Bacteroides forsythus has emerged as a crucial periodontal pathogen with possible implications for systemic disease. The aim of this study was to isolate the S-layer from B. forsythus and examine its virulence potential as a part of efforts to characterize virulence factors of B. forsythus. The role of the S-layer in the haemagglutinating and adherent/invasive activities was evaluated. It was observed that the S-layer alone was able to mediate haemagglutination. In adherent and invasive studies, transmission electron microscopy clearly revealed that B. forsythus cells were able to attach to and invade KB cells, showing the formation of a microvillus-like extension around adherent and intracellular bacteria. The quantitative analysis showed that five different B. forsythus strains exhibited attachment (1?9-2?3 %) and invasion (0?4-0?7 %) capabilities. It was also observed through antibody inhibition assays that adherent/invasive activities of B. forsythus are mediated by the S-layer. Furthermore, an in vivo immunization study adopting a murine abscess model was used to prove that the S-layer is involved in the infectious process of abscess formation. While mice immunized with purified S-layer and B. forsythus whole cells did not develop any abscesses when challenged with viable B. forsythus cells, unimmunized mice developed abscesses. Collectively, the data obtained from these studies indicate that the S-layer of B. forsythus is a virulence factor.

The ultrastructure of three strains of water Leptospira was studied by negative staining, thin sectioning, and freeze-etching. The cells possessed a triple-layered sheath which covered two independent axial filaments, one inserted subterminally in each end of the cell. The protoplasmic cylinder was surrounded by a triple-layered cell wall and possessed ribosomes, lamellar structures, and a typical procaryotic nuclear region. The axial filament was comprised of several component structures. An axial fibril, with a diameter of 20 to 25 nm, consisted of a solid inner core (13 to 16 nm in diameter) surrounded by a coat. A terminal knob (40 to 70 nm in length) was connected to a series of disc insertion structures at the terminal end of the axial fibril. The axial fibril was surrounded by a helical outer coat (35 to 60 nm in diameter) which was composed of a continuously coiled fiber, 3 to 4 nm in diameter, embedded in an electron-dense material. A procedure for the purification of the axial fibrils was presented and their ultrastructural, physical, and chemical properties were determined. Similarities in ultrastructural, physical, and chemical properties were noted between the axial fibrils and bacterial flagella. A schematic model of the leptospiral axial filament is presented, and a mechanism is proposed for its function as a locomotor organelle.

35 ANUG patients were examined and compared clinically and demographically. Plaque removed from ulcerated sites in 20 patients was cultured using quantitative anaerobic procedures and examined by electron and darkfield microscopy. Patients were classified as having ANUG when presenting with ulceration and necrosis of interproximal papillae, pain and bleeding. The clinical symptoms of fetid odor, pseudomembrane formation, lymphadenopathy and elevated body temperature were present in 97%, 85%, 61% and 39% of the ANUG patients, respectively. 83% of the patients were smokers. The ANUG patients demonstrated a lower average age (24 years) than the general clinic population (32 years). There was a slightly higher % of male (54%) than female (46%) and the % of Caucasian (51%) and black (49%) ANUG patients were almost equal. Cultural studies revealed that gram-negative rods were the predominant cultivable micro-organisms present in the plaque, representing 78.2% of the total recoverable count. Of these, nearly half were strict anaerobes with Bacteroides gingivalis and Fusobacterium nucleatum accounting for 7.8% and 3.4%, respectively. Anaerobic and facultative gram-positive cocci (15.5%), gram-negative cocci (3.5%) and gram-positive rods (2.8%) were also isolated. Microscopic analysis of the morphologic composition of plaque revealed that rods (43%) constituted the greatest % of the total microorganisms observed followed by spirochetes (30%) and cocci (27%). 8 distinct types of spirochetal periplasmic flagellar arrangement were observed by electron microscopy, the "2-4-2" periplasmic flagellar arrangement being most numerous.

Segments of the ospA gene of Borrelia burgdorferi were amplified by the polymerase chain reaction (PCR). Oligonucleotide primers used in the reaction flank a 309-base-pair segment within the ospA gene. After 35 cycles of amplification, the product could be detected by agarose gel electrophoresis or dot hybridization with a 32P-labeled probe. This segment was amplified in all strains of B. burgdorferi tested, but it was not detected in other bacterial species. An additional primer pair which has a broad specificity for conserved 16S rRNA sequences that are present in eubacteria amplified a 215-base-pair fragment in all organisms tested. The sensitivity of PCR for the detection of B. burgdorferi in clinical samples was evaluated by seeding blood and urine specimens with B. burgdorferi and subjecting them to amplification. We were able to detect 10 organisms per ml of blood or urine, using PCR with dot hybridization detection. DNA extraction is not required for sample preparation. Blood and urine specimens were obtained from canines with clinical and serologic evidence of Lyme disease and subjected to PCR analysis. Of 17 clinical specimens from 15 animals, one blood specimen showed reactivity in the PCR.

The enzyme-linked immunosorbent assay (ELISA) was used for the detection of circulating antibodies in human sera to antigens of three serotypes of Treponema denticola. The antigens used in the assay were obtained by a sodium deoxycholate extraction procedure. Sera obtained from totally edentulous patients, patients with advanced periodontitis, as well as dental students who were in good to excellent periodontal health were tested for antibodies to these antigens by ELISA. Significantly elevated antibody levels were detected in iO/U patients with advanced periodontitis towards the antigens of all 3 serotypes. In contrast, 11/ 13 edentulous patients were negative for antibodies to serotypes W and TT and 9/13 were negative against serotype 11. In the mild gingival inflammation group, 11/14 individuals were negative against serotypes W and 11, and all were found to be negative against serotype TT. Adsorption studies carried out with 4 highly reactive sera indicated that the IgG antibodies detected by ELISA were specific for these antigens of T. denticola. The results of these studies are at variance with those of previous investigators who reported the total absence of detectable antibodies to T. denticola in sera from patients with advanced periodontal disease.