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Pediatric osteomyelitis: III. anaerobic microorganisms.

Primary osteomyelitis consequent to obligate anaerobic microorganisms represents an infrequently encountered type of infection in pediatric patients. Unlike osteomyelitis caused by more common microorganisms such as Staphylococcus, children with osseous lesions due to anaerobic microorganisms are frequently minimally symptomatic and rarely present the classic signs of fulminant osteomyelitis. Radiographically, the lesions may mimic malignant osseous tumors. Fastidious microbiologic analysis of the material obtained at surgery is necessary to isolate obligate anaerobes. Basic treatment, comprising surgical drainage and appropriate antimicrobial agents, does not differ from that for osteomyelitis caused by aerobic or by facultative anaerobic microorganisms.

Adolescent↗

Detection of bacteria in equine synovial fluid by use of the polymerase chain reaction.

Equine synovial fluid aliquots were inoculated with Salmonella enteritidis, Escherichia coli, Actinobacillus equuli, Staphylococcus aureus, and Streptococcus zooepidemicus to obtain approximate concentrations of 1000, 100, 10, and 1 colony forming U/mL. Synovial fluid aliquots were also inoculated with an unquantitated inoculum of Bacteroides fragilis and Clostridium perfringens. Inoculated synovial fluid was incubated in trypticase-soy broth or Columbia broth for approximately 12 hours. Then aliquots were removed for DNA extraction and polymerase chain reaction (PCR) analysis for detection of a 531 base-pair segment of bacterial DNA corresponding to a region of the 16S ribosomal gene. Duplicate samples of inoculated synovial fluid were prepared for microbial culture. Bacteria were detected in all samples inoculated with bacteria but not in control synovial fluid samples. Under experimental conditions there was no difference between microbial culture and PCR analyses for detection of bacteria. Experimentally, PCR was able to detect bacteria in synovial fluid within 24 hours of inoculation.

Actinobacillus↗

Intermittent hyperbaric oxygen therapy for reduction of mortality in experimental polymicrobial sepsis.

Hyperbaric oxygen therapy has a marked beneficial effect in experimental intraabdominal sepsis. Two rat models involving implantation of either rat fecal material or a mixture of pure cultures of Escherichia coli, Streptococcus faecalis, and Bacteroides fragilis were used in this study. A death rate of 100% was obtained in control animals implanted with fecal material; with intermittent hyperbaric oxygen treatment, a death rate of only 8% was observed (P less than .005). With a mixture of pure cultures of clinical pathogens, the death rate in control animals was 79%, and intermittent hyperbaric oxygen treatment reduced the rate to 23% (P less than .005). Data from cultures of blood indicated that the efficacy of hyperbaric oxygen was not related to antibacterial activity.

Animals↗

Aerobic and anaerobic microbiology of necrotizing fasciitis in children.

Specimens obtained from eight children with necrotizing fasciitis (NF) were cultured for aerobic and anaerobic bacteria. A total of 21 isolates were recovered, 13 anaerobic and 8 aerobic or facultatives. The facultative organism Streptococcus pyogenes was present alone in two (25%) instances, and mixed aerobic and anaerobic bacteria were isolated in six (75%). The predominant isolates were Peptostreptococcus spp. (6 isolates, including 3 Peptostreptococcus magnus). S. pyogenes (4), Bacteroides fragilis group (3), Clostridium perfringens (2), Escherichia coli (2), and Prevotella spp. (2). Organisms similar to the ones isolated from the NF aspirates were recovered in the blood of all patients except one. These included S. pyogenes (3 isolates). B. fragilis group (2), E. coli (1), and P. magnus (1) and Clostridium perfringens (1). All patients underwent surgical fasciotomy, and four required skin grafting. Antimicrobials were administered to all children. Despite extensive resection and intense supportive therapy, three patients died from sepsis accompanied by shock acidosis and disseminated intravascular coagulation. These findings illustrate the polymicrobial aerobic-anaerobic flora of NF in children.

Acidosis↗

Histopathology of mixed anaerobic intraabdominal infection in mice.

This study characterized acute peritonitis and chronic abscess formation resulting from experimental mixed anaerobic infection with Bacteroides melaninogenicus and Fusobacterium necrophorum. At intervals after infection liver and spleen samples were obtained, fixed, and processed for histological examination. An acute to chronic infection progressed in mice infected with this mixture of anaerobic bacteria, whereas, no infection resulted when either organism was injected alone. Acute inflammatory cell infiltrates were noted in tissue samples at 12 h postinfection. Small, discrete areas of liver cell necrosis with neutrophilic infiltrates were observed as early as 24 h. By 48 h after infection the liver parenchyma was infiltrated with both acute and chronic inflammatory cells, with moderate to severe hepatocyte degeneration recognized at 72 h. Large intrahepatic abscesses were present in the subphrenic (upper lobe) area 2 to 6 weeks after experimental infection.

Animals↗

Comparison of in vitro activities of DU-6859a and other fluoroquinolones against Japanese isolates of anaerobic bacteria.

The in vitro activity of DU-6859a, a new fluoroquinolone, was compared with those of other fluoroquinolones against clinical isolates of anaerobic bacteria and Gardnerella vaginalis. DU-6859a was the most active agent; it inhibited 90% of isolates of almost all species tested, including Bacteroides fragilis at < or = 0.39 micrograms/mL. Although the other quinolones tested were active against gram-positive anaerobes, inhibiting their growth at < or = 1.56 micrograms/mL, these agents were less active against the B. fragilis group and Prevotella bivia (90% of which were inhibited at > or = 6.25 micrograms/mL). Mobiluncus species and G. vaginalis, which are well associated with bacterial vaginosis, were inhibited by DU-6859a at 0.1 microgram/mL. These results suggest that DU-6859a is a promising oral agent for the treatment of bacterial infections due to anaerobic bacteria; however, further studies, including determination of vaginal levels of this compound, should be performed to study the role of DU-6859a in the treatment of bacterial vaginosis.

Administration, Oral↗

The capsular polysaccharide complex of Bacteroides fragilis induces cytokine production from human and murine phagocytic cells.

To stimulate early-phase immunologic events following Bacteroides fragilis infection in the peritoneal cavity, we examined the cytokine response of several cell types to purified capsular polysaccharide complex (CPC) and lipopolysaccharide (LPS) of this organism. Cytokines were produced from murine resident peritoneal (MRP) cells as well as human peripheral blood leukocytes. MRP cells cocultured with either B. fragilis CPC of LPS in vitro produced tumor necrosis factor alpha and interleukin-1alpha (IL-1alpha). In addition, MRP cells challenged with CPC produced IL-10. Human peripheral blood monocytes and polymorphonuclear leukocytes secreted IL-8 when cultured in the presence of CPC.

Animals↗

Chloramphenicol: recent developments and clinical indications.

Recent developments (including more accurate assays) that have led to revised recommendations for route of administration, dosage, and indications for chloramphenicol are reviewed. Chloramphenicol is most bioavailable by the oral route; doses of 75 mg/kg/day provide adequate therapeutic concentrations for most clinical indications. Serum concentrations of the drug should be monitored to ensure adequate therapeutic concentrations and to avoid toxicity. This is particularly important in newborns, in patients with liver dysfunction, and in those receiving concomitant drugs that may influence free chloramphenicol concentrations. The indications for chloramphenicol therapy evaluated are: Haemophilus influenzae infections, anaerobic infections, salmonellosis, Rocky Mountain spotted fever, and eye infections. Chloramphenicol is useful in the treatment of invasive Haemophilus influenzae infections resistant to ampicillin, in selected anaerobic and ocular infections, and for rickettsioses in patients under the age of eight years. Because the rare but life-threatening complication of chloramphenicol (aplastic anemia) persists, indications for the drug's clinical use are narrowing once again with the advent of third-generation cephalosporins highly active against gram-negative bacilli including ampicillin-resistant H. influenzae. Similarly, metronidazole or clindamycin may be preferred for some anaerobic infections.

Bacteroides Infections↗

A multiplex polymerase chain reaction-based diagnostic method for bacterial vaginosis.

OBJECTIVE: To develop a polymerase chain reaction (PCR)-based diagnostic method for bacterial vaginosis using bacterial vaginosis-associated anaerobes. METHODS: A multiple PCR assay was developed using primers specific to 16S ribosomal deoxyribonucleic acid (DNA) (Mobiluncus mulieris and Mobiluncus curtisii), nanH (Bacteroides fragilis), and an internal spacer region of ribosomal DNA (Gardnerella vaginalis). The vaginal swabs from pregnant and nonpregnant women were examined by Gram stain-based Nugent scoring system. One hundred seventy-two samples of 853 Gram stain-interpretable samples were randomly selected and subjected to multiplex PCR assay. RESULTS: The sensitivity of the PCR assay ranged from 10 to 10 colony-forming units per vaginal swab. The prevalence of the bacterial vaginosis, intermediate, and normal categories was found by Nugent scoring system to be 21.6% (184/853), 26.0% (222/853), and 52.4% (447/853), respectively. By the multiplex PCR-based diagnostic method, 20.3% (35/172) of the samples were identified as bacterial vaginosis. The diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of multiplex PCR in comparison with Gram stain examination were 78.4% (95% confidence interval [CI] 65.1%, 91.6%), 95.6% (95% CI 92.1%, 99.0%), 82.9% (95% CI 70.4%, 95.4%), and 94.2% (95% CI 90.3%, 98.1%), respectively. CONCLUSION: This multiplex PCR can be used as a diagnostic or screening test for bacterial vaginosis.

Adult↗

Use of Tn KPK2 for sequencing a 10.6-kb PstI DNA fragment of Bradyrhizobium japonicum and for the construction of aspA and ndvA mutants.

Transposon Tn KPK2 was used to saturate a randomly cloned Bradyrhizobium japonicum PstI fragment and the insertions were used as starting points for the sequence determination. The first gene of the 10.6-kb DNA insert encodes a homologue to ndvA, the product of which is known to be involved in the formation of periplasmic cyclic glucans. Selected Tn KPK2 insertions were introduced into the B. japonicum wild-type strain. The resulting mutants were subsequently tested for their symbiotic interactions with soybeans. As in Sinorhizobium meliloti, a B. japonicum ndvA mutant was affected in salt-stress tolerance and exhibited symbiotic defects in that it induced the formation of ineffective soybean nodules. The central nodule tissue was infected by bacteroids, but within the infected cells the mutant was not properly maintained. Another gene was found to be highly similar to bacterial aspartases and thus was named aspA. The putative function of the product of this gene was confirmed by genetic complementation of aspartase-less Escherichia coli strain TK237. The symbiotic phenotype of a B. japonicum aspA:Tn KPK2 mutant consisted of enlarged symbiosomes that made the system ineffective. In general, Tn KPK2 is a suitable means for fast sequencing. In combination with pJQ200SK, the resulting recombinant plasmids can be directly used to create genetically defined mutants.

ATP-Binding Cassette Transporters↗

The International Infections in Pregnancy (IIP) study: variations in the prevalence of bacterial vaginosis and distribution of morphotypes in vaginal smears among pregnant women.

OBJECTIVE: The objective of the study was to determine the prevalence of bacterial vaginosis and the distribution of associated morphotypes among asymptomatic pregnant women in different countries. STUDY DESIGN: In 8 institutions participating in the Global Network for Perinatal and Reproductive Health (www.gnprh.org) from July 1999 to September 2001, 1466 women were enrolled. Vaginal smears were Gram stained and scored with Nugent's method at a reference laboratory. The prevalence of bacterial vaginosis and bacterial morphotype distributions were compared. RESULTS: Overall, 12.3% of women had bacterial vaginosis according to Nugent's criteria. Zimbabwe had the highest prevalence (24.4%) when compared with all other sites, except Myanmar (P < .05). Among bacterial vaginosis cases, 98.9% of vaginal smears had more than 30 Gardnerella/Bacteroides morphotypes present per oil immersion field. Individual centers showed significant differences in the number of Mobiluncus and lactobacillus morphotypes (P < .01). CONCLUSION: The prevalence of bacterial vaginosis and distribution of bacterial morphotypes in vaginal smears among asymptomatic pregnant women vary significantly in populations from different countries.

Actinomycetales Infections↗

Early treatment of cavernositis resulted in erectile function preservation.

INTRODUCTION: Some of the more common complications of intracavernous injection (ICI) therapy include pain and prolonged erections. Rare reported complications include intracorporeal needle breakage and postinjection cavernositis. AIM: We report a case of early management of postinjection cavernositis resulting in preservation of erectile function with no angulation. METHODS: A 53-year-old male with no history of diabetes mellitus presented to our department with a small painful penile swelling 36 hours after ICI of 15 microg prostaglandin E1. Clinical, laboratory, and ultrasound assessments suggested the occurrence of cavernositis. Surgical debridement, abscess drainage, and antibiotics resulted in marked improvement. Follow-up showed normal erectile function with minimal scarring. CONCLUSIONS: Timely management of cavernositis can help prevent loss of erectile function. Stringent supervision of patients, after adequate ICI training, together with drug prescription, is essential.

Alprostadil↗

The bacterial causation of postoperative osteitis pubis.

Five patients with a bacterial causation of postoperative osteitis pubis were treated during a three year period. In each patient, the bacterial cause was suggested by a known infectious process adjacent of the symphysis pubis. It is suggested that a bacterial causation be considered for the clinical and roentgenographic syndrome of osteitis pubis if a contigous infectious process is present.

Aged↗

Different antimicrobial agents in treatment and prophylaxis of experimentally induced intraabdominal sepsis.

A reproducible experimental model of intraabdominal infection in rats has been worked out in order to simulate intraabdominal sepsis seen in humans and to test different antimicrobial agents in treatment and prophylaxis of intraabdominal infections. This experimental model was used to evaluate the efficacy of benzylpenicillin, benzylpenicillin plus sulbactam, cefoxitin, thienamycin, clindamycin, tinidazole, netilmicin, clindamycin plus netilmicin, and tinidazole plus netilmicin in the treatment of intraabdominal sepsis. Sixty-five per cent of the untreated animals died within two days. Within four days, 43% of the animals receiving clindamycin alone, 22% receiving tinidazole alone and 46% receiving netilmicin alone died. Animals treated with piperacillin, clindamycin plus netilmicin or tinidazole plus netilmicin showed a significantly decreased mortality and increased cure rates during the experimental period. Sixty-five per cent of the untreated animals and the animals given sulbactam alone died within 48 h. Over 90% of the animals given benzylpenicillin died within five days. Animals treated with benzylpenicillin plus sulbactam, cefoxitin or thienamycin had a significantly decreased mortality. Within four days 22% of the animals receiving tinidazole alone, 43% receiving clindamycin alone and 46% receiving netilmicin alone died. Animals treated with tinidazole plus netilmicin or clindamycin plus netilmicin had a significantly decreased mortality and increased cure rates during the experimental period. Only 5% of these animals died. In the prophylaxis experiment the following agents were tested: cefoxitin, doxycycline, tinidazole plus netilmicin, clindamycin plus netilmicin, and trimethoprim-sulfa plus tinidazole. One dosage of the antimicrobial(s) reduced the mortality rate significantly.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Sustained infection induces 2 distinct microvascular mechanisms in the splanchnic circulation.

BACKGROUND: Altered intestinal blood flow during systemic inflammation leads to organ dysfunction. Mucosal ischemia occurs during sepsis despite an increase in portal blood flow. We hypothesized that separate mechanisms are active in the large resistance and small mucosal microvessels to account for this dichotomy. METHODS: Chronic infection was induced in rats by bacterial inoculation (Escherichia coli and Bacteroides fragilis) of an implanted subcutaneous sponge. Separate groups were studied at 24 and 72 hours after a single inoculation of bacterium or 24 hours after a second inoculation (ie, 72 hours of sepsis). Time-matched controls were used for each group. Intravital microscopy of the terminal ileum was used to assess endothelial-dependent vasodilation to acetylcholine (10(-9) to 10(-5) mol/L) in resistance (A(1)) and premucosal (A(3)) arterioles. Threshold sensitivity (-log of 20% response dose) was calculated from dose response curves for each animal. RESULTS: Vasodilator sensitivity to acetylcholine in A(1) arterioles was significantly decreased at 24 hours, and these changes persisted up to 72 hours after a single bacterial inoculation. There was no change in the dilator sensitivity of A(3) arterioles after a single inoculation. When there was a challenge with a second bacterial inoculation, there was a reversal of the A(1) dilator response and an increase in A(3) sensitivity. CONCLUSIONS: An initial septic event results in a decrease in dilator reactivity in the resistance A1 arterioles that persists for at least 72 hours. A sustained septic challenge results in increased dilator reactivity in both A(1) and A(3) vessels. This enhanced sensitivity during sepsis suggests that more than 1 therapeutic approach to preservation of intestinal blood flow will be necessary.

Acetylcholine↗

Thrombosis and infection: a case of transient anti-cardiolipin antibody associated with pylephlebitis.

Infections with Bacteroides species have been noted to occur in association with cases of thrombophlebitis. This association has led to the speculation that the microorganisms themselves may contribute to the pathogenesis of thrombus formation through elaborated enzymes, including heparinases, or by interactions between the clotting cascade and the unique structure of the Bacteroides lipopolysaccharide. Anti-phospholipid antibodies have been linked with hypercoagulable states and thrombus formation. Although a number of infections have been associated with the transient production of anti-cardiolipin antibodies, the effect the antibodies may have in contributing to thrombus formation is not well understood. The occurrence of Bacteroides species infection with transient anti-cardiolipin antibody has not been previously reported.

Antibodies, Anticardiolipin↗