The purpose of this study was to assess whether an intensive oral hygiene regimen practised during pregnancy results in a clinically healthy gingival state, and to assess whether experimentally-induced gingivitis differs in severity during pregnancy as compared to post-partum. In addition, levels of black-pigmented Gram negative anaerobes at subgingival and oral mucosal sites and plasma concentrations of free estrogens and prosterone were determined. These parameters were studied during a 14-day episode of experimental gingivitis induced in the 25th week of pregnancy, and again 6 months post-partum. The subjects were selected on shallow pockets < or = 4 mm and interproximal loss of attachment not exceeding 2 mm. As a result of controlled oral hygiene, the gingival condition improved both during pregnancy and post-partum. At day 0 during pregnancy, however, gingival swelling, redness, and bleeding on probing were found to be higher than post-partum. Free plasma levels of estrogens and progesterone were found to be normal throughout the study. It was hypothesized that the increase in severity of gingival symptoms during pregnancy reflect microvascular physiologic effects of increased levels of these hormones. During pregnancy, more swelling, redness and bleeding on probing developed during experimental gingivitis than post-partum, whereas the amount of plaque was similar in both phases. This suggests that as a result of dental plaque accumulation, gingival inflammation develops superimposed on pregnancy-associated physiologic alterations. Microbiological evaluation showed that the mean proportions of Prevotella intermedia in subgingival plaque increased during experimental gingivitis performed during pregnancy, whereas no increase of this micro-organism was found post-partum.

Today, 10 black‐pigmented Bacteroides (BPB) species are recognized. The majority of these species can be isolated from the oral cavity. BPB species are involved in anaerobic infections of oral and non‐oral sites. In the oral cavity. BPB species are associated with gingivitis, periodontitis, endodontal infections and odontogenic abscesses. Cultural studies suggest a specific role of the various BPB species in the different types of infection. Bacteroides gingivalis is closely correlated with destructive periodontitis in adults as well as in juveniles. Bacteroides intermedius seems to be less specific since it is found in gingivitis, periodontitis, endodontal infections and odontogenic abscesses. The recently described Bacteroides endodontalis is closely associated with endodontal infections and odontogenic abscesses of endodontal origin. There are indications that these periodontopathic BPB species are only present in the oral cavity of subjects suffering from periodontal breakdown, being absent on the mucosal surfaces of subjects without periodontal breakdown. BPB species associated with healthy oral conditions are Bacteroides melaninogenicus, Bacteroides denticola and Bacteroides loescheii. There are indications that these BPB species are part of the normal indigenous oral microflora. Many studies in the past have documented the pathogenic potential and virulence of BPB species. This virulence can be explained by the large numbers of virulence factors demonstrated in this group of microorganisms. Among others, the proteolytic activity seems to be one of the most important features. Several artificial substrates as well as numerous biological proteins are degraded. These include anti‐inflammatory proteins such as alpha‐2‐macroglobulin, alpha‐1‐antitrypsin, C3 and C5 complement factors and immunoglobulins. B. gingivalis is by far the most proteolytic species, followed by B. endodontalis. Like other bacteria, the lipopolysaccharide of B. gingivalis has shown to be active in bone resorption in vitro and is capable in stimulating interleukin‐1 production in human peripheral monocytes. Based on the well documented association with periodontal disease and the possession of relevant virulence factors. BPB species must be considered as important micro‐organisms in the etiology of oral infections. B. gingivalis seems to be the most pathogenic and virulent species.

The virulence of different black‐pigmented Bacteroides strains (BPB) was studied using monoinfections in an experimental animal model. Bacterial suspensions were injected subcutaneously in the back of mice. After four days the mice were killed and the induced inflammations were examined histologically. Bacteroides gingivalis strains, which are mostly isolated from subgingival plaque associated with adult destructive periodontitis. were characterized by inducing a spreading type of inflammation, mostly resulting in a gravity abscess or a phlegmonous abscess. Strains of Bacteroides melaranogenicus subspecies intermedius, isolated from gingivitis or periodontitis, always induced a localized abscess at the site of injection. Most Bacteroides asaccharolyticus strains, which are isolated from nonoral sites, caused a localized inflammation, and Bacteroides melaninogenicus subspecies melaninogenicus strains caused only minimal inflammation.

The present study primarily aimed at investigating the oral microbiota in smokers and non-smokers with established gingivitis and monitoring its composition during experimental gingivitis. Secondly, it aimed at examining whether the composition of the microbiota is associated with different levels of gingival inflammation during this experimental gingivitis trial. For this purpose, 25 non-dental university students with gingivitis were recruited. 11 subjects were smokers and 14 were non-smokers. After achieving gingival health, they entered a 14-day experimental gingivitis trial. Plaque and bleeding were assessed before entering into the study (intake), at day 0, day 5 and at day 14 of the experiment. Microbiological samples from mucosal sites and dental plaque (taken at intake, day 0, and day 14) were analysed for the presence of Actinomyces species, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Fusobacterium nucleatum, Peptostreptococcus micros, Porphyromonas gingivalis, Prevotella intermedia and Streptococcus species. At day 14 of the experimental period, the level of plaque formation was not different between smokers and non-smokers, but bleeding scores were lower in smokers than in non-smokers (15% and 30% respectively, p=0.01). The change from natural gingivitis to a state of gingival health and a subsequent change from gingival health to experimentally induced gingivitis was accompanied by quantitative alterations in the cultivable microbiota in both groups. Changes were most prominent in the transition from gingival health to experimental gingivitis and were found in dental plaque for Actinomyces species, C. rectus, F. nucleatum, and P. intermedia. Within the group of non-smokers, a distinction was made between subjects with a 'weak' or 'strong' inflammatory response. No relationship with a single bacterial species could be established which would likely explain the differences in levels of inflammation. It is concluded that differences in response to experimental gingivitis are not caused by major differences in the composition of the oral microbiota.

In order to gain insight into the relative importance of several virulence factors of Bacteroides gingivalis, 8 strains with a varying virulence were studied. The virulence of B. gingivalis was determined in a mouse model. Strains HG 66, HG 76 and HG 184 were very virulent causing phlegmonous abscesses with lesions and necrosis. The strains HG 405 and HG 462 caused phlegomonous abscesses with pus. Strains HG 91, HG 94 and HG 185 were less virulent and induced gravity abscesses. In vitro strains HG 66, HG 76 and HG 184 induced low amounts of chemiluminescence by polymorphonuclear leucocytes. All other strains including HG 405 and HG 462 caused a relatively high chemiluminescence. Most strains displayed a high sensitivity to the bactericidal activity of fresh serum except for the highly virulent strains HG 66, HG 76 and HG 184. No differences in extracellular proteolytic activity on Azocoll, production of volatile fatty acids and ammonia were found between the B. gingivalis strains studied. In conclusion, differences in virulence were shown within the species B. gingivalis; the relative importance of several virulence factors was investigated.