had been observed that 1g of dental plaque
contains more than 1011 microorganism [23].The biofilm present in the gingival crevice, and
later in the periodontal pocket, is extremely diverse, with up to 180 culturable species from a
single pocket [4].Since such a diverse flora is present, trying to identify the particular species
responsible for disease initiation and progression is a very complex, and difficult undertaking.
Therefore, it is to be expected that a progressively more diverse and anaerobic flora will be
isolated during disease progression. Moore and Moore presented large numbers of anaerobes
increase in their overall proportions during disease progression and, conversely, aerobe and
facultative species, decrease (changes in the microflora of the biofilm as a function of
periodontitis severity) [27,28].The present study demonstrated a different microbiota in
periodontal pockets. The finding presented here that Vellionella sp (7.7) Fusibcterium varium
(5.5%)
,
Campylobacter
gracilis
(1.6)
,Capnocytophaga
sp.(1.1)
,Prevotella
intermedia(0.5%), Prevotella melani (0.5%) , Prevotella disiens (0.5%) ,Bifidobacterium sp.
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(0.5%) ,Fusibacterium nucleatum(0.5%),and Wolinella sp.(0.5%) were more frequent as
isolated from periodontal pocket as showed in Table 11 .It was ,however, interesting to note
that gram positive anaerobic bacteria, especially Peptostreptococcus spp. were isolated in
high rates in periodontal pocket which includes Peptostreptococcus prevotii (8.3%),
Peptostreptococcus magnus(1.1%), Peptostreptococcus micros (1.6%), Peptostreptococcus
tetadius(0.5%), Peptostreptococcus niger (2.2%)
, and Peptostreptococcus anaerobius
(2.7%) .Moreover , Staphylococcus saccharolyticus , Streptococcus consellatus, and Gemella
morbillorum were isolated in the present study in percentages of 2.7%,2.2%,and 1.6%
respectively. These results were almost similar to those of Daniluk et al who found that the
most
common
periodontal
pathogens
were
Veillonella
species,
Fusobacterium,
Staphylococcus saccharolyticus ,Streptococcus consellatus, Gemella morbillorum ,
Peptostreptococcus spp. and Prevotella intermedia[28].Moreover, these results were similar
to those of Spartt , Winkelhoff et al , Piovano , Haffajee and Socransky who found that
Veillonella
species,
Fusobacterium,
Staphylococcus
saccharolyticus
consellatus, Gemella morbillorum, Peptostreptococcus spp.
,Streptococcus
Prevotella intermedia,
Campylobacter gracilis and Bifidobacterium sp.[29]. Furthermore , Mohammad et al found
that Actinobacillus actinomycetemcomitans (26.8%), Porphyromonas gingivalis (21.9%),
Capnocytophaga sputigena (16.7%), Eikenella corrodens (13.2%), Prevotella intermedia
(10.5%), Prevotella disiens (3.1%), Capnocytophaga gingivalis (2.2%), Peptostreptococcus
micros (2.9%), Prevotella corporis (1.8%), Peptostreptococcus magnus
(1.3%) , and
Fusobacterium nucleatum (0.4%)were most common isolates[30].Spratt et al was found that
Capnocytophga sp. involvement in some forms of periodontitis[31].According to Gürsoy et
al Prevotella intermedia is associated with periodontal disease [32].Mayorga-Fayad et al
revealed that the frequency increases Prevotella intermedia in patients with periodontitis[33].
The present results were different from those of
Yacoubi et al who found that
Aggregatibacterium actinomycetemcomitans , Eikenella corrodens are primary causative
agent in periodontitis[14].Moreover , Cisar et al and Ximenez-Fyvie et al found that
actinomyces were the most common component of periodontal disease[34].The data from the
investigation suggest that there was heterogeneity in the subgingival periodontopathogenic
bacteria among subjects. However, as many bacteria in the oral cavity cannot be cultured, it
was likely that these still uncharacterized bacteria could play a role in the initiation and
progression of periodontal disease.
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4.2. Antibiotics Sensitivity Testing of Periodontal Pathogens
During several years, an increasing resistance to different antimicrobial agent in anaerobic
bacteria has been observed[35].The present study demonstrated that 100% of anaerobic
periodontal pathogens isolates were resistant to kanamycin and metronidazole (Table 18)
.The imipeneme ,erythromcin , doxycycline, ampicillin ,pipracillin, rodgyle ,spiromycin
,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin were the most effective .
In the present study Prevotella intermedia was highly sensitive (100%) to doxycycline ,
imipeneme, amoxycillin ,and augmentine .On the other hand , Prevotella melani was also
sensitive to all antimicrobial agents used except kanamycin and metronidazole. Prevotella
disiense and Bifidobacterium sp. were 100% sensitive to amoxycillin , doxycycline ,
imipeneme, amoxycillin, augmentine, cephalthin ,and pipracillin. Whereas Fusibacterium
nucleatum only sensitive to doxycycline, imipenem, and pipracillin . Fusibacterium varium
was mostly sensitive to amoxycillin, doxycycline, imipenem, ampicillin, cephalothin, and
pipracillin. Vellionella sp. was sensitive to amoxycillin ,doxycycline, imipeneme
,augmentin,cephalothin,and pipracillin while 50% of its isolates were sensitive to rodogyle
and spyromycin.These results were almost similar to those of
Brook who found that
Vellionella sp. was generally susceptible to betalactam but resistant to tetracycline and
erythromycin[36]. Campylobacter gracilis were 100% sensitive to erythromycin , but 50%
sensitive to clindamycin,cephalothin and pipracillin. Wollinella sp. isolates were sensitive to
doxycycline ,clindamycin,cephalothin,and pipracillin. Furthermore , Capnocytophaga sp
revealed a sensitivity to cephalexin , rodogyle ,spiromycin, erythromycin,clindamycin ,and
cephalothin, and 100% of its isolates were sensitive to doxycycline , tetracycline and
pipracillin .Moreover , Peptostreptococcs spp. were more sensitive to doxycycline,
amoxycillin, cephalothin and pipracillin .Staphylococcus saccharolyticus were sensitive to
doxycycline , imipeneme,cephalothin and pipracillin 100% but 50% of its isolates were
sensitive to amoxycillin,augmentin and clindamycin . Streptococcus conselatus and Gemella
morbillorum were sensitive to doxycycline , imipenem,cephalothin,and pipracillin as showed
in Table 18. These results were almost similar to those of Thomas who found that Prevotella
sp , Fusibacterium sp, Campylobacter sp. and Streptococcus conselatus were sensitive to
amoxycillin, ampicillin ,doxycycline, and pipracilin and also anaerobic periodontal pathogens
resist to metronidazole [37]. Sally et al found that metronidazole ,pipracillin ,cephalexin,
amoxycilin, augmentin remain good empirical choices for anaerobic bacteria[38]. Nyfors et
al
found
that
Fusibacterium
sp.
was
susceptible
to
penicillin
,amoxycillin
,augmentin,cephalexin, and metronidazole[39].In addition , pierre et al found that anaerobic
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periodontal pathogens were susceptible to spiromycin and rodogyle[40]. Maria et al found
that 70% of Prevotella sp.and 30% of Fusibacterium sp. produce b-lactamases are making
penicillin inactive [41] . Boyanova et al found that 21% of gram positive anaerobic cocci
were resistant to penicillin and 16% of them were resistant to clindamycin
[42]. These
results were different from those of Schwach et al who found that metronidazole most
common selective efficacy against obligate anaerobic periodontal pathogens[8]. Hagashi et al
and Sauvetre
et al found that clindamycin used for treatment periodontal disease [43] .
Moreover , Louise et al found that 50% of the isolates were inhibited by metronidazole when
tested anearobically[44]
.Selcuk et al found that metronidazole had a good effect on
anaerobes[45]. Ahmed found that metronidazole was not inhibiting aerobic facultative,
microearophillic organisms such as Actinomyces, Streptococcus , and Capnocytophaga[46].
The diffusability of the antimicrobial agents depends on it’s composition , hydrophilicity
and its rate of release from soluble matrix in which bound. Therefore , the size of the
inhibition zone may not be related entirely to the differences in the chemical effect of
different antimicrobial agents[47].
5.CONCLUSION
The present study revealed that 19 different anaerobic species were isolated from periodontal
infection site and the most dominant types were Peptostreptococcus prevotii, Vellionella sp.
and Fusibacterium varium and their frequencies of isolation were 8.3,7.7 and 5.5
respectively.Fifteen different antibiotics were tested against the different isolates and
Imipeneme ,erythromcin , doxycycline, amoxycillin ,pipracillin, rodgyle ,spiromycin
,clindamycin , cephalexin ,cephalothin ,tetracycline and augmentin were the most effective
drugs for eradication of these periodontal pathogens.
REFERENCES
1. Jayamathi G , Pamela E and Puvanakrishnan R. Therapeutic effect of proanthocyanidins on
the pathogenesis of periodontitis . Indian J Experim Biol.2011;49:83-93.
2. Kesic L, Milasin J, Igic M and Obradovic R. Microbial etiology of periodontal disease.
Med Biol. 2008;15: 1 – 6.
3. Socransky SS, and Haffajee AD. The bacterial etiology of destructive periodontal disease:
Current concepts. J. Periodontol. 1992;63: 322-331.
4. Haffajee AD, and Socransky SS. Microbial etiological agents of destructive periodontal
diseases. J.Periodontol. 2000; 5: 78-111.
1730
Vol 3, Issue 6, 2014.
www.wjpps.com
Al-Jebouri et al.
World Journal of Pharmacy and Pharmaceutical Sciences
5. Zambon JJ. Periodontal diseases: Microbial factors. Ann Periodontol.1996; 1: 879-925.
6. Avila-Campos MJ. PCR detection of four periodontopathogens from subgingival clinical
samples. Braz. J .Microbiol. 2003;34(1): 81-84.
7.Jovanovic Lj. Komparativno proucavanje promenauparodontalnom dzepu dijabeticara tip III. Doktorska disertacija, Nis .1997.
8. Schwach-Abdellaouia K. Vivien-Castionib N. and,
Gurnya R.
.Local delivery of
antimicrobial agents for the treatment of periodontal diseases.Europ J Pharm Biopharm
.2000; 50: 83-99.
9. Brook I. Treatment of anaerobic infection. Expert Rev Anti Infect Ther. 2007 ;5:991-1006.
10. Condorelli F, Scalia G, Calì G, Rossetti B, Nicoletti G, and Bue A M. Isolation of
Porphyromonas gingivalis and Detection of Immunoglobulin A Specific to Fimbrial
Antigen in Gingival Crevicular Fluid. J. Clin. Microbiol. 1998; 36(8): 2322–2325.
11. Fukui K, Kato N, Kato H, Watanabe K, and Tatematsu N. Incidence of Prevotella
intermedia and Prevotella nigrescens Carriage among Family Members with Subclinical
Periodontal Disease. J Clin Microbiol. 1999; 37(10): 3141–3145.
12. Chandra C, Valavalkar N, and Vandana KL.The comparitive evaluation of xanthan gel
with chlorhexidin (chlosite)in smoker and non smokers:A clinical and microbiological
assessment.2011.
13.L. Boyanova L,Setchanova G, Gergova T, Kostyanev D, Yordanov
Chr,Popova K,
Kotsilkov and Mitov I. Microbiological diagnosis of the sever chronic periodontitis. J
IMAB .2009; 7:56-62.
14. Amel Y, Bouziane D, Leila M and Ahmed B. Microbiological Study of Periodontitis in
the West of Algeria. World J Med Sci . 2010;5 (1): 07-12.
15. Forbes B A,.Sahm D F, and
Weissfeld A S.Baily and Scotts Diagnostic
Microbiology.12th ed.2010.
16 Murray PR., Baron EJ., Pfaller MA., Tenover FC., and Yolker RH. Manual of clinical
microbiology.7th ed. 1999.
17. Mory F, Alauzet C, Matuszeswski C, Riegel P and Lozniewski A. Evaluation of the
New Vitek 2 ANC Card for Identification of Medically Relevant Anaerobic Bacteria . J
Clin Microbiol. 2009 ; 47(6): 1923–1926.
18. Reynolds J. Kirby- Bauer test for antibiotic susceptibility . Richland College.2011.
19. Brown D, Perry J , Wheat P, Gemmell C, Winstanley T , Damani N and MacGowan
A . Disc Diffusion Method for Antimicrobial Susceptibility Testing . 2004.
20.International chart of national committee for clinical laboratory standards, subcommittee
1731
Vol 3, Issue 6, 2014.
www.wjpps.com
Al-Jebouri et al.
World Journal of Pharmacy and Pharmaceutical Sciences
on antimicrobial susceptibility testing .1995.
21.Spratt D. Dental plaque and bacterial colonization. In: Medical biofilms. Jass J, Surman S,
Walker J, editors, John Wiley and Sons Ltd, 2003: 175-98.
22.Zaremba ML, Borowski J. Mikrobiologia lekarska. Warszawa, Wydawnictwo Lekarskie
PZWL, wyd. 2001.
23. Meurman JH. Dental infections and general health. Quintessence Int.1997; 28: 807-11.
24. Debelian GJ, Olsen I, and Tronstad L. Systemic diseases caused by oral microorganisms.
Endod .Dent .Traumatol. 1994; 10: 57-65.
25. Piovano S. Bacteriology of most frequent oral anaerobic infections. Anaerobe.1999; 5:
221-7.
26. Marsh PD, and Bradshaw DJ. Dental plaque as a biofilm. J Ind Microbiol.1995; 15: 16975.
27. Moore WEC, and Moore LVH. The bacteria of periodontal disease.J Periodontol.2000; 5:
66-77.
28.Daniluk T, Tokajuk G, Cylwik-Rokicka D, Rokiewicz D,Zaremba ML and Stokowska
W.Aerobic and anaerobic bacteria in subgingival and supragingival plaques of adult
patients. 2006.
29. Socransky SS, Haffajee AD, Cugini MA, Smith C, and
Kent RL. Jr. Microbial
complexes in subgingival plaque. J .Clin. Periodontol. 1998;25:134–44.
30. Salari MH, and Kadkhoda Z. Rate of cultivable subgingival periodontopathogenic
bacteria in chronic periodontitis. J Oral Science. 2004;46 (3 )157-161.
31. Spratt D.A, Greenman J and Schaffer A G . Capnocytophaga gingivalis: effects of
glucose
concentration
on
growth
and
hydrolytic
enzyme
production.
Microbiol.1996;142(8): 2161-4.
32. Gürsoy M , Haraldsson G, Hyvönen M, Sorsa T, Pajukanta R and Könönen E . Does
the frequency of Prevotella intermedia increase during pregnancy? Oral. Microbiol.
Immunol. 2009;24(4): 299-303.
33. Mayorga-Fayad I, Lafaurie GI, Contreras A, Castillo D M , Barón A and Aya Mdel R.
Subgingival microbiota in chronic and aggressive periodontitis in Bogotá, Colombia: an
epidemiological approach Biomedica .2007;27(1): 21-33.
34. Ximenez-Fyvie L A , Haffajee A D and Socransky S S . Microbial composition of supra
and subgingival plaque in subjects with adults periodontitis. J Clin Periodontol. 2000;
27: 722-732.
1732
Vol 3, Issue 6, 2014.
www.wjpps.com
Al-Jebouri et al.
35. Rams T E,
World Journal of Pharmacy and Pharmaceutical Sciences
Dujardin S, Jacqueline D ,
John E. , Arie J ,and Winkelhoff V.
Spiramycin resistance in human periodontitis microbiota . 2011.
36. Brook I. Treatment of anaerobic infection. Expert Rev Anti Infect Ther. 2007 ;5:991-1006.
37. Haukioja A. Probiotics and Oral Health.Eur. J .Dent. 2010 ; 4(3): 348–355.
38. Sally A, Keith P, Susan D, David H and Arthur J. Antimicrobial susceptibility of
anaerobic bacteria in New Zealand :1999-2003.J antimicrobial chemother.2013;68:3440.
39. Nyfros S, Kononen E, and Syrjanen R. Emergence of penicillin resistance among
fusibacterium nucleatum populations of commensal oral flora during early childhood.J.
Antimicrob .Chemther.2003;51:107-112.
40. Poulet P, Duffaut D, Barthet P and Brumpt I. Concentrations and in vivo antibacterial
activity of spiramycin and metronidazole in patients with periodontitis treated with highdose metronidazole and the spiramycin/metronidazole combination.J Antimicrob
Chemother. 2013; 55( 3 ): 347-351.
41. Hedberg M
and arl Nord C E. Anaerobic bacteria.Infect Dis
Antimicrob
Agents.2013;34:76-81.
42. Boyanova L, Djambazov V, Gergova G,Lotov D,Petrov D ,Osmanliev D ,Minchev Z and
Mitov I. Anaerobic microbiology in 198 cases of pleural empyema:a Bulgarian
study.Anaerobe. 2004; 10:261-267.
43. Higashi K, Matsushita M, Morisaki K, Hayashi S I and Mayumi T. Local drug delivery
systems for the treatment of periodontal disease.J. Pharmacobio. Dyn. 1991;14 :72-81.
44. Louise CS , Jayshree D , Philip AC , and John H. Antibiotic resistance in general dental
practice-a cause for concern? J antimicrob agents and chemother .2004;53:567-576.
45. Selckuk MO , Sinan ME , Amet O , Aziz S , and Mustafa K. Microbiological
abscesses (pain and swelling) and the antimicrobial susceptibility of isolated bacteria.
J
clinical pain. 2006;18(4):327-337.
46. Ahmed MA. The clinical effectiveness of treatment of chronic periodonititis. BDS.
Thesis. University of Baghdad.2001.
47. Stevens RH, and Grossman LI. Antimicrobial effects of root canal cements on obligate
an aerobic organisms.J
1733
endodental .1981 ; 45:266 -272.
Vol 3, Issue 6, 2014.
www.wjpps.com