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The gram-negative bacillary pneumonias.

With increasing age, chronic underlying disease, and debility, the oropharyngeal flora are colonized with aerobic gram-negative bacilli. In this debilitated population, gram-negative bacillary pneumonias (GNBP) are increasingly common. GNBP account for two of every three pneumonia deaths today. As a group, the mortality of GNBP is about 50%. Although the original epidemiologic surveys were done 15 years ago, there is little evidence for an improving case fatality rate despite the appearance of aminoglycoside antibiotics, carbenicillin, and cephalosporins. In susceptible patients, GNBP pneumonias occur both in the community and as nosocomial infections. Recognition of the dangers of contaminated reservoir nebulizers or other similar devices used in inhalation therapy has led to epidemiologic measures within hospitals that have markedly decreased the incidence of this nosocomial GNBP. The role of Gram stain and culture of expectorated sputum and similar examinations of specimens obtained by transtracheal aspiration, fiberoptic bronchoscopy, and lung biopsy in the diagnosis of GNBP are discussed in this review (see Criteria for Diagnosis). In the presence of pulmonary emphysema, congestive heart failure, mixed gram-negative bacillary infections, or the use of immunosuppressive drugs, typical characteristics of individual GNBP may not be apparent. Typical features of Pseudomonas aeruginosa, Escherichia coli, Enterobacter, Proteus, Hemophilus, and anaerobic pulmonary infections are described. Early recognition and institution of appropriate antibacterial agents are emphasized, particularly in GNBP caused by Pseudomonas aeruginosa, Escherichia coli, or Friedländer's bacillus, where the mortality approaches 70%. The mortality of GNBP, including Enterobacter, Proteus, Hemophilus, and anaerobic GNBP, is about 20%. The latter figure is the same as the mortality of pneumococcal pneumonia in similar patients.

Adolescent↗

Activity of Novispirin G-10, a novel antimicrobial peptide against Chlamydia trachomatis and vaginosis-associated bacteria.

We tested the activity of Novispirin G-10, a novel antimicrobial alpha-helical octadecapeptide structurally related to cathelicidins and other innate immunity peptides, against Chlamydia trachomatis serovars L2, D, and E and three organisms associated with bacterial vaginosis (BV). The peptide's activity against C. trachomatis was measured in 48-h shell vial assays with McCoy cell targets. Exposure to 100 micro g/ml of Novispirin G-10 reduced the infectivity of serovars D and E by 99.4-100% and serovar L2 by 91.7-99.1%. At the same concentration of 100 micro g/ml, Novispirin G-10 rapidly killed >99% of Mobiluncus curtisii, Gardnerella vaginalis, and Prevotella bivia, in standard colony-forming unit (CFU) assays. Given its simple structure and relative lack of cytotoxic and hemolytic activity, Novispirin G-10 may be a useful component of microbicide preparations designed to prevent chlamydial infection and/or remediate the abnormal vaginal flora associated with BV.

Anti-Bacterial Agents↗

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↗

Occurrence of Prevotella nigrescens and Prevotella intermedia in infections of endodontic origin.

The occurrence of Prevotella intermedia and Prevotella nigrescens in endodontic infections was studied using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole cell protein to distinguish between the species. Previous studies have shown an association between black-pigmented bacteria (BPB) and endodontic infections and that Prevotella intermedia (previously known as Bacteroides intermedius) was the most commonly isolated BPB. Recently, however, strains identified as P. intermedia were shown to in fact be composed of two separate species, P. intermedia and P. nigrescens. Fifty-six strains of BPB isolated from endodontic infections and previously identified as P. intermedia were used in this study. Following SDS-PAGE, P. nigrescens showed a unique 18.6 kDa band that was used to differentiate P. nigrescens from P. intermedia. Of the 56 strains of BPB, 41 (73.2%) were identified as P. nigrescens and 15 (26.8%) as P. intermedia. This study confirms that P. nigrescens, and not P. intermedia, is the BPB most often isolated from infections of endodontic origin.

Amino Acid Sequence↗

Modulation of chronic enteric infection by distant oral infection.

This investigation determined the effects of a distant oral infection (Porphyromonas gingivalis) on a concurrent local enteric infection (Echerichia coli). A modified murine subcutaneous tissue chamber model was used. Subcutaneously implanted chambers with different microbial makeups were monitored for sloughing and their contents assayed for prostaglandin E2. Bilaterally implanted chamber experiments revealed that a distant "chronic" (immunization with heat-killed organism, followed by live challenge) P. gingivalis infection offered protection against a local chronic E. coli infection, as evidenced by delaying the time for 50% of the chambers to reject from day 19 to day 25 and a statistically significant prostaglandin E2 decrease from 529.4 +/- 176.6 ng/ml to 191.5 +/- 100.9 ng/ml (p < 0.01) (Mann-Whitney test). An acute (live challenge only) distant P. gingivalis infection or immunization with the heat-killed organism alone had no effect on "chronic" E. coli infection in this model. These data suggest that the presence of low-grade chronic oral infection may modify the responses to other infectious challenges.

Animals↗

Survey for collagenase gene prtC in Porphyromonas gingivalis and Porphyromonas endodontalis isolated from endodontic infections.

Collagenase is a potential virulence factor shown to be expressed by Porphyromonas gingivalis associated with periodontal disease. The purpose of this study was to use the polymerase chain reaction (PCR) to detect the presence of the collagenase gene (prtC) in 21 strains of Porphyromonas species isolated from endodontic infections. Type strains for P. gingivalis (ATCC 33277), P. endodontalis (ATCC 35406), Prevotella intermedia (ATCC 25611), and Prevotella nigrescens (ATCC 33563) were used as controls. When PCR primers specific for the 16S ribosomal RNA gene of P. gingivalis or P. endodontalis were used, 16 of the strains were identified as P. gingivalis, and five strains were identified as P. endodontalis. The presence of the prtC gene for collagenase was detected using PCR. Amplicons were analyzed by agarose gel electrophoresis, with an 815 bp amplicon representing the presence of the collagenase gene. Type strain ATCC 33277 and all 16 clinical isolates of P. gingivalis produced the collagenase gene amplicon. Neither type strain ATCC 35406 nor the five strains from clinical isolates of P. endodontalis produced the collagenase gene amplicon. These results indicate that P. gingivalis from endodontic infections possesses the prtC gene. P. endodontalis does not seem to exhibit prtC. The virulence of P. gingivalis may be related to its production of collagenase.

Bacterial Proteins↗

Soft-tissue abscess involving Actinomyces odontolyticus and two Prevotella species in an intravenous drug abuser.

Skin and soft-tissue infections in intravenous users comprise a variety of microorganisms and anaerobic bacteria are frequently involved in these suppurative infections. A case of subcutaneous abscess into anterior femoral muscles involving Actinomyces odontolyticus and two Prevotella species (Prevotella buccae and Prevotella melaninogenica) in an intravenous drug abuser is presented. This combination of microorganisms has not previously been described in soft-tissue infections. The patient volunteering that he licked his hypodermic needle prior to cocaine injection supports that the implicating bacteria originated from the oral cavity. Eventually, the patient recovered and at a 6-month follow-up a gradual improvement of his subcutaneous infection was noticed.

Abscess↗

Severe sepsis caused by Mobiluncus curtisii subsp. curtisii in a previously healthy female: case report and review.

We present the case of a 54-year-old female with life-threatening septicaemia due to Mobiluncus curtisii subsp. curtisii. Her admission was complicated by septic shock, renal failure, disseminated intravascular coagulation, the adult respiratory distress syndrome and spontaneous splenic rupture. The patient survived with full intensive care support and intravenous ceftriaxone. Extra-genital infection with Mobiluncus species is rarely diagnosed and has been confined to breast abscesses and non-life-threatening bacteraemia. A review of extra-genital infections with Mobiluncus species is presented.

Bacteroidaceae Infections↗

Quantitative reverse transcription polymerase chain reaction analysis of Porphyromonas gingivalis gene expression in vivo.

An etiological relationship between periodontitis, a significant oral health problem, and the anaerobe Porphyromonas gingivalis may be related to the expression of a variety of putative virulence factors. The objective of the experiments described here was to develop a quantitative reverse transcription polymerase chain reaction (QRT-PCR) method to examine P. gingivalis gene expression in human dental plaque from periodontitis subjects. PCR primers and probes for six target genes representing putative virulence factors were chosen and evaluated in vitro for specificity. A potential cross-reactivity level of only 10 copies/10(7) whole genomic equivalents was occasionally observed with non-P. gingivalis microbes. P. gingivalis cells stressed in vitro by a 5 degrees C temperature increase showed a rapid rise in the mRNA associated with the molecular chaperons (htpG, dnaK, groEL), SOD (sodA) and gingipain (rgp-1) genes. We examined the stability of bacterial RNA in plaque specimens and found no significant difference in the amount of RNA obtained before or after storage 3 months in a stabilizing buffer (p=0.786, t-test). Sixty-five percent of plaque samples obtained from two clinical locations contained P. gingivalis; there was a mean level of gene expression (fold increase) for all samples tested for groEL, dnaK, htpG, sodA, PG1431 and rgp-1 of 0.84+/-2.03 to 7.85+/-10.0. ANOVA showed that the levels of stress gene transcription for dnaK and htpG were significantly elevated (p<0.05) at diseased sites; groEL gene transcription approached statistically significant elevation (p=0.059). We found correlations between probing depth and increased transcription of groEL, htpG and rgp-1 and between attachment loss and htpG. When sorted by disease status, we detected correlations between disease status and elevated expression of dnaK and htpG.

Adhesins, Bacterial↗

Over-expression of the Saccharomyces cerevisiae exo-beta-1,3-glucanase gene together with the Bacillus subtilis endo-beta-1,3-1,4-glucanase gene and the Butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene in yeast.

The EXG1 gene encoding the main Saccharomyces cerevisiae exo-beta-1,3-glucanase was cloned and over-expressed in yeast. The Bacillus subtilis endo-1,3-1,4-beta-glucanase gene (beg1) and the Butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene (end1) were fused to the secretion signal sequence of the yeast mating pheromone alpha-factor (MF alpha 1S) and inserted between the yeast alcohol dehydrogenase II gene promoter (ADH2P) and terminator (ADH2T). Constructs ADH2P-MF alpha 1S-beg1-ADH2T and ADH2P-MF alpha 1S-end 1-ADH2T designated BEG1 and END1, respectively, were expressed separately and jointly with EXG1 in S. cerevisiae. The construction of fur 1 ura3 S. cerevisiae strains allowed for the autoselection of these multicopy URA3-based plasmids in rich medium. Enzyme assays confirmed that co-expression of EXG1, BEG1 and END1 enhanced glucan degradation by S. cerevisiae.

Amino Acid Sequence↗

Sinusitis in neurologically impaired children.

The microbiologic features of infected sinus aspirates in nine children with neurologic impairment were studied. Anaerobic bacteria, always mixed with aerobic and facultative bacteria, were isolated in 6 (67%) aspirates and aerobic bacteria only in 3 (33%). There were 24 bacterial isolates, 12 aerobic or facultative and 12 anaerobic. The predominant aerobic isolates were Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus (2 each) and Proteus mirabilis, Pseudomonas aeruginosa, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae (1 each). The predominant anaerobes were Prevotella sp. (5), Peptostreptococcus sp. (4), Fusobacterium nucleatum (2), and Bacteroides fragilis (1). Beta-lactamase-producing bacteria were isolated from 8 (89%) patients. Organisms similar to those recovered from the sinuses were also isolated from tracheostomy site and gastrostomy wound aspirates in five of seven instances. This study demonstrates the uniqueness of the microbiologic features of sinusitis in neurologically impaired children, in which, in addition to the organisms known to cause infection in children without neurologic impairment, facultative and anaerobic gram-negative organisms that can colonize other body sites are predominant.

Adolescent↗

[Liver abscess and Crohn' disease. Report of 3 cases].

Liver abscess is a rare complication of Crohn's disease. Its prevalence and mortality are higher in patients with Crohn's disease than in the general population. Owing to its nonspecific clinical presentation, which may be mistaken for reactivation of Crohn's disease or be masked by simultaneous steroid therapy, a high index of suspicion is required for an early diagnosis and prompt treatment. We report 3 cases of Crohn's disease complicated with liver abscess in which the only common features were the absence of clinical or even endoscopic activity of Crohn's disease at diagnosis and the presence of an anastomotic leak due to right ileocolectomy in the previous year. In all patients, outcome was satisfactory with antibiotic therapy and percutaneous catheter drainage.

Adult↗

Pulmonary mucormycosis in diabetic renal allograft recipients.

Renal allograft recipients are prone to opportunistic infections due to their need of immunosuppression to prevent rejection. Mucormycosis is a rare opportunistic infection caused by a fungi of the order Mucorales. Risk factors predisposing to this disease include prolonged neutropenia, chelation therapy for iron or aluminum overdose, diabetes, and patients who are immunosuppressed. Life-threatening infections can occur, as this fungi has the propensity to invade blood vessel endothelium, resulting in hematologic dissemination. Early diagnosis and prompt aggressive therapy is imperative to achieve an improved outcome. We present two cases of pulmonary mucormycosis in diabetic renal allograft recipients who were treated successfully with amphotericin B and surgical resection of the lesions with preservation of their allograft function. In this era of intensified immunosuppression, we may see an increased incidence of mucormycosis in our transplant population.

Adult↗