OBJECTIVETo set up a rapid, efficient, reliable and accurate method for separation of Bacteriodes forsythus proteins.

METHODSBacteroides 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).

RESULTS1. 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).

CONCLUSIONHorizontal two-dimension electrophoresis is a useful method for separating B. forsythus proteins.

Bacterial Proteins ; isolation & purification ; Bacteroides ; isolation & purification ; Electrophoresis, Gel, Two-Dimensional ; methods

Primary aortoenteric fistula (PAEF) is a rare but catastrophic cause of massive gastrointestinal bleeding. Diagnosis of PAEF is difficult to make and is frequently delayed without strong clinical suspicion. Timely surgical intervention is essential for patient's survival. We report on a case of an 86-year-old woman with no history of abdominal surgery, who presented with abdominal pain. Initially, computed tomography scan showed an intra-abdominal abscess, located anterior to the aortic bifurcation. However, she was discharged without treatment because of spontaneous improvement on a follow-up computed tomography scan, which showed a newly developed right common iliac artery aneurysm. One week later, she was readmitted due to recurrent abdominal pain. On the second day of admission, sudden onset of gastrointestinal bleeding occurred for the first time. After several endoscopic examinations, an aortoenteric fistula bleeding site was found in the sigmoid colon, and aortography showed progression of a right common iliac artery aneurysm. We finally concluded that intra-abdominal abscess induced an infected aortic aneurysm and enteric fistula to the sigmoid colon. This case demonstrated an extremely rare type of PAEF to the sigmoid colon caused by an infected abdominal aortic aneurysm, which has rarely been reported.
Abdominal Abscess/*diagnosis/microbiology ; Aged, 80 and over ; Aorta, Abdominal/radiography ; Aortic Aneurysm, Abdominal/*diagnosis/etiology ; Bacteroides/isolation & purification ; Bacteroides fragilis/isolation & purification ; Colon, Sigmoid/radiography ; Colonoscopy ; Enterococcus/isolation & purification ; Female ; Fistula/*diagnosis ; Humans ; Tomography, X-Ray Computed

OBJECTIVETo observe the prevalence of Porphyromonas gingivalis(Pg), Treponema denticola(Td) and Bacterides forsythus(Bf) subgingivally in diseased sites of chronic periodontitis (CP) patients.

METHODSSamples of subgingival plaque were detected by 3 kinds of oligonucleotide DNA probe from 60 sites of CP patients and 10 healthy sites of 10 healthy people.

RESULTSThe positive rates of Pg, Bf and Td in patients were 91.67%, 90.00% and 95.67% respectively; Pg, Bf and Td were detected simultaneously in 83.33% patients and Pg, Bf and Td were found to be related with each other(P < 0.01).

CONCLUSIONPg, Bf and Td were prominent periodontopathic bacteria and related each other, might exist in complexes in subgingival plaque and coaggretate together.

Adult ; Aged ; Bacteroides ; isolation & purification ; DNA Probes ; Dental Plaque ; microbiology ; Female ; Humans ; Male ; Middle Aged ; Oligonucleotides, Antisense ; Periodontitis ; microbiology ; Porphyromonas gingivalis ; isolation & purification ; Treponema ; isolation & purification

We report three cases of recently named Bacteroides spp. isolates, two B. faecis isolates and one B. intestinalis isolate from clinical specimens of inpatients at a Korean tertiary-care hospital in 2011. All isolates were susceptible to piperacillin-tazobactam, imipenem, meropenem, chloramphenicol, and metronidazole.
Aged ; Anti-Bacterial Agents/pharmacology ; Bacteroides/drug effects/*isolation & purification ; Female ; Humans ; Intestines/*microbiology ; Male ; Microbial Sensitivity Tests ; Middle Aged

OBJECTIVE: To analyze the characteristics of intestinal flora in Crohn's disease (CD) and intestinal tuberculosis (ITb), and to find potential identification features to differentiate these 2 diseases. METHODS: Fifteen CD patients, 23 ITb patients, and 21 healthy volunteers (controls) were enrolled from June 2007 to November 2009. Selective culturing was used for the enumeration of bacteria count. RESULTS: The intestinal flora was mainly composed of Bifidobacteria, Bacteroids, Escherichia coli and Staphylococcus aureus both in CD and ITb patients. Lactobacillus and Bifidobacteria decreased obviously but Bacteroid increased in CD patients compared with the control (P<0.01). Lactobacillus, Bifidobacteria and Escherichia coli decreased obviously (P<0.05), but Bacteroid increased in ITb patients compared with the control (P<0.01). Bacteroid increased in ITb patients compared with CD patients (P<0.05). No difference in Enterococcus, Staphylococcus aureus and Saccharomycete was found among the 3 groups (P>0.05). CONCLUSION Intestinal flora disorder occurred in either CD or ITb patients. The alteration of Bacteroid and Escherichia coli can help differentiate the 2 diseases.
Adult ; Bacteroides ; isolation & purification ; Crohn Disease ; diagnosis ; microbiology ; Diagnosis, Differential ; Escherichia coli ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; Male ; Tuberculosis, Gastrointestinal ; diagnosis ; microbiology

OBJECTIVETo examine the differential levels of fecal Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile between patients with hepatic cirrhosis and healthy controls. Fecal samples were collected from 29 patients with hepatic cirrhosis treated in the Department of Digestive Diseases at Zunyi Hospital between March and December of 2010.

METHODSFecal samples were collected from 13 healthy college students for use as controls. All samples were assessed by pH measurement, bacterial culture for turbidity, fluorescence in situ hybridization, and laser scanning confocal microscopy. The t-test and rank correlation test were used to determine statistical significance of intergroup differences in each tested parameter.

RESULTSThe feces of patients with hepatic cirrhosis had higher pH than that of healthy controls (6.79+/-0.64 vs. 6.18+/-0.74, P less than 0.05). The bacterial turbidity was not significantly different between the feces of hepatic cirrhosis patients and healthy controls (1.15+/-0.59 vs. 1.39+/-1.01, P more than 0.05). The numbers of Bifidobacterium, Bacteroides, Eubacterium rectale-Clostridium, Escherichia coli, Enterococcus, and Clostridium difficile in feces of patients with hepatic cirrhosis were significantly lower than those of the controls (all P less than 0.01). No significant correlation was found between the number or ratio of bacteria species and the severity of hepatic cirrhosis (Child-Pugh scores; P more than 0.05).

CONCLUSIONThe total quantity of intestinal bacteria in patients with hepatic cirrhosis is not significantly different from that in healthy patients. However, the profile of intestinal bacteria is different, which may explain the increased pH of fecal samples from patients with hepatic cirrhosis, but the differential profile is not correlated to cirrhosis pathogenesis.

Adult ; Aged ; Aged, 80 and over ; Bacteroides ; isolation & purification ; Bifidobacterium ; isolation & purification ; Case-Control Studies ; Clostridium ; isolation & purification ; Enterobacteriaceae ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Liver Cirrhosis ; microbiology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the changes in fecal flora and its correlation with the occurrence and progression of inflammatory bowel disease (IBD).

METHODSWe collected fresh fecal specimens from 167 IBD patients (including 113 with ulcerative colitis and 54 with Crohn's disease) and 54 healthy volunteers. The fecal flora was analyzed by gradient dilution method and the data of inflammatory markers including WBC, PLT, CRP and ESR were collected to assess the association between the fecal flora and the inflammatory markers.

RESULTSThe species Enterrococcus (6.60∓0.23, P<0.01), Saccharomyces (2.22∓0.27, P<0.05), Bacteriodes (5.57∓0.28, P<0.001), Bifidobacterium (5.08∓0.30, P<0.01), Peptococcus (6.22∓0.25, P<0.001), Lactobacillus (6.00∓0.26, P<0.001), and Clostridium (3.57∓0.30, P<0.05) all increased significantly, while Eubacterium (1.56∓0.24, P<0.01) reduced markedly in patients with ulcerative colitis compared with those in the control subjects. Enterrococcus (6.93∓0.28, P<0.01), Saccharomyces (2.73∓0.37, P<0.01), Bacteriodes (4.32∓0.52, P<0.05), Bifidobacterium (4.88∓0.42, P<0.05), Peptococcus (6.19∓0.32, P<0.01) and Lactobacillus (4.73∓0.47, P<0.001) all increased significantly and Eubacterium (1.01∓0.29, P<0.01) and Clostridium (0.87∓0.31, P<0.01) decreased in patients with Crohn's disease. The positivity rates of bacterial culture were consistent with the results of quantitative analysis of the fecal flora. The changes in fecal flora did not show a significant correlation with these inflammatory markers.

CONCLUSIONIBD patients have fecal flora imbalance compared with the healthy controls, and this imbalance may contribute to the occurrence and progression of IBD. The decline of Eubacterium contributes to the occurrence and development of IBD.

Adult ; Bacteria ; isolation & purification ; Bacteroides ; isolation & purification ; Bifidobacterium ; isolation & purification ; Biomarkers ; analysis ; Clostridium ; isolation & purification ; Colitis, Ulcerative ; microbiology ; Crohn Disease ; microbiology ; Enterococcus ; isolation & purification ; Eubacterium ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; Inflammatory Bowel Diseases ; etiology ; microbiology ; Lactobacillus ; isolation & purification ; Male ; Peptococcus ; isolation & purification ; Saccharomyces ; isolation & purification

Oral lichen planus (OLP) is a chronic inflammatory disease that is frequently detected in oral tissues. The aim of our study was to identify the prevalence of the detection of periodontopathogenic microorganisms (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia and Treponema denticola in OLP patients and to compare with this prevalence of periodontopathogenic microorganisms in healthy non-OLP patients. Our study included 27 (18 chronic periodontitis (OLPP) and 9 gingivitis (OLPG)) patients diagnosed with OLP along with 26 (13 chronic periodontitis (HP) and 13 gingivitis (HG)) healthy non-OLP patients. The multiplex polymerase chain reaction (PCR) with subsequent reverse hybridization method (micro-IDent) was used for identifying periodontopathogenic microorganisms present in subgingival plaque samples. The percentages of detection for A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia and T. denticola in subgingival plaque samples taken from OLP patients (OLPG and OLPP) were 18.5%, 85.1%, 81.4%, 88.8% and 74%, respectively. Meanwhile, in the non-OLP patients (HG and HP), these values were 7.6%, 50%, 46.1%, 73% and 57.7%, respectively. Thus, comparing the non-OLP groups with the OLP groups, the periodontopathogens' percentages of detection in the OLP groups were higher than those in the non-OLP groups. According to our study results, OLP patients have higher levels of infection with A. actinomycetemcomitans, P. gingivalis, P. intermedia, T. forsythia and T. denticola than non-OLP patients. We argue that the high percentages in patients with OLP may help identify the importance of periodontopathogenic microorganisms in the progress of periodontal diseases of OLP.
Actinobacillus Infections ; diagnosis ; Adult ; Aggregatibacter actinomycetemcomitans ; isolation & purification ; Bacteroidaceae Infections ; diagnosis ; Bacteroides ; isolation & purification ; Bacteroides Infections ; diagnosis ; Chronic Periodontitis ; microbiology ; Dental Plaque ; microbiology ; Dental Plaque Index ; Female ; Gingivitis ; microbiology ; Gram-Negative Bacteria ; isolation & purification ; Humans ; Lichen Planus, Oral ; microbiology ; Male ; Middle Aged ; Periodontal Attachment Loss ; microbiology ; Periodontal Index ; Periodontal Pocket ; microbiology ; Porphyromonas gingivalis ; isolation & purification ; Prevotella intermedia ; isolation & purification ; Treponema denticola ; isolation & purification ; Treponemal Infections ; diagnosis

BACKGROUND: Bacteroides fragilis group organisms are the most frequently isolated anaerobes in human infections. Increasing resistance to various antimicrobial agents is a significant problem in choosing appropriate antimicrobial agents to treat anaerobic infections. Periodic monitoring of the regional resistance trends of B. fragilis group isolates is needed. METHODS: A total of 466 nonduplicate clinical isolates of B. fragilis group organisms (276 B. fragilis, 106 Bacteroides thetaiotaomicron, and 84 other B. fragilis group organisms) were collected during the 8-yr period from 1997 to 2004 in a Korean university hospital. Minimum inhibitory concentrations to various antimicrobial agents were determined by the CLSI agar dilution method. RESULTS: Eight isolates were resistant to imipenem. Additionally, the resistance rates to cefotetan were decreased in B. thetaiotaomicron, while those for clindamycin were significantly increased compared to the rates found in previous studies. Depending on species, resistance rates were 1-4% for imipenem, 1-6% for piperacillin-tazobactam, 4-11% for cefoxitin, 33-49% for piperacillin, 14-60% for cefotetan, and 51-76% for clindamycin. No isolates were resistant to chloramphenicol or metronidazole. CONCLUSIONS: Piperacillin-tazobactam, cefoxitin, imipenem, chloramphenicol, and metronidazole are still active against B. fragilis group isolates, while clindamycin no longer has a value as an empirical therapeutic agent in Korea. Furthermore, this study identified the first imipenem-resistant B. fragilis group isolates in Korea.
Anti-Bacterial Agents/pharmacology ; Bacteroides/classification/*drug effects/isolation & purification ; Bacteroides fragilis/drug effects/isolation & purification ; Cefoxitin/pharmacology ; Chloramphenicol/pharmacology ; *Drug Resistance, Multiple, Bacterial ; Humans ; Imipenem/pharmacology ; Metronidazole/pharmacology ; Microbial Sensitivity Tests ; Penicillanic Acid/analogs & derivatives/pharmacology ; Piperacillin/pharmacology ; Republic of Korea

BACKGROUND: Periodic monitoring of antimicrobial resistance trends of clinically important anaerobic bacteria such as Bacteroides fragilis group organisms is required. We determined the antimicrobial susceptibilities of clinical isolates of B. fragilis group organisms recovered from 2009 to 2012 in a tertiary-care hospital in Korea. METHODS: A total of 180 nonduplicate clinical isolates of B. fragilis group organisms were collected in a tertiary care hospital. The species were identified by conventional methods: the ATB 32A rapid identification system (bioMerieux, France) and the Vitek MS matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (bioMerieux). Antimicrobial susceptibility was determined by the CLSI agar dilution method. RESULTS: Imipenem and meropenem resistance rates were 0-6% for B. fragilis group isolates. The rate of resistance to piperacillin-tazobactam was 2% for B. fragilis and 0% for other Bacteroides species, but 17% for B. thetaiotaomicron isolates. High resistance rates to piperacillin (72% and 69%), cefotetan (89% and 58%), and clindamycin (83% and 69%) were observed for B. thetaiotaomicron and other Bacteroides spp. The moxifloxacin resistance rate was 27% for other Bacteroides spp. The MIC50 and MIC90 of tigecycline were 2-4 microg/mL and 8-16 microg/mL, respectively. No isolates were resistant to chloramphenicol or metronidazole. CONCLUSIONS: Imipenem, meropenem, chloramphenicol, and metronidazole remain active against B. fragilis group isolates. Moxifloxacin and tigecycline resistance rates are 2-27% and 8-15% for B. fragilis group isolates, respectively.
Anti-Infective Agents/*pharmacology ; Bacteroides Infections/*microbiology/pathology ; Bacteroides fragilis/*drug effects/isolation & purification ; Drug Resistance, Multiple, Bacterial ; Humans ; Imipenem/pharmacology ; Inhibitory Concentration 50 ; Microbial Sensitivity Tests ; Penicillanic Acid/analogs & derivatives/pharmacology ; Piperacillin/pharmacology ; Republic of Korea ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tertiary Care Centers ; Thienamycins/pharmacology