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Production of menaquinones by intestinal anaerobes.

Ninety intestinal organisms (71 isolates from fecal samples of neutropenic patients with cancer or from various sites in patients with intraabdominal infections and 19 control strains) were examined by reverse-phase thin-layer chromatography for their ability to produce menaquinones in vitro. Menaquinones were found in all of 24 organisms of the Bacteroides fragilis group. Two other Bacteroides species, Bacteroides disiens and Bacteroides bivius, also produced menaquinones. A single isolate of Bacteroides species lacked menaquinones. These constituents were found in all of five strains of Escherichia coli, all of four strains of Klebsiella pneumoniae, five of eight strains of Propionibacterium species, two of five strains of Eubacterium species, and the one strain each of Arachnia propionica and Veillonella parvula tested. No menaquinones were detected in organisms of the genera Fusobacterium, Clostridium, Bifidobacterium, Lactobacillus, Actinomyces, Peptococcus, or Peptostreptococcus. These findings suggest that E. coli, Bacteroides species, and some gram-positive, anaerobic, non-spore-forming bacilli produce menaquinones that may be a source of vitamin K in patients who are deprived of exogenous vitamin K1.

Actinomycetaceae↗

Bactericidal effects of photoradiation therapy with hematoporphyrin derivative.

Hematoporphyrin derivative (Hpd) localizes selectively in malignant and rapidly metabolizing tissues and undergoes a cytotoxic reaction when exposed to light of a specific wavelength. Hpd has been studied extensively with regard to the diagnosis and treatment of tumors but not with regard to bactericidal activity. This investigation assessed the effect of light-activated Hpd on various microorganisms, on human polymorphonuclear leukocytes, and on the interactions of polymorphonuclear leukocytes and bacteria. Light-activated Hpd was 99.9% bactericidal against Staphylococcus aureus, Streptococcus faecalis, Bacteroides fragilis, Streptococcus M-G intermedius, Streptococcus mutans, Peptostreptococcus anaerobius, Peptococcus magnus, and Clostridium perfringens, but did not affect Escherichia coli, Pseudomonas aeruginosa, or Candida albicans. Photoactivation of Hpd rendered 98% of polymorphonuclear leukocytes inviable. Combinations of light-activated Hpd and polymorphonuclear leukocytes had a 4-log (99.99%) bactericidal effect on both intra- and extracellular S. aureus. The ability of Hpd to localize in inflammatory tissues may have therapeutic applications in the treatment of abscesses.

Bacteria↗

Polymicrobial early postpartum endometritis with facultative and anaerobic bacteria, genital mycoplasmas, and Chlamydia trachomatis: treatment with piperacillin or cefoxitin.

A protected, triple-lumen transcervical culture method was used to recover organisms from the endometrium. At least one facultative or one anaerobic species of bacteria was recovered from 82% of the patients, and genital mycoplasmas were recovered from 76% of the women with endometritis. Bacteria together with genital mycoplasmas were present in 61% of the women, bacteria alone were present in 20%, genital mycoplasmas alone were present in 16%, and Chlamydia trachomatis was isolated from 2% of the patients. The most common organisms included Gardnerella vaginalis, Peptococcus spp., Bacteroides spp., Staphylococcus epidermidis, group B Streptococcus, and Ureaplasma urealyticum. A randomized, double-blind regimen of either piperacillin or cefoxitin was equally successful in treating the postpartum endometritis.

Bacteriuria↗

Interaction between penicillin, clindamycin or metronidazole and gentamicin against species of clostridia and anaerobic and facultatively anaerobic gram-positive cocci.

Seven anaerobic and facultative Gram-positive cocci and 12 clostridial species were tested for in-vitro and in-vivo susceptibilities to penicillin, clindamycin, and metronidazole, used singly or in combination with gentamicin. The in-vitro tests consisted of determination of minimal inhibitory concentration (MIC), done without or with constant amounts of gentamicin. When used alone or in combination with penicillin or metronidazole, gentamicin had negligible effects on the bacteria. When used with clindamycin, gentamicin significantly reduced the MIC for one strain each of Peptococcus magnus and Clostridium difficile. The in-vivo tests were carried out in mice and consisted of studying the bacterial contents of abscesses induced by subcutaneous injection of bacterial suspensions. Synergy between gentamicin and penicillin, clindamycin or metronidazole was shown respectively in five, three and one strain. Consistency between in-vitro and in-vivo findings was present in the above mentioned strains only between gentamicin and clindamycin. The synergy between penicillin, clindamycin or metronidazole and gentamicin in Gram-positive anaerobic and facultative organisms may have clinical implications.

Animals↗

Evaluation of imipenem/cilastatin against nosocomial infections and multiresistant pathogens.

Thirty-five patients suffering from soft tissue infections (12), upper UTIs (6), bronchopneumonia (6), septicaemia (2), chronic osteomyelitis (2), intra-abdominal abscess (2), liver abscess (1), lung abscess (1), acute cholangitis (1), thoracic empyema (1) and chronic prostatitis (1) were given imipenem/cilastatin for 6-21 days. In 22 patients several aggravating factors coexisted, while infection in 16 patients was polymicrobial. The following pathogens were implicated: Pseudomonas aeruginosa (21), Escherichia coli (15), Enterobacter cloacae (6), Proteus spp. (3), Klebsiella pneumoniae(3), Citrobacter freundii (1), Salmonella enteritidis (1), Acinetobacter spp. (4), Haemophilus influenzae (2), Bacteroides fragilis (1) and Peptococcus saccharolyticus (1) with MICs to imipenem ranging between 0.5 and 8 mg/l. A successful clinical response was observed in 91.4% of the patients, while pathogens were eradicated in 75.9%, persisted in 24.2% and recurred, in 9.1% of patients, with development of resistance to imipenem in two Ps. aeruginosa strains. Against 150 multiresistant strains of Ps. aeruginosa, 40% of which were resistant to amikacin, 86.4% and 88.9% were found sensitive to ceftazidime and imipenem respectively. It is concluded that imipenem provides the possibility of treating successfully multiresistant and polymicrobial infections with a single antimicrobial.

Adult↗

Microbiological and clinical aspects of aspiration pneumonia.

Aspiration pneumonia is characterized by a pneumonitis in a dependent segment of the lung with typical necrosis or abscess-formation in the parenchyma. Observed aspiration or predisposition to aspiration, cavitation or abscess formation, with or without empyema fluid and isolation of distinctive micro-organisms are important clues to the diagnosis. Diagnostic procedures to collect anaerobic uncontaminated secretions are transtracheal aspiration, blood cultures, pleural fluid aspiration, fibreoptic bronchoscopic protected investigations and percutaneous transthoracic aspiration. Leading pathogens in more than 90% are anaerobic bacteria, mostly species of Bacteroides, Fusobacterium, Peptococcus and Peptostreptococcus; aerobic bacteria include Staphylococcus aureus and Gram-negative bacilli, mainly Klebsiella spp. and Pseudomonas aeruginosa. Treatment depends on bacteriological results; penicillin G and clindamycin are the most useful antibiotics against anaerobes and should be administered over a long period of time (4-12 weeks), adjusted to the clinical course of the individual patient.

Humans↗

In-vitro activity of a new penem FCE 22101.

The in-vitro activity of a new penem FCE 22101 was evaluated in comparison with other antimicrobial agents against 966 aerobic and anaerobic Gram-negative and Gram-positive bacteria. FCE 22101 inhibited 90% of Enterobacteriaceae, Haemophilus influenzae, H. parainfluenzae, H. ducreyi, Neisseria gonorrhoeae, Branhamella catarrhalis, Staphylococcus aureus, Staph. epidermidis and group B streptococcus at concentrations of 0.12-4.0 mg/l. Pseudomonas aeruginosa was resistant with MIC90 of greater than 32 mg/l. Streptococcus faecalis displayed MIC90 of 16 mg/l. Bacteroides fragilis, Clostridium spp., Peptococcus spp. and Peptostreptococcus spp. were inhibited at concentrations of 0.25-2.0 mg/l. beta-Lactamase-producing or methicillin resistant bacteria did not have significantly elevated MICs. FCE 22101 was generally less active than imipenem but more active than other beta-lactam antibiotics examined.

Anti-Bacterial Agents↗

Treatment of bone and soft tissue infections with teicoplanin.

The use of teicoplanin was evaluated in a group of 33 patients, which included 18 prosthetic infections, seven with osteomyelitis, five skin and soft tissue infections, and three cases of septic arthritis. The bacterial pathogens isolated were 27 isolates of Staphylococcus aureus (methicillin sensitive 18; methicillin resistant 9), Staph. hominis (2), Staph. epidermidis (1), Staph. haemolyticus (2), Enterococcus faecalis (1), Streptococcus pyogenes (1), Peptococcus spp. (1). Eleven patients had polymicrobial infections (Gram-positive plus Gram-negative and/or anaerobic bacteria). In ten cases (nine polymicrobial infections) teicoplanin was used in combination with other antibiotics. Teicoplanin was administered intravenously 400 mg every 24 h in 26 cases, 200 mg every 24 h in six cases and 600 mg every 24 h in one case. The duration of the treatment ranged from 9 to 73 days with a mean of 27.3. Efficacy was evaluable in 30 patients, because in three patients treatment was stopped because of adverse reactions. Cure or improvement was noted in 28 patients (93%). Bacteriological cure occurred in 25 patients (83.3%).

Adolescent↗

The protein responsible for center A/B in spinach photosystem I: isolation with iron-sulfur cluster(s) and complete sequence analysis.

The 9 kDa polypeptide from spinach photosystem I (PS I) complex was isolated with iron-sulfur cluster(s) by an n-butanol extraction procedure under anaerobic conditions. The polypeptide was soluble in a saline solution and contained non-heme irons and inorganic sulfides. The absorption spectrum of this iron-sulfur protein was very similar to those of bacterial-type ferredoxins. The amino acid sequence of the polypeptide was determined by using a combination of gas-phase sequencer and conventional procedures. It was composed of 80 amino acid residues giving a molecular weight of 8,894, excluding iron and sulfur atoms. The sequence showed the typical distribution of cysteine residues found in bacterial-type ferredoxins and was highly homologous (91% homology) to that deduced from the chloroplast gene, frxA, of liverwort, Marchantia polymorpha. The 9 kDa polypeptide is considered to be the iron-sulfur protein responsible for the electron transfer reaction in PS I from center X to [2Fe-2S] ferredoxin, namely a polypeptide with center(s) A and/or B in PS I complex. It is noteworthy that the 9 kDa polypeptide was rather hydrophilic and a little basic in terms of the primary structure. A three-dimensional structure was simulated on the basis of the tertiary structure of Peptococcus aerogenes [8Fe-8S] ferredoxin, and the portions in the molecule probably involved in contacting membranes or other polypeptides were indicated. The phylogenetic implications of the structure of the present polypeptide as compared with those of several bacterial-type ferredoxins are discussed.

Amino Acid Sequence↗

Endometrial histopathology in patients with culture-proved upper genital tract infection and laparoscopically diagnosed acute salpingitis.

To define and quantitate histologic changes in the endometrium that best correlate with documented upper genital tract infection (UGTI) and laparoscopically diagnosed acute salpingitis, we studied endometrial biopsy specimens from 69 consecutive patients with clinically suspected acute pelvic inflammatory disease (PID) who underwent microbiological evaluation for UGTI and laparoscopic examination for acute salpingitis. Both UGTI and acute laparoscopically confirmed salpingitis were present in 37 patients (54%), UGTI without salpingitis in 1 (1%), salpingitis without UGTI in 11 (16%), and neither UGTI nor salpingitis in 20 (29%). Chlamydia trachomatis or Neisseria gonorrhoeae UGTI was found in 34 women, Escherichia coli in two patients, Peptococcus magnus in one woman, and with Streptococcus agalactiae in one woman. The following features were correlated both with UGTI and with salpingitis: presence of any neutrophils in the endometrial surface epithelium; neutrophils within gland lumens; dense subepithelial stromal lymphocytic infiltration; any stromal plasma cells; and germinal centers containing transformed lymphocytes. The simultaneous presence of five or more neutrophils per X 400 field in endometrial surface epithelium, together with one or more plasma cell per X 120 field in endometrial stroma, was the best predictor of UGTI plus salpingitis. This combination had a sensitivity of 92% and a specificity of 87% for predicting the diagnosis of both UGTI and laparoscopically confirmable acute salpingitis. Prospective studies are needed to assess the usefulness of these criteria.

Acute Disease↗

Studies on the endogenous flora of the human breast.

Identification of the endogenous microbiological flora of the human breast and its role in breast infections following subglandular augmentation or reduction mammaplasty was undertaken. A total of 231 cultures were performed on 59 breasts in 30 patients. Patients were followed for 12 months. No fungus was cultured from any specimen. Of the breasts cultured 53% were positive for coagulase-negative staphylococcus. Other aerobes found included diphtheroids, lactobacillus, D-enterococcus, micrococcus, and alpha-hemolytic streptococcus. Propionibacterium acne was the most frequent anaerobic bacteria cultured. Other anaerobes included peptococcus and clostridium sporogenes. There was no correlation with respect to the type of bacterium and the depth within the breast where the culture specimens were taken. Postoperative wound infections developed in 2 of 19 patients undergoing reduction mammaplasty. Bacteria identical to those cultured at the time of surgery were again cultured from the wound. Twenty subglandular augmentation mammaplasties were performed with a 25% capsular rate at one year. Two capsules were associated with no bacterial growth at the time of mammaplasty surgery, whereas three were associated with coagulase-negative staphylococcus, Propionibacterium acne, and diphtheroids, respectively. Of the 15 breasts with no capsular contracture after one year, operative culture revealed coagulase-negative staphylococcus in 8 and no bacterial growth in 7. Even breast tissue located deep within the gland away from the nipple contains a flora that is similar to that of normal skin. Cases of infection in which the endogenous bacteria were correlated with later infection was documented.

Bacteria↗

Aerobic and anaerobic bacterial flora of burns in children.

Aerobic and anaerobic bacterial flora of burn sites in 180 children were monitored. Specimens were obtained twice a week; each patient had between one and 21 cultures taken (mean 2.4). A total of 392 specimens were collected over 2 years, of which 319 were positive. Aerobic bacteria alone were present in 225 specimens (71%) and anaerobic bacteria alone were present in 26 (8%). Mixed aerobic and anaerobic bacteria were present in 68 burn specimens (21%). A total of 580 isolates (448 aerobes and 132 anaerobes) were recovered, accounting for 1.8 isolates per specimen (1.4 aerobes and 0.4 anaerobes). The predominant aerobic isolates were S. epidermidis, S. aureus, alpha hemolytic streptococcus, Pseudomonas sp., and Group D streptococcus. The predominant anaerobic isolates were: P. acnes, anaerobic Gram-positive cocci, and Bacteroides sp. Blood cultures were drawn from 45 children: four showed bacterial growth of one of each of the following isolates: S. aureus, E. coli, Peptococcus asaccharolyticus, and B. fragilis. The numbers of isolates per specimen were higher in the oral and anal area (3.2 and 2.8) than in the extremities and trunk (1.8 and 0.9). Gram-negative enteric rods and Group D streptococci were more frequently recovered from the anal area. S. aureus, S. epidermidis, and P. acnes were more frequently recovered from extremities. Bacteroides sp. and Fusobacterium nucleatum were more frequently recovered from the anal and oral areas. All children were treated with local application of silver sulfadiazine, and antimicrobial therapy was used in 128 children. Statistical analysis showed no correlation between the bacteria isolated and use of antimicrobial agents. These data suggest a role for anaerobic bacteria in the colonization of burns in children, especially in burns of the anal and oral areas.

Adolescent↗

Use of parenteral antibiotic therapy to eradicate bacterial colonization of amniotic fluid in premature rupture of membranes.

The successful eradication of bacterial colonization of the amniotic fluid with parenteral maternal antibiotic therapy in a preterm gestation with premature rupture of membranes is reported. Amniotic fluid, obtained by amniocentesis, of a pregnancy complicated by premature rupture of membranes at 29 weeks was found to have three different bacterial species (Bacteroides bivius, Veillonella parvula, and Peptococcus). Because of prematurity and clinical quiescence, parenteral antibiotics were administered to the mother. Repeat amniocentesis was performed during treatment and 48 hours after the cessation of antibiotics failed to demonstrate bacterial growth in the amniotic fluid. Fetal lung maturity was attained, and an induction of labor was performed when antepartum heart rate testing was equivocal. This was accomplished without maternal or fetal/neonatal mishap.

Adult↗

Anaerobic spondylodiscitis: case series and systematic review.

BACKGROUND: Bacterial spondylodiscitis is rarely caused by anaerobic organisms. We describe two patients with lumbar vertebral osteomyelitis and discitis caused by anaerobic bacteria, including an unusual occurrence after an endodontic procedure, and review the salient clinical features and outcomes of 31 previously reported cases. METHODS: Case reports and review of the literature. RESULTS: Median age at presentation was 65 years, with a male-to-female ratio of 2:1. The most common presenting symptoms were back pain, fever, and neurologic deficits. The lumbar spine was most frequently involved (43%); an equal number of cases involved contiguous extension or hematogenous spread. Causative anaerobes were recovered from disk space or vertebrae (13), blood (4), and/or soft tissue abscess and included Bacteroides species (12), Propionibacterium acnes (7), Peptococcus species (4), Peptostreptococcus species and Clostridium species (3 each), Corynebacterium diphtheroides and Fusobacterium species (2 each), and unspecified anaerobes (3). CONCLUSIONS: Apart from specific antibiotic selection, medical treatment and outcomes for anaerobic spondylodiscitis are similar to those for aerobic vertebral disk infection.

Aged↗

Effect of temperature, humidity and exposure to oxygen on the survival of anaerobic bacteria.

The relative effect of humidity, temperature and exposure to oxygen on the survival of 11 strains of Bacteriodes spp., Fusobacterium necrophorum, Eubacterium lentum, Clostridium ramosum, Peptococcus asaccharolyticus and Peptostreptococcus anaerobius was evaluated. Suspensions of the test strains were applied to membrane filters either in empty sterile petri dishes or on plates of non-nutrient agar to provide dry or moist conditions. The filters were stored aerobically or anaerobically in an atmosphere containing 10% CO2 at either 4 degrees C or 35 degrees C. After holding periods of 0.375-384 h, the filters were transferred to nutrient plates inside an anaerobic glove box. After incubation the growth on the filters was compared with that on control filters that had not been stored. Survival was better at 4 degrees C than at 35 degrees C. For the gram-negative organisms, survival was better under moist than under dry conditions, whereas for gram-positive organisms this finding was sometimes reversed. The results indicate that, generally, anaerobic bacteria kept under aerobic conditions survive best under moist conditions at 4 degrees C.

Anaerobiosis↗

The laboratory identification of gram-positive anaerobic cocci.

A collection of 256 clinical strains and 40 reference strains of gram-positive anaerobic cocci (GPAC) was studied, to characterise the recognised species more fully and to define groups of strains which might correspond to previously undescribed species. The methods used were: gas-liquid chromatography (GLC) for the detection of volatile fatty acids (VFAs); determination of the pre-formed enzyme profile with a commercially available kit, ATB 32A; microscopic appearance; colonial morphology; and antibiotic sensitivity tests. Strains were placed in one of five VFA groups according to their GLC profile; 96% of strains were further assigned to 12 groups by their enzyme profile. There was less than 99% agreement between the two methods. Of 111 clinical strains in the VFA-negative group, 110 gave one of three distinct enzyme profiles corresponding to Peptostreptococcus magnus, P. micros and P. heliotrinreducens. The assignment of strains to groups based on their microscopic appearance and colonial morphology agreed well with groupings according to enzyme profile. Identification of butyrate-producing GPAC was unsatisfactory because it relied heavily on the enzyme profile; testing for indole production was of limited discriminative value. Most strains of P. asaccharolyticus and P. indolicus were very similar in enzyme profile, microscopic appearance and colonial morphology, but a sub-group of P. asaccharolyticus could be distinguished. A further indole-positive group corresponding to Hare group III was also noted. Strains of P. prevotii and P. tetradius were very similar, but easily distinguished from other butyrate-producing GPAC. However, 45% of the butyrate-producing cocci could not be assigned to recognised species; most of these were assigned to one of two new groups, the ADH group and the bGAL group, by their enzyme profile, microscopic appearance and smell. Four strains that produced a terminal VFA peak of isovaleric acid formed a new group designated 'ivoricus'. Reliable features for the identification of P. anaerobius were GLC (all GPAC that produced isocaproic acid were identified as P. anaerobius), enzyme profile and sensitivity to SPS. Two clinical strains that produced caproci acid were identified as Hare group VIII; they were distinguished from Peptococcus niger by their enzyme profile and colonial morphology. A phenotypic classification based on GLC and enzyme profile is presented, with a method for the identification of most strains of GPAC within 48 h of primary isolation.

Bacteria, Anaerobic↗

A numerical taxonomic study of the gram-positive anaerobic cocci.

Clinical (101) and collection (26) strains of gram-positive anaerobic cocci were examined in conventional tests and pyrolysis mass spectrometry (PMS). Numerical classifications based upon conventional test reaction patterns (CTRPs) and PMS showed 27 and 22 clusters, respectively. Cross-tabulation of cluster membership in the two classifications showed excellent correlation, with the combined classifications showing clear groups corresponding to the currently recognised species Peptostreptococcus anaerobius, P. heliotrinreducens, P. hydrogenalis, P. indolicus, P. lactolyticus, P. magnus, P. micros and Peptococcus niger. Strains of P. prevotii and P. tetradius clustered together in a heterogeneous group of saccharolytic organisms. However, strains previously identified as P. asaccharolyticus were divided into three distinct groups in PMS, two of which differed only in indole-associated pyrolysis products. A further four groups and several single-member clusters were distinct from these species. PMS data supported the validity of identification by pre-formed enzyme profiles and confirmed that Hare group III is synonymous with P. hydrogenalis, Hare group IV with P. magnus, and the "ADH group" with P. vaginalis. There is clearly a need for a taxonomic revision of the genus Peptostreptococcus, which probably encompasses several generic groups.

Bacteria, Anaerobic↗

Structure at pH 6.5 of ferredoxin I from Azotobacter vinelandii at 2.3 A resolution.

Ferredoxin I from Azotobacter vinelandii (AvFdI) is an iron-sulfur protein composed of 106 amino acids, seven Fe atoms and eight inorganic S* atoms. A crystallographic redetermination of its structure showed the originally reported structure to be incorrect. We report here the crystal structure of AvFdI at pH 6.5. Extensive refinement has led to a final R value of 0.170 for all 6986 non-extinct reflections in the range 10-2.3 A using a solvent model which includes 98 discrete solvent atoms with occupancies between 0.3 and 1.0 and an average B value of 22.5 A(2). The first half of the peptide chain closely resembles that of the 55-residue ferredoxin from Peptococcus aerogenes (PaFd), while the remainder consists of three turns of helix and a series of loops which form a cap over part of the molecular core. Despite the similarities in structure and surroundings, the corresponding 4Fe4S* clusters in PaFd and AvFdI have strikingly different redox potentials; a possible explanation has been sought in the differing hydration models for the two molecules.

Journal Article↗