Scand.J. Dent. fl«. 19 7 8: 86: 17 4 - 1 8 1 (Key words: bacteria; gingivitis; miaobiology, oral)
Microbiota of gingivitis in man J0RGEN SLOTS, DIANA M O E N B O , J0RGEN LANGEBAEK AND ASGER FRANDSEN Departments of Periodontology and Microbiology, Royal Dental College, Copenhagen, Denmark
ABSTRACT - A Study on the predominant cultivable microorganisms inhabiting gingival crevices affected with a chronic gingivitis was carried out using the roll tube culture technique. Samples were obtained from nine individuals 25—42 years of age. Gram-positive rods made up 26.1% of the isolates and included mainly Actinomyces naeslundii, Actinomyces israelii, and Actinomyces viscosus. Streptococcus mitis and Streptococcus sanguis together made u p
26.8% of the cultivable organisms. Feptostreptococcus averaged 3.0% of the organisms recovered. Gram-negative anaerobic rods constituted 25.0% of the total isolates with Fusobacterium nucleatum, Bacteroides melaninogenicus ss. intermedius, Bacteroides ochraceus, other Bacteroides species, Selenomonas sputigena, and Campylobacter sputorum as the most predominant
isolates. Haemophilus parainfiuenzae averaged about 14% and Veillonella species 4.3% of the cultivable microflora. The data presented indicate that the subgingival microflora of a chronic gingivitis differs from those of healthy periodontium and advanced adult and juvenile periodontitis. This might suggest that different infectious processes may be operative in various clinical entities of periodontal disease. (Accepted for publication 18 February 1978)
Gingivitis in man has been shown to be initiated by dental plaque (LOE, THEILADE &; JENSEN 1965) and is considered to be an inevitable antecedent to loss of periodontal attachment. However, in some patients gingivitis can persist for a considerable period of time without progression to periodontitis, whereas gingivitis in other patients will rapidly progress into a destructive periodontitis. Similar variations in the rate of progression of periodontal disease can be found between tooth surfaces in the same patient and have also been observed in an experimental periodontitis study in Beagle dogs (LiNDHE, HAMP &: LOE 1975). The reason for these variations may be differences in host response to the causative bacteria and/or various degrees
of periodontopathic potentials of dental plaque (PAGE &: SCHROEDER 1976). Recent findings indicate that different clinical entities of periodontal disease may reflect not only variations in host response, but also characteristic variations of the infectious flora. Thus, a scanty microflora dominated by Gram-positive organisms (8596), usually Actinomyces and Streptococcus species, was found in healthy gingival sulci (SLOTS 1977b). In the apical part of deep periodontal pockets in adults. Gram-negative anaerobic rods, mainly Bacteroides melaninogenicus ss. asaccharolyticus and Fusobacterium nucleatum made up about 7596 of the isolates (SLOTS
1977a). The front bacteria of deep pockets in patients with juvenile periodontitis also had a strong component of Gram-
MICROBIOTA OF GINGIVITIS
-iff:
mm and with GI= 1 (patients: K.S., E.H., L.J., A.S., J.S., T.S.) and GI = 2 (patients: I.M., D.M., H.T.). No dental calculus or tooth Bacteroides species, such as B. ochraceus restoration was present in the sampling (Capnocytophaga), and other organisms of region. The gingivitis was diagnosed at least unknown species (NEWMAN, SOCRANSKY, 2-3 months before the time of sampling and the dental history suggested that the inflammaSAVITT, PROPAS & CRAWFORD 1976, SLOTS tion had remained for a long time. Efforts 1976, NEWMAN & SOCRANSKY 1977). were made to remove subgingival material Few data are available about the from the most apical part of the pockets cultivable periodontal pocket microflora without contamination from other sources. associated with gingivitis without loss of Prior to sampling, saliva, debris, and plaque attachment. Recently, HELDERMAN (1975) were carefully removed from the gingival studied the occurrence of five groups of margin and the supragingival tooth surface with sterile gauze. Then, the gingival papilla Gram-negative anaerobes in noninflamed was deflected from the tooth surface by and inflamed gingival crevices and found appropriate dental instruments in order to an increase in the proportions of allow a small periodontal sealer to freely enter the base of the pocket for the sample collecCampylobacter sputorum, F. nucleatum, B. tion. Treatment of the samples and partial ochraceus and Selenomonas sputigena with incharacterization of the microorganisms creasing gingival index score. However, followed essentially the procedures previously
negative organisms (about 65%), but of a different nature. They included strains of
the proportions of B. melaninogenicus and other Gram-negative organisms were not determined due to the inability of the culture medium to grow these organisms (HELDERMAN & WINKLER 1975). The present investigation attempted to characterize the predominant cultivable microorganisms inhabiting gingival crevices affected with a chronic gingivitis. In order to further elucidate the kinds of microorganisms related to the development of periodontal disease, the microflora described in the present study was compared with that of healthy periodontium and advanced juvenile and adult periodontitis.
Material and methods Seven females and two males were included in the study. Their ages ranged from 25 to 42 years with a mean of 33 years. Their gingival inflammation index (Gl according to LOE 8C SiLNESS 1963) ranged from 0.74 to 1.48 (mean 1.18). No individuals had teeth with more than 4-mm pocket depth. From each subject, subgingival material was collected from one approximal pocket with a pocket depth of 2-3
described (SLOTS 1977a).
Results
The total number of cultivable microorganisms sampled from the nine pockets examined ranged from 6.9x10^ to 2.6 X 10^ with a mean of 1.5 x 10^ Table 1 lists the colony counts on various selective media and shows B. melaninogenicus to
average 7.696 of the organisms recovered by deep agar growth in MM 10-roll tubes. The predominant cultivable pocket flora associated with gingivitis is presented in Table 2. From each individual, 36-54 bacterial strains were subcultured from MM 10-roll tubes. A total of 403 strains were examined from the nine samples: 173 (42.9%) isolates were anaerobes, 176 (43.7%) were Gram-negative, and 264 (65.5%) were categorized as rods. Gram-negative
anaerobic rods
averaged
25.0% of the total isolates and predominated in samples L.J., A.S., J.S. and T.S. (Table 2). About 35% of the Gramnegative anaerobic rods were fusiform
SLOTS,
176
ose
s ^ ;| .^ s
MOENBO,
^ •
O '
X
CM CO CO CO CO
1 CO
d
^ s o
e o
organisms which produced butyric acid with variable amounts of acetic, propionic, lactic, and succinic acids in glucose broth cultures. These strains were generally non-fermentative, indol positive, esculin-hydrolysis negative, and produced little or no gas, indicating that they were strains of F. nucleatum. Among a total of 31 strains of B. melaninogenicus recovered, 23 were glucolytic, produced indol, hydrolized starch and gelatin, and exhibited no hydrolysis of esculin and casein. These strains were identified as B.
• i fc 2
o o o '~*
LANGEBAEK AND ERANDSEN
Q
o o
o 'X'
d CO d
eil
, ;
o
u
y
1 lO
cri CM
>
3
melaninogenicus
O ^~.
2 \
>
o o o
_ OJ
strains
o1
J CO
bo rt
\,
o o X
JJ
CO
65.2
•s
^^
^-^ CM
o
1
1
s •'-.^ 2
d
•
3 X) O
'Si " > 3 S
:s '§ ~g s S
CO
o o o ^-*
od
X
T CO
c/i
oo
1 s s CJ
oj
o
o
icus bo 1
s
>'a.
o CO
o oo
T o
O rt „
2
[J
G O
e
1 05
1
-O
'5 J [^
JS "^
- O bo . cu rt CQ
^ O IJ
X
s
ill S ^
rt XI
O p.
> 2
.2 o
intermedius.
Five
saccharolytic isolates were indol negative and hydrolyzed esculin, starch, and gelatin, indicating that they might be
•
Microbiota of gingivitis in man J0RGEN SLOTS, DIANA M O E N B O , J0RGEN LANGEBAEK AND ASGER FRANDSEN Departments of Periodontology and Microbiology, Royal Dental College, Copenhagen, Denmark
ABSTRACT - A Study on the predominant cultivable microorganisms inhabiting gingival crevices affected with a chronic gingivitis was carried out using the roll tube culture technique. Samples were obtained from nine individuals 25—42 years of age. Gram-positive rods made up 26.1% of the isolates and included mainly Actinomyces naeslundii, Actinomyces israelii, and Actinomyces viscosus. Streptococcus mitis and Streptococcus sanguis together made u p
26.8% of the cultivable organisms. Feptostreptococcus averaged 3.0% of the organisms recovered. Gram-negative anaerobic rods constituted 25.0% of the total isolates with Fusobacterium nucleatum, Bacteroides melaninogenicus ss. intermedius, Bacteroides ochraceus, other Bacteroides species, Selenomonas sputigena, and Campylobacter sputorum as the most predominant
isolates. Haemophilus parainfiuenzae averaged about 14% and Veillonella species 4.3% of the cultivable microflora. The data presented indicate that the subgingival microflora of a chronic gingivitis differs from those of healthy periodontium and advanced adult and juvenile periodontitis. This might suggest that different infectious processes may be operative in various clinical entities of periodontal disease. (Accepted for publication 18 February 1978)
Gingivitis in man has been shown to be initiated by dental plaque (LOE, THEILADE &; JENSEN 1965) and is considered to be an inevitable antecedent to loss of periodontal attachment. However, in some patients gingivitis can persist for a considerable period of time without progression to periodontitis, whereas gingivitis in other patients will rapidly progress into a destructive periodontitis. Similar variations in the rate of progression of periodontal disease can be found between tooth surfaces in the same patient and have also been observed in an experimental periodontitis study in Beagle dogs (LiNDHE, HAMP &: LOE 1975). The reason for these variations may be differences in host response to the causative bacteria and/or various degrees
of periodontopathic potentials of dental plaque (PAGE &: SCHROEDER 1976). Recent findings indicate that different clinical entities of periodontal disease may reflect not only variations in host response, but also characteristic variations of the infectious flora. Thus, a scanty microflora dominated by Gram-positive organisms (8596), usually Actinomyces and Streptococcus species, was found in healthy gingival sulci (SLOTS 1977b). In the apical part of deep periodontal pockets in adults. Gram-negative anaerobic rods, mainly Bacteroides melaninogenicus ss. asaccharolyticus and Fusobacterium nucleatum made up about 7596 of the isolates (SLOTS
1977a). The front bacteria of deep pockets in patients with juvenile periodontitis also had a strong component of Gram-
MICROBIOTA OF GINGIVITIS
-iff:
mm and with GI= 1 (patients: K.S., E.H., L.J., A.S., J.S., T.S.) and GI = 2 (patients: I.M., D.M., H.T.). No dental calculus or tooth Bacteroides species, such as B. ochraceus restoration was present in the sampling (Capnocytophaga), and other organisms of region. The gingivitis was diagnosed at least unknown species (NEWMAN, SOCRANSKY, 2-3 months before the time of sampling and the dental history suggested that the inflammaSAVITT, PROPAS & CRAWFORD 1976, SLOTS tion had remained for a long time. Efforts 1976, NEWMAN & SOCRANSKY 1977). were made to remove subgingival material Few data are available about the from the most apical part of the pockets cultivable periodontal pocket microflora without contamination from other sources. associated with gingivitis without loss of Prior to sampling, saliva, debris, and plaque attachment. Recently, HELDERMAN (1975) were carefully removed from the gingival studied the occurrence of five groups of margin and the supragingival tooth surface with sterile gauze. Then, the gingival papilla Gram-negative anaerobes in noninflamed was deflected from the tooth surface by and inflamed gingival crevices and found appropriate dental instruments in order to an increase in the proportions of allow a small periodontal sealer to freely enter the base of the pocket for the sample collecCampylobacter sputorum, F. nucleatum, B. tion. Treatment of the samples and partial ochraceus and Selenomonas sputigena with incharacterization of the microorganisms creasing gingival index score. However, followed essentially the procedures previously
negative organisms (about 65%), but of a different nature. They included strains of
the proportions of B. melaninogenicus and other Gram-negative organisms were not determined due to the inability of the culture medium to grow these organisms (HELDERMAN & WINKLER 1975). The present investigation attempted to characterize the predominant cultivable microorganisms inhabiting gingival crevices affected with a chronic gingivitis. In order to further elucidate the kinds of microorganisms related to the development of periodontal disease, the microflora described in the present study was compared with that of healthy periodontium and advanced juvenile and adult periodontitis.
Material and methods Seven females and two males were included in the study. Their ages ranged from 25 to 42 years with a mean of 33 years. Their gingival inflammation index (Gl according to LOE 8C SiLNESS 1963) ranged from 0.74 to 1.48 (mean 1.18). No individuals had teeth with more than 4-mm pocket depth. From each subject, subgingival material was collected from one approximal pocket with a pocket depth of 2-3
described (SLOTS 1977a).
Results
The total number of cultivable microorganisms sampled from the nine pockets examined ranged from 6.9x10^ to 2.6 X 10^ with a mean of 1.5 x 10^ Table 1 lists the colony counts on various selective media and shows B. melaninogenicus to
average 7.696 of the organisms recovered by deep agar growth in MM 10-roll tubes. The predominant cultivable pocket flora associated with gingivitis is presented in Table 2. From each individual, 36-54 bacterial strains were subcultured from MM 10-roll tubes. A total of 403 strains were examined from the nine samples: 173 (42.9%) isolates were anaerobes, 176 (43.7%) were Gram-negative, and 264 (65.5%) were categorized as rods. Gram-negative
anaerobic rods
averaged
25.0% of the total isolates and predominated in samples L.J., A.S., J.S. and T.S. (Table 2). About 35% of the Gramnegative anaerobic rods were fusiform
SLOTS,
176
ose
s ^ ;| .^ s
MOENBO,
^ •
O '
X
CM CO CO CO CO
1 CO
d
^ s o
e o
organisms which produced butyric acid with variable amounts of acetic, propionic, lactic, and succinic acids in glucose broth cultures. These strains were generally non-fermentative, indol positive, esculin-hydrolysis negative, and produced little or no gas, indicating that they were strains of F. nucleatum. Among a total of 31 strains of B. melaninogenicus recovered, 23 were glucolytic, produced indol, hydrolized starch and gelatin, and exhibited no hydrolysis of esculin and casein. These strains were identified as B.
• i fc 2
o o o '~*
LANGEBAEK AND ERANDSEN
Q
o o
o 'X'
d CO d
eil
, ;
o
u
y
1 lO
cri CM
>
3
melaninogenicus
O ^~.
2 \
>
o o o
_ OJ
strains
o1
J CO
bo rt
\,
o o X
JJ
CO
65.2
•s
^^
^-^ CM
o
1
1
s •'-.^ 2
d
•
3 X) O
'Si " > 3 S
:s '§ ~g s S
CO
o o o ^-*
od
X
T CO
c/i
oo
1 s s CJ
oj
o
o
icus bo 1
s
>'a.
o CO
o oo
T o
O rt „
2
[J
G O
e
1 05
1
-O
'5 J [^
JS "^
- O bo . cu rt CQ
^ O IJ
X
s
ill S ^
rt XI
O p.
> 2
.2 o
intermedius.
Five
saccharolytic isolates were indol negative and hydrolyzed esculin, starch, and gelatin, indicating that they might be
•
