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Objective: The antimicrobial effect of photoactivated disinfection (PAD) using toluidine blue and an LED lamp was tested on endodontic pathogens in planktonic suspension and after inoculation into extracted teeth. Irradiation time was limited to 30 seconds., Study Design: The effect of PAD on planktonic suspensions of Escherichia coli, Candida albicans, Enterococcus faecalis, Fusobacterium nucleatum, and Streptococcus intermedius was analyzed using Poisson regression. Moreover, cultures of S. intermedius were inoculated into prepared root canals of extracted molars. The effect of PAD performed immediately after inoculation or after overnight bacterial incubation was determined by a 2-sample t test., Results: Photoactivated disinfection yielded significant reductions (P < .001) in the viable counts of all organisms in planktonic suspension. The PAD treatment of S. intermedius in root canals yielded a mean log10 reduction of 2.60 (P < .001) immediately after inoculation and of 1.38 (P < .001) after overnight incubation., Conclusion: Photoactivated disinfection using a conventional light source strongly reduces the number of viable endodontic pathogens in planktonic suspension and in root canals., (Copyright 2010 Mosby, Inc. All rights reserved.)

Bacteria of the genus Capnocytophaga form part of the resident oral flora in children and adults. They are recognized as opportunistic pathogens of various extra-oral infections. The significance of individual species in periodontal and extra-oral diseases is unclear, due to the inability of conventional phenotypic tests to identify clinical isolates to species level. Aiming at a clear distinction between species, we undertook a phylogenetic study of a collection of 102 Capnocytophaga strains including 62 oral isolates from children and 40 reference strains from oral and extra-oral infections representing the five known, human, oral Capnocytophaga species. The phylogeny was estimated on the basis of multilocus enzyme electrophoresis (MLEE) of 12 intracellular, housekeeping enzymes and by partial 16S rRNA gene sequencing, and was compared to phenotypic characteristics. The clustering profiles in the MLEE and sequence-based dendrograms were concordant and allowed identification of isolates to species level, based on co-clustering with reference strains. The study confirmed Capnocytophaga ochracea and Capnocytophaga sputigena as separate species, and underlined the problems of distinguishing between them by conventional phenotypic tests. The presence of two distinct clusters of oral isolates from children indicated the existence of novel species, supported by analysis of near-full-length 16S rRNA gene sequences and by DNA-DNA hybridization results. One cluster of weakly saccharolytic isolates without the ability to ferment sucrose is proposed as Capnocytophaga leadbetteri sp. nov. (type strain AHN8855(T)=CCUG 51857(T)=NCTC 13375(T)). Another cluster not phenotypically distinguishable from C. ochracea and C. sputigena is designated Capnocytophaga genospecies AHN8471 (represented by strain AHN8471=CCUG 51856=NCTC 13374).

The association of Porphyromonas gingivalis to periodontal disease is not clearly understood. Similar proportions of P. gingivalis may be cultivated from both inactive and actively degrading periodontal pockets. Differences in virulence among strains of P. gingivalis exist, but the molecular reason for this remains unknown. We examined the population structure of P. gingivalis to obtain a framework in which to study pathogenicity in relation to evolution. Phylogenetic trees derived from the sequencing of fragments of four housekeeping genes, ahp, thy, rmlB, and infB, in 57 strains were completely different with no correlation between clustering of strains in the four dendrograms. Combining the various alleles of the four gene fragments sequenced resulted in 41 different sequence types. The index of association, I(A), based on a single representative of each sequence type was 0.143 +/- 0.202, indicating a population at linkage equilibrium. Inclusion of all isolates for the calculation of I(A) resulted in a value of 0.206 +/- 0.171. This suggests an epidemic population structure supported by the finding of genetically identical strains in different parts of the world. We observed a random distribution of two virulence-associated mobile genetic elements, the ragB locus and the insertion sequence IS1598, among 132 strains tested. In conclusion, P. gingivalis has a nonclonal population structure characterized by frequent recombination. Our study suggests that particular genotypes, possibly with increased pathogenic potential, may spread successfully in the human population.

The causes of bracket loss during the late phases of orthodontic treatment have hitherto been considered to be of mechanical nature (e.g. trauma, high forces applied). Recently, it was hypothesized that bacterial degradation of composite resin could be a reason for late bracket failure. This was based on the observation of apparently degraded composite on bracket bases sent to a recycling company, and on a weight loss of 0.1-1% of a limited number of composite resin foils incubated with bacteria. To further elucidate the basis for this hypothesis, we examined brackets, immediately after debonding, for signs of potential decay and tested the ability of selected bacterial species to degrade composites. Out of a total of 1056 brackets collected, 6.2% displayed signs of decay indicative of inclusion of air bubbles, corrosion of the bracket base and discoloration of the composite and comparable to the decay observed on brackets received from the recycling company. Composite discs of two different brands were each incubated with either of five strains representing different bacterial species. The incubation lasted 3 months or until the bacteria had died. There was no significant difference in weight change of the discs incubated with bacteria and control discs incubated without bacteria. Thus, our results do not support that bacterial degradation of composite resin bonding materials is a likely cause of bracket loss.

Currently, the Prevotella intermedia group includes three biochemically and phylogenetically related species: Prevotella intermedia, Prevotella nigrescens, and the newly described Prevotella pallens. The two first-named species are mentioned with varying emphasis in connection with periodontal diseases, while such a connection of P. pallens is not known. Mothers serve as a plausible source of bacteria to their children, and conceivably, a mother with periodontitis as a recurrent reservoir of periodontally infecting organisms. In the present study, 23 mothers and their young children were examined for the presence of the P. intermedia group organisms in relation to maternal periodontal status (I: periodontal health, II: initial periodontitis, and III: advanced periodontitis). Species differentiation was based on established biochemical methods, electrophoretic mobility patterns, SDS-PAGE, and DNA hybridization. P. intermedia was not recovered from children but nearly exclusively from mothers in group III, thus confirming its association with periodontitis. P. nigrescens and P. pallens were frequently found in mothers and children. To determine bacterial transmission between a mother and her child, 72 isolates from 13 mother-child pairs were analyzed by arbitrarily primed PCR (AP-PCR). Similar AP-PCR types of P. nigrescens and/or P. pallens were recovered from 3/4 pairs in group I, 2/5 pairs in group II, and none in group III. Our results indicate that different species within the P. intermedia group have a different colonization pattern in childhood and that the periodontal status reflects qualitatively their presence in maternal saliva. Intra-familial transmission of P. nigrescens and P. pallens can occur in early childhood, however similar AP-PCR types were most obvious within periodontally healthy mother-child pairs.

Periodontal diseases affect millions of people world wide. Prevention and treatment of these diseases require considerable attention from the individual as well as society and cause great expenses. Understanding disease etiology and mechanisms of pathogenesis is a prerequisite for optimal treatment strategies. The highly variable speed of periodontal destruction and in some sites persistence for years of deep pockets without further periodontal destruction points to the significance of individual bacterial species in the complex subgingival microflora for pathogenesis. Destruction of periodontal tissue occurs when the load of bacterial virulence factors overcomes the local immune defense. One way of doing this is by bacterially-induced degradation of IgA which is considered to mediate its protective functions in an anti-inflammatory way and to down-regulate inflammation through inhibition of IgG- and IgM-mediated activities. A competent IgA system may be of particular significance in chronic inflammatory diseases, as periodontal diseases, where the inflammatory reaction in itself probably is the main cause of destruction. In these cases, degradation of IgA may serve the purpose of immune evasion for the bacteria and at the same time may induce a relatively increased activity in the inflammation-stimulating part of the immune system which may aggravate periodontal destruction. Both gram-positive rods, streptococci, and Veillonella species from the subgingival microflora induce an altered immunoelectrophoretic mobility of IgA1 indicative of removal of terminally positioned sialic acid. Quantitative determination of residual carbohydrate content of IgA1 after incubation with bacterial cells of Gram-positive rods has confirmed that they remove sialic acid, and in addition to that, only minor amounts of carbohydrates. Apart from serving a nutritional purpose, desialylation of IgA may also serve a purpose of immune evasion. Glycosylation and, in particular sialic acid protects glycoproteins, including immunoglobulins, against proteolytic enzymes and deglycosylation of antibodies increase their sensitivity to proteolytic degradation and inhibit the Fc-mediated effector functions that mediate antigen disposal. Extensive proteolytic degradation of IgA1 is induced by a number of bacterial species often associated with periodontal diseases, including P. gingivalis, Pr. intermedia, and Pr. nigrescens. These species produce enzymes of broad proteolytic activity, that also may degrade immunoglobulins of other isotopes, complement factors, iron-containing plasma proteins etc. Extensive hydrolysis of immunoglobulins induced by these bacteria serve a nutritional purpose and is essential for growth of other bacteria in mixed cultures. It also has an obvious detrimental effect on the defence potential of the specific humoral immune system. These bacteria seem to be essential for the transmissibility of experimental infections in animals with mixtures of oral bacteria and a likely reason is their ability to provide the other bacteria with amino acids, peptides, and iron for growth and their ability to inhibit the immune defence. The hinge region of IgA1 is relatively resistant to proteolysis because of a high proline content and presence of several oligosaccharide side chains. It is therefore interesting that a number of taxonomically unrelated bacteria, including both commensals and overt pathogens, have evolved the capability to specifically cleave human IgA1 in the hinge region. These so-called IgA1 proteases leave Fab and Fc fragments intact for which reason a direct nutritional purpose of the enzymes may be excluded. In the oral cavity, specific IgA1 proteases are produced by the streptococcal species that constitute a considerable proportion of initial dental plaque and the flora on buccal and pharyngeal mucosa. In all three cases the flora is sparse and contact with saliva, including S-IgA1 antibodies is intimate. (ABSTRACT TRUNCATED)

Thirty-three previously non-typable faintly pigmented Gram-negative anaerobic bacterial isolates, biochemically most closely related to Prevotella intermedia and Prevotella nigrescens, were analysed for enzymic reactions, cellular fatty acid (CFA) composition, electrophoretic mobility of malate and glutamate dehydrogenases, hybridization with P. intermedia and P. nigrescens species-specific oligonucleotide probes and, for genetic heterogeneity, by arbitrarily primed PCR (AP-PCR). P. intermedia ATCC 25611T and P. nigrescens ATCC 33563T were run in parallel for comparison. Twenty-nine isolates originated from the normal oral flora of 18 subjects (including five mother-child pairs), and four isolates from various infections. Except for a negative lipase reaction, enzymic profiles of the test isolates were similar to those of P. intermedia and P. nigrescens. Clustering of CFAs, electrophoretic mobility patterns, hybridization with DNA probes for P. intermedia and P. nigrescens, and AP-PCR band patterns of the test isolates differed from those of the type strains of P. intermedia and P. nigrescens, suggesting the existence, in humans, of a new anaerobic species of pigmented, moderately saccharolytic, indole-positive Gram-negative rods.

Complete 16S rRNA gene sequences of three representative strains of anaerobic, Gram-negative, pigmented, moderately saccharolytic, indole-positive bacteria isolated from the oral cavity of humans were determined. According to comparative analyses of the rRNA sequence data, this organism represents a previously unknown species within the genus Prevotella. In addition, 22 representative strains and 21 reference strains (including 11 Prevotella intermedia and 10 Prevotella nigrescens strains) were subjected to multilocus enzyme electrophoretic analysis. The strains were consistently separated into three clearly distinct groups, corresponding to their previous entities. On the basis of the present phylogenetic results that confirmed our biochemical and genetic data, we propose a new species, Prevotella pallens. The type strain is NCTC 13042 (= AHN 10371).

Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region into intact Fab and Fc fragments. To investigate whether these enzymes are subject to inhibition in vivo in humans, we tested 34 sera from periodontally diseased and healthy individuals in an enzyme-linked immunosorbent assay for the presence and titers of inhibition of seven Prevotella and Capnocytophaga proteases. All or nearly all of the sera inhibited the IgA1 protease activity of Prevotella buccae, Prevotella oris, and Prevotella loescheii. A minor proportion of the sera inhibited Prevotella buccalis, Prevotella denticola, and Prevotella melaninogenica IgA1 proteases, while no sera inhibited Capnocytophaga ochracea IgA1 protease. All inhibition titers were low, ranging from 5 to 55, with titer being defined as the reciprocal of the dilution of serum causing 50% inhibition of one defined unit of protease activity. No correlation between periodontal disease status and the presence, absence, or titer of inhibition was observed. The nature of the low titers of inhibition in all sera of the IgA1 proteases of P. buccae, P. oris, and P. loescheii was further examined. In size exclusion chromatography, inhibitory activity corresponded to the peak volume of IgA. Additional inhibition of the P. oris IgA1 protease was found in fractions containing both IgA and IgG. Purification of the IgG fractions of five sera by passage of the sera on a protein G column resulted in recovery of inhibitory IgG antibodies against all three IgA1 proteases, with the highest titer being for the P. oris enzyme. These finding indicate that inhibitory activity is associated with enzyme-neutralizing antibodies.

The humoral immune response, especially the production of IgG and IgA, is considered to have a protective role in the pathogenesis of periodontal disease, but the precise mechanisms are still unknown. In order to determine local IgG and IgA production, we investigated the presence of human IgG and IgA subclass mRNA-bearing plasma cells within periodontal tissue by in situ hybridization using digoxigenin-labelled oligonucleotide probes in 24 gingival biopsy samples (pocket depth > or = 5 mm) which were obtained from eight patients with adult periodontitis. Furthermore, we examined IgG and IgA subclass proteins and digested IgA1 Fab portions in the gingival crevicular fluid (GCF), corresponding to the sites from which the tissues were taken, by ELISA. IgG and IgA subclass mRNA-expressing cells were detected in all serial formalin-fixed/paraffin-embedded gingival tissue sections sampled. Plasma cells showed strong cytoplasmic staining with a high contrast and a good retention of morphology with these probes. IgG1 mRNA-expressing cells were predominant (mean 63%) and IgG2 mRNA-expressing cells were present at around 23% of total IgG plasma cells, while IgG3 and IgG4 mRNA-expressing cells were present to a much lesser extent (3% and 10%, respectively). Similar proportions of IgG subclass proteins in GCF were detected, which were also consistent with 'normal' serum levels. In terms of IgA subclass, IgA1 mRNA-positive cells were predominant (mean 65.1%, P < 0.001). In contrast, IgA2 protein in the GCF samples were detected at higher concentrations than IgA1 (P < 0.001). The ratio of total IgG to IgA mRNA-positive plasma cells was approximately 7.5:1. There was a good correlation between the amounts of IgG subclass proteins in GCF and the number of IgG subclass mRNA-positive cells in the same sites, but not between IgA subclass proteins and the number of IgA subclass mRNA-positive cells. These data suggest that IgG and IgA subclass proteins can be locally produced in the periodontitis gingiva. In addition, as we detected IgA1 Fab fragments in GCF, this is further confirmation that secreted IgA1 protein in GCF may be digested by periodontal bacteria.

IgA1 protease activity, which allows bacteria to cleave human IgA1 in the hinge region, represents a striking example of convergent evolution of a specific property in bacteria. Although it has been known since 1979 that IgA1 protease is produced by the three leading causes of bacterial meningitis in addition to important urogenital pathogens and some members of the oropharyngeal flora, the exact role of this enzyme in bacterial pathogenesis is still incompletely understood owing to lack of a satisfactory animal model. Cleavage of IgA1 by these post-proline endopeptidases efficiently separates the monomeric antigen-binding fragments from the secondary effector functions of the IgA1 antibody molecule. Several in vivo and in vitro observations indicate that the enzymes are important for the ability of bacteria to colonize mucosal membranes in the presence of S-IgA antibodies. Furthermore, the extensive cleavage of IgA sometimes observed in vivo, suggests that IgA1 protease activity results in a local functional IgA deficiency that may facilitate colonization of other microorganisms and the penetration of potential allergens. It has been hypothesized that IgA1 protease activity of Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae, under special immunological circumstances, allows these bacteria to take advantage of specific IgA1 antibodies in a strategy to evade other immune factors of the human body. The decisive factor is the balance between IgA antibodies against surface antigens of the respective bacteria and their IgA1 protease. Recent studies have shown that serine-type IgA1 proteases of H. influenzae, meningococci, and gonococci belong to a family of proteins used by a diverse group of Gram-negative bacteria for colonization and invasion.

Immunoglobulin A1 (IgA1) proteases secreted by oral Prevotella and Capnocytophaga species specifically cleave IgA1 at the same peptide bond in the hinge region, leaving intact monomeric Fab and Fc fragments. Assuming that Prevotella- and Capnocytophaga-induced Fab fragments of IgA1 expose a specific immunogenic neoepitope at the cleavage site, we established an enzyme-linked immunosorbent assay to measure human serum antibodies to this neoepitope as indirect evidence of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases. The assay used a monoclonal antibody with specificity for the neoepitope, and the ability to block binding of the monoclonal antibody to the neoepitope was investigated. Absorption of sera with Prevotella melaninogenica-induced Fab fragments of IgA1 resulted in removal of antibodies blocking binding of the monoclonal antibody, whereas absorption with Fab fragments induced by bacterial IgA1 proteases of other cleavage specificities did not remove blocking antibodies. Consequently, we assume that the antibodies detected had been induced by a neoepitope an the Fab fragment of IgA1 exposed exclusively after cleavage with IgA1 proteases from Prevotella and Capnocytophaga, indicating in vivo activity of these IgA1 proteases. Evidence, though indirect, of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases was present in 42 of 92 sera examined and in a significantly higher proportion of sera from adults with periodontal disease compared with control individuals. No correlation with disease was observed for the juvenile periodontitis groups.

The elevation of the two genotypes of Prevotella intermedia to species rank as P. intermedia and Prevotella nigrescens has increased the need for reliable differentiation between the two taxa. In this study, 53 strains, including strains whose species affiliations were known as well as fresh dental plaque isolates, were subjected to a multilocus enzyme electrophoretic analysis, DNA analyses in which we used whole genomic DNA, rRNA sequences, and an oligonucleotide specific for the former P. intermedia genotype II (P. nigrescens) as probes, and a sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of soluble cellular proteins. All of these tests consistently separated the strains into the same two distinct groups corresponding to P. intermedia and P. nigrescens, confirming that the two species constitute two distinct populations of bacteria. Each of the tests used independently provided reliable identification to the species level. A previously reported heterogeneity in the pattern of human immunoglobulin A1 (IgA1) degradation was not confirmed. No differences between species were observed. All of the strains induced total degradation of IgA1 within 48 h, a property that may be a virulence factor in periodontal disease development. The enzymes responsible for IgA1 degradation were not inactivated by the physiological proteinase inhibitors alpha 2-macroglobulin and alpha 1-proteinase inhibitor.

Bacterial deglycosylation of immunoglobulin Al (IgA1), the dominant isotype of antibody in the oral cavity, probably provides both nutrition as well as protection to the oral bacterial community. Representative strains of oral gram-positive rods were tested for their ability to remove carbohydrates from IgA1. Detection of sialic acids was performed by means of high-performance liquid chromatography (HPLC) separation (Aminex HPX-87H) and ultraviolet light absorption at 190 nm, and neutral carbohydrates were measured by HPLC separation (Capcell Pak C-18 SG 120) and ultraviolet light absorption at 245 nm after derivatization. Four strains of Actinomyces naeslundii, two strains of Corynebacterium matruchotii and one of two strains of Actinomyces odontolyticus partially or totally removed sialic acid, while two strains of Propionibacterium propionicus and the other strain of A. odontolyticus did not. Complete correlation was observed between sialic acid removal, neuraminidase activity measured with fluorogenic substrate and with one exception, altered immunoelectrophoretic mobility of IgA1. Only limited removal of other carbohydrates was observed with poor correlation to exoglycosidase activities measured with chromogenic substrates. Desialylation increases the susceptibility of glycoproteins, including IgA1, to proteolysis. Therefore, the desialylation of IgA1 by oral gram-positive rods may facilitate the proteolytic activities of other oral bacteria, and the concerted action may positively influence the survival of the bacteria in the oral community.

The effect of periodontal healing after guided tissue regeneration (GTR) in association with local application of metronidazole gel was evaluated in this study. Twelve patients with one pair of vertical periodontal bone defects of comparable size and configuration participated in the study. In a matched paired design, the test defects were treated by GTR using expanded polytetrafluoroethylene (ePTFE) membranes in combination with local application of metronidazole gel. The controls were treated in the same way except for application of metronidazole. During the first month of membrane implantation, no statistically significant differences between test and control surfaces were noted with respect to inflammation of the marginal gingiva. Six weeks following insertion, the membranes were removed. Six months after removal of the membrane, the median gain in probing attachment level as a percentage of the initial defect depth was 92% for test defects and 50% for control defects (P = .001). No statistically significant differences were found between test and control sites regarding plaque, bleeding on probing, reduction in pocket depth, gain in bone height, or recession of the gingival margin. In conclusion, the results of the present study indicate that local application of metronidazole gel has a beneficial effect on healing of periodontal vertical defects treated by guided tissue regeneration.

The bacteriological colonization of healing periodontal defects was investigated after treatment with guided tissue regeneration using expanded polytetrafluoroethylene membranes together with local metronidazole gel (25%, 250 mg/g). Twelve patients, each with one pair of comparable defects, had the test defect treated with the membrane plus metronidazole gel and the control defect treated with the membrane alone. Thirty weeks after removal of the membrane, the median gain in probing attachment level as a percentage of the initial defect depth was 92% for the test defects and 50% for the control defects (P = 0.001). The median number of cultivable bacteria decreased from 1.2 x 10(6) at the presurgical examination to 3.0 x 10(5) at the one week examination in the test group (P = 0.02), whereas an increase was observed in the control group. Similarly, a lower median proportion of black-pigmented Gram-negative anaerobic rods was observed one week postsurgically in the test group (0.004%) compared to the control group (3.5%) (P = 0.02). Two weeks after membrane insertion, and at all following examinations, no microbiological differences between test and control group were observed. Consequently, the influence of the metronidazole gel on the treatment result appears to have been confined to the initial regeneration phase. Despite the good clinical results in the test group, all membranes from both test and control pockets were heavily colonized with bacteria at the time of removal. To ensure maximal periodontal regeneration with formation of bone, future research in this area should concentrate on reducing the microbial colonization of the wound area.

Recently published taxonomic studies of viridans streptococci have resulted in several changes in the nomenclature and definition of oral streptococcal species. With this background, the ecology of streptococci in the oropharyngeal cavities was reinvestigated. The results based on the examination of 1426 streptococcal isolates confirmed and extended earlier findings. Apart from mature supragingival plaque, which contained a mixture of all orally encountered streptococci, each site showed a characteristic streptococcal flora. Initial dental plaque formation is primarily associated with Streptococcus sanguis, Streptococcus mitis biovar 1 and Streptococcus oralis. Our investigation showed that S. sanguis and S. mitis biovar 1 were the most prominent streptococci, also on buccal mucosa. In contrast, S. oralis was almost exclusively found in initial dental plaque. Streptococcus gordonii, formerly part of S. sanguis, was found in small numbers on the oropharyngeal mucosa and in mature supragingival plaque. The dorsum of the tongue was dominated by S. mitis biovar 2 and Streptococcus salivarius, the latter of which was predominant also on the pharyngeal mucosa. Streptococcus anginosus was by far the most predominant streptococcus in subgingival plaque. Immunoglobulin A1 (IgA1) protease-producing streptococci were primarily isolated from initial dental plaque and from the buccal mucosa. This lends further support to the concept of IgA1 proteases being important for the ability of streptococci to evade the local immune defence during their initial colonization of certain oral surfaces.

The ecology and identity of IgA1 protease-producing streptococci in the human oral cavity were studied in an attempt to obtain insight into the significance of IgA1 proteases for host-parasite relationships. Contrary to previous observations, a detailed taxonomic analysis of oral streptococci revealed that all strains of S. sanguis and S. oralis ("S.mitior") produced IgA1 protease. In addition, IgA1 protease activity was observed in some isolates of S.mitis. Of the streptococci that initiate plaque formation on dental enamel 88% (median value) had IgA1 protease activity. Low proportions of the streptococci that colonize the tongue and oropharyngeal mucosae produced IgA1 protease, in contrast to 60% of streptococci isolated from buccal mucosa. The IgA1 proteases from S. sanguis I-III, S. sanguis IV, S. oralis, and S.mitis were distinct as revealed by studies using enzyme-neutralizing antibodies.

Bacteroides and Capnocytophaga species have been implicated as periodontal pathogens. Some of these species possess immunoglobulin A1 (IgA1) proteases that are capable of cleaving the human IgA1 molecule in the hinge region, leaving intact Fc alpha and Fab alpha fragments. The purpose of this study was to characterize this activity. In addition to IgA1 protease activity in already known species, IgA1 protease activity was a feature of Bacteroides buccalis, Bacteroides oralis, Bacteroides veroralis, Bacteroides capillus, and Bacteroides pentosaceus. Results of immunoelectrophoretic and sodium dodecyl sulfate-polyacrylamide gel electrophoretic analyses suggested that all species cleave the alpha-chain at the same peptide bond, i.e., the prolyl-seryl bond between residues 223 and 224 in the hinge region. The Bacteroides proteases could be classified as thiol proteases, which were at the same time dependent on metal ions, while the Capnocytophaga proteases were metallo enzymes. None of the proteases were inhibited by the physiologic proteases inhibitors alpha 2-macroglobulin and alpha 1-proteinase inhibitor. Investigations with enzyme-neutralizing antibodies raised in rabbits against protease preparations from the respective type strains revealed that, despite otherwise identical characteristics, the IgA1 protease of each Bacteroides species was antigenically distinct. Bacteroides buccae and the two later synonymous species B. capillus and B. pentosaceus produced identical proteases. In contrast, IgA1 proteases from Capnocytophaga ochracea and Capnocytophaga sputigena strains were apparently identical, while Capnocytophaga gingivalis had a protease that differed from those of the other Capnocytophaga species.