J Clin Periodontol, 2005 Jan, 32(1), 21 - 8 Clinical effects of scaling and root planing on untreated teeth; Pawlowski AP et al.; Pawlowski AP, Chen A, Hacker BM, Mancl LA, Page RC, Roberts FA: Clinical effects of scaling and root planing on untreated teeth . J Clin Periodontol 2004; doi: 10.1111/j.1600-051X.2004.00626.x . (c) Blackwell Munksgaard, 2004 . Abstract Objective: The aim of this report is to examine whether scaling and root planing (SRP) in one area of the mouth may affect periodontal improvement in untreated areas in the same patient, possibly through systemic effects of treatment . Material and Methods: Twenty patients diagnosed with generalized aggressive periodontitis were randomized into treatment (n=11) and no treatment (n=9) groups . Within the treatment group, three quadrants were treated by SRP at week 0, 3, 12, and 24, while a single experimental quadrant remained untreated throughout the study . The outcome for all teeth was assessed using clinical parameters, subtraction radiography, and pathogenic bacteria levels in the subgingival flora over the 24-week study period . Results: Compared with sites in no treatment patients, the treated sites in the treated patients showed a 1 mm decrease in probing depth (PD) (p<0.01) and a 0.5 mm increase in bone height (p<0.01) by 24 weeks . In untreated sites within treated subjects, however, PDs tended to improve (p=0.09) but at a reduced rate compared with treated sites . The levels of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythensis (Bacteroides forsythus) remained unchanged in untreated sites while levels of Prevetolla intermedia and Treponema denticola tended to decrease as compared with controls but did not reach significance . Conclusions: This study indicates that untreated sites in treated periodontitis patients show a trend towards clinical improvement and exhibit reductions in some but not all periodontopathic bacterial species tested. Antibiot Khimioter, 2004, 49(6), 25 - 9 {Clinical and bacteriological substantiation of the use of cefoperazone/sulbactam in complex therapy of patients with pyo-destructive forms of lower respiratory tract infection (LRTI)}; Pneumocephalus associated with Bacteroides fragilis meningitis; University of Ottawa, Timmins & District Hospital, Timmins, Ontario P4N 8R1, Canada . atbeat@ntl.sympatico.caGas within the intracranial cavity (pneumocephalus) commonly results from trauma or after surgery and rarely from infection by gas-forming organisms . The presence of pneumocephalus in the absence of injury or surgery should raise the suspicion of anaerobic infection of the central nervous system . I present a case of pneumocephalus associated with Bacteroides fragilis meningitis where the diagnosis was suspected after CT findings become available . Bacteroides fragilis meningitis is rare and often occurs in premature infants and neonates; only few cases are reported in adults . Pneumocephalus associated with Bacteroides fragilis meningitis is not described in the literature . This case also illustrates the absence of classic findings of meningeal irritation in the elderly . The literature is reviewed. FEMS Microbiol Lett, 2005 Jan 15, 242(2), 339 - 44 Interaction between H(2)-producing and non-H(2)-producing cellulolytic bacteria from the human colon; Chassard C et al.; The cellulose-degrading species recently isolated from the human colon showed diverse ability to degrade and ferment cellulose . In the present study, the nature of the inter-relation existing between one H(2)-producing cellulolytic isolate (Ruminococcus sp . nov.) and one non-H(2)-producing cellulose-degrading species (Bacteroides sp . nov.) was investigated in vitro . Coculture experiments revealed synergism in cellulose degradation between these two cellulolytic species . An increase in total bacterial population was measured in the coculture, Bacteroides sp . being the predominant organism . As a result, a large decrease in H(2) production from cellulose fermentation was observed . Predominance of Bacteroides sp . might thus contribute to limit gas produced from fibre fermentation in the gut. FEMS Microbiol Lett, 2005 Jan 15, 242(2), 297 - 303 Molecular phylogenetic diversity of bacteria associated with the leachate of a closed municipal solid waste landfill; Huang LN et al.; A 16S rDNA-based molecular study was performed to determine the nature of the bacterial constituents of the leachate from a closed municipal solid waste landfill . Total community DNA was extracted and bacterial 16S rRNA genes were subsequently amplified and cloned . Recombinant rDNA clones in the library were randomly selected, and they were sequenced for a single run and then grouped . A total of 76 sequence types representing 138 randomly selected nonchimeric clones were identified . Full-length sequencing and phylogenetic analysis of the sequence types revealed that more than 90% of the screened clones were affiliated with low-G+C gram-positive bacteria (38.4%), Proteobacteria (35.5%), the Cytophaga Flexibacter Bacteroides group (11.6%), and Spirochaetes (5.1%) . Minor portions were affiliated with Verrucomicrobia (2.9%), candidate division OP11 (2.2%), and the green nonsulfur bacteria, Cyanobacteria and the Deinococcus Thermus group (each <1.0%) . Although some rDNA sequences clustered with genera or taxa that were classically identified within anaerobic treatment systems and expected with known functions, a substantial fraction of the clone sequences showed relatively low levels of similarity with any other reported rDNA sequences and thus were derived from unknown taxa . These results suggest that bacterial communities in landfill environment are far more complex than previously expected and remain largely unexplored. Pediatr Emerg Care, 2004 Sep, 20(9), 636 - 40 Anaerobic pulmonary infections in children; Brook I; Pulmonary infections due to anaerobic bacteria usually occur in children prone to aspiration . The source of the anaerobic bacteria is the oropharyngeal bacterial flora, where these organisms outnumber aerobic and facultative organisms in a 10:1 ratio . The most common lower respiratory tract infections where anaerobic bacteria are recovered mixed with aerobic organisms are aspiration pneumonia, lung abscess, and empyema . The predominant isolated anaerobic bacteria are Peptostreptococcus, Fusobacterium, pigmented Prevotella, and Porphyromonas spp . and Bacteroides fragilis group . Management of these infections includes the administration of antimicrobials effective against the anaerobic as well as the aerobic pathogens. Clin Ther, 2004 Oct, 26(10), 1564 - 77 Use of ciprofloxacin in the treatment of hospitalized patients with intra-abdominal infections; Madan AK; BACKGROUND: Numerous combination and single-agent antimicrobial regimens are available for the treatment of intra-abdominal infections . Selection of empiric agents must be directed at providing reliable activity against endotoxin-generating Escherichia coli, other gram-negative facultative bacteria, and anaerobes such as Bacteroides fragilis . Safety profiles, pharmacokinetic profiles, and cost-effectiveness must also be considered . Use of fluoroquinolones for the treatment of intra-abdominal infections has recently been advocated . METHODS: We review 2 prospective, comparative clinical trials conducted between 1992 and 2002 that evaluated the efficacy and safety of IV ciprofloxacin in patients with intra-abdominal infections . Separate pharmacoeconomic analyses conducted for each study are also reviewed . RESULTS: A total of 4 ciprofloxacin studies (2 clinical, 2 pharmacoeconomic) comprise the database . The combination of ciprofloxacin plus metronidazole was at least as effective as imipenem/cilastatin and clinically more effective than piperacillin/tazobactam therapy, based on clinical success end points . In 1 trial, treatment success for the clinically valid population was reported for 84% (93/111) of patients treated with IV ciprofloxacin/metronidazole, 86% (91/106) of those treated with IV/oral ciprofloxacin/metronidazole, and 81% of those treated with IV imipenem/cilastatin (91/113) . The IV/oral ciprofloxacin/metronidazole regimen had a statistically significant lower mean infection-related cost than the IV only ciprofloxacin/metronidazole plus imipenem groups (difference of approximately 1100 US dollars; P = 0.029) . In the second clinical trial, clinical resolution rates were statistically different for patients receiving IV/oral ciprofloxacin/metronidazole (74%) versus IV piperacillin/tazobactam therapy (63%; P = 0.047) . Ciprofloxacin/metronidazole was more cost-effective compared with piperacillin/tazobactam (2200 US dollars-3600 US dollars lower cost-effective ratios per patient) regardless of whether the patient had a diagnosis of appendicitis or whether a switch to an oral drug was permissible . CONCLUSIONS: In the studies reviewed herein, the combination of ciprofloxacin plus metronidazole was an effective and safe regimen for the treatment of intra-abdominal infections . This regimen has potential advantages over exclusively IV regimens, including the option of sequential IV/oral therapy, patient convenience, cost savings, and reduced hospital stay. Nippon Rinsho, 2004 Dec, 62(12), 2330 - 6 {Incidence, treatment and outcome of anaerobic bacteremia}; Watanabe K et al.; In this paper, we discussed the incidence and clinical significance of blood cultures positive for anaerobes . Anaerobic bacteria is uncommon, accounting for 0.5-13% of all positive cultures . The incidence depend on the institution . Recent findings of a multicenter prospective observational trials showed that in vitro activity of the agents directed at Bacteroides species reliably predicts outcome . When a positive anaerobic blood culture was noted, appropriate action must be taken, since negligence in this respect may seriously impair the outcome of patients. Biochem Biophys Res Commun, 2005 Jan 21, 326(3), 607 - 13 Bacteroides fragilis interferes with iNOS activity and leads to pore formation in macrophage surface; Vieira JM et al.; Bacteroides fragilis is the anaerobe most commonly recoverable from clinical specimens . The wide genetic diversity of this bacterium related with virulence potential is still an open question . In this study, we analyzed the morphological aspects and microbicide action of MO during interactions with B . fragilis . A filamentous cytoplasm content release and a different actin organization colocalized with iNOS were detected . It was also possible to observe the reduction of NO production in the same conditions . The scanning electron microscopy showed the formation of pore-like structures in the surface of macrophages in the bacterial presence and by transmission electron microscopy we could observe the extrusion of cytoplasm contents as well as the condensation of chromatin in the nucleus periphery . These data suggest the existence of an inhibitory mechanism developed by B . fragilis strains for one of the macrophage microbicide actions. J Clin Microbiol, 2004 Dec, 42(12), 5565 - 70 "Bacteroides nordii" sp . nov . and "Bacteroides salyersae" sp . nov . isolated from clinical specimens of human intestinal origin; Song YL et al.; Two groups of unknown bacteria, which phenotypically resemble members of the Bacteroides fragilis group but phylogenetically display >5% 16S rRNA gene sequence divergence from their nearest validly described species, Bacteroides thetaiotaomicron, were characterized by phenotypic and molecular taxonomic methods . Phylogenetically and phenotypically, the unidentified bacteria displayed a relatively close association with each other . However, a 16S rRNA gene sequence divergence of approximately 4% between the two unknown bacteria, as well as distinguishable biochemical characteristics, demonstrates that these organisms are genotypically and phenotypically distinct, and each group may represent a previously unknown subline within the Bacteroides phylogenetic cluster . Subsequent DNA-DNA hybridization studies confirmed that the two novel organisms were indeed distinct from each other . The previously described species closest to both of them is B . thetaiotaomicron (approximately 94% sequence similarity), but they can be differentiated easily from B . thetaiotaomicron by virtue of not utilizing trehalose . DNA-DNA pairing studies also documented the separateness of the unknown species and B . thetaiotaomicron . Based on the phenotypic and phylogenetic findings, two new species, "Bacteroides nordii" sp . nov . and "Bacteroides salyersae" sp . nov, are proposed . The G+C content of the DNA is 41.4 mol% for Bacteroides nordii and 42.0 mol% for Bacteroides salyersae . The type strains of Bacteroides nordii and Bacteroides salyersae are WAL 11050 (ATCC BAA-998 or CCUG 48943) and WAL 10018 (ATCC BAA-997 or CCUG 48945), respectively. Res Microbiol, 2004 Dec, 155(10), 843 - 6 Plasmid-related beta-lactamase production in Bacteroides fragilis strains; Nakano V et al.; Twenty Bacteroides fragilis group species isolated from children with and without diarrhea were analyzed . Antibiotic susceptibility was performed using an agar dilution method; beta-lactamase production was determined using a nitrocefin method, and plasmids were extracted using a commercial Miniprep System . MIC values ranged from 16 to 256 microg/ml for penicillin, 4-128 microg/ml for amoxicillin/clavulanic acid, 0.25-256 microg/ml for clindamycin, and 16-256 microg/ml for penicillin . beta-Lactamase was detected in all isolates . Only five isolates harbored plasmids varying from 7.8 to 1.8 kb . Loss of 6.4- and 3.8-kb plasmids in B . fragilis C68c was related to antibiotic resistance . Low molecular weight plasmids of 2.8-1.8 kb were stable . PCR amplification of cfiA and cepA genes was observed using total DNA, and the cfiA gene was also amplified from the 6.4-kb plasmid. Antonie Van Leeuwenhoek, 2004 Oct, 86(3), 263 - 81 16S rDNA library-based analysis of ruminal bacterial diversity; E Edwards J et al.; Bacterial 16S rDNA sequence data, incorporating sequences > 1 kb, were retrieved from published rumen library studies and public databases, then were combined and analysed to assess the diversity of the rumen microbial ecosystem as indicated by the pooled data . Low G+C Gram positive bacteria (54%) and the Cytophaga-Flexibacter-Bacteroides (40%) phyla were most abundantly represented . The diversity inferred by combining the datasets was much wider than inferred by individual studies, most likely due to different diets enriching for bacteria with different fermentative activities . A total of 341 operational taxonomic units (OTU) was predicted by the Chao1 non-parametric estimator approach . Phylogenetic and database analysis demonstrated that 89% of the diversity had greatest similarity to organisms which had not been cultivated, and that several sequences are likely to represent novel taxonomic groupings . Furthermore, of the 11% of the diversity represented by cultured isolates (> 95% 16S rDNA identity), not all of the bacteria were of ruminal origin . This study therefore reinforces the need to reconcile classical culture-based rumen microbiology with molecular ecological studies to determine the metabolic role of uncultivated species. Bioinformatics . 2004 Nov 11; {Epub ahead of print} Operon prediction without a training set; Westover BP et al.; MOTIVATION: Annotation of operons in a bacterial genome is an important step in determining an organism's transcriptional regulatory program . While extensive studies of operon structure have been carried out in a few species, such as E . coli, fewer resources exist to inform operon prediction in newly sequenced genomes . In particular, many extant operon finders require a large body of training examples to learn the properties of operons in the target organism . For newly sequenced genomes, such examples are generally not available; moreover, a model of operons trained on one species may not reflect the properties of other, distantly related organisms . We encountered these issues in the course of predicting operons in the genome of Bacteroides thetaiotaomicron, a common anaerobe that is a prominent component of the normal adult human intestinal microbial community . RESULTS: We describe an operon predictor designed to work without extensive training data.We rely on a small set of a priori assumptions about the properties of the genome being annotated that permit estimation of the probability that two adjacent genes lie in a common operon . Predictions integrate several sources of information, including intergenic distance, common functional annotation, and a novel formulation of conserved gene order . We validate our predictor both on the known operons of E . coli and on the genome of B . theta, using expression data to evaluate our predictions in the latter . AVAILABILITY: software is available online at http://www.cse.wustl.edu/~jbuhler/research/operons. Folia Microbiol (Praha), 2004, 49(4), 423 - 9 Soil microbial counts and identification of culturable bacteria in an extreme by arid zone; Su J et al.; Sixteen samples of two soil cores (about 550 and 180 cm in depth) were drilled at intervals in the lower reach of Heihe river basin (northwest of China) in order to illustrate soil microbial characteristics and diversity of culturable bacteria in an extreme by arid environment . Soil water content, organic matter, total nitrogen, pH, direct cell counts, and culturable microorganism counts were evaluated . The total cell concentration was 19-1120/microg (i.e . 0.19-11.2 x 10(8) per g) soil, the culturable bacteria count being 0.2-10.9 per microg (i.e . 2 x 10(5)-10.9 x 10(6) CFU/g) soil . The number of direct cell counts obtained by 4',6-diamidino-2-phenylindole-staining or the cound of culturable microbes after enrichment with different media were statistically significantly correlated with soil organic matters, total nitrogen content, soil water content and surface vegetation; this partly explained the larger number in the deeper first core than in the shallower one . As part of identification of 228 colonies isolated from the two cores, thirty-two were selected for 16S rDNA amplification, sequencing and molecular identification . These 32 isolates were affiliated to 5 major groups of bacteria: alpha-Proteobacteria, 5-Proteobacteria, gamma-Proteobacteria, the high-G+C G+-bacteria, the low-G+C G- -bacteria, and the Cytophaga-Flexibacter-Bacteroides group . Twenty-eight were rod- or short-rod shaped, which accounted for >87.5% of all species; only 4 of 32 species were cocci (<12.5%). J Obstet Gynaecol, 1997, 17(3), 290 - 2 The determination of phospholipase A2 enzyme activity in the vaginal secretions of pregnant and non-pregnant women with bacterial vaginosis-and in culture exudates of its causative organisms; M Jones And S Al-Mushrif B; Summary Bacterial vaginosis (BV) is presently being cited as a probable cause of premature labour, where it is thought that an abnormal excess of phospholipase A2 enzyme (PLA2 ), generated by infecting organisms, prematurely liberates prostaglandins, which trigger-off the labour process . PLA levels of pregnant and non-pregnant women, with and without BV infection were compared . The in vitro concentrations of PLA2 in broth cultures of infecting organisms were also measured . Mean PLA2 level in non-infected pregnant women was 777 units per mg but was raised to 1226 U/mg in those with BV ( P=0.001) . Mean level in non-infected normal women was 21 U/mg, but was raised to 97 U/mg in those having BV ( P=0.001) . PLA2 concentrations in broth cultures of the causative organisms showed that most Bacteroides strains produced the enzyme, having a mean concentration of 95 U/mg, but that it was generated by only 34% of Gardnerella vaginalis strains, their mean concentration being 32 U/mg. J Oral Sci, 2004 Sep, 46(3), 149 - 56 Intra-familial distribution of nine putative periodontopathogens in dental plaque samples analyzed by PCR; Okada M et al.; It is of great importance to understand the distribution of periodontopathogens within family members when considering the risk of periodontitis in children . The purpose of this study was to investigate the distribution of periodontopathogens among family members . We used the polymerase chain reaction method to test 4,8, and 7 probands with healthy gingiva, gingivitis, and periodontitis, respectively, and their 60 immediate family members . Plaque samples were collected from all erupted teeth sites using a sterile toothbrush . In 161 of the 165 positive cases, if a child harbored one of the periodontopathogens then at least one of the parents was also positive for the same bacterium . The prevalence of parent-child co-infection was 42.9% for Actinobacillus actinomycetemcomitans, 21.4% for Porphyromonas gingivalis, 29.2% for Treponema denticola, 59.5% for Tannerella forsythensis (Bacteroides forsythus) and 16.7% for Prevotella intermedia . Our results indicate that parents could be an important source of periodontopathogens for the colonization that occurs in their children. Antimicrob Agents Chemother, 2004 Nov, 48(11), 4195 - 9 Pharmacodynamics of pulse dosing versus standard dosing: in vitro metronidazole activity against Bacteroides fragilis and Bacteroides thetaiotaomicron; Ibrahim KH et al.; Pulse dosing is a novel approach to dosing that produces escalating antibiotic levels early in the dosing interval followed by a prolonged dose-free period . Antibiotic is frontloaded by means of four sequential bolus injections, after which antibiotic levels are allowed to diminish until the next dose . This study compares standard thrice-daily dosing and pulse dosing of metronidazole against Bacteroides spp . in an in vitro model . Two American Type Culture Collection Bacteroides fragilis isolates (metronidazole MIC for each organism = 1 mg/liter) were exposed to metronidazole for 48 or 96 h . Human pharmacokinetics were simulated for an oral 500-mg dose given every 8 h (maximum concentration of drug {C(max)} = 12 mg/liter; half-life = 8 h; area under the curve {AUC} = 294 mg . h/liter) and for pulse dosing . Pulses, each producing an increase in metronidazole concentration of 9 mg/liter, were administered at times 0, 2, 4, and 6 h of each 24-h cycle, with a targeted half-life of 8 h (AUC = 347 mg . h/liter) . A metronidazole-resistant B . fragilis strain (metronidazole MIC = 32 mg/liter) was exposed to both dosing regimens and, additionally, to a regimen of 1,500 mg administered once daily (C(max) = 36 mg/liter; AUC = 364 mg . h/liter) . Furthermore, regimens against one B . fragilis isolate and one B . thetaiotaomicron isolate corresponding to one-fourth and one-eighth of the thrice-daily and pulse dosing regimens, mimicking peak metronidazole concentrations achieved in abscesses, were simulated in 48-h experiments (metronidazole MIC = 1 mg/liter) . Time-kill curves were generated for each experiment and analyzed for bactericidal activity, defined as a bacterial burden reduction >/= 3 log(10) CFU/ml . The results of paired (Wilcoxon matched-pair signed-rank test) and nonpaired (Mann-Whitney test) statistical analyses conducted on time to 3 log(10) kill data and area under the kill curve data from each of the thrice-daily dosing experiments versus each of the pulse dosing experiments were considered not significant for a given isolate-dosing regimen combination . The thrice-daily dosing, pulse dosing, and once-daily dosing regimens all exhibited bactericidal activity . Metronidazole administered in standard or pulse dosing fashion was highly active against both susceptible and resistant strains of Bacteroides spp. Turk J Pediatr, 2004 Jul-Sep, 46(3), 279 - 82 Pyogenic liver abscesses in a child spreading to pulmonary and subcutaneous tissues: case report; Arslan N et al.; Pyogenic liver abscess is a rare and life-threatening disease in children . Our case is noteworthy because of the rapid advancement of liver abscesses without any other systemic disorder . A 16-year-old girl was admitted to the hospital with fatigue, pallor, weight loss and high fever . In physical examination a fluctuating mass was observed under the scapular area and hepatosplenomegaly was found . In computed tomography, three septated cystic lesions which looked like abscesses were demonstrated in the liver . The abscess was drained through percutaneous route . Right pleural empyema with clinical features of adult respiratory distress syndrome appeared after the first day of treatment . Bacteroides sp . was isolated from pus . On the twentieth day of the therapy, control abdominal computed tomography revealed two new abscesses in the liver . They were drained and the antibiotic therapy was continued with ticarcillin-clavulanate, fluconazole and levofloxacin . By the end of the first week of the therapy, the fever of the patient had abated . This therapy was continued for four weeks; 15 days after the end of the therapy there was prominent healing of the liver lesions with only one necrotic remnant 2 cm in diameter on abdominal computed tomography. J Biol Chem, 2004 Dec 31, 279(53), 55840 - 9 Epub 2004 Oct 18. Structural basis of 5-nitroimidazole antibiotic resistance: the crystal structure of NimA from Deinococcus radiodurans; Leiros HK et al.; 5-Nitroimidazole-based antibiotics are compounds extensively used for treating infections in humans and animals caused by several important pathogens . They are administered as prodrugs, and their activation depends upon an anaerobic 1-electron reduction of the nitro group by a reduction pathway in the cells . Bacterial resistance toward these drugs is thought to be caused by decreased drug uptake and/or an altered reduction efficiency . One class of resistant strains, identified in Bacteroides, has been shown to carry Nim genes (NimA, -B, -C, -D, and -E), which encode for reductases that convert the nitro group on the antibiotic into a non-bactericidal amine . In this paper, we have described the crystal structure of NimA from Deinococcus radiodurans (drNimA) at 1.6 A resolution . We have shown that drNimA is a homodimer in which each monomer adopts a beta-barrel fold . We have identified the catalytically important His-71 along with the cofactor pyruvate and antibiotic binding sites, all of which are found at the monomer-monomer interface . We have reported three additional crystal structures of drNimA, one in which the antibiotic metronidazole is bound to the protein, one with pyruvate covalently bound to His-71, and one with lactate covalently bound to His-71 . Based on these structures, a reaction mechanism has been proposed in which the 2-electron reduction of the antibiotic prevents accumulation of the toxic nitro radical . This mechanism suggests that Nim proteins form a new class of reductases, conferring resistance against 5-nitroimidazole-based antibiotics. J Periodontal Res, 2004 Dec, 39(6), 442 - 6 Periodontal pathogens in atheromatous plaques . A controlled clinical and laboratory trial; Cairo F et al.; OBJECTIVE: A possible relationship between periodontitis and cardiovascular disease has been suggested . The aims of this controlled clinical study were: (i) to ascertain the presence of periodontal bacteria DNA {Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis (formerly Bacteroides forsythus)} in carotid atheromatous plaques and (ii) to assess the concomitant presence of the same periodontal bacteria DNA, if any, in periodontal pockets and in carotid atheroma in the same patient . METHODS: A total of 52 patients scheduled for carotid endarderectomy were enrolled in this study . The test group consisted of 26 dentate patients; the control group included 26 edentulous patients . A complete periodontal examination, including radiographic orthopanoramic and subgingival plaque sample, was performed in the test population . Oral and X-ray examinations were performed in the control group . Atheromatous plaques were harvested during surgical procedure for each dentate and edentulous patient and then sent to the microbiological laboratory . Subgingival plaque samples and carotid specimens were examined using the polymerase chain reaction (PCR) technique by means of specific primers for periodontal bacteria . Amplification of extracted DNA was tested using human beta-globin specific-primers . RESULTS: Out of 52 endarterectomy samples, 12 (seven dentate, five edentulous patients) were excluded as negative to DNA amplification . In subgingival plaque samples of 19 test patients, T . forsythensis (79%), F . nucleatum (63%), P . intermedia (53%), P . gingivalis (37%) and A . actinomycetemcomitans (5%) were found . No periodontal bacteria DNA was detected by PCR in any of the carotid samples in either patient group . CONCLUSION: The presence of periodontal bacteria DNA in atheromatous plaques could not be confirmed by this study and thus no correlation could be established between species associated with periodontal disease and putative bacteria contributing to atheromatous plaques . (c)Blackwell Munksgaard 2004 Dig Dis Sci, 2004 Sep, 49(9), 1425 - 32 Are Helicobacter species and enterotoxigenic Bacteroides fragilis involved in inflammatory bowel disease? Basset C, Holton J, Bazeos A, Vaira D, Bloom S. The aim of this study was to determine if either Helicobacter or enterotoxigenic Bacteroidesfragilis (ETBF) was linked to inflammatory bowel disease (IBD), using PCR . We analyzed the luminal washings and colonic biopsies of 35 patients with IBD and 37 control patients . The presence of Helicobacter was confirmed in the luminal washing of one IBD patient and three control patients and in the biopsies of two IBD patients . Ten of 28 control patients and 8 of 32 IBD patients had a positive luminal washing for the enterotoxin gene . Six of 33 control patients and 4 of 32 IBD patients had positive biopsies . The prevalence of the enterotoxin gene was higher in IBD patients with active disease compared with patients with inactive disease, although it did not achieve statistical significance . In conclusion, Helicobacter was not associated with IBD in our population of patients, although ETBF may be associated with active disease. An Med Interna, 2004 Sep, 21(9), 425 - 32 {Anaerobic bacteriemias: clinical and epidemiological characteristics of anaerobic bacteremias in ten years}; Ruiz-Giardin JM et al.; BACKGROUND: The anaerobic bacteremia incidence is decreasing during the last years, and although it carries a high mortality rate there are studies that ask about the possibility of avoid anaerobic blood cultures thinking about the clinical prediction capacity of them . The objective of this study is the analysis of incidence and clinical characteristics of anaerobic bacteremias in two periods with 10 years of difference and empiric antibiotic treatment received, and if it was changed when microbiological results were received . METHODS: Prospective study of anaerobic bacteremias during 1985-86 and 1996-97, at university hospital analyzing clinical characteristics, incidence, analysis, evolution and empiric antibiotic treatment received . The statistical analysis was performed with the chi square test or exact Fisher test with statistical significance with p<0.05, talking about variables with p<0.10 . RESULTS: The incidence of anaerobic bacteremias was higher in 1996-97 with 24 cases (5.08%), that during 1985-86 with 22 cases (4.24%) . The variables with statistical significative differences with p<0.10 were: intrahospitalary adquisition (p<0.10); genitourinary and vascular manipulation (p=0.02 and p=0.06), and hypotension (p=0.034) more frequent during the first period than during the second one . There isn t statistical difference in evolution to cure although the percentage was higher during the second period (62.5%), that during the first one (54.6%), being the empiric treatment very high in both periods . Bacteroides fragilis was the most frequent microorganism and abdominal origin the most frequent one in both periods . CONCLUSION: Although there is a low anaerobic bacteremia incidence this one hasn't diminished in ten years . Anaerobic bacteremias have a high mortality index although the most part of empirical antibiotic treatments are correct . It could ask about the possibility of avoid anaerobic blood cultures or ask about them only in clinic suspicion of them. Proc Natl Acad Sci U S A, 2004 Oct 12, 101(41), 14919 - 24 Epub 2004 Oct 04. Genomic analysis of Bacteroides fragilis reveals extensive DNA inversions regulating cell surface adaptation; Kuwahara T et al.; Bacteroides are predominant human colonic commensals, but the principal pathogenic species, Bacteroides fragilis (BF), lives closely associated with the mucosal surface, whereas a second major species, Bacteroides thetaiotaomicron (BT), concentrates within the colon . We find corresponding differences in their genomes, based on determination of the genome sequence of BF and comparative analysis with BT . Both species have acquired two mechanisms that contribute to their dominance among the colonic microbiota: an exceptional capability to use a wide range of dietary polysaccharides by gene amplification and the capacity to create variable surface antigenicities by multiple DNA inversion systems . However, the gene amplification for polysaccharide assimilation is more developed in BT, in keeping with its internal localization . In contrast, external antigenic structures can be changed more systematically in BF . Thereby, at the mucosal surface, where microbes encounter continuous attack by host defenses, BF evasion of the immune system is favored, and its colonization and infectious potential are increased. Appl Environ Microbiol, 2004 Oct, 70(10), 5955 - 62 Bacterial community associated with black band disease in corals; Frias-Lopez J et al.; Black band disease (BBD) is a virulent polymicrobial disease primarily affecting massive-framework-building species of scleractinian corals . While it has been well established that the BBD bacterial mat is dominated by a cyanobacterium, the quantitative composition of the BBD bacterial mat community has not described previously . Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to characterize the infectious bacterial community of the bacterial mat causing BBD . These analyses revealed that the bacterial composition of the BBD mat does not vary between different coral species but does vary when different species of cyanobacteria are dominant within the mat . On the basis of the results of a new method developed to identify organisms detected by T-RFLP analysis, our data show that besides the cyanobacterium, five species of the division Firmicutes, two species of the Cytophaga-Flexibacter-Bacteroides (CFB) group, and one species of delta-proteobacteria are also consistently abundant within the infectious mat . Of these dominant taxa, six were consistently detected in healthy corals . However, four of the six were found in much higher numbers in BBD mats than in healthy corals . One species of the CFB group and one species of Firmicutes were not always associated with the bacterial communities present in healthy corals . Of the eight dominant bacteria identified, two species were previously found in clone libraries obtained from BBD samples; however, these were not previously recognized as important . Furthermore, despite having been described as an important component of the pathogenetic mat, a Beggiatoa species was not detected in any of the samples analyzed . These results will permit the dominant BBD bacteria to be targeted for isolation and culturing experiments aimed at deciphering the disease etiology. Appl Environ Microbiol, 2004 Oct, 70(10), 5853 - 8 Characterization of plasmid pOR1 from Ornithobacterium rhinotracheale and construction of a shuttle plasmid; Jansen R et al.; The bacterium Ornithobacterium rhinotracheale has been recognized as an emerging pathogen in poultry since about 10 years ago . Knowledge of this bacterium and its mechanisms of virulence is still very limited . Here we report the development of a transformation system that enables genetic modification of O . rhinotracheale . The system is based on a cryptic plasmid, pOR1, that was derived from an O . rhinotracheale strain of serotype K . Sequencing indicated that the plasmid consisted of 14,787 nucleotides . Sequence analysis revealed one replication origin and several rep genes that control plasmid replication and copy number, respectively . In addition, pOR1 contains genes with similarity to a heavy-metal-transporting ATPase, a TonB-linked siderophore receptor, and a laccase . Reverse transcription-PCR demonstrated that these genes were transcribed . Other putative open reading frames exhibited similarities with a virulence-associated protein in Actinobacillus actinomycetemcomitans and a number of genes coding for proteins with unknown function . An Escherichia coli-O . rhinotracheale shuttle plasmid (pOREC1) was constructed by cloning the replication origin and rep genes from pOR1 and the cfxA gene from Bacteroides vulgatus, which codes for resistance to the antibiotic cefoxitin, into plasmid pGEM7 by using E . coli as a host . pOREC1 was electroporated into O . rhinotracheale and yielded cefoxitin-resistant transformants . The pOREC1 isolated from these transformants was reintroduced into E . coli, demonstrating that pOREC1 acts as an independent replicon in both E . coli and O . rhinotracheale, fulfilling the criteria for a shuttle plasmid that can be used for transformation, targeted mutagenesis, and the construction of defined attenuated vaccine strains. J Biol Chem, 2004 Dec 10, 279(50), 52346 - 52 Epub 2004 Dec 10. TetX is a flavin-dependent monooxygenase conferring resistance to tetracycline antibiotics; Yang W et al.; The tetracycline antibiotics block microbial translation and constitute an important group of antimicrobial agents that find broad clinical utility . Resistance to this class of antibiotics is primarily the result of active efflux or ribosomal protection; however, a novel mechanism of resistance has been reported to be oxygen-dependent destruction of the drugs catalyzed by the enzyme TetX . Paradoxically, the tetX genes have been identified on transposable elements found in anaerobic bacteria of the genus Bacteroides . Overexpression of recombinant TetX in Escherichia coli followed by protein purification revealed a stoichiometric complex with flavin adenine dinucleotide . Reconstitution of in vitro enzyme activity demonstrated a broad tetracycline antibiotic spectrum and a requirement for molecular oxygen and NADPH in antibiotic degradation . The tetracycline products of TetX activity were unstable at neutral pH, but mass spectral and NMR characterization under acidic conditions supported initial monohydroxylation at position 11a followed by intramolecular cyclization and non-enzymatic breakdown to other undefined products . TetX is therefore a FAD-dependent monooxygenase . The enzyme not only catalyzed efficient degradation of a broad range of tetracycline analogues but also conferred resistance to these antibiotics in vivo . This is the first molecular characterization of an antibiotic-inactivating monooxygenase, the origins of which may lie in environmental bacteria. Infect Immun, 2004 Oct, 72(10), 5832 - 9 Bacteroides fragilis enterotoxin induces intestinal epithelial cell secretion of interleukin-8 through mitogen-activated protein kinases and a tyrosine kinase-regulated nuclear factor-kappaB pathway; Wu S et al.; Enterotoxigenic Bacteroides fragilis (ETBF) secretes a 20-kDa metalloprotease toxin termed B . fragilis toxin (BFT) . ETBF disease in animals is associated with an acute inflammatory response in the intestinal mucosa, and lethal hemorrhagic colitis may occur in rabbits . In this study, we confirmed recent reports (J . M . Kim, Y . K . Oh, Y . J . Kim, H . B . Oh, and Y . J . Cho, Clin . Exp . Immunol . 123:421-427, 2001; L . Sanfilippo, C . K . Li, R . Seth, T . J . Balwin, M . J . Menozzi, and Y . R . Mahida, Clin . Exp . Immunol . 119:456-463, 2000) that purified BFT stimulates interleukin-8 (IL-8) secretion by human intestinal epithelial cells (HT29/C1 cells) and demonstrate that stimulation of IL-8 production is dependent on biologically active BFT and independent of serum . Induction of IL-8 mRNA expression occurs rapidly and ceases by 6 h after BFT treatment, whereas IL-8 secretion continues to increase for at least 18 h . Our data suggest that BFT-stimulated IL-8 secretion involves tyrosine kinase-dependent activation of nuclear factor-kappaB (NF-kappaB) as well as activation of the mitogen-activated protein kinases (MAPKs), p38 and extracellular signal-related kinase . Simultaneous activation of NF-kappaB and MAPKs appears necessary for secretion of IL-8 by HT29/C1 cells treated with BFT. J Clin Microbiol, 2004 Sep, 42(9), 4127 - 9 Isolation of metronidazole-resistant Bacteroides fragilis carrying the nimA nitroreductase gene from a patient in Washington State; Schapiro JM et al.; Members of the Bacteroides fragilis group are among the most common anaerobic bacterial isolates in clinical specimens . Metronidazole, a 5-nitroimidazole, is often used as empirical therapy for anaerobic infections . Susceptibility testing is not routinely performed because of nearly universal susceptibility of Bacteroides spp . to this agent . We report a case of metronidazole-resistant Bacteroides fragilis in the United States and demonstrate the presence of the nimA gene, encoding a nitroreductase previously shown to mediate resistance to 5-nitroimidazole antimicrobial agents in B . fragilis strains from Europe and Africa . Because clinical failures in Bacteroides infections have been associated with the use of inactive antimicrobial agents, clinicians need to be aware of the possibility of metronidazole-resistant B . fragilis strains in the United States and the importance of susceptibility testing in selected situations. Eur J Obstet Gynecol Reprod Biol, 2004 Oct 15, 116(2), 152 - 6 Relationship between a toll-like receptor-4 gene polymorphism, bacterial vaginosis-related flora and vaginal cytokine responses in pregnant women; Genc MR et al.; OBJECTIVE: The relationship between a single nucleotide polymorphism (TLR4 896 A > G) in the toll-like receptor-4 (TLR4) gene, qualitative and quantitative changes in vaginal micro-flora and vaginal interleukin (IL)-1beta and IL-1 receptor antagonist (IL-1ra) concentrations in pregnant women were evaluated . STUDY DESIGN: Qualitative and quantitative microbial methods were used to characterize vaginal micro-flora of 238 women at 18-22 weeks gestation . Polymerase chain reaction was used to determine TLR4 genotype . IL-1beta and IL-1ra concentrations in vaginal lavage samples were measured by ELISA . RESULTS: The TLR4 variant was identified in 10.3% of women . Carriage of this variant was associated with a median increase in vaginal pH (P = 0.05), a greater than 10-fold increase in vaginal Gardnerella vaginalis levels (P < 0.0001) and a 10-fold increase in the vaginal concentration of three species of anaerobic Gram-negative rods, Prevotella, Bacteroides, and Porphyromonas (P = 0.08 ) . Colonization with G . vaginalis and/or the anaerobic Gram-negative rods resulted in elevated vaginal IL-1 (P = 0.01) and IL-1ra (P < 0.0002) concentrations in women who were TLR4 896A homozygotes, but not in TLR4 896G carriers . CONCLUSION: The TLR4 896 A > G polymorphism contributes to inter-individual differences in the vaginal immune defense against G . vaginalis and anaerobic Gram-negative rods. Am J Orthod Dentofacial Orthop, 2004 Sep, 126(3), 363 - 6 Clinical and microbiologic changes after removal of orthodontic appliances; Sallum EJ et al.; The goal of this study was to evaluate the clinical and microbiological factors associated with orthodontic appliances during an episode of gingival inflammation and the impact of appliance removal on periodontal health . This prospective study included 10 patients, aged 12 to 20 years, with clinical signs of gingival inflammation at the final phase of orthodontic treatment (appliance removal) . Plaque index, gingival index, and probing depth were evaluated, and microbiological samples were collected from teeth 16, 11, and 26 at 2 times: during the gingival inflammation (baseline) and 30 days after the removal of the appliance and professional prophylaxis . Polymerase chain reaction analysis was used to detect Porphyromonas gingivalis, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Prevotella nigrescens . A statistically significant improvement of the plaque and gingival indexes was seen, as well as a reduction in probing depth (P <.05) . Periodontal pathogens were associated with gingival inflammation during orthodontic treatment . The improvement in periodontal health at 30 days was concomitant with a reduction of sites positive for A . actinomycetemcomitans and B . forsythus (P <.05) . Periodontal pathogens associated with gingival inflammation during orthodontic treatment can be significantly reduced by orthodontic appliance removal and professional prophylaxis. J Bacteriol, 2004 Sep, 186(18), 6077 - 92 The Bacteroides fragilis pathogenicity island is contained in a putative novel conjugative transposon; Franco AA; The genetic element flanking the Bacteroides fragilis pathogenicity island (BfPAI) in enterotoxigenic B . fragilis (ETBF) strain 86-5443-2-2 and a related genetic element in NCTC 9343 were characterized . The results suggested that these genetic elements are members of a new family of conjugative transposons (CTns) not described previously . These putative CTns, designated CTn86 and CTn9343 for ETBF 86-5443-2-2 and NCTC 9343, respectively, differ from previously described Bacteroides species CTns in a number of ways . These new transposons do not carry tetQ, and the excision from the chromosome to form a circular intermediate is not regulated by tetracycline; they are predicted to differ in their mechanism of transposition; and their sequences have very limited similarity with CTnDOT or other described CTns . CTn9343 is 64,229 bp in length, contains 61 potential open reading frames, and both ends contain IS21 transposases . Colony blot hybridization, PCR, and sequence analysis indicated that CTn86 has the same structure as CTn9343 except that CTn86 lacks a approximately 7-kb region containing truncated integrase (int2) and rteA genes and it contains the BfPAI integrated between the mob region and the bfmC gene . If these putative CTns were to be demonstrated to be transmissible, this would suggest that the bft gene can be transferred from ETBF to nontoxigenic B . fragilis strains by a mechanism similar to that for the spread of antibiotic resistance genes. Plasmid, 2004 Sep, 52(2), 119 - 30 Factors required in vitro for excision of the Bacteroides conjugative transposon, CTnDOT; Sutanto Y et al.; Four genes have been found to be essential for excision of the Bacteroides conjugative transposon CTnDOT in vivo: intDOT, orf2c, orf2d, and exc . The intDOT gene encodes an integrase that is essential for integration and excision . The function of the other genes is still uncertain . Previously, we developed an in vitro system for the integration reaction . We have now developed an in vitro system for excision . In this system, the left and right junctions of CTnDOT, attL, and attR, are provided on separate plasmids . The excision reaction produced a cointegrate which contained the attDOT (the joined ends of CTnDOT) and attB (the chromosomal target site) . Cointegrate formation was observed after electroporation of Escherichia coli with the assay mixture and was also detected directly in the assay mixture by Southern hybridization . The highest reaction frequencies (10(-3)) were obtained with a mixture that contained purified IntDOT and a cell extract from Bacteroides thetaiotaomicron 4001, which contained the excision region of CTnDOT carried on a plasmid . An unexpected finding was that the addition of purified Exc, which is essential for excision in vivo, was not required for excision in vitro, nor did it increase the frequency of cointegrate formation. Acta Microbiol Pol, 2004, 53(1), 35 - 9 Enterotoxigenic Bacteroides fragilis (ETBF) strains isolated in The Netherlands and Poland are genetically diverse; Obuch-Woszczatynski P et al.; Gram-negative anaerobic rods isolated in The Netherlands and Poland from extraintestinal and intestinal sources were identified as Bacteroides fragilis (n = 210) on the basis of Gram staining, growth on selective Bacteroides Bile Esculine medium as black colonies, and biochemical characteristics . PCR-mediated assessment of the presence of the B . fragilis enterotoxin (fragilysin) gene in all strains identified 12 so-called enterotoxin-positive B . fragilis (ETBF) strains (15%) among the Dutch strains and 16 ETBF among the Polish strains (13%) . NotI Pulsed Field Gel Electrophoresis (PFGE) analysis revealed that these strains are genetically heterogeneous . Among the Dutch strains an identical pair and a set of four indiscriminate strains were identified . This suggests that limited nosocomial spread of ETBF can be observed . However, there was no identity obeserved when strains from The Netherlands were compared to their Polish counterparts . The antimicrobial susceptibility testing revealed that one Polish strain isolated from a patient with antibiotic associated diarrhoeae (AAD) was simultaneously highly resistant to clindamycin and cefoxitin (MIC>256 mg/L) . Two other strains appeared to be clindamycin resistant . All resistant strains had different PFGE patterns, suggesting that resistance development occurred at independent occassions. Infez Med, 2004 Mar, 12(1), 44 - 50 Efficacy of Telithromycin in the treatment of experimental Bacteroides fragilis intraabdominal abscess in the senescent mice; Thadepalli H et al.; The efficacy of telithromycin (HMR 3647), a new ketolide, in the treatment of experimental Bacteroides fragilis intraabdominal abscess in young and senescent mice was evaluated . Two different age groups of mice, young (2-3 months) and senescent (18-24 months) were used in this study . Telithromycin (50mg/kg/bid) was compared with clindamycin and metronidazole, both administered in 100 mg/kg/bid doses . Telithromycin cured the infection in 74% of the young and 67% of the old mice but this difference was not significant . Telithromycin efficacy was comparable to that of clindamycin which cured 82% of the young and 75% of the old, but was superior to the efficacy of metronidazole, which cured 61% of the young and 50% of the senescent mice . Young animals that were not cured by any of the three antibiotics showed decrease in the viable bacterial cell counts by two logs while the senescent mice had a one log difference . Serum, pus and tissue concentrations of telithromycin were five-fold higher in the old mice than in the young . Age by itself had no adverse effect on therapeutic outcome of any of the three antibiotics used. Int Endod J, 2004 Sep, 37(9), 588 - 92 Induction of vascular endothelial growth factor expression in human pulp fibroblasts stimulated with black-pigmented Bacteroides; Yang LC et al.; AIM: To investigate the effect of black-pigmented Bacteroides on the expression of vascular endothelial growth factor (VEGF) gene in human pulp fibroblasts . METHODOLOGY: The supernatants of Porphyromonas endodontalis, Porphyromonas gingivalis and Prevotella intermedia were used to evaluate VEGF gene expression in human pulp fibroblasts . The levels of mRNAs were measured by the quantitative reverse-transcriptase polymerase chain reaction analysis . RESULTS: Black-pigmented Bacteroides induced significantly high levels of VEGF mRNA gene expression in human pulp fibroblasts (P < 0.05) . In addition, the expression of VEGF depended on the bacteria tested . CONCLUSIONS: Black-pigmented Bacteroides may be involved in developing pulpal disease through the stimulation of VEGF production that would lead to the expansion of the vascular network coincident to progression of the inflammation. Res Microbiol, 2004 Sep, 155(7), 522 - 4 Non-toxigenic pattern II and III Bacteroides fragilis strains: coexistence in the same host; Antunes EN et al.; Diarrhoeic stool samples from 334 0-5-year-old children were analysed with respect to the incidence of Bacteroides fragilis as well as other enteropathogens . B . fragilis was recovered in 9.3% (31/334) of the samples, and 79 strains were examined for the presence of the bft gene or the BfPAI flanking region using polymerase chain reaction assays . No enterotoxigenic B . fragilis strains were detected . In 29% (9/31) of the samples the coexistence of both II and III non-toxigenic B . fragilis (NTBF) patterns could be seen . In 51.6% (16/31) of the samples there existed a pattern II NTBF only, and in 19.4% (6/31) only pattern III could be detected . Strains from the same patient representing different patterns were submitted to pulsed-field gel electrophoresis assays . Fingerprints obtained by this technique showed that there was strong heterogeneity among strains from different individuals . However, different patterns from the same individual shared 100% similarity. Acta Crystallogr D Biol Crystallogr, 1997 Jul, 53(Pt 4), 485 - 7 Purification, crystallization and preliminary X-ray analysis of Bacteroides fragilis Zn(2+) beta-lactamase; Carfi A; The Zn(2+) beta-lactamase from Bacteroides fragilis (E.C . 3.5.2.6) was overexpressed in Escherichia coli using an isopropylthiogalactoside-inducible T7 RNA polymerase expression system . Crystallization trials by the hanging-drop vapour-diffusion method have yielded two different crystal forms from two slightly different conditions . Crystals of form I belong to the monoclinic space group C2 with unit-cell dimensions a = 56.03, b = 43.98, c = 105.32 A, beta = 112 degrees and diffracted only up to 4.0 A . Crystals of form II are orthorhombic, space group P2(1)2(1)2(1) with unit-cell dimensions a = 48.10, b = 98.05, c = 111.76 A, diffract to at least 2.0 A and are suitable for high-resolution structural analysis. Hua Xi Kou Qiang Yi Xue Za Zhi, 2004 Jun, 22(3), 177 - 9 {Separation of Bacteroides forsythus ATCC43037 proteins by horizontal two-dimensional gel electrophoresis}; Huang DM et al.; OBJECTIVE: To set up a rapid, efficient, reliable and accurate method for separation of Bacteriodes forsythus proteins . METHODS: Bacteroides forsythus ATCC43037 cells were harvested by centrifugation, washed in Tris buffer to remove excess medium, and lysed by sonication . The sonicated lysis proteins were extracted step by step with ReadyPrep Sequential Extraction Kit (Bio-Rad) . The supernatant of B . forsythus proteins were used for two-dimensional gel electrophoresis . The first dimension IEF (isoelectric focusing) was run with Immobiline DryStrip (pH 3-10) and the second dimension SDS-PAGE was run with Excelgel SDS, gradient 8-18 precast gel and buffer strips . The separated proteins were stained with Coomassie Brilliant Blue and silver staining kit (Plusone Silver Staining Kit, Protein, Pharmacia Biotech) . RESULTS: 1 . The protein spots were clear when sample cups were used in the middle of IPG strip during IEF . 2 . B . forsythus proteins were separated clearly by horizontal two-dimensional electrophoresis and most of B . forsythus whole-cell proteins were acidic proteins (P13-7) . CONCLUSION: Horizontal two-dimension electrophoresis is a useful method for separating B . forsythus proteins. Int J Antimicrob Agents, 2004 Aug, 24(2), 135 - 43 A Bacteroides thetaiotamicron porin that could take part in resistance to beta-lactams; Behra-Miellet J et al.; The aim of this study was to investigate porin absence or deficiency in two Bacteroides thetaiotaomicron strains resistant to amoxicillin combined with clavulanic acid . Their outer membrane protein (OMP) extracts and those of two susceptible strains were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and compared to detect differences between the strains . A protein band of interest at around 70 kDa electro-eluted for each strain, was tested in a liposome swelling assay . A decrease in initial absorbency was noted for the two susceptible strains but not for the two resistant strains . The liposome swelling of the two susceptible strains was directly visualized by photon microscopy and then photographed . This suggested a B . thetaiotaomicron porin of around 70 kDa could take part in resistance to beta-lactams. Mem Inst Oswaldo Cruz, 2004 May, 99(3), 319 - 24 Epub 2004 Jul 19. Survey of antimicrobial susceptibility patterns of the bacteria of the Bacteroides fragilis group isolated from the intestinal tract of children; Nakano V et al.; The bacteria of the Bacteroides fragilis group are considered important clinical pathogens and they are the most common anaerobes isolated from human endogenous infections . In this study, the susceptibility patterns to antibiotics and metals of 114 species of the B . fragilis group isolated from children with and without diarrhea were determined . Susceptibility was assayed by using an agar dilution method with Wilkins-Chalgren agar . All B . fragilis strains were resistant to lead and nickel, but susceptible to metronidazole and imipenem . beta-lactamase production was detected by using biological and nitrocefin methods, respectively, in 50% and 90.6% of the isolates of children with diarrhea and in 60% and 90% of the isolates of children without diarrhea . Our results show an increase of antibiotics and metals resistance in this microbial group, and a periodic evaluation of the antimicrobial susceptibility is needed . In Brazil, the contamination for antibiotics or metal ions is often observed, and it is suggested an increase the antimicrobial resistance surveillance of this microbial group, mainly those isolated from children's diarrhea. Mem Inst Oswaldo Cruz, 2004 May, 99(3), 307 - 12 Epub 2004 Jul 19. Virulence markers and antimicrobial susceptibility of bacteria of the Bacteroides fragilis group isolated from stool of children with diarrhea in São Paulo, Brazil; Nakano V et al.; Bacteroides fragilis has been isolated from several human and non-human monomicrobial and mixed infections . In this study, some virulence markers and the antimicrobial susceptibility of bacteria of the B . fragilis group isolated from children's stools were evaluated . All the 64 isolates showed the following characteristics: capsulated, beta-hemolytic, hydrophilic, and serum-resistant . Only, 24 (37.5%) strains were resistant at 60 masculine C, for 30 min, and among them, 12 (18.75%) were resistant at 60 masculine C, for 60 min . Also, none strain was resistant at 100 masculine C . Four strains were able to hemagglutinate erythrocytes and D-mannose, D-galactose, D-arabinose, and D-xylose inhibited hemagglutination in 2 B . fragilis strains (p76a, p76b) . The hemagglutination in the strain B . uniformis p3-2 was inhibited by D-xylose and D-galactose . The bft gene detection and the enterotoxin production were observed only in 13 EF-enterotoxigenic species . Fragilysin activity was confirmed on HT-29 cells . The antimicrobial determination confirmed that both imipenem and metronidazole were efficient against B . fragilis species; all the strains were resistant to lead and nickel . Plasmids of 2.9, 4.4, 4.8, and 8.9 kb were observed in 6 tested strains . These results show the values of the species identification from clinical infections, as well as of the periodic evaluation of the resistance patterns of the B . fragilis group at Brazilian medical institutions. J Adolesc Health, 2004 Aug, 35(2), 156 - 8 Allergic symptoms and microflora in schoolchildren; Fukuda S et al.; We studied 867 junior high school children and administered a questionnaire documenting allergic symptoms and environmental variables, and measured Immunoglobulin E serum levels and the immunoglobulin G titers of serum antibody to microflora . A total of 716 subjects were ultimately used for statistics; those with at least two of the following allergic symptoms: asthma, rhinitis, eczema, or food allergy, showed significantly higher IgG titers to Bactroides vulgatus than other groups . This finding suggests that a species of the Bacteroides genus of the intestinal microflora tends to affect the gut issues, but further studies are needed to clarify this. Microbiology, 2004 Jul, 150(Pt 7), 2125 - 34 Transcriptional regulation of the Bacteroides fragilis ferritin gene (ftnA) by redox stress; Rocha ER et al.; This study shows that the iron-storage protein ferritin is a component of the redox-stress response in the obligate anaerobe Bacteroides fragilis . It is up-regulated at transcriptional level under aerobic conditions but constitutively expressed at low levels under anaerobic conditions . Northern hybridization and primer extension analysis revealed that ftnA is transcribed as a monocistronic mRNA of approximately 600 nt . Under reduced anaerobic conditions, ftnA mRNA levels were not dependent on the iron content of the culture medium . Following oxygen exposure ftnA message increased about 10-fold in iron-replete medium compared to a fourfold increase under low-iron conditions . Addition of the oxidant potassium ferricyanide induced expression of ftnA mRNA anaerobically, suggesting that the oxidation of the medium affected expression of ftnA . Two transcription initiation start sites were identified . Both transcripts were expressed constitutively under anaerobic conditions but one promoter was induced by oxidative stress or the addition of the oxidant potassium ferricyanide . The effect of redox stress on ftnA expression was further investigated by addition of diamide, a thiol-oxidizing agent, which induced ftnA mRNA levels anaerobically, suggesting that an unbalanced cellular redox state also affects ftnA expression . Induction by hydrogen peroxide and oxygen was decreased in an oxyR deletion mutant but some oxygen induction still occurred . This strongly suggests that ftnA is regulated by both the peroxide response transcriptional activator, OxyR, and another unidentified oxygen-dependent regulator . Taken together, these data show that ftnA mRNA levels are controlled by both iron and oxidative stress; this coordinated regulation may be important for survival in an adverse aerobic environment. Folia Microbiol (Praha), 2004, 49(2), 151 - 5 The effects of plant extracts on microbial community structure in a rumen-simulating continuous-culture system as revealed by molecular profiling; Ferme D et al.; An in vitro study in dual-flow continuous-culture fermentors was conducted with two different concentrations of monensin, cinnamaldehyde or garlic extract added to 1:1 forage-to-concentrate diet in order to determine their effects on selected rumen bacterial populations . Samples were subjected to total DNA extraction, restriction analysis of PCR amplified parts of 16S rRNA genes (ARDRA) and subsequent analysis of the restriction profiles by lab-on-chip technology with the Agilent's Bioanalyser 2100 . Eub338-BacPre primer pair was used to select for the bacteria from the genera Bacteroides, Porphyromonas and Prevotella, especially the latter representing the dominant Gram-negative bacterial population in the rumen . Preliminary results of HaeIII restriction analysis show that the effects of monensin, cinnamaldehyde and garlic extract on the BacPre targeted ruminal bacteria are somewhat different in regard to targeted populations and to the nature of the effect . Garlic extract was found to trigger the most intensive changes in the structure of the BacPre targeted population . Comparison of the in silico restriction analysis of BacPre sequences deposited in different DNA databanks and of the results of performed amplified ribosomal DNA restriction analysis showed differences between the predicted and obtained HaeIII restriction profiles, and suggested the presence of novel, still unknown Prevotella populations in studied samples. Int J Antimicrob Agents, 2004 Jul, 24(1), 53 - 8 Determinants of resistance in Bacteroides fragilis strains according to recent Brazilian profiles of antimicrobial susceptibility; Paula GR et al.; Susceptibility profiles of 99 Bacteroides fragilis strains for 9 antimicrobial agents were defined by using an agar dilution method . The isolates were uniformly susceptible to imipenen and metronidazole . All isolates were resistant to ampicillin . The resistance rates to amoxicillin/clavulanate, cefoxitin, cefotaxime, chloramphenicol, clindamycin and tetracycline were 3.0, 12.1, 15.1, 1.0, 18.2 and 75.7%, respectively . Sixteen strains showed reduced susceptibility to metronidazole (MIC 2-4 mg/L) but none had nim genes using PCR . All strains were also investigated for the presence of cepA, cfiA, cfxA, ermF and tetQ genes by PCR methodology and 92.9, 4.9, 24.2, 2 and 64.6% of the strains were respectively found positive . These results reflect the importance of surveys of susceptibility profiles and the relevance of detecting major genetic determinants to monitor the dissemination of these genes. J Chemother, 2004 Apr, 16(2), 151 - 5 Comparison of systemic flurithromycin therapy and clinical procedures in the treatment of periodontal diseases; Blandino G et al.; The purpose of the present investigation was to evaluate, in 20 periodontal patients, the microbial and clinical effects of flurithromycin therapy plus scaling and root planning (SRP) in comparison with SRP alone . Clinical assessments of plaque, bleeding on probing and pocket depth were made prior to SRP alone and SRP plus flurithromycin therapy (375 mg twice daily for 5 days) and after both treatments . Subgingival plaque samples (n . 180) were taken prior to and after both treatments and analyzed by conventional bacteriological procedures . Differences in pocket depth and prevalence of bacterial species were analyzed pre- and post-therapies using statistical analyses . A significant decrease (p<0.001) was seen for pocket depth post SRP alone and post SRP plus flurithromycin . After two treatments, Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Prevotella melaninogenica were eradicated from all tested sites . If we compare the prevalence of the species isolated after SRP alone and after SRP plus flurithromycin statistically significant differences were detected for P . gingivalis and for Fusobacterium nucleatum (p<0.05 and p<0.01, respectively) . Flurithromycin can be considered a useful adjunct to mechanical periodontal treatment since it is more efficient in eliminating periodontal pathogens. Lett Appl Microbiol, 2004, 38(4), 327 - 32 Fermentation of pectin and glucose, and activity of pectin-degrading enzymes in the rabbit caecal bacterium Bacteroides caccae; Sirotek K et al.; AIMS: To compare fermentation pattern in cultures of Bacteroides caccae supplied with pectin and glucose, and identify enzymes involved in metabolism of pectin . METHODS AND RESULTS: A strain KWN isolated from the rabbit caecum was used . Fermentation pattern, changes of viscosity and enzyme reactions products were determined . Cultures grown on pectin produced significantly more acetate and less formate, lactate, fumarate and succinate than cultures grown on glucose . Production of cell dry matter and protein per gram of substrate used was the same in pectin- and glucose-grown cultures . The principal enzymes that participated in the metabolism of pectin were extracellular exopectate hydrolase (EC 3.2.1.67), extracellular endopectate lyase (EC 4.2.2.2) and cell-associated 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase (EC 4.1.2.14) . The latter enzyme is unique to the Entner-Doudoroff pathway . Activities of pectinolytic enzymes in cultures grown on glucose were low . Activity of KDPG aldolase was similar in pectin- and glucose-grown cells . CONCLUSIONS: Metabolites and activities of pectin-degrading enzymes differed in cultures of B . caccae KWN grown on pectin and glucose . Yields of dry matter and protein were the same on both substrates . SIGNIFICANCE AND IMPACT OF THE STUDY: Information on metabolism of pectin in animal strains of Bacteroides is incomplete . This study extends the knowledge on metabolism in bacteria from the rabbit caecum. Eur J Immunol, 2004 Jul, 34(7), 2052 - 9 NOD2 mediates anti-inflammatory signals induced by TLR2 ligands: implications for Crohn's disease; Netea MG et al.; Mutations of the NOD2 gene have been associated with an increased susceptibility to Crohn's disease, but the pathogenetic mechanisms mediated by NOD2 remain elusive . In the present study, we demonstrate that the 3020insC frameshift-mutation in the NOD2 gene associated with Crohn's disease results in defective release of IL-10 from blood mononuclear cells after stimulation with the Toll-like receptor (TLR)2 ligands, peptidoglycan and Pam3Cys-KKKK, but not with bacterial LPS, a TLR4 ligand . The potential pathophysiological significance of this finding in patients with Crohn's disease and who are homozygous for this NOD2 mutation was substantiated by the finding of decreased anti-inflammatory cytokine release when cells from these patients were stimulated with different species of Bacteroides, an enteric microorganism implicated in the pathogenesis of Crohn's disease . In conclusion, defective NOD2 function results in a pro-inflammatory cytokine bias after stimulation of mononuclear cells with TLR2 stimuli, and this could contribute to the overwhelming inflammation seen in Crohn's disease. Chemotherapy, 2004 Jun, 50(2), 76 - 80 Therapeutic efficacy of moxifloxacin, a new quinolone, in the treatment of experimental intra-abdominal abscesses induced by Bacteroides fragilis in mice; Thadepalli H et al.; Moxifloxacin, a new quinolone, is effective in vitro against several anaerobic bacteria including Bacteroides fragilis, but its in vivo activity against anaerobic infections is not known . In this study, we evaluated the in vivo activity of moxifloxacin in the treatment of experimentally induced intra-abdominal abscesses (IAA) caused by B . fragilis . For comparison, clindamycin, metronidazole, and levofloxacin were used, and saline for control . Absence of bacteria (sterile) in the abscess pus was required to call it a cure . Mice were intraperitoneally injected with B . fragilis plus sterile rat feces and barium sulfate . Animals were treated with moxifloxacin (40 mg/kg/b.i.d.), clindamycin (75 mg/kg/b.i.d.), levofloxacin (40 mg/kg/b.i.d.) or metronidazole (75 mg/kg/b.i.d.) for 10 days . The cure rate was 12% in controls on saline therapy, 57% on metronidazole, 67% on levofloxacin, 73% on moxifloxacin and 79% on clindamycin . The therapeutic efficacy of moxifloxacin in this B . fragilis infection was not significantly different from that observed with clindamycin . By virtue of its established efficacy on gram-negative aerobic bacteria and the observed in vivo efficacy on B . fragilis, moxifloxacin can be evaluated in the treatment of clinical anaerobic infections . Eur J Biochem, 2004 Jul, 271(13), 2670 - 81 Molecular and biochemical characterization of D-phosphoglycerate dehydrogenase from Entamoeba histolytica . A unique enteric protozoan parasite that possesses both phosphorylated and nonphosphorylated serine metabolic pathways; Ali V et al.; A putative phosphoglycerate dehydrogenase (PGDH), which catalyzes the oxidation of d-phosphoglycerate to 3-phosphohydroxypyruvate in the so-called phosphorylated serine metabolic pathway, from the enteric protozoan parasite Entamoeba histolytica was characterized . The E . histolytica PGDH gene (EhPGDH) encodes a protein of 299 amino acids with a calculated molecular mass of 33.5 kDa and an isoelectric point of 8.11 . EhPGDH showed high homology to PGDH from bacteroides and another enteric protozoan ciliate, Entodinium caudatum . EhPGDH lacks both the carboxyl-terminal serine binding domain and the 13-14 amino acid regions containing the conserved Trp139 (of Escherichia coli PGDH) in the nucleotide binding domain shown to be crucial for tetramerization, which are present in other organisms including higher eukaryotes . EhPGDH catalyzed reduction of phosphohydroxypyruvate to phosphoglycerate utilizing NADH and, less efficiently, NADPH; EhPGDH did not utilize 2-oxoglutarate . Kinetic parameters of EhPGDH were similar to those of mammalian PGDH, for example the preference of NADH cofactor, substrate specificities and salt-reversible substrate inhibition . In contrast to PGDH from bacteria, plants and mammals, the EhPGDH protein is present as a homodimer as demonstrated by gel filtration chromatography . The E . histolytica lysate contained PGDH activity of 26 nmol NADH utilized per min per mg of lysate protein in the reverse direction, which consisted 0.2-0.4% of a total soluble protein . Altogether, this parasite represents a unique unicellular protist that possesses both phosphorylated and nonphosphorylated serine metabolic pathways, reinforcing the biological importance of serine metabolism in this organism . Amino acid sequence comparison and phylogenetic analysis of various PGDH sequences showed that E . histolytica forms a highly supported monophyletic group with another enteric protozoa, cilliate E . caudatum, and bacteroides. J Biol Chem, 2004 Aug 20, 279(34), 36103 - 11 Epub 2004 Jun 15. 15-deoxy-delta12,14-prostaglandin J2-mediated ERK signaling inhibits gram-negative bacteria-induced RelA phosphorylation and interleukin-6 gene expression in intestinal epithelial cells through modulation of protein phosphatase 2A activity; Ruiz PA et al.; We have previously shown that non-pathogenic Gram-negative Bacteroides vulgatus induces transient RelA phosphorylation (Ser-536), NF-kappaB activity, and pro-inflammatory gene expression in native and intestinal epithelial cell (IEC) lines . We now demonstrate that 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) but not prostaglandin E(2) inhibits lipopolysaccharide (LPS) (B . vulgatus)/LPS (Escherichia coli)-induced RelA phosphorylation and interleukin-6 gene expression in the colonic epithelial cell line CMT-93 . This inhibitory effect of 15d-PGJ(2) was mediated independently of LPS-induced IkappaBalpha phosphorylation/degradation and RelA nuclear translocation as well as RelA DNA binding activity . Interestingly, although B . vulgatus induced nuclear expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in native epithelium of monoassociated Fisher rats, PPARgamma-specific knock-down in CMT-93 cells using small interference RNA failed to reverse the inhibitory effects of PPARgamma agonist 15d-PGJ(2), suggesting PPARgamma-independent mechanisms . In addition, 15d-PGJ(2) but not the synthetic high affinity PPARgamma ligand rosiglitazone triggered ERK1/2 phosphorylation in IEC, and most importantly, MEK1 inhibitor PD98059 reversed the inhibitory effect of 15dPGJ(2) on LPS-induced RelA phosphorylation and interleukin-6 gene expression . Calyculin A, a specific phosphoserine/phospho-threonine phosphatase inhibitor increased the basal phosphorylation of RelA and reversed the inhibitory effect of 15d-PGJ(2) on LPS-induced RelA phosphorylation . We further demonstrated in co-immunoprecipitation experiments that 15d-PGJ(2) triggered protein phosphatase 2A activity, which directly dephosphorylated RelA in LPS-stimulated CMT-93 cells . We concluded that 15d-PGJ(2) may help to control NF-kappaB signaling and normal intestinal homeostasis to the enteric microflora by modulating RelA phosphorylation in IEC through altered protein phosphatase 2A activity. J Antimicrob Chemother, 2004 Jul, 54(1), 109 - 16 Epub 2004 Jun 09. Metronidazole resistance in Bacteroides spp . carrying nim genes and the selection of slow-growing metronidazole-resistant mutants; Gal M et al.; OBJECTIVES: Human clinical isolates of Bacteroides spp . originating from patients in the UK were investigated for the presence of metronidazole resistance determinants (nim genes) and their presence was related to the MIC of metronidazole for the isolates . METHODS: Isolates were screened for susceptibility to a metronidazole disc and had their MIC determined by the Etest method . They were investigated for the presence of nim genes by PCR . An experiment to determine the effect of prolonged exposure to metronidazole was applied to nim-positive isolates with MICs below the therapeutic breakpoint . RESULTS: Fifty of 206 isolates (24%) were found to possess nim genes and these had MICs of metronidazole ranging from 1.5 to >256 mg/L with 24 (11.6%) above the therapeutic breakpoint of 16 mg/L . The remaining 26 nim-gene-positive isolates had MICs that were still below the therapeutic breakpoint, ranging from 1.5 to 6.0 mg/L . nim genes were not found in 156 (76%) isolates, and all but seven of these were susceptible to a 5 microg disc of metronidazole . Ten members of the group for which the MICs were below the therapeutic level were found to have slow-growing sub-populations with metronidazole MICs ranging from 8.0 to >256 mg/L that became evident after prolonged exposure to metronidazole in vitro . This resistance selection process was sometimes reversible after passage in the absence of metronidazole; however, seven of the 10 slow-growing mutants converted to stable high-level resistance (MIC >256 mg/L) . CONCLUSIONS: Although the presence of nim genes per se does not always equate to therapeutic resistance, and other metronidazole resistance mechanisms may exist, this study has shown that prolonged exposure of nim-gene-carrying Bacteroides spp . to metronidazole can select for therapeutic resistance. J Antimicrob Chemother, 2004 Jul, 54(1), 100 - 8 Epub 2004 May 18. Differential gene expression in a Bacteroides fragilis metronidazole-resistant mutant; Diniz CG et al.; OBJECTIVES: The current work focused on molecular changes in a spontaneous Bacteroides fragilismutant selected by low concentrations of metronidazole as an adaptive response to the drug . METHODS: A metronidazole-resistant strain derived from B . fragilis ATCC 25285 was selected by passage in the presence of drug using 0-4 mg/L gradient plates . Using a combination of proteomics for identification of differentially expressed proteins by two-dimensional electrophoresis and selected mutational analyses by single cross-over insertion and an allelic exchange, we have identified genes involved in the adaptive response to metronidazole . RESULTS: There are significant changes in the protein profiles of resistant strains . These changes appeared to affect a wide range of metabolic proteins including lactate dehydrogenase (up-regulated) and flavodoxin (down-regulated), which may be involved in electron transfer reactions . Also, the enzymic activity of the pyruvate-ferredoxin oxidoreductase (PorA) complex was impaired . Mutant strains lacking the genes for flavodoxin and PorA were less susceptible to metronidazole than the sensitive parent, and a double flavodoxin/PorA mutant had even less susceptibility but none of the mutants were as resistant as the spontaneous metronidazole-resistant strain . CONCLUSIONS: Overall, the data indicated that there were global changes in the regulation of the physiology of the metronidazole-resistant strain . In addition, flavodoxin was identified as an important contributor to metronidazole sensitivity in B . fragilis. Immunology, 2004 Jun, 112(2), 310 - 20 Differential effect of immune cells on non-pathogenic Gram-negative bacteria-induced nuclear factor-kappaB activation and pro-inflammatory gene expression in intestinal epithelial cells; Haller D et al.; We have previously shown that non-pathogenic Gram negative bacteria induce RelA phosphorylation, nuclear factor (NF)-kappaB transcriptional activity and pro-inflammatory gene expression in intestinal epithelial cells (IEC) in vivo and in vitro . In this study, we investigated the molecular mechanism of immune-epithelial cell cross-talk on Gram-negative enteric bacteria-induced NF-kappaB signalling and pro-inflammatory gene expression in IEC using HT-29/MTX as well as CaCO-2 transwell cultures Interestingly, while differentiated HT-29/MTX cells are unresponsive to non-pathogenic Gram negative bacterial stimulation, interleukin-8 (IL-8) mRNA accumulation is strongly induced in Escherichia coli- but not Bacteroides vulgatus-stimulated IEC cocultured with peripheral blood (PBMC) and lamina propria mononuclear cells (LPMC) . The presence of PBMC triggered both E . coli- and B . vulgatus-induced mRNA expression of the Toll-like receptor-4 accessory protein MD-2 as well as endogenous IkappaBalpha phosphorylation, demonstrating similar capabilities of these bacteria to induce proximal NF-kappaB signalling . However, B . vulgatus failed to trigger IkappaBalpha degradation and NF-kappaB transcriptional activity in the presence of PBMC . Interestingly, B . vulgatus- and E . coli-derived lipopolysaccharide-induced similar IL-8 mRNA expression in epithelial cells after basolateral stimulation of HT-29/PBMC cocultures . Although luminal enteric bacteria have adjuvant and antigenic properties in chronic intestinal inflammation, PBMC from patients with active ulcerative colitis and Crohn's disease differentially trigger epithelial cell activation in response to E . coli and E . coli-derived LPS . In conclusion, this study provides evidence for a differential regulation of non-pathogenic Gram-negative bacteria-induced NF-kappaB signalling and IL-8 gene expression in IEC cocultured with immune cells and suggests the presence of mechanisms that assure hyporesponsiveness of the intestinal epithelium to certain commensally enteric bacteria. Gastroenterology, 2004 May, 126(5), 1358 - 73 DNA from probiotic bacteria modulates murine and human epithelial and immune function; Jijon H et al.; BACKGROUND & AIMS: The intestinal epithelium must discriminate between pathogenic and nonpathogenic bacteria and respond accordingly . The aim of this study was to examine whether bacterial DNA can serve as the molecular basis for bacterial recognition . METHODS: HT-29 monolayers were treated with various bacterial DNA and interleukin (IL)-8 secretion measured by enzyme-linked immunosorbent assay, nuclear factor kappaB activation by electrophoretic mobility shift assay and reporter assays, and IkappaB levels by Western blotting . Cytokine secretion in response to bacterial DNA was measured in murine colonic segments and splenocytes . IL-10-deficient mice were fed DNA from VSL probiotic compound daily for 2 weeks . Colons were removed and analyzed for cytokine production and inflammation . RESULTS: HT-29 cells responded with IL-8 secretion to bacterial DNA in a differential manner . In the presence of proinflammatory stimuli, VSL3 DNA inhibited IL-8 secretion, reduced p38 mitogen-activated protein kinase activation, delayed nuclear factor kappaB activation, stabilized levels of IkappaB, and inhibited proteasome function . VSL3 DNA inhibited colonic interferon (IFN)-gamma secretion in mouse colons and also attenuated a Bacteroides vulgatus-induced IFN-gamma release from murine splenocytes . In mice, VSL3 DNA attenuated a systemic release of tumor necrosis factor alpha in response to Escherichia coli DNA injection . Treatment of IL-10-deficient mice with oral VSL3 DNA resulted in a reduction in mucosal secretion of tumor necrosis factor alpha and IFN-gamma and an improvement in histologic disease . CONCLUSIONS: DNA from probiotic bacteria can limit epithelial proinflammatory responses in vivo and in vitro . Systemic and oral administration of VSL3 DNA ameliorates inflammatory responses. J Clin Microbiol, 2004 May, 42(5), 2338 - 40 Brain abscess due to Gemella haemolysans; Lee MR et al.; We present a case of brain abscess due to Gemella haemolysans and Bacteroides species in a 60-year-old-immunocompetent man who underwent dental procedures . The patient completely recovered following intravenous therapy with ampicillin and metronidazole for 6 weeks. J Antimicrob Chemother, 2004 Jun, 53(6), 1039 - 44 Epub 2004 May 05. Surveillance of susceptibility patterns in 1297 European and US anaerobic and capnophilic isolates to co-amoxiclav and five other antimicrobial agents; Koeth LM et al.; In vitro susceptibility data were collected for co-amoxiclav and other antimicrobial agents against 1297 recent anaerobe isolates collected in Europe and the USA . The co-amoxiclav (amoxicillin/clavulanic acid) MIC(50/90)s (amoxicillin/clavulanic acid concentration in a ratio of 2:1, expressed in terms of amoxicillin concentration in mg/L) were 0.5/4 for Bacteroides fragilis, </=0.125/1 for Prevotella species, </=0.125/0.25 for Fusobacterium nucleatum, 0.5/1 for Eikenella corrodens, 0.25/8 for Peptostreptococcus anaerobius, </=0.125/0.5 for Micromonas (Peptostreptococcus) micros, </=0.25/0.5 for Fingoldia (Peptostreptococcus) magna, and </=0.125/0.125 for Porphyromonas species . The co-amoxiclav susceptibility rate for B . fragilis was 94.6%, for P . anaerobius 84.3% and for all other species tested 100% . These data indicate that co-amoxiclav remains an effective drug for the antimicrobial treatment and prophylaxis of many anaerobic infections . Among the comparator drugs, metronidazole was very active against all bacterial species (>96% susceptible) except E . corrodens (MIC(50/90) of >32/>64 mg/L), which is a capnophilic organism . Imipenem was also highly active against all species (>98% susceptible) . Levofloxacin and clindamycin were the least potent agents tested, particularly against Bacteroides, Prevotella and Peptostreptococcus (levofloxacin susceptibility rates: Bacteroides 72.7%, Prevotella 71.5%, F . magna 72.4%; clindamycin susceptibility rates: Bacteroides 79.5%, Prevotella 92.1%, F . magna 84.7%). Appl Environ Microbiol, 2004 May, 70(5), 2867 - 79 Seasonal changes in an alpine soil bacterial community in the colorado rocky mountains; Lipson DA et al.; The period when the snowpack melts in late spring is a dynamic time for alpine ecosystems . The large winter microbial community begins to turn over rapidly, releasing nutrients to plants . Past studies have shown that the soil microbial community in alpine dry meadows of the Colorado Rocky Mountains changes in biomass, function, broad-level structure, and fungal diversity between winter and early summer . However, little specific information exists on the diversity of the alpine bacterial community or how it changes during this ecologically important period . We constructed clone libraries of 16S ribosomal DNA from alpine soil collected in winter, spring, and summer . We also cultivated bacteria from the alpine soil and measured the seasonal abundance of selected cultured isolates in hybridization experiments . The uncultured bacterial communities changed between seasons in diversity and abundance within taxa . The Acidobacterium division was most abundant in the spring . The winter community had the highest proportion of Actinobacteria and members of the Cytophaga/Flexibacter/Bacteroides (CFB) division . The summer community had the highest proportion of the Verrucomicrobium division and of beta-PROTEOBACTERIA: As a whole, alpha-Proteobacteria were equally abundant in all seasons, although seasonal changes may have occurred within this group . A number of sequences from currently uncultivated divisions were found, including two novel candidate divisions . The cultured isolates belonged to the alpha-, beta-, and gamma-Proteobacteria, the Actinobacteria, and the CFB groups . The only uncultured sequences that were closely related to the isolates were from winter and spring libraries . Hybridization experiments showed that actinobacterial and beta-proteobacterial isolates were most abundant during winter, while the alpha- and gamma-proteobacterial isolates tested did not vary significantly . While the cultures and clone libraries produced generally distinct groups of organisms, the two approaches gave consistent accounts of seasonal changes in microbial diversity. Med Dosw Mikrobiol, 2003, 55(4), 365 - 70 {Biological activity of lipopolysaccharides from clinical Bacteroides fragilis strains isolated in Poland determined in reaction with limulus amoebocyte lysate}; Rokosz A et al.; The aim of this study was to determine a biological activity of lipopolysaccharides (LPS) from clinical Bacterioides fragilis strains isolated in Poland by means of quantitative, photometric BET (LAL) method with Limulus polyphemus amoebocyte lysate and chromogenic substrate S-2423 . Lipopolysaccharides were extracted from nine clinical B . fragilis strains by the procedure of Westphal and Jann (1965) . Crude LPS preparations were purified with ultracentrifugation . Biological activities of bacterial endotoxins were determined by quantitative BET method with chromogenic substrate S-2423 (ENDOCHROME kit) . Tests were performed according to the recommendations of the producer (Charles River Endosafe Ltd., USA) . E . coli O55:B5 LPS and LPS preparations from reference B . fragilis strains were applied to compare the results of examinations . Activities of endotoxins from clinical B . fragilis strains isolated in Poland determined in reaction with Limulus amoebocyte lysate were differentiated . Among endotoxins of clinical B . fragilis strains the most active was the preparation from strain cultured in the case of pancreatic ulcer (B . fragilis 80/81 LPS) . Lipopolysaccharides of examined B . fragilis strains were less active in BET test than E . coli O55:B5 LPS. J Med Microbiol, 2004 May, 53(Pt 5), 413 - 9 Molecular characterization of imipenem-resistant, cfiA-positive Bacteroides fragilis isolates from the USA, Hungary and Kuwait; Soki J et al.; Fifteen Bacteroides fragilis isolates from the USA, Hungary and Kuwait were examined for carbapenem resistance, for carbapenemase activity and, with the use of various PCR-based methods and nucleotide sequencing, for cfiA genes and activating insertion sequence (IS) elements . All the B . fragilis isolates were cfiA-positive, 10 of the cfiA genes being upregulated by IS elements that are already known . Of these 10, one was of a novel type (designated IS943) and two further ones (IS614B and IS614C) were suspected hybrids of IS612, IS614 and IS942 . There were five cfiA-positive imipenem-resistant B . fragilis isolates with elevated imipenem MICs (minimal inhibitory concentration) that harboured no IS insertion upstream of the cfiA gene, but produced carbapenemase; these isolates might possess a novel activation mechanism . On the basis of the available phenotypic and genotypic evidence, the present data suggest that there are at least two cfiA activation mechanisms among B . fragilis isolates. Acta Microbiol Pol, 2003, 52(4), 361 - 72 Influence of clindamycin, metronidazole and polymyxin B on the expression of cell adhesion molecules stimulated by endotoxin and enterotoxin of Bacteroides fragilis; Meisel-Mikolajczyk F et al.; The aim of this study was to compare the influence of antimicrobials (clindamycin, metronidazole and polymyxin B) on the expression of adhesion molecules (VCAM-1, ICAM-1 and E-selectin) on the HMEC-1 cell line stimulated by LPS and enterotoxin of B . fragilis . LPS was extracted from two reference: ATCC 43858 and NCTC 11295 and one isolated in our laboratory (W2) enterotoxigenic strains, and one nonenterotoxigenic reference strain--IPL E 323 . Enterotoxin preparations (Tox 1 and Tox 2) were isolated from supematant of B . fragilis ATCC 43858 culture and purified . HMEC-1 cell line was stimulated with bacterial preparations at concentration of 10 mg/ml . For measuring the expression of adhesion molecules we used ELISA test . Clindamycin, metronidazole and polymyxin B supressed the ICAM-1 expression when endothelium was stimulated with B . fragilis LPS and augmented ICAM-1 expression by Tox 1 and Tox 2 . The expression of VCAM-1 was augmented by antimicrobials when endothelium was stimulated with LPS or enterotoxin preparations . The expression of E-selectin was differentiated. J Bacteriol, 2004 May, 186(9), 2548 - 57 Regulation of a Bacteroides operon that controls excision and transfer of the conjugative transposon CTnDOT; Wang Y et al.; CTnDOT is a conjugative transposon (CTn) that is found in many Bacteroides strains . Transfer of CTnDOT is stimulated 100- to 1,000-fold if the cells are first exposed to tetracycline (TET) . Both excision and transfer of CTnDOT are stimulated by TET . An operon that contains a TET resistance gene, tetQ, and two regulatory genes, rteA and rteB, is essential for control of excision and transfer functions . At first, it appeared that RteA and RteB, which are members of a two-component regulatory system, might be directly responsible for the TET effect . We show here, however, that neither RteA nor RteB affected expression of the operon . TetQ, a ribosome protection type of TET resistance protein, actually reduced operon expression, possibly by interacting with ribosomes that are translating the tetQ message . Fusions of tetQ with a reporter gene, uidA, were only expressed at a high level when the fusion was cloned in frame with the first six codons of tetQ . However, out of frame fusions or fusions ending at the other five codons of tetQ showed much lower expression of the uidA gene . Moreover, reverse transcription-PCR amplification of tetQ mRNA revealed that despite the fact that the uidA gene product, beta-glucuronidase (GUS), was produced only when the cells were exposed to TET, tetQ mRNA was produced in both the presence and absence of TET . Computer analysis of the region upstream of the tetQ start codon predicted that the mRNA in this region could form a complex RNA hairpin structure that would prevent access of ribosomes to the ribosome binding site . Mutations that abolished base pairing in the stem that formed the base of this putative hairpin structure made GUS production as high in the absence of TET as in TET-stimulated cells . Compensatory mutations that restored the hairpin structure led to a return of regulated production of GUS . Thus, the tetQ-rteA-rteB operon appears to be regulated by a translational attenuation mechanism. Ann Surg, 2004 May, 239(5), 733 - 8; discussion 738-40 Small intestinal submucosa for vascular reconstruction in the presence of gastrointestinal contamination; Jernigan TW et al.; INTRODUCTION: Surgical options for vascular reconstruction in a contaminated field are limited and include prosthetic reconstruction or ligation with extra-anatomic bypass . With prosthetic insertion, rates of graft infection and failures (pseudoaneurysms and thrombosis) are high . In the emergent situations, extra-anatomic bypass is time-consuming and complex, and it produces marginal long-term results . Small intestinal submucosa (SIS) is a cell-free collagen matrix derived from porcine small intestine . Preliminary studies have demonstrated its ability to be remodeled into host tissue . In this study, we compared SIS to polytetrafluoroethylene (PTFE) as a vascular patch for arterial repair in the presence of massive gastrointestinal contamination to evaluate graft patency, incorporation, infection, and aneurysm formation . METHODS: Adult mongrel pigs underwent general anesthesia with Isoflurane and were then randomized to 1 of 3 groups: control, contamination (colon puncture with stool contamination of the pelvis), or shock + contamination (40% blood volume for 1 hour, then resuscitation with shed blood and crystalloid, plus contamination) . All groups then underwent a left common iliac arteriotomy and further randomized to a 1 x 3-cm patch angioplasty with either SIS or PTFE . All received cefotetan for 24 hours . All animals were sacrificed between 2 and 4 weeks, and necropsy was performed . Grafts were cultured, and microscopic analysis with hematoxylin and eosin and trichrome was performed . Outcomes included pulse quality (normal or diminished) compared with opposite side, graft infection, and pseudoaneurysm; all were determined by a blinded investigator . RESULTS: Forty animals were randomized, and 1 died of abdominal sepsis . All control animals had normal distal pulses, no pseudoaneurysms, and no patch infections . The pseudoaneurysm rate for the contaminated PTFE patches was 25% compared with 0% in the SIS group (P = 0.09) . Patch infection occurred in 73% of all PTFE patches compared with 8% of SIS patches (P < 0.03) . Organisms present in the infected grafts included Escherichia coli, Bacteroides species, and other Gram-negative enterics . Histopathology demonstrated the presence of neointima in both SIS and PTFE . Only SIS was completely incorporated, with infiltration of collagen fibrils and lymphocytes . CONCLUSIONS: SIS was associated with improved graft patency, less infection, complete incorporation, and no false aneurysm formation when compared with PTFE . This may be due to its ability to provide a durable scaffold for cellularization and tissue remodeling . This material may offer a superior alternative to more complex vascular reconstruction techniques in contaminated fields. Med Princ Pract, 2004 May-Jun, 13(3), 147 - 52 Molecular characterization of nitroimidazole resistance in metronidazole-resistant bacteroides species isolated from hospital patients in Kuwait; Jamal WY et al.; OBJECTIVES: The aim of this study was to screen for infections caused by metronidazole (MTZ)-resistant Bacteroides spp., and to characterize the genes that encode the MTZ resistance . MATERIALS AND METHODS: A total of 7 MTZ-resistant Bacteroides spp . were isolated from 5 patients with MTZ-resistant infections . These organisms were investigated for carriage of genes that encode MTZ resistance . The presence of these genes was investigated by PCR and the PCR products were subjected to PCR-RFLP analysis . RESULTS: The strains were MTZ-resistant with minimum inhibitory concentrations of > 32 microg/ml . The presence of nim genes was indicated by PCR in all 7 strains . PCR-RFLP analysis of the nim gene products demonstrated two of the five reported resistance genes, nimA-nimE . These two resistance genes were nimE in 5 of the 7 isolates and nimA in 2 strains . CONCLUSION: MTZ-resistant Bacteroides spp . have been isolated from patients in Kuwait . Nim genes, specifically nimE and nimA, mediate the drug resistance in these isolates . The methods used in detecting these genes are rapid, accurate and relatively inexpensive and could be adopted easily to help in monitoring emergence of MTZ resistance determinants in Kuwait . J Clin Microbiol, 2004 Apr, 42(4), 1727 - 30 Identification of Bacteroides thetaiotaomicron on the basis of an unexpected specific amplicon of universal 16S ribosomal DNA PCR; Teng LJ et al.; We applied a set of commonly used universal primers (primers RW01 and DG74) to amplify partial fragments of 16S ribosomal DNA for bacterial identification and found an unexpected amplicon (547 bp), in addition to the expected 362-bp product, in samples containing Bacteroides thetaiotaomicron . It was demonstrated that the internal sequence (508 bp, excluding the primers) of the 547-bp amplicon was identical to the genomic sequence from nucleotide positions 165800 to 166307 of B . thetaiotaomicron type strain VPI-5482 by a BLAST search of the sequences in the GenBank database . The existence of this unexpected yet specific amplicon strongly indicated the presence of B . thetaiotaomicron in the sample, and it was found that it could be used to discriminate B . thetaiotaomicron from closely related species . Another set of PCR primers specific for B . thetaiotaomicron was developed on the basis of the sequence of this 547-bp genomic fragment . Both PCR-based assays showed the same sensitivity (88%) and specificity (100%). Zhonghua Shao Shang Za Zhi, 2004 Feb, 20(1), 34 - 6 {Effects of succinic acid on the function of in vitro cultured human fibroblasts}; Ren LC et al.; OBJECTIVE: To explore the mechanism of injurious effect of succinic acid on human fibroblast and it's role in bacteroides fragilis infection . METHODS: In vitro cultured human fibroblasts were challenged by succinic acid in concentrations of 5, 10, 20 and 30 mmol/L (pH5.5), respectively . The cellular activity, apoptosis rate, the collagen synthesis in the supernatant of the cell culture, and the activity of caspase-3 were determined 24 hours after challenge . Isotonic saline challenged fibroblast were employed as control and the changes in the indices before and after succinic acid challenge were observed . RESULTS: Along with the increase in the concentration of succinic acid, the fibroblast proliferation rate was decreased and so was the collagen synthesis . But the apoptosis rate and caspase-3 activity were increased . The activity of caspase-3 was markedly higher than that in normal control when the succinic acid concentration was 10-30 mmol/L . The cellular activity and collagen synthesis were significantly lower and the apoptosis rate was obviously higher than those in control group when the succinic acid concentration was 20 or 30 mmol/L (P < 0.05) . CONCLUSION: The proliferation and collagen synthesis in fibroblast culture could be significantly inhibited and the cellular apoptosis could be promoted by succinic acid . The process of wound healing of the wounds infected by bacteroides fragilis would be delayed due to the production of succinic acid by the bacteria. Shanghai Kou Qiang Yi Xue, 1999 Sep, 8(3), 129 - 31 {The chemical degradation of lipopolysaccharides from oral anaerobes}; Xie DY et al.; OBJECTIVE:To evaluate the effect of drug on chemical degradation of lipopolysaccharide (LPS) from oral anaerobes.METHODS:LPSs from porphyromonas gingivalis (Pg),bacteroides fragilis (Bf) and fusobacteria nucleatum (Fn) were extracted by the hot phenol-water method and purified by the phenol-chloroform-petroleum ether procedure.1.0 ml (200microg) various LPSs were incubated with 2.0 ml various drugs at 37degrees centigrade for 15 or 30 minutes in vitro,respectively.The chemical degradation of LPS was quantitated by limulus synthetic chromogenic substrated method after dialysis.RESULTS:The order of degradation was 30% hydrogen peroxide (H),50% citric acid (C),garlic guice (G),1:1 diluted G,25% C and 3% H,and their efffcts were dose dependent and were time dependent except H but the effect of lysozyme was minimal.CONCLUSION:The study may imply that the dose and the mechanism of various drugs on LPS degradation are different and remains to be elucidated. Antimicrob Agents Chemother, 2004 Apr, 48(4), 1344 - 6 Role of topoisomerase mutations and efflux in fluoroquinolone resistance of Bacteroides fragilis clinical isolates and laboratory mutants; Ricci V et al.; Twelve laboratory mutants and 32 ciprofloxacin-resistant isolates of Bacteroides fragilis were examined for the mechanism(s) of fluoroquinolone resistance . Five mutants had mutations in gyrA . One mutant and two clinical isolates contained a mutation in gyrB . Eight mutants and five clinical isolates accumulated significantly less ciprofloxacin than did wild-type isolates; the mutants and clinical isolates were restored to wild-type characteristics when carbonyl cyanide m-chlorophenylhydrazone was used. Clin Orthop, 2004 Jan, (418), 222 - 4 Infection of a total hip arthroplasty with Prevotella loeschii; Steingruber I et al.; Infection is a serious complication of total hip replacement . It has been proposed that 6% of all infections after total hip arthroplasty may be of dental origin through hematogenous spread . However, no conclusive evidence that the mouth is a definitive source for infection of a total hip replacement has been reported . In the current case, Prevotella loeschii, a pigmented bacteroides species was identified in a total hip replacement . Prevotella loeschii is an organism which exclusively inhabits the dental region . Hematogenous spread of Prevotella loeschii may occur after penetration of the mucosal barrier in cases of endodontic or periodontic lesions, pericoronitis, or complications of tooth extraction . The involvement of Prevotella loescheii in an infection in a patient who had a total hip arthroplasty is strong evidence for the mechanism of a hematogenous infection from a dental source. J Virol Methods, 2004 Apr, 117(1), 19 - 25 Standardised evaluation of the performance of a simple membrane filtration-elution method to concentrate bacteriophages from drinking water; Mendez J et al.; The bacteriophage elution procedure described further after adsorption to acetate-nitrate cellulose membrane filters allows better recovery of phages concentrated from 1l of water than elution procedures used previously . The improvement is due to the combined effect of the eluent (3% (w/v) beef extract, 3% (v/v) Tween 80, 0.5M NaCl, pH 9.0) and the application of ultrasound instead of agitation or swirling . Average recovery of somatic coliphages, 82 +/- 7%, was the greatest, and that of phages infecting Bacteroides fragilis, 56 +/- 8%, the lowest, with intermediate values for F-specific and F-specific RNA bacteriophages . Thus, the method allowed recovery of over 56% for all the phages suggested as surrogate indicators . The method was then validated according to an International Standardisation Organisation validation standard procedure and implemented in routine laboratories, which obtained reproducible results. Drugs Exp Clin Res, 2003, 29(4), 153 - 5 Characteristics of anaerobes from skin specimens; Higaki S et al.; Anaerobes isolated from skin specimens from 1999 to 2001 were examined . The most common type was Peptostreptococcus spp., especially P . magnus and P . assaccharolyticus and Bacteroides fragilis . Dominance was seen for P . magnus, P . acnes and P . prevotii . Peptostreptococcus spp . and P . acnes showed high susceptibility to four antimicrobial agents . Prevotella spp . and B . fragilis showed low or no susceptibility to ampicillin, while B . fragilis showed low susceptibility to ceftizoxime . Evaluation of anaerobes is important for the balance of skin flora as well as for the choice of antimicrobial agents, when the anaerobes are pathogenic. J Clin Periodontol, 2004 Feb, 31(2), 141 - 8 Quadrant root planing versus same-day full-mouth root planing . II . Microbiological findings; Apatzidou DA et al.; OBJECTIVES: The aim of this study was to test the hypothesis that over a period of 6 months, same-day full-mouth scaling and root planing (FM-SRP) resulted in greater reductions in the detection frequency of five putative periodontal pathogens compared with quadrant scaling and root planing (Q-SRP) in chronic periodontitis patients . MATERIALS AND METHODS: Forty patients were recruited into this study . Subjects were randomised into two groups . The FM-SRP group received full-mouth scaling and root planing completed within the same day, while the Q-SRP group received quadrant root planing at 2-weekly intervals over four consecutive sessions . Selected-site analyses were performed on the deepest site in each quadrant before and after therapy, at approximately 3 and 6 months from baseline (R1 and R2) and clinical indices were recorded with an electronic pressure-sensitive probe . In addition, subgingival plaque samples were collected from these sites at baseline (BAS), at reassessment 1 (R1), approximately 6 weeks after the completion of therapy and at reassessment 2 (R2), 6 months from baseline . Polymerase chain reaction (PCR) was used to determine the presence of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Treponema denticola and Bacteroides forsythus in plaque . RESULTS: Both therapies resulted in significant improvements in all clinical indices both at R1 and R2 . A marked reduction in the presence of all candidate periodontal pathogens was noted after both treatment modalities, reaching statistical significance for the majority of the test organisms . These improvements were maintained over a period of 6 months . When the two treatment groups were compared, a significantly higher percentage of Q-SRP patients was positive for P . intermedia at R1 compared with FM-SRP patients (p<0.05) . In addition, a greater reduction in the patient prevalence for T . denticola was found for the FM-SRP group than the Q-SRP group at R1 and R2 from baseline (p<0.005), but the significance of this is questionable given the skewed detection frequency of this organism at baseline between the two treatments (p<0.01) . CONCLUSION: This study failed to confirm that same-day FM-SRP resulted in greater microbiological improvements compared with Q-SRP at 2-weekly intervals over a 6-month period, as determined by PCR . Copyright Blackwell Munksgaard, 2004. J Med Microbiol, 2004 Apr, 53(Pt 4), 273 - 80 Experimental swine dysentery: comparison between infection models; Jacobson M et al.; The aim of the present study was to develop a reproducible porcine infection model with Brachyspira hyodysenteriae . The influence of different factors was evaluated, namely, age, a diet containing large quantities of soybean meal, housing and administration of cortisol or antacids . Furthermore, the synergistic effect of additional bacteria (Escherichia coli O141, Bacteroides vulgatus or a mixture of Bacteroides fragilis, a field isolate of Bacteroides and Fusobacterium necrophorum) was studied . Experimental infection resulted in an increase in the serum concentrations of the acute-phase proteins serum amyloid A and haptoglobin and the percentages of neutrophils and monocytes . These alterations were specifically related to haemorrhagic diarrhoea . Inoculation combined with feeding of large quantities of soybean meal and group-housing induced swine dysentery in all experimental animals . If the pigs were fed soybean meal, kept in single pens and circulated between the pens, five out of nine developed disease. J Clin Periodontol, 2004 Mar, 31(3), 166 - 72 Debridement and local application of tetracycline-loaded fibres in the management of persistent periodontitis: results after 12 months; Aimetti M et al.; BACKGROUNDS, AIMS: The aim of our study was to evaluate the clinical, radiological and microbiological response to the local delivery of tetracycline (TE) of sites with persistent periodontal lesions . MATERIALS AND METHODS: The study was conducted in a split-mouth design . Nineteen patients with at least four bilateral pockets 4-5 mm and bleeding on probing (BOP) were treated with scaling and root planing (SRP) plus TE fibres (test sites) or with SRP alone (control sites) . Clinical and radiological measurements were taken at baseline, 6 months and 12 months post-operatively . Subgingival plaque samples were collected at baseline, at fibres removal, 6 and 12 months following treatment and analysed by polymerase chain reaction . RESULTS: Both treatments yielded a statistically significant (p<0.05) reduction of probing depth (2.05 and 1.21 mm), gain of clinical attachment level (1.71 and 0.53 mm) and reduction of BOP scores (23.68% and 57.89%) for TE and SRP groups, respectively, when comparing 12-month data with baseline . The differences between two groups were significant . The prevalence of Treponema denticola and Bacteroides forsythus decreased after therapy in both groups but only in the test sites Actinobacillus actinomycetemcomitans and Prevotella intermedia were not yield detected . The pathogens could be eliminated from five periodontal pocke