Cent Afr J Med, 1989 Mar, 35(3), 344 - 51 Vaginal flora of women admitted to hospital with signs of sepsis following normal delivery, cesarean section or abortion . The Puerperal Sepsis Study Group; Mason PR et al.; The microbial flora of the genital tract of 95 women who developed clinical signs of infection within 48 hr of vaginal delivery, Cesarean section delivery or abortion were compared with 111 women who delivered at the same hospital during the same time period but who showed no signs of sepsis . While there were no significant differences in the prevalence of most organisms in the lower genital tract of women with and without sepsis, there was evidence of a higher prevalence of gonococcal, chlamydial and anaerobic infection in the former . Gonococci were isolated from over 20 percent of untreated women with sepsis, more than three times the prevalence in controls . A third of the isolates were penicillinase-producing and another third showed in vitro resistance to penicillin . Chlamydial antigen was detected in 16-20 percent of women with sepsis following vaginal delivery or abortion, compared with 6 percent of controls . Neither gonococcal nor chlamydial infections were significantly associated with sepsis following Cesarean section delivery . Clue cells, indicative of G . vaginalis infection were noted in 20 percent of patients with sepsis compared with 7 percent of controls while amongst the other anaerobes only pigment producing Bacteroides were associated with sepsis . These findings suggest that antepartum investigations for clue cells, chlamydial antigen, gonococci and pigment producing anaerobes may identify patients most at risk from obstetric sepsis in Harare, and identify those for whom prophylactic administration of antibiotics may be of benefit. Res Vet Sci, 1989 Mar, 46(2), 147 - 52 Environmental influences on the progression of clinical and microbiological parameters of sheep periodontal disease; Frisken KW et al.; The responses of some clinical and microbiological parameters of periodontal disease (PD) in sheep were examined subsequent to transferring animals between PD-affected and PD-free farms . Previously healthy animals showed transient deterioration in some clinical, but not microbiological parameters, which suggests either that a different microbiota to the one studied may be more important in the initiation of the disease, or that sampling did not intercept periods of destructive disease activity in the early lesions . In sheep with established disease, those parameters indicative of periodontitis which included pocket depth and bleeding on probing as well as the proportions of black-pigmented Bacteroides species were not significantly altered by environmental changes . This observation suggests that once the disease is established on PD-affected farms, the hand, some clinical signs of the disease including lengthening and mobility of incisor teeth increased in sheep on the PD-affected farm relative to the PD-free farm . This suggests that the disease may have a complex aetiology. Diagn Microbiol Infect Dis, 1989 Mar-Apr, 12(2), 165 - 70 Comparison of the bactericidal activity of cefotaxime and desacetylcefotaxime alone and in combination against Bacteroides fragilis group organisms; Aldridge KE et al.; Through the use of time-kill kinetic studies, the bactericidal activity of cefotaxime (CTX) and desacetylcefotaxime (dCTX) alone and in combination against 18 strains of Bacteroides fragilis group was studied . Each isolate was tested at subinhibitory, inhibitory, and suprainhibitory concentrations of each drug as determined from the MIC values . Overall CTX was more bactericidal than dCTX at each of the three concentration levels tested . The combination of CTX and dCTX showed comparable bactericidal activity to CTX at the subinhibitory and inhibitory concentrations, even though each component was present at only one-half the concentration of CTX alone . At suprainhibitory concentrations, the combination of CTX/dCTX appeared synergistic since the combination with each component at a concentration of 1 x MIC was as bactericidal as CTX at a concentration of 4 x MIC . CTX and dCTX alone and in combination exhibited comparable bactericidal activity against test isolates with high (greater than or equal to 32 micrograms/ml) or low (less than or equal to 16 micrograms/ml) MICs . Thus, in vitro the combination of CTX and its naturally occurring metabolite dCTX interacts to produce an additive or synergistic effect against strains of B . fragilis group . Whether the in vitro testing of the combinations is more relevant to clinical outcome than testing CTX alone needs further study. Plasmid, 1989 Mar, 21(2), 151 - 4 Transferable 5-nitroimidazole resistance in the Bacteroides fragilis group; Breuil J et al.; We report the characterization of a strain of Bacteroides vulgatus, BV17, that exhibits a moderate resistance to 5-nitroimidazoles and carries plasmids of 4.5, 5, 7.7, and 56 kb . A genetic determinant involved in this resistance is carried by the 7.7 +/- 0.2-kb plasmid (pIP417) . This plasmid can be introduced and replicated in a sensitive strain of B . fragilis 638R by transformation or by conjugation . In the latter case, the transfer may involve mobilization by the 56-kb conjugative plasmid (pIP418) regularly found in transconjugants but not in transformants. Can J Microbiol, 1989 Mar, 35(3), 416 - 22 Development of the cellulolytic microflora in the rumen of lambs transferred into sterile isolators a few days after birth; Fonty G et al.; Seven lambs, separated from their dam 24 h after birth, were kept in a conventional environment until transferred to sterile isolators between 1 and 9 days of age: two on day 1 (IA and IB), two on day 4 (IVA and IVB), one on day 8 (VIIIA), and two on day 9 (IXA and IXB) . The lambs were reared in these isolators until 120 days of age . Lambs IA, IB, IXA, and IXB were free of cellulolytic bacteria when they were placed in the isolators . They were then inoculated with Bacteroides succinogenes S85 which became established in the four lambs . Until the age of 2 months, the population of this strain fluctuated and then stabilized at a high level (10(8)-10(9) cells/mL) . Cellulolytic bacteria were present in the rumen of lambs IVA, IVB, and VIIIA when they were transferred to the isolators . In IVA, and IVB, the cellulolytic population slowly increased with the animal age . In contrast, in VIIIA, the cellulolytic bacteria disappeared within a few days . Bacteroides succinogenes S85 inoculated thereafter became established rapidly and reached a level comparable to that observed in lambs IA and IB . The total number of viable rumen bacteria in the isolated lambs was similar to that observed in conventionally raised animals, but differences were observed in the selective enumeration of bacteria utilizing specific energy sources. Antimicrob Agents Chemother, 1989 Mar, 33(3), 394 - 5 Comparative in vitro antimicrobial susceptibilities of a special panel of 74 Bacteroides fragilis group isolates in Wilkins-Chalgren agar with and without sheep blood; Gill CJ et al.; Seventy-four cefoxitin-resistant Bacteroides fragilis group isolates were tested by the serial twofold agar dilution method for susceptibility to imipenem and other agents in medium with and without 5% sheep blood . Imipenem (MIC for 90% of strains tested, 1 microgram/ml) and metronidazole (MIC for 90% of strains tested, 2 micrograms/ml) were the two most active agents . The addition of 5% sheep blood to the medium had little or no effect on the activity of the antibiotics tested. FEMS Microbiol Lett, 1989 Mar, 49(1), 37 - 41 Degradation of intestinal glycoproteins by Bacteroides vulgatus; Ruseler-van Embden JG et al.; Bacteroides vulgatus, isolated from a patient with Crohn's disease, produced in gnotobiotic rats 7 constitutive enzymes that might be concerned with the degradation of intestinal glycoproteins . Furthermore Bacteroides vulgatus caused an almost complete loss of blood group antigenicity of the intestinal glycoproteins . Enzymes with the potency to release toxic compounds from hepatic conjugates and plant glycosides, beta-glucuronidase and beta-glucosidase, respectively, were only detectable in small amounts . These findings indicate that Bacteroides vulgatus, which accounts for 40% of the total flora of patients with Crohn's disease, may play a role in the pathogenesis of Crohn's disease, by increasing the break-down of the mucus layer and therefore damaging its protective function. Vet Microbiol, 1989 Mar, 19(3), 275 - 81 Bacteroides species from the oral cavity and oral-associated diseases of cats; Love DN et al.; One hundred and sixty-seven strains of Bacteroides were isolated from 71 subcutaneous fight-wound abscesses of cats, 21 cases of feline pyothorax, normal gingival margins from 10 cats and 6 cases of feline gingivitis . Bacteroides species constituted (as a proportion of all anaerobic isolates examined) 44.5% from subcutaneous abscesses, 33.7% from pyothoraxes, 37.5% from normal gingiva and 27.7% from diseased gingiva . Bacteroides tectum comprised 43.7% or 73 of 167 strains, followed by the black- or brown-pigmented asaccharolytic feline species of B . gingivalis, B . salivosus and Group B, comprising 32.3% or 54 of 167 strains . B . heparinolyticus (some 10% or 17 of 167 strains) was the next most common species described . The remainder consisted of two strains of B . fragilis and 21 unspeciated strains . Bacteroides tectum was frequently isolated from subcutaneous abscesses (43.7%) and pyothoraxes (46.6%), and it constituted some 33% of anaerobic isolated from normal gingiva . Bacteroides heparinolyticus was more commonly encountered in purulent lesions (abscesses and pyothoraxes) than in the oral cavity. Nippon Shishubyo Gakkai Kaishi, 1989 Mar, 31(1), 43 - 54 {Correlation between subgingival microflora and peptidase activity in periodontal pockets measured with synthetic substrates}; Miyazawa Y; The activity of peptidases in periodontal pockets of patients were examined by using four different synthetic enzyme substrates, and surveyed their correlation with the microbial populations of subgingival plaque and clinical symptoms . The substrates were (1) alpha-N-benzoyl-DL-arginine-beta-naphthylamide (BANA), (2) N-carbobenzoxy-glycylglycyl-arginine-4-methoxy-beta-naphthylamide, (3) alpha-N-benzoyl-L-arginylglycyl-L-phenylalanyl-L-proline-4-methoxy -beta-naphthylamide and (4) N-carbobenzoxy-prolyl-L-alanylglycyl-L-proline-4-methoxy-beta naphthylamide . Whereas substrates (1) and (2) were hydrolyzed more specifically by peptidases of mainly Bacteroides gingivalis and Treponema denticola, substrates (3) and (4) were susceptible to most of the strains of black-pigmented Bacteroides, Capnocytophaga and T . denticola . Correlation between the peptidase activity and the level of Spirochetes in the plaques were observed with the substrates (1), (2) and (3) . Substrate (3) had the strongest correlation also with the level of the black-pigmented Bacteroides and with the depth of the periodontal pockets, suggesting that it is a better substrate for enzymatic diagnosis than BANA which is currently used as an indicator of oral Spirochetes and the black-pigmented Bacteroides. Nippon Shishubyo Gakkai Kaishi, 1989 Mar, 31(1), 29 - 42 {Microbial flora of periodontitis and monitoring of the effect of initial preparation by immunofluorescence microscopy}; Seida K; The effect of initial preparation in adult periodontitis was evaluated by changes in clinical parameters and immunofluorescence microscopic counts of periodontal disease associated bacteria . Subgingival plaque samples were taken with sterilized paper points from 10 sites of 5 periodontally healthy persons and 44 sites of 23 adult periodontitis patients . Twenty-one diseased sites were periodically examined after plaque control and scaling, root planing . The direct immunofluorescence technique was used to detect Bacteroides gingivalis, Eikenella corrodens, Fusobacterium nucleatum and Treponema denticola, Bacteroides intermedius, Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Wolinella recta were counted by the indirect immunofluorescence technique . The proportions of B . gingivalis, B . forsythus, F . nucleatum, E . corrodens, T . denticola and W . recta in periodontitis lesions were significantly higher than those in healthy sites . The proportions of B . gingivalis and T . denticola were significantly related to GI, PlI, BI and PD, those of B . forsythus and W . recta to GI, PlI and BI, E . corrodens to GI and PlI, and F . nucleatum to BI . Reduced proportions of T . denticola were found in samples taken after establishment of proper plaque control . Subgingival scaling and root planing resulted in the reduction of proportions of B . gingivalis, E . corrodens, T . denticola and B . forsythus in the samples . The samples from positive bleeding sites contained higher proportions of B . gingivalis, T . denticola, B . forsythus and W . recta than did resolved sites . The present study shows that the immunofluorescence technique which detects B . gingivalis, T . denticola and B . forsythus is useful in monitoring the efficacy of initial preparation. Nippon Shishubyo Gakkai Kaishi, 1989 Mar, 31(1), 13 - 28 {Microbiological evaluation of initial preparation for adult periodontitis patients}; Nakagawa T; The microbial flora from 46 adult periodontitis lesions of 23 patients and 18 sites in 9 healthy persons were examined and levels of serum IgG antibody to gram negative periodontal disease-associated bacteria were measured by enzyme-linked immunosorbent assay . Plaque samples and serum samples were taken 40-50 days after initial preparation consisting of scaling and root planing . To evaluate the effects of the therapy on 10 patients with adult periodontitis, changes in clinical parameters were compared with alterations of the microbial flora and serum IgG antibody levels . Black-pigmented Bacteroides species, mainly Bacteroides gingivalis, were found to be predominant in periodontitis lesions . A significant relationship was found between the prevalence of B . gingivalis and elevated titers of serum IgG antibody against the microorganism . No relationships between Bacteroides intermedius, Actinobacillus actinomycetemcomitans and elevated titers of serum IgG antibody to them were detected . The fact that there was no marked reduction of serum IgG antibody to B . gingivalis after initial preparation suggests that a more extended, longitudinal study is required . Although brushing resulted in a significant reduction of the number of total cultivable organisms in samples from periodontal pockets, no significant proportional changes in gram-negative bacteria in the lesional flora were found . Initial preparation was not effective in eliminating gram-negative bacteria from deep periodontal pockets . However, the microbiological shifts, especially the reduction in the proportion and frequency of detection of B . gingivalis in periodontal pockets, was paralleled by significant improvement in the clinical parameters. J Antimicrob Chemother, 1989 Mar, 23(3), 383 - 8 Effect of Bacteroides fragilis on mortality induced by Escherichia coli in an experimental infection treated with cefotaxime, aztreonam or gentamicin; Soriano F et al.; The possibility that beta-lactamase-producing strains of Bacteroides fragilis can protect Escherichia coli from cefotaxime was studied in an in-vivo model of peritoneal infection in rats . The protective effect of cefotaxime, aztreonam and gentamicin in peritonitis induced by E . coli alone or combined with B . fragilis was evaluated by analysing mortality at 24 and 48 h after bacterial inoculation and treating the animals with two doses of each antibiotic . Comparisons, by drugs, at 24 and 48 h revealed that a statistically significant high mortality rate was obtained at 48 h when mixed infections were treated with cefotaxime, a drug very active in the infection caused by E . coli alone . Infections by mixed flora or E . coli alone treated with aztreonam or gentamicin did not show any significant difference in mortality rate analysed at 24 or 48 h . These in-vivo results confirm previous in-vitro studies and suggest that cefotaxime could be inactivated in mixed infections if a beta-lactamase-producing strain, such as B . fragilis, is involved in a clinical infection. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1989 Mar, 270(4), 492 - 502 Rapid diagnosis of intestinal Bacteroides species by means of the immunofluorescence; Hohne C et al.; Rapid methods serving the detection of intestinal Bacteroides species in clinical specimens are of great significance . For this purpose, an identification of the species is possible at present only by means of immunofluorescence (IF) . Using preparations of antigens of different intestinal Bacteroides species (B . fragilis, B . thetaiotaomicron, B . uniformis, B . ovatus, and B . distasonis) as well as immune sera prepared against them, the presented results have shown the possibility of specific identification of the species involved by means of direct and indirect IF, respectively . However, it could be demonstrated that the direct IF results in a more brilliant appearance of the bacteria than the indirect procedure . Convincing results could be obtained also in the investigation of clinical specimens by means of the direct IF using a pool of conjugated immunoglobulin fractions (sensitivity 88%, specificity 100%) . Therefore, the direct method to detect intestinal Bacteroides species by IF should be preferred. J Bacteriol, 1989 Mar, 171(3), 1294 - 302 Cloning and characterization of a Bacteroides conjugal tetracycline-erythromycin resistance element by using a shuttle cosmid vector; Shoemaker NB et al.; The Bacteroides conjugal tetracycline resistance (Tcr) elements appear not to be plasmids . In many cases, resistance to erythromycin (Emr) is cotransferred with Tcr . Using a newly constructed shuttle cosmid, pNJR1, we cloned 44 to 50 kilobase pairs of a conjugal Tcr Emr element on overlapping cosmid clones . Cosmid libraries were made in Escherichia coli with DNA from the original clinical Bacteroides thetaiotaomicron DOT strain containing Tcr Emr-DOT or from a Bacteroides uniformis Tcr Emr-DOT transconjugant strain . The cosmid clones were mobilized from E . coli into B . uniformis in groups of 10 to 20 per filter mating, with selection for Tcr or Emr transconjugants . The Tcr and Emr genes were cloned both separately and together on 30-kilobase-pair fragments . Several of the Tcr clones also contained transfer genes that permitted self-transfer of the cosmid from B . uniformis donors to E . coli or B . uniformis recipients . Neither the Tcr nor the Emr gene conferred resistance on E . coli, and the transfer-proficient clones did not self-transfer out of E . coli . Southern blot analysis was used to compare DNA from independently isolated Bacteroides strains carrying conjugal Tcr or Tcr Emr elements and their respective B . uniformis transconjugants . Results of these analyses indicate that there are large regions of homology, including regions outside the Tcr and Emr genes, but that the elements are not identical . Some Tcr clones contained a region which hybridized to chromosomal DNA from the wild-type B . uniformis recipient strain that did not carry the Tcr Emr-DOT element . This region of homology appeared not to be a junction fragment . It was not required in a Bacteroides recipient for successful transfer of the Tcr Emr element . Although we are not sure we have cloned a junction fragment between the Tcr Emr-DOT element and the B . uniformis chromosome, the preliminary function and restriction map appears to be linear. Nippon Shishubyo Gakkai Kaishi, 1989 Mar, 31(1), 156 - 65 {Effects of periodontopathic bacterial components on phagocytic activity of rat peritoneal macrophages . Examination using ELISA}; Ichimura K et al.; This study examined the phagocytic activity of rat peritoneal resident macrophages to determine the movement of macrophages in local inflammation in periodontal disease . We studied phagocytic activity by enzymelinked immunosorbent assay (ELISA) and used the peroxidase-anti-peroxidase soluble complex (PAP; soluble immune complex) as a marker in . We also determined the basic conditions of this examination and studied the effects of bacterial components and the supernatants of sonicated periodontopathic bacterias . We obtained the number of applied macrophages, the concentration of PAP to use and the incubation time . The phagocytic activity of macrophages was enhanced significantly by the bacterial components lipopolysaccharide (LPS) and muramyldipeptide (MDP) . Phagocytic activity was also enhanced by the addition of the supernatant of sonicated Bacteroides gingivalis at 40 micrograms/ml (concentration of protein) and significantly suppressed at 320 micrograms/ml . Moreover, activity was significantly enhanced by the supernatant of sonicated Capnocytophaga suputigena at 40 micrograms/ml and 160 micrograms/ml, and suppressed by the supernatant of Fusobacterium nucleatum at a low concentration of protein (5 micrograms/ml) . These results suggested that LPS of gram-negative bacteria's endotoxicity and MDP on pivotal structure of peptidoglycans, which are bacterial cell surface components, exerted an effect on phagocytic activity . It was further indicated that the phagocytic activity of macrophages varied with the effects of each periodontopathic bacteria. Nippon Shishubyo Gakkai Kaishi, 1989 Mar, 31(1), 1 - 12 {Effects of anti-Bacteroides gingivalis (Bg), and anti-Actinobacillus actinomycetemcomitans (Aa) antibodies on Bg and Aa}; Horino K et al.; The purpose of this study was to determine the effects of anti-Bacteroides gingivalis (Bg) and anti-Actinobacillus actinomycetemcomitans (Aa) antibodies on Bg and Aa respectively . Bg and Aa were resistant to complement-mediated bactericidal activity, and killing by PMN . However a significant reduction in CFU of Bg and Aa was found when these bacteria were incubated with the antibodies and not agitated . This would suggest that the antibody was capable of aggregating or phagocytizing, but not killing, these bacteria when complement and PMN were added . Moreover the antibodies had no effect on bacterial proliferation . Anti Aa antibody also had no effects on killing of Aa or PMN infiltration in experimentally established periodontal pockets in dogs. Oral Microbiol Immunol, 1989 Mar, 4(1), 6 - 11 Experimental infections by Bacteroides gingivalis in non-immunized and immunized rabbits; Dahlen G et al.; The interactions between Bacteroides gingivalis and systemic antibodies were studied in tissue cages implanted in the backs of New Zealand white rabbits . Infectivity was evaluated according to clinical signs and to leukocyte and bacterial counts in material aspirated from the tissue cages . Pre- and post-inoculation antibody levels to sonicated whole bacterial cells were determined by enzyme-linked immunosorbent and agar immunodiffusion assays . Rabbits immunized against B . gingivalis and then challenged with pure cultures of B . gingivalis revealed complete elimination or markedly lower postinoculation bacterial counts and considerably weaker tissue reactions than non-immunized animals . B . gingivalis co-inoculated with Actinobacillus actinomycetemcomitans caused significantly more severe infections than observed in monoinfected animals . The present results suggest that the immune system acting through systemic antibodies and/or cellular mechanisms may modulate the pathogenic potential of infecting periodontal pathogens. Oral Microbiol Immunol, 1989 Mar, 4(1), 24 - 9 Immunomodulatory effects of Bacteroides products on in vitro human lymphocyte functions; Shenker BJ et al.; Bacteroides spp . have been implicated in the pathogenesis of several diseases, including periodontal diseases . In this study sonic extracts of 6 Bacteroides spp . were examined for their abilities to alter human lymphocyte function . We found that soluble extracts from Bacteroides intermedius, Bacteroides endodontalis, Bacteroides asaccharolyticus, Bacteroides melaninogenicus, and to a lesser degree Bacteroides loescheii, caused dose-dependent inhibition of human lymphocyte responsiveness to both mitogens and antigens . Suppression involved altered DNA, RNA and protein synthesis as well as immunoglobulin production . In contrast, Bacteroides gingivalis did not suppress these responses; instead, it stimulated lymphocyte proliferation and enhanced immunoglobulin production . It has been proposed that impaired host defense may play a pivotal role in the pathogenesis of many infections . The data presented in this paper suggest that microbial mediated immunosuppression may conceivably alter the nature and consequences of host-parasite interactions in periodontal disease. Oral Microbiol Immunol, 1989 Mar, 4(1), 19 - 23 Studies on the virulence properties and metabolism of pleiotropic mutants of Porphyromonas gingivalis (Bacteroides gingivalis) W50; Shah HN et al.; Porphyromonas gingivalis (Bacteroides gingivalis) strain W50 and variants isolated from continuous culture designated W50/BP1 (black pigmented), W50/BR1 (brown pigmented) and W50/BE1 (beige or non-pigmented) were previously shown to lose virulence with the loss of pigmentation . Major properties which may affect the virulence and metabolism of P . gingivalis were compared amongst the 4 strains . The non-pigmented strain lost the ability to hemagglutinate sheep erythrocyte, had a reduced hydrophobicity and possessed lower levels of proteolytic activity . Defects in the electron transport system occurred at the level of cytochrome b but not menaquinone synthesis and resulted in an altered metabolic end product profile of the non-pigmented strain. Oral Microbiol Immunol, 1989 Mar, 4(1), 12 - 8 Further studies on the degradation of immunoglobulins by black-pigmented Bacteroides; Grenier D et al.; The ability of several species of black-pigmented Bacteroides to degrade immunoglobulins A and G was confirmed in this study . The cleavage products from IgG strongly stimulated the growth of bacteria degrading IgG . Growth of Bacteroides gingivalis on limiting media supplemented with IgG paralleled growth on complete medium . The degradation of IgG and IgA by black-pigmented Bacteroides appeared to occur in 2 stages . The molecules were broken into large fragments which were subsequently degraded into small peptides not visible on SDS-PAGE . B . gingivalis degraded IgG to peptides with Mrs of 33,000 and 11,000 whereas Bacteroides asaccharolyticus, Bacteroides intermedius and Bacteroides loescheii formed only the 33,000 Mr peptide . Electrophoresis in polyacrylamide gels containing covalently linked IgG, IgA and bovine serum albumin revealed that B . gingivalis elaborated 8 electrophoretically distinct proteolytic activities . The proteases protected the cell from reaction with anti-B . gingivalis antibody and were capable of hydrolyzing antibody bound to the bacterial cell surface. J Gen Microbiol, 1989 Mar, 135 ( Pt 3), 557 - 64 Production of cell-bound and vesicle-associated trypsin-like protease, alkaline phosphatase and N-acetyl-beta-glucosaminidase by Bacteroides gingivalis strain W50; Minhas T et al.; Bacteroides gingivalis strain W50 was grown in batch and continuous culture on complex medium with haemin . In batch culture, cell-bound levels of trypsin-like protease (EC 3.4.21.4), alkaline phosphatase (EC 3.1.3.1) and N-acetyl-beta-glucosaminidase (EC 3.2.1.30) increased during the exponential phase of growth . These enzyme activities were also detected in extracellular vesicles and in extracellular soluble forms in the supernatant fluid, but in lower amounts per unit biomass compared to cell-bound levels . In continuous culture, at high relative growth rates (0.7-0.9 murel), the highest proportions of enzyme activities were cell-bound . In contrast, at low relative growth rates (0.1-0.2 murel), highest enzyme levels were detected in the extracellular vesicle fraction . Levels of extracellular soluble enzymes were always low compared to cell-bound or extracellular vesicle levels, but were highest at low relative growth rates . All three enzymes appeared to be relatively stable in their soluble forms . Vesicle production appeared to be associated with actively growing cells but was influenced by growth rate . The results are consistent with the hypothesis that cell-bound 'periplasmic' enzymes are encapsulated into vesicles which are subsequently released by the cells . Therefore, levels of total extracellular enzyme (extracellular vesicle plus extracellular soluble) may depend on the rate of vesicle formation superimposed on the rates of production of 'periplasmic' enzymes in the cell. J Periodontal Res, 1989 Mar, 24(2), 96 - 105 Microbiota and crevicular fluid collagenase activity in the osseointegrated dental implant sulcus: a comparison of sites in edentulous and partially edentulous patients; Apse P et al.; The soft tissues adjacent to osseointegrated dental implants (OII) were investigated using clinical, biochemical and microbiological methods . Tooth and implant crevices were compared in 15 partially edentulous patients, examining 28 peri-implant and 19 periodontal sites, and in 6 edentulous patients, examining 13 implant sites . Sites were classified by standard periodontal indices; the crevicular fluid flow determined; crevicular fluid was collected for collagenase assays; and the subgingival bacterial flora was examined and cultured . Differences in clinical parameters were noted in that implants had significantly less keratinized gingiva and deeper probing depths . Crevicular fluid was present in the OII sulcus but the crevicular fluid flow did not differ from that observed from tooth sites either in the partially edentulous or edentulous patients . Tissue collagenase activity and collagenase inhibitor were detected in the implant crevicular fluid and, as in periodontal sites, a strong inverse relationship was found between the levels of active collagenase and collagenase inhibitor . Microbiology included darkfield microscopy, anaerobic culturing for total colony forming unit counts and identification of black pigmented Bacteroides (BPB) . Few differences were observed between implants and teeth in partially edentulous patients, indicating that crevices around teeth may act as reservoirs of bacteria which can colonize implant sites . A higher percentage of BPBs and wet spreaders (Capnocytophaga) was noted at partially edentulous implant sites when compared with edentulous implant sites, perhaps reflecting the lower numbers of periodontal pathogens present in edentulous mouths . Overall, the characteristics of implant sulci appear to be similar to periodontal sulci with respect to crevicular fluid flow and microflora. Infect Immun, 1989 Mar, 57(3), 745 - 53 A soluble Bacteroides by-product impairs phagocytic killing of Escherichia coli by neutrophils; Rotstein OD et al.; The effect of Bacteroides culture filtrate on killing of Escherichia coli by neutrophils was examined as a potential mechanism for E . coli-Bacteroides microbial synergy . A low-molecular-weight heat-stable factor present in the 22-h culture filtrate of Bacteroides fragilis 9032 impaired neutrophil killing function . To determine whether short-chain fatty acids present in the filtrate could account for the inhibition, the fatty acid content of the culture filtrate was determined and sterile medium supplemented with measured concentrations of fatty acids was tested for its effect on neutrophil function . Succinic and acetic acids were measured in high concentrations, while lactic, formic, and fumaric acids were present in lower concentrations . Reconstituted media mimicked the inhibitory effect of B . fragilis filtrate on neutrophil killing capacity . In further support of the hypothesis that short-chain fatty acids were responsible for the inhibition, the filtrates of other Bacteroides strains were found to be inhibitory only after bacterial growth had entered the stationary phase, a period during which fatty acid production is maximized . Further studies investigating the mechanism of impaired neutrophil killing showed that B . fragilis 9032 culture filtrate inhibited both phagocytosis of {3H}thymidine-labeled E . coli by neutrophils and the intrinsic microbicidal functions of the neutrophil . Impairment of neutrophil superoxide production was mediated via the ability of short-chain fatty acids present in B . fragilis filtrate to reduce neutrophil cytoplasmic pH . These studies suggest that Bacteroides strains capable of reaching stationary phase in vivo may contribute to the pathogenesis of mixed infections by direct inhibition of neutrophil function. J Periodontal Res, 1989 Mar, 24(2), 155 - 60 Bactericidal concentrations of chlorhexidine-digluconate, amine fluoride gel and stannous fluoride gel for subgingival bacteria tested in serum at short contact times; Oosterwaal PJ et al.; In vitro inhibitory and bactericidal concentrations in serum of chlorhexidine-digluconate, amine fluoride gel, stannous fluoride gel, stannous fluoride, metronidazole and amoxicillin were determined against Bacteroides gingivalis, Bacteroides intermedius, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans and Capnocytophaga sputigena . The minimal inhibitory concentration was assessed by the agar dilution technique . The killing curves and minimal bactericidal concentration of the antimicrobial agents in inactivated bovine serum were determined after 5, 10, 20 and 60 minutes contact time . The minimal inhibitory concentration varied amongst the tested bacteria . A concentration of 128 micrograms/ml chlorhexidine digluconate, 20 mg/ml amine fluoride gel, 1 mg/ml stannous fluoride, 128 micrograms/ml metronidazole and 4 micrograms/ml amoxicillin inhibited the growth of the tested species . The minimal bactericidal concentration in serum for B . gingivalis, B . intermedius, F . nucleatum, A . actinomycetemcomitans and C . sputigena after 10 min contact time was 5 mg/ml for chlorhexidine digluconate and 100 mg/ml for amine fluoride gel . A concentration of 200 mg/ml stannous fluoride gel in serum was bactericidal for the tested species after 10 min contact time, with exception of F . nucleatum. J Periodontal Res, 1989 Mar, 24(2), 113 - 20 Prevalence of Bacteroides forsythus and Bacteroides gingivalis in subgingival plaque of prosthodontically treated patients on short recall; Gmur R et al.; The prevalence of Bacteroides forsythus and Bacteroides gingivalis in subgingival plaque of patients on short recall was analyzed in relation to the probing depth of the test sites . The subjects had excellent oral hygiene and therefore were unlikely to suffer from active periodontal destruction . Sixty-four subgingival plaque samples, taken from gingival or periodontal pockets with probing depths ranging from 1 to 8 mm, were quantitatively assessed for the presence of the two species using species-specific monoclonal antibodies in conjunction with a very sensitive indirect immunofluorescence technique . Both organisms were encountered in probes from sites as shallow as 2 mm, but the percentage of positive samples clearly rose in relation to the probing depth of the test sites . Overall, B . forsythus was found to colonize lesions earlier, that is at smaller probing depths, than B . gingivalis . Interestingly, whenever a sample was found to be positive for B . gingivalis it was also positive for B . forsythus . The numbers of B . forsythus and B . gingivalis and the total bacterial cell number found in the pockets were significantly correlated to the probing depth . However, with advancing probing depth the increase of the total cell numbers of the two Bacteroides species was considerably more pronounced than the increase of the total subgingival plaque cell number . The recall interval neither affected the frequency of sites positive for B . forsythus or B . gingivalis nor influenced significantly the proportions of the two species in subgingival plaque.(ABSTRACT TRUNCATED AT 250 WORDS) Clin Exp Immunol, 1989 Feb, 75(2), 245 - 51 Limit dilution analysis of peripheral blood T lymphocytes specific to periodontopathic bacteria; Mahanonda R et al.; Limit dilution analysis (LDA) was used to determine the presence and frequency of periodontopathic-bacteria-specific T cells in the peripheral blood of patients with chronic inflammatory periodontal disease . Twelve adult periodontitis (AP), 13 marginal gingivitis (MG) and 12 healthy control subjects took part in the study . Bacteroides gingivalis and Actinomyces viscosus were used as test organisms, while tetanus toxoid was used as the control antigen . The median PTL-p frequencies to B . gingivalis were 46.33 x 10(-6), 45.33 x 10(-6) and 58.83 x 10(-6) in the control, gingivitis and AP groups respectively, while the median PTL-p frequencies to A . viscosus were 13.8 x 10(-6), 17.33 x 10(-6) and 11.5 x 10(-6), again in the control, gingivitis and AP groups . There were no statistically significant differences between the groups . All subjects displayed 'single-hit' kinetics with the control tetanus toxoid antigen and, with three exceptions, 'single-hit' kinetics was also found with the two test organisms . One control subject displayed a 'saw-tooth' curve with A . viscosus and a 'suppressor' curve with B . gingivalis, while two MG subjects had a 'saw-tooth' curve with B . gingivalis . These complex curves suggest that, in some subjects, more than one limiting cell type may exist in the cultures . Nevertheless, the results of the present study illustrate that lymphocytes specific to periodontopathic bacteria exist in the peripheral blood of both diseased and non-diseased subjects. Can J Microbiol, 1989 Feb, 35(2), 313 - 7 Effects of methanol on the growth of gastrointestinal anaerobes; Caldwell DR; The effects of methanol on the growth of representative, predominant, anaerobic gut bacteria were studied . Growth yields and rates were determined in a base medium to which methanol was added to produce media with methanol concentrations varying, in twofold steps, over a concentration range of 0.01 to 25%, by volume . The growth of many of the organisms was completely inhibited by a methanol concentration equal to, or less than, 6.2% . Isolates representing cellulolytic species were completely inhibited at a methanol concentration of 3.1%, and inhibitory effects on the yield of some cellulolytic isolates were found at a methanol concentration as small as 0.01% . Although most of the organisms studied were inhibited at relatively small methanol concentrations, isolates of Selenomonas ruminantium, Bacteroides ovatus, and Fusobacterium necrophorum were relatively methanol resistant . A methanol concentration of 12.5% was required to completely inhibit S . ruminantium . Substantial growth of B . ovatus was obtained in media containing 12.5% methanol, and for F . necrophorum, substantial growth occurred in media containing 25% methanol . The yields of F . necrophorum strain B85 and S . ruminantium strain PC18 were enhanced by relatively small methanol concentrations and reduced with further methanol concentration increase Anaerobic, nonsporing gut bacteria exhibit a diversity of responses to methanol. Antimicrob Agents Chemother, 1989 Feb, 33(2), 242 - 4 Macrolide accumulation by Bacteroides fragilis ATCC 25285; Muto Y et al.; The accumulation of macrolide antibiotics in Bacteroides fragilis ATCC 25285 was increased in the order erythromycin, josamycin, and rokitamycin, depending on hydrophobicity . The half-times of efflux were also prolonged in the same order . Furthermore, MICs of the antibiotics were correlated with the extent of hydrophobicity . These findings suggest that the macrolide antibiotics are accumulated in B . fragilis by means of their hydrophobic properties, and the efficient accumulation of the drugs may explain the susceptibility of this gram-negative bacterium to macrolides. Zh Mikrobiol Epidemiol Immunobiol, 1989 Feb, (2), 20 - 4 {A study of the features of E . coli bacteroid infection in experiments on mice}; Komarovskaia TP et al.; Experiments on the intraperitoneal infection of F1 (CBA X C57BL/6) mice with bacteroids of the group Fragilis in combination with E . coli have revealed the presence of a synergic effect leading to more pronounced (greater frequency of abscess formation) and even qualitatively new (necrotic lesions of the liver) pathological changes in comparison with those observed in monoinfections . Bacteroides fragilis strain 1/1087 gamma has been shown to have greater virulence than that of Bacteroides vulgaris strain 18; their virulence was determined from the capacity of microorganisms for survival in the abdominal cavity of mice, as well as from their capacity for inducing the formation of necrotic lesions and abscesses. J Periodontol, 1989 Feb, 60(2), 96 - 103 Antibodies to Bacteroides gingivalis in patients with treated and untreated periodontal disease; Murray PA et al.; It is proposed that the development of periodontal disease is associated with rising levels of serum and gingival crevice fluid (GCF) IgG antibodies to specific organisms, while treatment of periodontal disease is associated with a decline in specific IgG antibodies . This study examined the immune response to Bacteroides gingivalis, a suspected periodontal pathogen, in serum and GCF of patients with adult periodontitis . Three groups of subjects were studied: (1) patients with untreated adult periodontitis, (2) patients with treated adult periodontitis, and (3) patients with gingivitis (controls) . An enzyme-linked immunosorbent assay was employed using whole formalinized B . gingivalis (ATCC 33277) as antigen . Results showed that the untreated adult periodontitis patients had a humoral immune response to B . gingivalis, producing significantly higher serum levels of IgG antibody to that organism than did patients with treated adult periodontitis (p less than or equal to 0.01) or gingivitis (p less than or equal to 0.005) . The untreated patients also demonstrated a local immune response to B . gingivalis in that their GCF levels of IgG antibody to that organism were also significantly higher than levels in treated adult periodontitis patients (p less than or equal to 0.005) and gingivitis patients (p less than or equal to 0.001) . These results are consistent with reports by other investigators . However, ratios of GCF antibody to serum antibody in the untreated adult periodontitis group were not significantly higher than ratios in the other two groups.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1989 Feb, 57(2), 338 - 43 Induction of macrophage procoagulant activity by Bacteroides fragilis; Rosenthal GA et al.; Fibrin deposition in the peritoneal cavity during acute peritonitis appears to predispose the host to abscess formation by providing an environment for bacterial proliferation protected from host defenses . The purpose of the present study was to determine whether the potent abscess-inducing anaerobe Bacteroides fragilis could promote fibrin deposition by inducing mononuclear cells to express procoagulant activity (PCA) . B . fragilis stimulated PCA in a dose-dependent fashion, achieving a maximum at 10(7) CFU/ml . Heat-killed B . fragilis induced comparable levels of PCA, while a nonspecific phagocytic stimulus, latex beads, was not stimulatory . B . fragilis was capable of inducing PCA even when phagocytosis was blocked by preexposure of cells to latex beads . The results suggested that phagocytosis was neither necessary nor sufficient for the generation of PCA . Cell separation studies showed that PCA was solely produced by macrophages and that lymphocytes did not augment its production . These studies suggest one potential mechanism by which B . fragilis might initiate abscess formation. Surg Gynecol Obstet, 1989 Feb, 168(2), 112 - 4 No effect of topical ampicillin prophylaxis in elective operations of the colon or rectum; Raahave D et al.; Whether or not topical application of ampicillin is necessary in patients undergoing elective colorectal operations was investigated . After mechanical preparation, 193 patients received 2 grams of cefotaxime administered intravenously from the start of the operation; patients received two more doses within the next 12 hours . In addition, patients were randomized to receive or not receive prophylaxis against infection of 2 grams of ampicillin in the site of the incision at closure . Twenty-three patients did not complete the study . Wound infection occurred in five of 81 patients who had topical application of ampicillin compared with six of 89 patients who did not receive prophylaxis; the difference was not significant . There were no significant differences in rates of wound dehiscence, intra-abdominal abscess or anastomotic leakage . Escherichia coli and Bacteroides fragilis were the predominant microorganisms isolated . Thus, topical application of ampicillin did not lower the wound infection rate when there was a preoperative antibiotic administered intravenously. Endod Dent Traumatol, 1989 Feb, 5(1), 1 - 10 Bacteroides spp . in dental root canal infections; Haapasalo M; A summary of a series of bacteriological studies of endodontic infections is presented in this article . The bacteriology of 62 root canal infections was studied with special attention focused on the occurrence, role and taxonomy of Bacteroides spp . All infections except one were mixed infections dominated usually by anaerobic bacteria . Four to 6 different species were present in most canals . Species of the genus Bacteroides were found more frequently than species of any other genus . Seventy-eight Bacteroides strains were isolated from 45 canals . B . buccae, B . intermedius, B . denticola, B . oris, B . oralis, and B . gingivalis were the most common Bacteroides spp . At the beginning of the treatment 35 of 62 teeth caused acute symptoms . The results indicated that symptoms were a result of the synergistic action of the mixed anaerobic flora . The presence of B . gingivalis, B . endodontalis, and B . buccae was more often related to acute cases than other Bacteroides spp . Black-pigmented Bacteroides and a new Bacteroides-like organism, Mitsuokella dentalis, seemed to increase the probability that acute symptoms would persist one week after the beginning of the treatment . However, the treatment result assessed after 4 weeks and after 1 year was not affected by the composition of the mixed anaerobic flora . Calcium hydroxide was the only canal disinfectant used . Its efficacy was proved by a bacteriological sample at the second appointment in 10 cases . All teeth were asymptomatic at the third appointment . The susceptibility of the isolated Bacteroides strains to penicillin G was also studied . Only 2 B . buccae strains and 2 B . denticola strains were resistant at a concentration of 2.4 micrograms/ml . The patients were randomly divided into 3 groups which received 1) no antibiotics, 2) penicillin V (650 mg x 3) for 7 days, or 3) for 12 weeks . There was no difference between the 3 groups in the healing of the periapical lesion after one year . All patients attended the 1-year control . Fifty cases showed complete healing, partial healing was obtained in 11 cases and in 1 case no healing was observed. Infect Immun, 1989 Feb, 57(2), 566 - 73 Immunochemical identification and preliminary characterization of a nonfimbrial hemagglutinating adhesin of Bacteroides gingivalis; Mouton C et al.; A cell-bound hemagglutinating adhesin (HA-Ag2) of Bacteroides gingivalis was identified by crossed immunoaffinity electrophoresis as one of the common antigens of the species . A polyclonal antiserum with a restricted specificity for HA-Ag2 was produced by immunizing with the relevant immunoprecipitate excised from crossed-immunoelectrophoresis gels . The immunoglobulin G fraction of this monospecific antiserum inhibited hemagglutination . The antiserum was used against a cell surface extract of B . gingivalis in immunoblotting experiments, and we detected two antigens with apparent molecular masses of 33 and 38 kilodaltons in B . gingivalis ATCC 33277 and W83 . Monoclonal antibody, C1.17, produced in another laboratory against B . gingivalis 381 and characterized as showing reactivity with a hemagglutinin of this strain (Y . Naito, K . Okuda, T . Kato, and I . Takazoe, Infect . Immun . 50:231-235, 1985), was also used to produce immunoblots of extracts of strains ATCC 33277 and W83 . The apparent molecular masses of the major polypeptides recognized by monoclonal C1.17 in the immunoblots were the same as those detected by the polyclonal monospecific antiserum, i.e., 33 and 38 kilodaltons . Significantly, none of the polypeptides identified in this study corresponded to the polypeptide appearing in the 41- to 43-kilodalton region and identified by Yoshimura and co-workers (F . Yoshimura, K . Takahashi, N . Yoshinobu, and T . Suzuki, J . Bacteriol . 160:949-957, 1984) as the fimbrial protein characteristic of the species . Enzyme-linked immunosorbent assay inhibition experiments with the monospecific antiserum indicated that the cell surface extracts from strains ATCC 33277 and W83 were strong inhibitors, whereas the fimbria-enriched preparations from both strains failed to inhibit binding of antibodies to the cell surface antigens . As a whole, our study indicates that a nonfimbrial surface protein complex demonstrating erythrocyte-binding capacity, HA-Ag2, is common to three strains of B . gingivalis and is composed of at least two associated polypeptides with apparent molecular masses of 33 and 38 kilodaltons which share at least one antigenic determinant. Immunol Cell Biol, 1989 Feb, 67 ( Pt 1), 71 - 8 Monoclonal antibodies defining immunogenic regions of pili from Bacteroides nodosus strains 198 (A1), 265 (H1) and 336 (F1); Young D et al.; A total of 17 monoclonal antibodies (MoAb) were used to analyse the antigenic structure of pilus protein from three serogroups of Bacteroides nodosus . The four MoAb which agglutinated pili were serogroup (and subgroup) specific, and the agglutinating epitope was present on the pili monomer and dependent on the intra-chain disulfide bond . Non-agglutinating MoAb identified two further non-linear and serogroup-restricted epitopes on strain 198 (A1) pili and two linear epitopes on 336 (F1) and 265 (H1) pili . Three MoAb cross-reacted with pili from six of the eight major serogroups and recognized an epitope in the N-terminal region of the molecule . This panel of MoAb has therefore identified at least four epitopes on pilus protein and will facilitate serotopic analyses of the immunogenicity of each epitope in sheep during vaccination against footrot. J Biol Chem, 1989 Jan 15, 264(2), 872 - 83 Structures of neutral O-linked polylactosaminoglycans on human skim milk mucins . A novel type of linearly extended poly-N-acetyllactosamine backbones with Gal beta(1-4)GlcNAc beta(1-6) repeating units; Hanisch FG et al.; O-Linked oligosaccharides were isolated from human skim milk mucins and from mucin-derived glycopeptides by reductive beta-elimination . The released alditols were fractionated by DEAE-Sephadex chromatography and purified by high performance liquid chromatography on primary amine bonded phase . The structures of the major neutral oligosaccharide alditols could be established by fast atom bombardment and electron impact mass spectrometry, combined with methylation analysis, 500-MHz 1H nuclear magnetic resonance spectroscopy, and endo-beta-galactosidase (from Bacteroides fragilis, EC 3.2.1.103) digestion (where n = 0-3): (formula; see text) Major O-glycosidically linked oligosaccharides on skim milk mucins are of the Gal beta(1-3){GlcNAc beta(1-6)} GalNAc core type 2 and exhibit linearly extended backbone chains of the poly N-acetyllactosamine type comprizing up to at least four repeating units, which are linked by the hitherto unknown sequence GlcNAc-beta(1-6) Gal rather than GlcNAc beta(1-3)Gal . A considerable portion of neutral alditols is represented by branched isomers of the linear species, which are distinguished by their content of 3,6-disubstituted galactose and their partial resistance to endo-beta-galactosidase digestion. J Clin Periodontol, 1989 Jan, 16(1), 38 - 45 Treatment of recurrent periodontal disease by root planing and Ornidazole (Tiberal) . Clinical and microbiological findings; Mombelli A et al.; The purpose of this study was to evaluate the use of Ornidazole as an adjunct to root planing in the therapy of patients suffering from recurrent periodontal disease . In 10 individuals who had previously been treated with scaling, root planing and periodontal surgery and who had followed a regular maintenance program including recall visits every 3-5 months for 1-7 years, 2 sites with recurrent periodontitis and 1 shallow site were selected . Reinfected sites had a record of losing clinical attachment of more than 3 mm since the completion of initial therapy, were bleeding upon probing and had a mean pocket probing depth of 7.85 +/- 1.31 mm . They had been reinstrumented several times by a registered dental hygienist, when clinical signs of recurrence of disease had appeared and the root surfaces were judged to be smooth and free of deposits . Clinical parameters were recorded and microbial samples were collected twice prior to retreatment . Then, 500 mg Ornidazole, to be taken twice a day for 10 days, was administered, and the whole dentition was thoroughly scaled and root planed . At day 10 as well as 2, 5, 8 and 11 months thereafter, samples were again obtained . At baseline, reinfected sites showed over 20% spirochetes, over 20% motile rods and over 9% fusiform organisms in darkfield preparations of subgingival plaque samples . Culturally, over 1/10 of organisms were identified as black pigmenting Bacteroides and in 18% of all baseline samples collected, B . gingivalis was found.(ABSTRACT TRUNCATED AT 250 WORDS) J Med Microbiol, 1989 Jan, 28(1), 5 - 8 Cytotoxic activity of crude extracts of Bacteroides gingivalis; Eke PI et al.; The cytotoxic activities of culture supernates, crude cell extracts and cell-wall materials of Bacteroides gingivalis were investigated in vitro . Each component was cytotoxic to Vero cells and, to a lesser extent, Wi 38 cells . The cytotoxic agents had similar effects on the cell lines to butyric acid, propionic acid and a partially-purified trypsin-like protein extracted from a clinical isolate of B . gingivalis; the effects were eliminated by heat . Cytotoxic materials obtained from young cultures were more susceptible to heat than those from older cultures . The heart-labile substance inside and outside the bacterial cell in young cultures of B . gingivalis may contribute to its overall cytotoxic activity. Infection, 1989, 17 Suppl 1, S11 - 3 {Susceptibility of clinically important Bacteroides species against enoxacin-metronidazole and enoxacin-clindamycin combinations}; Heizmann WR et al.; To assess the activity of enoxacin, clindamycin and metronidazole, MICs of clinical isolates of saccharolytic intestinal Bacteroides spp . were determined, using the agar dilution method according to NCCLS guidelines . Checkerboard titrations of enoxacin-metronidazole and enoxacin-clindamycin were done on Wilkins-Chalgren agar; inoculation, incubation and reading of plates were as for determination of MICs . Metronidazole MIC 90s for Bacteroides fragilis (23 strains) and Bacteroides thetaiotaomicron (23 strains) were 0.5 mg/l, clindamycin MIC 90 for B . fragilis was 1 mg/l, and for B . thetaiotaomicron 8 mg/l, whereas enoxacin MIC 90 values were 16 mg/l and 64 mg/l, respectively . The evaluation of the inhibitory effects of the combination enoxacin-metronidazole for B . fragilis showed additional effects in eleven strains, indifference in seven strains and antagonism in one . The figures for B . thetaiotaomicron showed addition in five strains, indifference in 17 strains, antagonism in one . For B . fragilis the combination enoxacin-clindamycin showed addition in ten strains, indifference in six, and antagonism in one; for B . thetaiotaomicron synergism in one strain, addition in four strains, indifference in 17 strains . In conclusion, the absence of antagonism and the overall preponderance of additional and indifferent effects warrant the use of enoxacin in combination with metronidazole or clindamycin in clinical trials of treatment of anaerobic-aerobic mixed infections. Free Radic Res Commun, 1989, 6(6), 359 - 67 Free radical generating mechanisms in the colon: their role in the induction and promotion of colorectal cancer? Blakeborough MH, Owen RW, Bilton RF. A hypothesis is presented to account for the dietary induction and promotion of colorectal cancer . The principal agents are the secondary bile acids, lithocholic and deoxycholic acids, the vitamin K group and ferrous iron complexes . These metabolites may interact to subvert the normal free radical generating mechanisms involved in mucosal defence . Diets high in fat and red meat and low in fibre support a Bacteroides-dominated colonic microflora, which both synthesis and utilises vitamin K2 isoprenalogues or menaquinones as enzyme co-factors . Iron(II) complexes such as haemin from the breakdown of dietary haemoglobin and myoglobin also serve as growth factors for these bacteria and provide a rich source of haem-iron for intestinal uptake . Biliary secretion is stimulated by dietary fat and bile acids are essential for the intestinal uptake of vitamin K and possibly of iron complexes such as haemin . In the mature colonocyte, vitamin K and haemin may initiate redox cycling reactions which liberate superoxide (O2-.) . Bile acids can activate the membrane bound phospholipase to liberate arachidonate and diacylglycerol . This leads in turn to the production of more O2- . which can enter the microcirculation and acts as a potent chemoattractant for the neutrophils that line the lamina propria . The released diacylglycerol can activate protein kinase C in the neutrophil membrane to switch on the respiratory burst oxidase system generating yet more O2- . and may stimulate the proliferation of transformed stem cells by a similar protein kinase C mediated mechanism.(ABSTRACT TRUNCATED AT 250 WORDS) Int J Biochem, 1989, 21(6), 661 - 6 Detection of 2-keto-3-deoxyoctonate in endotoxins isolated from six reference strains of the Bacteroides fragilis group; Beckmann I et al.; 1 . Endotoxins isolated from six serotype specific reference strains of the Bacteroides fragilis group were dephosphorylated by treatment with aqueous 50% hydrofluoric acid . 2 . Mild acidic hydrolysis of the dephosphorylated endotoxins released 2-keto-3-deoxyaldonic acid, the presence of which was demonstrated by the colorimetric thiobarbituric acid assay (TBA) . 3 . Thin layer chromatography of the dephosphorylated lipopolysaccharide of B . fragilis IPL E 323 (serotype E2), after acidic hydrolysis, revealed a TBA-positive substance with the same Rf-value as authentical 2-keto-3-deoxyoctolusonic acid (KDO) . 4 . Quantification of 2-keto-3-deoxyoctonate-in the lipopolysaccharide of B . fragilis IPL E 323 by means of the TBA resulted in a KDO content of 15 nM mg-1 lipopolysaccharide. Life Sci, 1989, 44(1), 35 - 40 Phenytoin induces interleukin-1 production in vitro; Modeer T et al.; Human adherent mononuclear cells and subcloned cell lines established from the human histiocytic cell line U-937 were cultured with phenytoin (PHT) and/or lipopolysaccharide (LPS) purified from Bacteroides fragilis . After the cultivation period, the cell-free supernatants were tested for interleukin-1 (Il-1) activity . The results showed that PHT induces Il-1 activity and potentiates LPS-induced Il-1 production . In the monocytic cell line U-937, the induced Il-1 production was found to be clonally distributed indicating that the response to PHT may be exerted by a subpopulation of monocytes . The PHT-induced Il-1 activity may be of importance in the development of gingival overgrowth . The induced Il-1 could also contribute to other known side effects such as dermatologic reactions accompanied by transient fever seen in patients medicating the drug. Chemotherapy, 1989, 35(3), 153 - 9 Influence of environmental factors on the in vitro efficacy of ciprofloxacin against anaerobic bacteria; Thadepalli H et al.; The effects of alterations in pH, size of inoculum, addition of human serum, and repeated exposure to sub-minimum inhibitory concentration of ciprofloxacin against anaerobic bacteria were assessed . Increase in the size of the inoculum and human serum had no effect on the minimum inhibitory concentration, whereas decrease in pH and exposure to sub-MIC decreased the activity of ciprofloxacin against Bacteroides fragilis . We tested for synergism of ciprofloxacin with cefoxitin, clindamycin and metronidazole against B . fragilis. J Antimicrob Chemother, 1989 Jan, 23(1), 99 - 106 Pharmacokinetics and pharmacodynamics of total and unbound cefoxitin and cefotetan in healthy volunteers; Carver PL et al.; Cefotetan is a semisynthetic cephalosporin with a broad spectrum of Gram-negative and anaerobic activity . We compared the pharmacokinetics and serum inhibitory activity of 2 g doses of cefotetan and cefoxitin in six healthy volunteers . The half-life of cefotetan (176 min) was significantly longer than that of cefoxitin (49 min) . Despite higher protein binding (85% versus 50%), free cefotetan serum concentrations 12 h post dose (1.6 mg/l) were higher than free cefoxitin serum concentrations 6 h post dose (0.32 mg/l) . Total and free serum inhibitory titres for Escherichia coli were greater than 1:8 for 12 h post dose for cefotetan, but only 1 h post dose for cefoxitin . Against Bacteroides fragilis, neither cefoxitin nor cefotetan exhibited free serum inhibitory titres greater than 1:8 at the end of their respective dosing intervals . However, cefotetan titres were equivalent or superior to those of cefoxitin over the entire 6 or 12 h dosing interval . Despite relatively high protein binding, cefotetan demonstrates comparable or superior activity 12 h after dosing to cefoxitin 6 h after dosing . More studies comparing the clinical outcome of cefoxitin and cefotetan are needed but our results support the clinical recommendations that cefoxitin should be given every 6 h, and cefotetan every 12 h. Immunol Invest, 1989 Jan-May, 18(1-4), 225 - 37 Effects of immunization with B . macacae on induced periodontitis--preliminary findings; Nisengard R et al.; This preliminary study examined the effects of immunization with Bacteroides macacae, the monkey equivalent of the human species of B . gingivalis on ligature-induced periodontitis . During a 12 week immunization period, 8 out of the 12 Macacae fasicularis monkeys were immunized weekly with B . macacae washed cells and 4 were sham-immunized with saline . At the same time, all were scaled and pumiced weekly to establish gingival health . Following this period, the mandibular first molars were ligated in 8 out of the 12 monkeys to induce periodontitis . The immunized, ligated experimental group, the ligated, sham-immunized control group, and the immunized, non-ligated control group were then followed for a 6 months ligation period while plaque was allowed to accumulate . Gingival indices, attachment levels, pocket depths, plaque indices, radiographs, serum and crevicular fluid antibodies and subgingival bacteria were assessed . Immunization led to elevated antibody levels to B . macacae while ligation increased plaque, gingival inflammation, and bone loss . Following the 6 month ligation period, B . macacae comprised 1.7% of the cultivable flora in the immunized, non-ligated monkeys, 2.1% in the immunized, ligated monkeys, and 5.6% in the sham-immunized, ligated monkeys . Similar differences between the immunized, ligated and and the sham-immunized, ligated groups were not seen for B . intermedius, nor B . melaninogenicus . These results suggest a heightened humoral response to B . gingivalis reduces subgingival re-colonization by this organism and modulates the course of ligature-induced periodontitis. Immunol Invest, 1989 Jan-May, 18(1-4), 171 - 85 Antigens released from four oral bacteria in periodontitis; Tolo K et al.; Antigens fractionated from cultures of four oral bacteria were tested for binding of serum IgG, IgA and IgM from patients in early and established phase of periodontitis . Bacteroides gingivalis and A.actinomycetemcomitans released antigens that discriminated between serum from individuals with or without periodontitis . The discriminating antigens ranged from 10 to 43 kDa and included neutral sugar and protein but no lipids . Significantly increased levels of IgG and IgA antibodies to the antigens released from B . gingivalis were detected before bone loss was seen and predicted such disease progression. Pediatr Med Chir, 1989 Jan-Feb, 11(1), 93 - 6 {Post-traumatic inflammatory pseudotumor of the lung . Description of a case}; Freschi P et al.; Inflammatory pseudotumor (P.I.) of the lung is a rare benign lesion histologically characterized by a variety of inflammatory and mesenchymal cells (plasma cells, lymphocytes and histiocytes) . The etiology remains obscure . We report a case of P.I . occurred immediately after a thoracic trauma . The infectious etiology is suggested by the growth of an anaerobic microorganism (Bacteroides) in tissue culture. FEMS Microbiol Lett, 1989 Jan 1, 48(1), 93 - 6 Glycylprolyl dipeptidase activity of Bacteroides gingivalis W50 and the avirulent variant W50/BEI; Kay HM et al.; Glycylprolyl dipeptidase activity was measured in cells, extracellular vesicles (ECV) and the soluble extracellular protein fraction (EP) of batch cultures of strains W50 and W50/BEI . Total culture enzyme activity of W50 dropped with age whilst that of W50/BEI remained constant . Activity was highest in the cellular fraction, greater for W50/BEI than W50 and rose with culture age . Both strains showed similar ECV activities but these declined with culture age . The EP glycylprolyl dipeptidase activity of W50/BEI in older cultures rose to a level 13-fold greater than W50 . The majority of extracellular activity was represented by the ECV for strain W50 but by EP for W50/BEI . Variable but incomplete attenuation of activity was achieved by dithiothreitol . ECV and EP activities were associated with a high molecular mass fraction, but a smaller fraction (molecular mass 30,000) was detected in W50/BEI EP. Diagn Microbiol Infect Dis, 1989 Jan-Feb, 12(1), 45 - 50 Comparison of the in vitro action and interaction of cefotaxime and desacetylcefotaxime against clinical isolates of Bacteroides spp; Aldridge KE; Desacetylcefotaxime (dCTX), the in vivo metabolite of cefotaxime (CTX), possess significant in vitro antimicrobial activity similar to the parent compound against a variety of aerobic and anaerobic bacteria . In vitro susceptibility studies showed that CTX inhibited 86% of 473 strains of the Bacteroides fragilis at a concentration of 32 micrograms/ml while dCTX inhibited 91% of the test isolates at the same concentration . Strains of the B . fragilis species were significantly more susceptible to CTX than were the non-B . fragilis species . Susceptibility testing of CTX and dCTX in a 1:1 ratio produced significantly more inhibitory activity, especially against the non-B . fragilis strains . Synergy studies showed that the interaction of CTX and dCTX was either completely or partially synergistic against 85% of 92 test organisms . The presence of dCTX was also shown to lower the CTX MIC values four-fold or greater in 82% of the synergy studies . Synergy was noted against strains of the B . fragilis group, B . melaninogenicus group, B . bivius, B . disiens, and B . capillosus . Through the use of time-kill kinetics studies, the interaction of CTX and dCTX was shown to be additive at subinhibitory and inhibitory concentrations and suggestedly synergistic at suprainhibitory concentrations against strains of the B . fragilis group . These in vitro studies demonstrate that dCTX increases the inhibitory and bactericidal activity of CTX when tested in combination. Diagn Microbiol Infect Dis, 1989 Jan-Feb, 12(1), 39 - 43 Effectiveness of cefotaxime alone and in combination with desacetylcefotaxime against Bacteroides fragilis; Wasilauskas BL; The synergistic activity of the combination cefotaxime-desacetylcefotaxime (CTX/dCTX) was compared to the effectiveness of seven other antimicrobial agents: cefoxitin (CFOX), cefotetan (CTAN), ceftizoxime (CTIZ), chloramphenicol (CLOR), clindamycin (CLIND), metronidazole (METR), and ampicillin-sulbactam (A/S) tested against 100 clinical isolates belonging to the Bacteroides fragilis group . All tests were performed using the NCCLS reference agar-dilution method . The overall susceptibility of these organisms to CTX/dCTX was 84% compared to CFOX at 78% or CTAN at 66% . The other antimicrobials inhibited greater than 90% of these isolates . There was no difference between the susceptibility rates of CTX/dCTX and CTX with the B fragilis (85%) or B . distasonis (75%) strains . Bacteroides thetaiotaomicron showed a 11% greater susceptibility to CTX/dCTX than to CTX . Of the 100 isolates tested, 40% showed either synergy or partial synergistic interactions between CTX and dCTX . Most of the isolates showed indifference (52%), while 8% demonstrated antagonism; a relatively unique finding to date. Diagn Microbiol Infect Dis, 1989 Jan-Feb, 12(1), 33 - 7 Comparative in vitro activity of cefoxitin, cefotaxime alone, and in combination with desacetylcefotaxime against the Bacteroides species; Canawati HN; The agar dilution method was used to determine the inhibitory activity of cefotaxime (CTX) alone, desacetylcefotaxime (dCTX) alone, CTX plus dCTX, and cefoxitin against 74 clinical isolates of the Bacteroides species recovered from diabetic patients with foot ulcers . The study concluded that the addition of dCTX to CTX increased the inhibitory activity from 45% to 73% for all strains tested and from 50% to 81% among the 32 strains of Bacteroides fragilis . This synergistic interaction against B . fragilis resulted in a four- to nine-fold reduction in the MIC of seven strains (64-128 micrograms/ml, resistant category MICs) . While the lowest CTX MIC for B . fragilis was 2 micrograms/ml (four strains), the addition of dCTX also produced a remarkable reduction in susceptible range CTX MICs to 0.05-2 micrograms/ml in 16 strains (50%) . The overall susceptibility to cefoxitin and CTX plus dCTX was as follows: 100% and 100% for Bacteroides vulgatus, 50% and 66% for Bacteroides thetaiotaomicron, 100% and 33% for Bacteroides ovatus, and 83% and 82% for Bacteroides species other than the B . fragilis group. Appl Environ Microbiol, 1989 Jan, 55(1), 148 - 53 Ruminal cellulolytic bacteria and protozoa from bison, cattle-bison hybrids, and cattle fed three alfalfa-corn diets; Varel VH et al.; Ruminal cellulolytic bacteria and protozoa and in vitro digestibility of alfalfa fiber fractions were compared among bison, bison hybrids, and crossbed cattle (five each) when they were fed alfalfa and corn in a ratio of 100:0, 75:25, and 50:50, respectively . The total number of viable bacteria (2.16 x 10(9) to 5.44 x 10(9)/ml of ruminal fluid) and the number of cellulolytic bacteria (3.74 x 10(7) to 10.9 x 10(7)/ml) were not different among groups of animals fed each diet . The genera of protozoa in all of the animal groups were similar; however, when either the 100:0 or 50:50 diet was used the percentage of Entodinium sp . was lower and the percentage of Diplodiniinae was higher (P less than 0.05) in bison than in bison hybrids or cattle . Bacteroides succinogenes made up the largest number of cellulolytic isolates from bison (58 and 36%, respectively, on the 100:0 and 75:25 diets), which were more numerous (P less than 0.05) than those from bison hybrids (36 and 12%) and cattle (33 and 18%) . This was offset by a lower number of cellulolytic Butyrivibrio isolates . The numbers of Ruminococcus albus and R . flavefaciens isolates, in general, were similar among the bovid species, although R . flavefaciens generally made up less than 10% of the cellulolytic isolates . In vitro digestibility coefficients were greater (P less than 0.05) for the bison when the 75:25 diet was used and similar for the other two diets . The concentration of ruminal volatile fatty acids was larger (P less than 0.05) in bison than in bison hybrids and cattle when the 50:50 diet was used.(ABSTRACT TRUNCATED AT 250 WORDS) Scand J Infect Dis Suppl, 1989, 62, 15 - 24 Virulence determinants in nonsporeforming anaerobic bacteria; Hofstad T; The literature dealing with adherence, host-protective mechanisms and tissues damaging products of nonsporeforming anaerobes is reviewed . The adherence mechanisms are poorly understood . There is evidence for that encapsulation plays a role in the pathogenicity of Bacteroides fragilis and black-pigmented bacteroides . A leukocidin is produced by Fusobacterium necrophorum, and Ig protease, collagenase and a trypsin-like enzyme by some Bacteroides species . Some Bacteroides fragilis strains produce an enterotoxin . The pathogenetic role of endotoxin is unclear. Rev Belge Med Dent, 1989, 44(2), 9 - 30 {Bacterial invasion in periodontitis, is it important in periodontal treatment?}; Adriaens PA; During the progression of periodontal disease bacteria invade the epithelial lining of the gingival pocket and the underlying connective tissue . This invasion was demonstrated in acute necrotizing ulcerative gingivitis (ANUG), localized juvenile periodontitis (LJP), rapidly progressive periodontitis in adults (RPP) and in children with Papillon-Lefevre syndrome associated periodontitis . Using morphologic criteria or fluorescent antibody staining techniques spirochetes, Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, B . intermedius, Actinomyces species, Fusobacterium nucleatum and Mycoplasma have been identified in these tissue compartments . Recent light microscopic, electron microscopic and bacteriological studies demonstrated the presence of invading bacteria in the cementum and in the radicular dentin of teeth with periodontal disease . In 83% of 69 carries free periodontally diseased teeth bacteria were present in one of the dentin layer from the root . In 59% of these teeth the dental pulps contained bacteria . The composition of the flora resembled that of the subgingival flora . The use of chlorhexidine, fluoride, tetracyclines or metronidazole in the elimination of invading bacteria following mechanical periodontal therapy is discussed. Int Arch Allergy Appl Immunol, 1989, 90(1), 24 - 30 Modulation of human neutrophil adherence by periodontopathic bacteria: reversal by specific monoclonal antibodies; Seow WK et al.; Previous investigations by us have shown that direct interaction of Fusobacterium nucleatum with polymorphonuclear neutrophils (PMNs) results in the stimulation of PMN adherence whereas direct interaction with Bacteroides gingivalis results in PMN suppression . In the present study, panels of monoclonal antibodies (MAbs) raised against cell wall antigens of F . nucleatum and B . gingivalis were tested to determine their ability to block the modulatory effects of the bacteria in their interactions with PMNs . While no activity was demonstrated for any of the 9 MAbs raised against F . nucleatum, it was found that 2 of 16 MAbs raised against B . gingivalis were able to reverse the suppression of PMN adherence induced by the bacteria . Further studies on these 2 reactive MAbs showed that the effect of MAbs were abrogated by heat treatment as well as by trypsin proteolysis . Investigations into the nature of the reactive epitopes on the bacterial surface showed that they probably contain protein components susceptible to proteolytic attack by subtilisin . In addition, beta-galactose may be a component of the reactive epitopes for one of the MAbs, but sialic acid residues on the bacterial surface are probably not involved as their elimination by neuraminidase did not affect the binding of both MAbs . The results of the present study strongly validate our previous observations that direct specific interaction of B . gingivalis with human PMNs occurs, resulting in the suppression of PMNs. Appl Environ Microbiol, 1989 Jan, 55(1), 132 - 6 Molecular cloning and expression in Escherichia coli of a cellodextrinase gene from Bacteroides succinogenes S85; Gong JH et al.; A DNA fragment coding for a cellodextrinase of Bacteroides succinogenes S85 was isolated by screening of a pBR322 gene library in Escherichia coli HB101 . Of 100,000 colonies screened on a complex medium with methylumbelliferyl-beta-D-cellobioside as the indicator substrate, two cellodextrinase-positive clones (CB1 and CB2) were isolated . The DNA inserts from the two recombinant plasmids were 7.7 kilobase pairs in size and had similar restriction maps . After subcloning from pCB2, a 2.5-kilobase-pair insert which coded for cellodextrinase activity was isolated . The enzyme was located in the cytoplasm of the E . coli host . It exhibited no activity on carboxymethyl cellulose, Avicel microcrystalline cellulose, acid-swollen cellulose, or cellobiose but hydrolyzed p-nitrophenyl-beta-D-cellobioside and p-nitrophenyl-beta-D-lactoside . The Km (0.1 mM) for the hydrolysis of p-nitrophenyl-cellobioside by the enzyme expressed in E . coli was similar to that reported for the purified enzyme from B . succinogenes . Expression of the cellodextrinase gene was subjected to catabolite repression by glucose and was not induced by cellobiose . The origin of the DNA insert from B . succinogenes was confirmed by Southern blot analysis . Western blotting (immunoblotting) using antibodies raised against the purified B . succinogenes cellodextrinase revealed a protein with a molecular weight of approximately 50,000 in E . coli clones which comigrated with the native enzyme isolated from B . succinogenes . These data indicate that the cellodextrinase gene expressed in E . coli is fully functional and codes for an enzyme with properties similar to those of the native enzyme. J Bacteriol, 1989 Jan, 171(1), 148 - 53 Novel aerobic tetracycline resistance gene that chemically modifies tetracycline; Speer BS et al.; A tetracycline resistance gene that was found originally on the Bacteroides plasmid pBF4 confers resistance on Escherichia coli but only when cells are growing aerobically . When E . coli EM24 carrying this aerobic tetracycline resistance (*Tcr) gene is grown in medium containing tetracycline, the resulting spent medium is no longer toxic to tetracycline-sensitive (Tcs) E . coli EM24 (B.S . Speer and A.A . Salyers, J . Bacteriol . 170: 1423-1429, 1988) . To determine whether the *Tcr gene product modified tetracycline, we characterized the material resulting from incubation of E . coli (*Tcr) with tetracycline . When {7-3H(N)}tetracycline was added to cultures of E . coli (*Tcr), at least 90% of the label was recovered in the extracellular fluid . Therefore, tetracycline was not being sequestered by the cells . The labeled material behaved similarly to tetracycline with respect to solubility in various organic solvents . However, the UV-visible light spectrum had a single peak at 258 nm, whereas the tetracycline spectrum had a peak at 364 nm . The labeled material also had a faster migration rate than did tetracycline on thin-layer plates in a solvent system of butanol-methanol-10% citric acid (4:1:2, vol/vol/vol) and was separable from tetracycline by reverse-phase high-pressure liquid chromatography, using an acetronitrile-0.1% trifluoroacetic acid solvent system . These results demonstrate that the *Tcr gene product chemically modifies tetracycline . The *Tcr gene is the first example of a chemically modifying tetracycline resistance mechanism. Arch Oral Biol, 1989, 34(9), 679 - 83 Interleukin-1 and interleukin-1 inhibitor production by human adherent cells stimulated with periodontopathic bacteria; Walsh LJ et al.; This study examined the effect of the putative periodontopathic bacteria Bacteroides gingivalis and Fusobacterium nucleatum on the production of interleukin-1 (IL-1) and IL-1 inhibitors by human plastic-adherent mononuclear cells from normal donors . Fusobacterium mortiferum was used as a non-oral, non-pathogenic control organism . Unstimulated adherent cells spontaneously secreted an IL-1 inhibitor, whereas stimulation with B . gingivalis induced the synthesis and secretion of IL-1 . With both fusobacteria IL-1 was present in the intracellular environment, whereas the predominant secretory product was either IL-1 or an IL-1 inhibitor . These results suggest that bacteria are capable of modulating cytokine production by monocytes and may thereby alter the local immune response. J Endod, 1989 Jan, 15(1), 13 - 9 Prevalence of black-pigmented bacteroides species in root canal infections; Sundqvist G et al.; The prevalence of black-pigmented Bacteroides species in the root canals of 72 teeth with apical periodontitis was evaluated . Twenty-two of the canals contained one or more species of black-pigmented Bacteroides . Bacteroides intermedius (14 strains) and Bacteroides endodontalis (5 strains) were most common . Of the species Bacteroides gingivalis, Bacteroides loeschei, and Bacteroides denticola, 2, 3, and 1 strains, respectively, were isolated . The median number of bacterial cells recovered from the root canals containing black-pigmented Bacteroides was 2.8 x 10(5) and from the other canals 3.0 x 10(3) . The mean number of strains was 7.9 and 3.3, respectively . Sixteen of the 22 root canals containing black-pigmented Bacteroides species were associated with acute apical abscesses and purulent drainage through the root canal . The other six teeth with black-pigmented Bacteroides were asymptomatic . One additional abscess was present among the 72 cases . This root canal contained Actinomyces israelii and Actinomyces naeslundii. Arch Oral Biol, 1989, 34(7), 579 - 83 Collagenolytic activity of the extracellular vesicles of Bacteroides gingivalis W50 and an avirulent variant W50/BE1; Smalley JW et al.; The activities of the extracellular vesicle fractions of these two organisms were compared . Lytic activity against a native type I placental collagen substrate at 30 degrees C was assessed following sodium dodecyl sulphate-polyacrylamide gel electrophoresis and densitometry . A rapid rate of collagen depolymerization was achieved by the extracellular vesicle fraction of W50, yielding approx . 90% substrate degradation compared to 5% for W50/BE1 extracellular vesicles over 6 h incubation . The polypeptide digestion patterns produced by incubation with extracellular vesicle fractions of both organisms were identical, and similar to those yielded by incubation of substrate with whole W50 cells. J Pedod, 1989 Spring, 13(3), 222 - 9 DNA probe detection of key periodontal pathogens in juveniles; Kisby LE et al.; Recognition of juvenile forms of periodontitis have been shown to be directly linked with specific Gram-negative rods, primarily Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius . However, clinical application of these laboratory findings have generally been restricted to the research environment . The purpose of this study was to explore the relationship of these three pathogenic species in children using the reliable and accurate new technology of DNA probes . Results indicated that the pathogenic oral flora did not differ significantly between subjects with and without gingival inflammation . Therefore, tracking of pathogens using clinical parameters of inflammation are unreliable and accurate monitoring requires microbiological testing in order to properly identify the presence or absence of pathogens. Scand J Infect Dis Suppl, 1989, 62, 29 - 34 Immune mechanisms in the prevention of intra-abdominal abscess formation; Kasper DL et al.; Bacteroides fragilis is the most commonly isolated anaerobe from intraabdominal infections . In experimental models of intraabdominal sepsis, B . fragilis has been shown to be uniquely virulent . Some of these virulence traits are due to the capsular polysaccharide of this organism . Immunity to infection secondary to B . fragilis seems to involve both arms of the immune system . Humoral immunity (complement, antibody and PMNs) is critical to clearance of these bacteria from the blood stream . Cellular immune mechanisms predominate against intraabdominal abscess formation . Adoptive transfer experiments have shown that a CD8+, IJ+, non H2 restricted immune T cell or lysate from this T cell confers protection to immunocompetent, naive mice . An in vivo system has been developed to begin defining the mechanism of protection . B . fragilis placed inside a filter containment chamber within the peritoneum of immune mice are specifically killed over an 8-day period in the absence of white blood cells . This killing phenomenon was also observed inside filter chambers within mice receiving adoptively transferred immune T cells or lysates of these T cells . Furthermore, killing is specific to B . fragilis . These results support a T cell dependent mechanism for killing this bacteria and provide an interesting model for further exploration. Scand J Infect Dis Suppl, 1989, 62, 25 - 8 Mechanisms of antimicrobial resistance in anaerobic bacteria: the predictive approach; Baquero F et al.; Antibiotic resistance in anaerobic bacteria probably evolves from broadly undetected mechanisms of resistance, as the generally accepted breakpoints in susceptibility testing are mainly based on pharmacological grounds . Some of the strains considered susceptible by conventional techniques may harbour mechanisms of resistance . These resistant organisms should be screened using the antibiotic concentrations immediately above those inhibiting the fully susceptible population of a given species as breakpoints . The following break-points for bacteroides strains are suggested as definitions of the entirely susceptible population: piperacillin 8 mg/l; ceftizoxime 8 mg/l: cefoxitin 8-16 mg/l: imipenem 1 mg/l: ticarcillin-clavulanate 2 mg/l; clindamycin 1 mg/l: chloramphenicol 8 mg/l: tetracycline 4 mg/l: metronidazole 1 mg/l . Detection of an increased frequency of organisms presenting low-level resistance may be useful to predict and possibly control the appearance and spread of fully resistant anaerobic organisms. Microbiol Immunol, 1989, 33(1), 75 - 80 Possibility of Bacteroides gingivalis hemagglutinin possessing protease activity revealed by inhibition studies; Nishikata M et al.; Inhibition of hemagglutinin (HA) activity in a membrane fraction of Bacteroides gingivalis was examined using various compounds . Leupeptin and anti-pain inhibited the HA activity at nM order . This potency was lost when the aldehyde group of leupeptin was converted to an alcohol moiety . Irreversible protease inhibitors, tosyl-L-lysine chloromethyl ketone (TLCK), p-chloromercuribenzoate (PCMB), and N-ethylmaleimide (NEM) were also inhibitory . From the inhibition experiments, we speculate that the HA possesses protease activity and that the same site of the molecule participates in the erythrocyte binding and the substrate binding. J Med Microbiol, 1989 Jan, 28(1), 9 - 14 Aggregation by fragilis and non-fragilis Bacteroides strains in vitro; Blake M et al.; Bacteroides fragilis is associated with the formation of intra-abdominal abscesses, whereas other Bacteroides species are rarely involved . Since bacterial clumping may contribute to the survival of bacteria in the face of host defence mechanisms, the hypothesis has been put forward that differences in aggregation between fragilis and non-fragilis strains of Bacteroides may account for their differences in survival in vivo . All seven B . fragilis strains tested formed aggregates within 4 h, but strains not associated with intra-abdominal sepsis--B . vulgatus, B . thetaiotaomicron and B . distasonis--did not form aggregates in vitro . Aggregation occurred at 37 degrees C, but not at 4 degrees C or 20 degrees C . Treatment with pronase partially inhibited aggregation . Periodate treatment killed the cells and caused them to form clumps which were distinguishable from the control aggregates . Heat-killed B . fragilis cells formed similar distinct clumps, but cells killed by glutaraldehyde and formaldehyde did so to a lesser degree . No inhibition was found upon addition of carbohydrates, ethylenediaminetetraacetic acid or after treatment with trypsin . These results demonstrate that aggregate formation occurs with B . fragilis strains alone, and that surface proteins probably mediate this interaction. Infect Immun, 1989 Jan, 57(1), 213 - 8 A protease of Bacteroides gingivalis degrades cell surface and matrix glycoproteins of cultured gingival fibroblasts and induces secretion of collagenase and plasminogen activator; Uitto VJ et al.; To assess the direct effects of Bacteroides gingivalis on periodontal cells, human gingival fibroblasts were cultured in the presence of B . gingivalis extracts or a trypsinlike enzyme partially purified from the bacteria by chromatography on benzamidine-Sepharose and Sephacryl S-200 . Analysis of cell surface glycoproteins by the periodate-{3H}borohydride labeling technique combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-fluorography demonstrated that fibronectin and some other high-molecular-weight cell surface glycoproteins were degraded by a 35,000-Mr(35K) B . gingivalis protease . Immunostaining of the fibroblast cultures showed degradation of intercellular matrix fibronectin by the 35K protease . The pattern of fibronectin degradation was monitored by examining the reaction products with the SDS-PAGE-immunoblotting technique . The protease degraded fibronectin rapidly and more extensively than did corresponding amounts of pancreatic trypsin . Collagenase secretion by the fibroblasts was assayed by incubating cell culture medium with soluble type I {3H}collagen at 25 degrees C followed by SDS-PAGE-fluorography analysis of the reaction products . The medium was also assayed for plasminogen activator activity by using a casein-agarose diffusion plate assay . The fibroblasts cultured with the 35K protease secreted increased amounts of collagenase and plasminogen activator into the medium . The results suggest that periodontal infection by B . gingivalis causes proteolytic damage of the host cell surface structures . Concomitantly, B . gingivalis may induce the cells to degrade their pericellular matrix. Zahn Mund Kieferheilkd Zentralbl, 1989, 77(7), 659 - 67 {A latex agglutination test for the rapid identification of periodontally pathogenic microorganisms in subgingival plaque . The preliminary results}; Tsalikis L et al.; Latex agglutination tests (LA) have been evaluated for identifying periodontal pathogens . This preliminary study compared LA tests with indirect immunofluorescence (IF) for the rapid identification of A . actinomycetemcomitans, B . gingivalis, and B . intermedius in clinical subgingival plaque samples . A total of 58 sites from 12 patients suffering from advanced periodontitis, exhibiting recent attachment loss were examined before and after initial periodontal treatment . The results of the two examinations were pooled and a total of 116 sites were statistically evaluated . The overall agreement between LA and IF was 75.8% for black pigmented Bacteroides (BPB) and 74.1% for Aa . These results demonstrate the feasibility of LA as a rapid, inexpensive, and reliable tool for monitoring periodontal disease offered to the general practitioner. J Periodontal Res, 1989 Jan, 24(1), 20 - 7 Failure of Bacteroides gingivalis W83 to accumulate bound C3 following opsonization with serum; Schenkein HA; Our previous studies have demonstrated that strains of Bacteroides gingivalis are capable of proteolytic degradation and inactivation of complement proteins including the third component of complement C3 . Since a crucial step in the ability of complement to control bacterial infections is the binding of C3 fragments to the bacterial surface with subsequent enhancement of phagocytosis, further examination of the importance of the proteolytic capacity of Bacteroides in interactions with complement proteins was carried out by quantitating the amount of C3 bound to two proteolytic Bacteroides gingivalis strains . Pooled normal human serum (NHS) containing 125I-C3 was incubated with strains of B . gingivalis (W83 and ATCC 33277) and the non-proteolytic pathogen A . actinomycetemcomitans strain Y4, and samples of the reaction mixtures were removed at various time intervals for determination of bound C3 . B . gingivalis 33277 bound only half the number of C3 molecules as did A . actinomycetemcomitans, while B . gingivalis W83 bound very little C3 . A large increase in the number of C3 molecules bound to B . gingivalis W83 was noted in assays carried out in the presence of the protease inhibitor TLCK, indicating that bacterial proteases may be responsible for the lack of binding of C3 to strain W83 . TLCK treatment modestly increased the accumulation of C3 on strain 33277, but had no effect on A . actinomycetemcomitans . Analysis of 125I-C3 in supernatants from reaction mixtures of strain 33277, W83, or a proteolytic strain of B . intermedius demonstrated no qualitative differences in the C3 fragments amongst the tested strains or in the presence or absence of TLCK.(ABSTRACT TRUNCATED AT 250 WORDS) Eur J Clin Microbiol Infect Dis, 1989 Jan, 8(1), 83 - 5 Selective and differential medium for isolation of Bacteroides ureolyticus from clinical specimens; Eley A et al.; A new selective and differential medium for Bacteroides ureolyticus was compared with other commonly used selective media for anaerobes in a diagnostic trial with routine clinical specimens . All Bacteroides ureolyticus strains grew on nalidixic acid-teicoplaninurea agar, producing red colonies against a straw-coloured background . This medium was found to be the most selective for Bacteroides ureolyticus . In the diagnostic trial with 185 clinical specimens a 20% increase in the isolation rate of Bacteroides ureolyticus was obtained with this new medium. Arch Oral Biol, 1989, 34(11), 911 - 6 Cleavage action of a trypsin-like protease from Bacteroides gingivalis 381 on reduced egg-white lysozyme; Endo J et al.; Soluble reduced lysozyme was extensively digested by a trypsin-like protease purified from the culture supernatant of the bacterium . The digestion peptides were separated and purified by reversed-phase high-performance liquid chromatography, and were subjected to amino acid analysis . The fragments were identified by their amino acid composition, and the cleavage sites in the lysozyme chain were determined . Like mammalian trypsin, the enzyme from B . gingivalis split peptide bonds non-specifically at carboxyl sides of internal arginine and lysine residues, but the lysine present at the amino terminus of the lysozyme chain was not released . In addition, the enzyme cleaved the peptide linkage at the amino side of lysine and bonds between leucine-glycine, alanine-leucine and leucine-serine . Thus the trypsin-like protease from B . gingivalis has some cleavage actions on lysozyme different from those of mammalian trypsin. Meikai Daigaku Shigaku Zasshi, 1989, 18(2), 187 - 96 {Biological characterization of Bacteroides intermedius specific monoclonal antibody}; Satoh T et al.; One monoclonal antibody (BIF6) directed against Bacteroides intermedius Group I was developed by hybridoma technology . In the present study, we used culture medium from BIF6 antibody-producing cells to characterize the antibody . Its reactivity against B . intermedius (Group I, II), B . gingivalis, B . corporis, B . melaninogenicus, B . loescheii, and B . asaccharolyticus was detected by the enzyme-linked immunosorbent assay . BIF6 monoclonal antibody specifically reacted with B . intermedius Group I, but not with B . intermedius Group II . Also, the BIF6 did not react with other black-pigmented Bacteroides . Since BIF6 antibody reacted with some clinical isolates from subgingival plaques of adult periodontal patients, we suggest that this antibody may be useful as a tool in the clinic to identify B . intermedius Group I . Specific antigen of B . intermedius Group I recognized by BIF6 antibody was located on the outer membrane of the organism . And the specific antigen was inactivated by treatment for 20 min at 80 degrees C and at pH2 and 12 . Also the antigen was sensitive to trypsin treatment. Medicina (B Aires), 1989, 49(4), 357 - 9 {Bacteroides distasonis meningitis}; Santoianni JE et al.; A 50 year old woman while undergoing severe treatment for rheumatoid arthritis, developed anaerobic meningitis . The cerebrospinal fluid (CSF) sample was transported and cultivated aerobically and anaerobically . After 48 h at 37 degrees C the anaerobically incubated plate, the enriched fluid thioglycollate medium and the anaerobic culture medium yielded luxuriant growth of an anaerobic Gram negative bacillum . The biochemical and antimicrobial susceptibility patterns were consistent with those for Bacteroides distasonis . Most of the strains of the 5 species included in the Bacteroides fragilis group (B . fragilis, B . vulgatus, B . ovatus, B . thetaiotaomicron and B . distasonis) are resistant to penicillins, cephalosporins of first generation and aminoglycosides . Anaerobic polyresistant flora from an intraabdominal focus (chronic cholecystitis) might have been selected by treatment with gentamicin and cephalotin, and proliferated into meningeal dissemination . It is important that CSF from immunocompromised patients with acute or chronic pulmonary, intraabdominal or cranium-facial infectious processes be transported and cultured in aerobic and anaerobic conditions . These patients must be treated with an initial therapeutic scheme that includes an effective antibiotic for the anaerobic microorganism that may be involved. Acta Microbiol Pol, 1989, 38(3-4), 285 - 91 The cell-surface components of Bacteroides thetaiotaomicron as coating antigens in ELISA test; Rokosz A et al.; Cell-surface antigens were extracted out of three Bacteroides thetaiotaomicron strains of different origin . Lipopolysaccharides, their fractions, L1 preparations and capsular antigen were obtained . All substances were tested as coating antigens in ELISA test against antibacterial rabbit immune sera . The highest absorbances were observed with lipopolysaccharides (LPS), their polysaccharide fractions (PS) and capsular material (CPS) . Lipopolysaccharides after nuclease treatment (N-LPS), free from nucleic acids, were more active than crude phenol-water extracts (PW-LPS). Acta Microbiol Pol, 1989, 38(2), 171 - 6 Serological studies of Bacteroides vulgatus from normal gut flora; Meisel-Mikolajczyk F et al.; A serological classification scheme for 48 strains of Bacteroides vulgatus was established by agglutination technique . Bacterial strains were isolated in our laboratory from gut flora of healthy persons . Absorbed antisera to 11 strains of B . vulgatus were prepared . All 48 strains tested reacted with one or more antisera . Among these strains, 22 serogroups were formed; 11 of them contained only one group component and 11 were made up of multiple group components. Infect Immun, 1989 Jan, 57(1), 95 - 9 Characterization of sodium dodecyl sulfate-stable Bacteroides gingivalis proteases by polyacrylamide gel electrophoresis; Grenier D et al.; Profiles of the proteolytic activities found in Bacteroides gingivalis culture supernatants, outer membranes, vesicles, and cell extracts were analyzed in sodium dodecyl sulfate-polyacrylamide gels containing covalently bound bovine serum albumin . A total of eight distinct bands of proteolytic activity could be detected . Four of these were found in the culture supernatant (P1, P2, P3, and P4) . The outer membranes, vesicles, and the cell extract each contained seven major proteolytic bands (P1, P3, P4, P5, P6, P7, and P8) . No activity was found in the membrane-free extract, suggesting that the proteases were associated with the cell envelope . With the exception of P7 and P8, all the proteolytic bands were dependent on reducing agents for activity . The eight proteolytic bands were distributed in an identical manner in all four strains of B . gingivalis studied . The effects of protease inhibitors, pH, and heat were determined . Sulfhydryl group reagents and N-alpha-p-tosyl-L-lysine chloromethyl ketone reduced proteolytic activity . The optimum pH was found to be between 7 and 8 . A 30-min preincubation at 50 degrees C inactivated the P6, P7, and P8 proteolytic bands . All proteolytic activity was lost after the samples were heated at 75 degrees C for 30 min. J Chromatogr, 1988 Dec 28, 459, 331 - 5 Rapid identification of Bacteroides species by high-performance liquid chromatography; Radin L et al.; High-performance liquid chromatography was evaluated for the rapid identification of Bacteroides species of clinical interest . Each isolate was inoculated into a defined chemical medium containing primarily carbohydrates and was incubated aerobically at 37 degrees C for 1 h . After centrifugation, the supernatants were placed on ice to stop further enzymatic reactions . Specimens were injected into an Aminex HPX-87H column in order to determine carbohydrates and acid metabolic products . Peak areas of carbohydrates for each isolate were compared with those for uninoculated medium . As the utilization indexes of raffinose, lactose and arabinose were found to be particularly significant, the patterns of carbohydrate utilization could be used for the identification of Bacteroides species . This method can be adapted for diagnostic laboratory use and has good potential for automated microbial identification. Showa Shigakkai Zasshi, 1988 Dec, 8(4), 405 - 12 Purification and characterization of intracellular dextranase of Bacteroides oralis Ig4a; Igarashi T et al.; Multiple forms of dextranase were detected in both intra- and extracellular fractions of Bacteroides oralis Ig4a . The molecular weights of these enzymes varied from 52,000 to 260,000 by sodium dodecyl sulfate-polyacrylamide-blue dextran gel electrophoresis . The intracellular dextranases were fractionated by chromatography and gel filtration steps, and the dextranases IV and V were obtained . The former was only partially pure . The molecular weights of the dextranases IV and V were estimated to be 120,000 and 105,000, respectively, by SDS-PAGE . The dextranase V was further characterized and it was revealed that the pH- and temperature optima were 5.0, and 55 degrees C, respectively . The Km value was 6.7 x 10(-2) mM for dextran T-70 . The enzyme did not exhibit any metal ion requirements, but was inhibited by CoCl2 and HgCl2; lysine and alanine contents were especially high; it hydrolyzed the alpha-1,6-glucan by an exo-type mechanism, and was inactive toward glucans containing alpha-1,3-, alpha-1,4-, and beta-1,4-linkages. Nippon Shishubyo Gakkai Kaishi, 1988 Dec, 30(4), 1156 - 67 {Microbiological evaluation of topical application of the controlled release strips containing ofloxacin (PT-01) in the human periodontal pocket}; Kimura S et al.; The recognition that destructive periodontal diseases may be caused by specific microorganisms has led to an increased interest and usage of antimicrobial agents in periodontal therapy . In this study, the effect of topical application of ofloxacin (OFLX), a synthetic antibiotic, was microbiologically evaluated . The new developed controlled release strips containing OFLX (PT-01), in which there were structurally immediate- and sustained-releasing portions, were applied to the periodontal pockets of 27 adult subjects with periodontitis . Three different sites with a deep probing pocket depth (greater than or equal to 5 mm) were randomly selected in each patient, and were divided into three groups, i.e., PT-01 applied site (T), placebo-applied site (P) and control site (C) . Periodontal treatments consisted of oral hygiene instruction and supragingival scaling on day 0 and 7, and subgingival scaling and root planing on day 14 . PT-01 was weekly applied on day 0 to 35, and the subgingival plaque samples from each site were collected on day 0, 14, 21 and 42 . The dynamics of subgingival microflora was investigated by dark field microscopy for determination of the %s of spirochetes, motile rods and coccoid cells, and anaerobic and aerobic cultivations for the determinations of the total number of subgingival bacteria, black-pigmented Bacteroides (BPB), Fusobacterium species and Actinobacillus actinomycetemcomitans . The results showed that the significant reduction of %s of spirochetes and motile rods and significant increase of % coccoid cells were found in only PT-01 applied sites during first 14 days . In this period, the total number of cultivable bacteria, BPB and Fusobacterium species were also significantly reduced in T sites . While, after subgingival scaling and root planing were performed, significant changes in the proportions and numbers of the subgingival microflora were found in all sites . Especially, PT-01 applied site showed significant improvement in the %s of spirochetes and motile rods as well as the total number of the bacteria . Moreover, the further microbiological determinations of the each isolate revealed that no detectable amounts of A . actinomycetemcomitans could be found from any samples in this study . These results suggested that weekly application of PT-01 in the periodontal pocket could have significant effects on the qualitative and quantitative improvements in the subgingival microflora . It was also suggested that the application of PT-01 might have ameliorating effect as adjuncts of mechanical subgingival plaque control in the periodontal treatment. Antimicrob Agents Chemother, 1988 Dec, 32(12), 1848 - 53 Mechanism of action of cephalosporins and resistance caused by decreased affinity for penicillin-binding proteins in Bacteroides fragilis; Yotsuji A et al.; The susceptibilities of 52 clinical isolates of Bacteroides fragilis to five monoanionic cephalosporins were examined . Cefoperazone showed the highest antibacterial activity, followed by ceftezole, cefazolin, cefamandole, and cephalothin . There were two groups of resistant strains: one group (ca . 15%), of which B . fragilis G-232 was a typical sample, was resistant to ceftezole (MIC, 100 micrograms/ml), cefazolin (MIC, 100 micrograms/ml), and cephalothin (MIC, 200 micrograms/ml) but not cefoperazone (MIC, 6.25 micrograms/ml) or cefamandole (MIC, 25 micrograms/ml) . On the basis of studies of stability to beta-lactamase, outer membrane permeation, and affinity for penicillin-binding proteins (PBPs), we conclude that decreased affinity for PBP 3 may play an important role in the resistance to ceftezole, cefazolin, and cephalothin in B . fragilis G-232 . Another group (also ca . 15%), of which B . fragilis G-242 was a representative, was resistant to all five cephalosporins (MIC, 100 to 400 micrograms/ml) and produced a high amount of beta-lactamase . Similar broad-spectrum resistance was seen in a mutant of strain G-232 that had a greater-than-30-fold increase in beta-lactamase production. J Antimicrob Chemother, 1988 Dec, 22(6), 785 - 90 Permeability to beta-lactams in Bacteroides fragilis; Cuchural GJ et al.; Bacteroides fragilis strains TAL2480 and TAL3636 were used to assess outer membrane permeability to various beta-lactam compounds . These strains were chosen because they possess beta-lactamases capable of hydrolysing all commonly employed beta-lactams except monobactams . The beta-lactamases are located in the periplasmic space and could be released by sonication and osmotic shock . Permeability was calculated by the method of Zimmerman & Rosselet (1977, Antimicrobial Agents and Chemotherapy 12, 368-72) . The rank order of permeative ability (from fastest to slowest) was as follows: cephaloridine, imipenem, cefotaxime, cefoxitin, cefoperazone, nitrocefin, cephalothin, and latamoxef . Our results showed that ionic charge, hydrophobicity, and molecular weight influenced beta-lactam drug uptake by B . fragilis . These results are similar to findings for Escherichia coli, where increased negative charge and increased molecular weight are associated with decreased drug uptake . However, unlike E . coli, increased drug hydrophobicity, for a given charge and molecular weight of the drug, was associated with increased uptake by B . fragilis. Antimicrob Agents Chemother, 1988 Dec, 32(12), 1825 - 9 Comparative pharmacokinetics and serum inhibitory activity of clindamycin in different dosing regimens; Flaherty JF et al.; The comparative pharmacokinetics and serum inhibitory effects of clindamycin were evaluated in six healthy male subjects given multiple-dose infusions of the following regimens in a crossover fashion: 600 mg every 6 h, 900 mg every 8 h, and 1,200 mg every 12 h . Serial blood samples were obtained after the