Bacterial resistance to the cyclic glycopeptides.

Cyclic-glycopeptide antibiotics, such as vancomycin and teicoplanin, have been almost uniformly active against pathogenic Gram-positive bacteria since their discovery in the 1950s. Resistance is now emerging among enterococci and staphylococci by acquisition of novel genes or by mutation, respectively. The mechanism of resistance for enterococci appears to be synthesis of an altered cell-wall precursor with lower affinity for the antibiotics.

Complementary roles of mutations at positions 69 and 242 in a class A beta-lactamase.

Analysis of the three-dimensional structure of class A beta-lactamases suggests that deformation of the substrate binding site can be produced by changes in the hydrophobicity of residue 69 behind the beta-sheet and by outward movement of the B3 beta-strand by introduction of a non-glycine residue at position 242 on the B4 beta-strand. By site-directed mutagenesis Met69-IleGly242-Cys, a double mutant, of the OHIO-1 beta-lactamase, was constructed. The minimum inhibitory concentrations (MICs) of the double mutant compared with the wild type and each single mutant revealed an increased susceptibility to beta-lactams. Met69-IleGly242Cys hydrolyzed cephaloridine (Km = 213 microM) but had Km > 500 microM for other beta-lactams tested including cefotaxime, and demonstrated a higher apparent Ki for inhibitors (clavulanate Ki = 500 microM sulbactam = 434 microM, and tazobactam = 70 microM). In a competition experiment with cephaloridine, the apparent Ki values for penicillin and cefotaxime remained low, 21 microM and 0.7 microM, respectively. Since Ile is twice as hydrophobic as Met, the Met69-Ile mutation may result in partial collapse of the oxyanion hole. This would also increase the distance between Arg-244 and the carboxyl of clavulanic acid. The Gly242-Cys mutation opens the lower portion of the active site to bulky R groups of cephalosporins. Although these two mutations result in a catalytically impaired enzyme, they can be used to model the complementary role of two distinct residues, neither of which interacts directly with beta-lactam substrates or inhibitors.

beta-Lactamase mutations far from the active site influence inhibitor binding.

Analysis of the three dimensional structure of the class A beta-lactamases shows that Arg-244, a spatially conserved residue important for inactivation by clavulanic acid, is held in place by a hydrogen (H) bond from the residue at 276. An Asn276-Gly mutant of OHIO-1, an SHV family class A enzyme, was constructed to investigate the importance of that interaction. Compared to a strain expressing the wild type enzyme, OHIO-1, the MIC of the Asn276-Gly mutant strain was more resistant to clavulanate (0.25 vs. 2.0 micrograms/ml) in the presence of ampicillin (16 micrograms/ml) but was as susceptible to sulbactam or tazobactam plus ampicillin as the OHIO-1 bearing strain. No difference in MICs was observed when other beta-lactams were tested. Consistent with the susceptibility test results, the apparent Ki of clavulanate for the Asn276-Gly enzyme (4.5 microM) was 10-fold greater than OHIO-1 (0.4 microM). For sulbactam and tazobactam the apparent Ki decreased for Asn276-Gly enzyme (1.0 and 0.1 micrograms/ml, respectively) compared to the wild-type parent (17 and 0.7 micrograms/ml, respectively). Comparing the Asn276-Gly heta-lactamase with OHIO-1, the Vmax for most substrates except cephaloridine did not change substantially. There was a 2-15 fold decreased affinity (Km) and catalytic efficiency (Vmax/Km) for beta-lactam substrates. These data support the observation and emphasize the role for this H bonding residue in orienting Arg-244 towards the active site.

Class A beta-lactamases--enzyme-inhibitor interactions and resistance.

Beta-Lactamases of Ambler's Class A are the most commonly encountered mechanism of bacterial resistance to beta-lactam antibiotics. In the face of selective pressure arising from use of either newer cephalosporins or beta-lactam/beta-lactamase inhibitor combinations, mutations arose among Class A beta-lactamase genes, leading to resistance. Clavulanic acid, a naturally occurring clavam, and the penicillanic acid sulfones sulbactam and tazobactam are the inhibitors in clinical use. This review focuses on the mechanism of inhibition by these currently marketed beta-lactamase inhibitors and on the mechanism by which specific amino acid substitutions might lead to resistance. The key amino acid positions important for inhibitor-resistance include Met69, Ser130, Arg244, Arg275, and Asn276. Ser130 is vital to the chemical mechanism of inhibition. Arg244 appears to be coordinated to Arg275 and Asp276 by hydrogen bonds. Arg244 is involved in positioning beta-lactams, especially penicillins and beta-lactamase inhibitors, via their carboxyl groups. Site-directed mutagenesis studies confirm the role of Arg244 and its coordinating partners in beta-lactam turnover and in the reactions leading to enzyme inactivation. This mechanism is dependent on the donation of a proton via a water coordinated to Arg244 and Val216 to clavulanic acid to allow formation of a favorable leaving group. This proton donation is probably not required for formation of a favorable leaving group for the sulfone inhibitors sulbactam and tazobactam. Therefore, some amino acid substitutions have differing effects on inhibition by clavulanic acid compared with the penicillanic acid sulfones. Met69 may play a more structural role in beta-lactam positioning within the oxyanion hole.

Enterococcal bacteremia without endocarditis.

Seventy-four cases of enterococcal bacteremia without endocarditis were reviewed retrospectively for the years 1963 through 1977. Thirty-nine patients had hospital-acquired infection, 27 had serious underlying disease, and 18 had polymicrobial bacteremia. The organisms isolated from the primary source of infection were similar in patients with pure enterococcal or polymicrobial bacteremia. The most common source was the urinary tract, followed by the abdomen, infected burns, and soft-tissue infections other than burns. The overall mortality was 34%, with significantly higher mortality in immunocompromised patients, including those with infected burn wounds. Mortality also was higher in patients with hospital-acquired infection. Eleven of the 25 deaths were considered directly related to enterococcal septicemia.

Streptococcus mitis. A cause of serious infection in adults.

Twenty strains of Streptococcus mitis were isolated from blood or body fluids at the Cleveland Veterans Administration Medical Center from Jan 1, 1981, to April 30, 1984. Fifteen (75%) isolates were considered contaminants. Five (25%) were clinically important and associated with a serious infection of the oropharynx or gastrointestinal tract (three of five), endovascular system (one of five), or a prosthetic hip. Four of five patients required surgical intervention for treatment. Two of five died; one death was directly attributable to S mitis infection. Eighteen strains were available for detailed bacteriologic study. Three strains had a minimum inhibitory concentration of greater than 0.1 micrograms/mL of penicillin and six other strains were tolerant to penicillin. This review suggests that S mitis can be an important pathogen in adults and may cause infections other than endocarditis.

Nosocomial clonal dissemination of methicillin-resistant Staphylococcus aureus. Elucidation by plasmid analysis.

An outbreak of methicillin sodium-resistant Staphylococcus aureus (MRSA) infection and colonization, mainly centered in the vascular surgery service, occurred in a 1000-bed tertiary care center between December 1983 and December 1984. Methicillin-resistant S aureus isolated before and during the outbreak was studied by both bacteriophage typing and by restriction endonuclease digestion of bacterial plasmid DNA. Bacteriophage typing was discrepant in nine (56%) of the 17 repeated analyses compared with one (3.4%) of the 29 for plasmid profiling. These typing methods revealed that the epidemic strain was introduced to the hospital from the community 15 months before the outbreak. The outbreak was caused by cross-transmission of the epidemic strain by health care personnel and was controlled by treatment of colonized personnel, education of personnel, and institution of barrier precautions for colonized or infected patients. Plasmid profiling with restriction endonuclease digestion was easier, more rapid, and more specific than bacteriophage typing in the evaluation of this outbreak.

Serious infections caused by Bacillus species.

Thirty-eight patients with serious infections caused by organisms belonging to the genus Bacillus are described. Our experience, and that reported in the literature, indicates that, in most cases, isolated Bacillus bacteremia is not a particularly serious disease. Therefore, under most circumstances, empiric antibiotic therapy designed specifically for treatment of Bacillus is probably not necessary. Endocarditis can occur, but apparently follows bacteremia only infrequently. When these bacteria cause localized infection such as pneumonia, pan-ophthalmitis, visceral abscess, or musculoskeletal infections, tissue necrosis and profound morbidity are the rule. The frequency of these complications following bacteremia appears to be low but cannot be estimated from our experience or that reported in the literature reviewed. The role of intravascular devices and trauma as predisposing factors is emphasized. Immunocompromised hosts and intravenous drug abusers appear predisposed, but intravascular devices in the former group may play an important role in the pathogenesis of Bacillus infections. Antibiotics which appear especially useful in the treatment of Bacillus infections are clindamycin and vancomycin, to which the vast majority of strains are susceptible in vitro. Beta-lactam antibiotics, including the new cephalosporins and penicillins, are of little value in this setting.

Infections due to Lancefield group F and related Streptococci (S. milleri, S. anginosus).

We can no longer accept classification of streptococci solely on the basis of hemolytic reactions or Lancefield agglutinations. While the "viridans" streptococci cannot be serologically differentiated, physiological separation of the species offers a satisfactory method of classifying human isolates. We review the microbiology of Lancefield group F and related streptococci (S. milleri, S. anginosus), emphasizing microbial ecology and current taxonomic considerations. A series of 28 patients infected with these organisms is presented. There was a striking male predominance in the series (6:1) as well as an obvious association with underlying diseases and/or antecedent trauma. The most remarkable feature of these pathogens is their predilection for abscess formation, confirming their overdue recognition as the most common "viridans" streptococcus recovered from abscesses within internal organs. We observed purulent disease of the nervous and skeletal systems, oral cavity, lung and pleural space, abdomen and subcutaneous tissues. Microbial synergy (i.e. polymicrobic infection) was not a requisite for abscess formation. Four of the five deaths in this series occurred in patients 60 years of age of older. Some degree of variability in antimicrobial susceptibility was noted, so speciation alone may not always provide sufficient information on which to base a therapeutic decision.

Infections due to Lancefield group G streptococci.

The group G streptococcus has surfaced in the past 10 to 15 years as an important opportunistic and nosocomial pathogen. Although more precise organism recognition accounts for a portion of these cases, there can be little doubt that the group G streptococcus has become a more prevalent pathogen. Commercial kits, utilizing staphylococcal coagglutination or latex agglutination, are now available, affording all clinical laboratories the opportunity to identify this organism easily. Published reviews encompassing the experiences of a single institution or even several institutions affiliated with a single medical center, particularly as they were influenced by referral patterns, did not reflect the broad scope of infections that we discovered by extending our survey into the community, beyond the medical center complex and its immediate affiliated hospitals. Although malignancy is the single most obvious background factor, alcoholism and diabetes are also important host determinants of infection. Skin and soft-tissue infections (and surface sources of infection) are equally important among patients with or without the element of malignancy. Polymicrobial infection, including polymicrobial bacteremia, is an important feature, with S. aureus infections accounting for most of these cases, relating to the skin and soft tissue sources of infections so commonly seen. We saw a panorama of problems including endocarditis, septic arthritis, pleuropulmonary infections, bone and joint infections, puerperal sepsis and neonatal infection, peritonitis and ophthalmitis; we also saw a significant number of patients with bacteremia and no apparent primary source of infection. Response to antibiotic therapy was dictated by the nature of the underlying diseases, and individuals without a background of malignant disease did well, particularly those with skin and soft-tissue infections. While the literature suggests that patients with endocarditis and septic arthritis due to this organism respond poorly to antibiotic therapy, implying that such failures relate to in vitro antibiotic phenomena, we preferred to examine the problem from the viewpoint of the host(s) involved. Subacute endocarditis and acute endocarditis due to the group G streptococcus may be clinically separable, and thus require separate therapeutic approaches. In patients with septic arthritis, prosthetic devices, prior joint disease and immunosuppressive diseases and therapy often adversely influence the response to antibiotic therapy.(ABSTRACT TRUNCATED AT 400 WORDS)

Infections due to Lancefield group C streptococci.

Our experience with group C streptococcal infection over the past 15 years demonstrates an important and emerging role for this hemolytic organism as an opportunistic and nosocomial pathogen. Significant risk factors in this predominantly male population included chronic cardiopulmonary disease, diabetes, malignancy, and alcoholism. Bacteremia occurred in 74% of cases seen in our series. Nosocomial acquisition of infection was observed in 26%, and infection was frequently polymicrobial in nature with gram-negative enteric bacilli isolated most commonly along with group C streptococci. We observed a broad spectrum of infections including puerperal sepsis, pleuropulmonary infections, skin and soft-tissue infection, central nervous system infection, endocarditis, urinary tract infection, and pharyngeal infections. Several cases of bacteremia of unknown source were observed in neutropenic patients with underlying leukemia. New syndromes of infection due to group C streptococci observed in our series included intra-abdominal abscess, epidural abscess, and dialysis-associated infection. Response to therapy and outcome was related to the underlying disease. While the literature suggests that patients with group C endocarditis respond better to synergistic penicillin-aminoglycoside regimens, patient numbers are too small to draw definite conclusions. The clinical significance of antibiotic tolerant group C streptococci remains uncertain. In patients with serious group C infections including endocarditis, meningitis, septic arthritis, or bacteremia in neutropenic hosts, we advocate the initial use of cell-wall-acting agents in combination with an aminoglycoside.

Polymicrobial bacteremia in the late 1980s: predictors of outcome and review of the literature.

Although polymicrobial bacteremia has been described in several previous series, there has been no recent study of patients using rigorous statistical analysis. Our objective was to characterize a present-day patient population with polymicrobial bacteremia and to define factors prognostic of survival. Polymicrobial bacteremia accounted for 6% of all positive blood cultures at a university hospital and a Veterans Administration hospital over a 2 1/2 year period in the late 1980s. The majority of these patients were elderly with significant underlying diseases, notably malignancies, and 56% of all episodes were nosocomially acquired. Enterobacteriaceae have remained the most common organisms, though the frequency of gram-positive cocci isolated has increased compared to older studies. Gastrointestinal, genitourinary, and skin and soft-tissue sources were the most common, although the incidence of infections due to central venous catheters appeared to be increasing. The source of 25% of bacteremia was not identified despite newer diagnostic techniques. By univariate analysis, mortality, which was 36% overall, correlated with thrombocytopenia, respiratory failure, disseminated intravascular coagulation, encephalopathy, severity of underlying disease, hemolysis, adult respiratory distress syndrome, use of steroids, renal insufficiency, institution, presence of central lines, and nosocomial acquisition. Using stepwise logistic regression analysis, mortality was predicted by respiratory failure, severity of underlying disease, and hemolysis. We conclude that polymicrobial bacteremia remains an important entity in the present-day hospitalized population, with an increasing frequency of gram-positive organisms and catheter sources, and a large proportion of undiagnosed etiologies.

Wangiella dermatitidis endocarditis in an intravenous drug user.

Wangiella dermatitidis is an infrequently encountered dematiacious fungus that usually causes localized infections of the skin and subcutaneous tissues. This report presents the first well-documented case of natural valve infection caused by this organism as it occurred in intravenous drug abuser. His course has been complicated by relapsing infection of two aortic prostheses and dissemination to the vertebral spine. Treatment with a combination of amphotericin B, rifampin, and ketoconazole has arrested the progression of his infection. The microbiologic features and existing clinical information regarding this fungus are reviewed and in vitro susceptibility data for the present isolate are presented.

Toxic-shock syndrome associated with nasal packing: analogy to tampon-associated illness.

A pure culture of migmatoxin-producing Staphylococcus aureus was isolated from the purulent nasal packing removed from a 13-year-old premenarchal girl with toxic-shock syndrome after rhinoplasty. The similarities between this case and the tampon-associated illness are examined. An increased awareness of these similarities and the identification of predisposing factors may lead to earlier detection and treatment of toxic-shock syndrome.

Elastin fibers in the sputum of patients with necrotizing pneumonia.

We have observed five patients for whom the presence of fibers of elastin in potassium hydroxide (KOH) preparations of sputum represented the first evidence of necrotizing pulmonary disease. In four cases, the discovery of elastin fibers in sputum provided additional evidence supporting initiation or modification of antibiotic therapy. Necrotizing disease was confirmed in all cases by autopsy or by the development of cavitation on chest x-ray film. Cytochemical staining, electron microscopy, and elastase digestion all suggest that the refractile fibers seen on KOH wet mount of sputum are elastin. The test, first described in 1846, is simple to perform, requires little experience to read, and may be a valuable adjunct to the chest roentgenogram in the diagnosis of pulmonary parenchymal destruction.

Serious infectious complications of corticosteroid therapy for COPD.

We report seven elderly patients with COPD who developed serious infectious complications during prolonged treatment with high doses of corticosteroids. Infections included invasive pulmonary aspergillosis, Herpes simplex stomatitis and esophagitis, cytomegalovirus pneumonia, bacterial sepsis, fungemia and meningitis due to Cryptococcus neoformans. Each of the three patients who developed invasive aspergillus pneumonia died. The efficacy of prolonged therapy with high doses of corticosteroids in patients with COPD is not proven. These cases illustrate the potential for serious infections in patients with COPD treated with corticosteroids.

Sputum elastin fibers and the diagnosis of necrotizing pneumonia.

Potassium hydroxide (KOH) preparations for elastin fibers on sputum obtained from 80 patients seen over a four-month period at two Cleveland hospitals were performed. The results were compared with roentgenographic evidence of necrosis and case diagnosis. Sixty-one patients had neither elastin in sputum nor roentgenographic evidence of cavitation; 11 had positive results using both methods. Two patients had no elastin fibers in sputum but had parenchymal pulmonary cavities on chest x-ray film. Six patients had elastin observed in KOH preparations of sputum, but no cavitation roentgenographically. The presence of elastin in sputum was strongly correlated with roentgenographic evidence of pulmonary necrosis (p = 5.7 X 10(-8]. Including patients seen before, after, and during the prospective study, we have observed a total of nine with positive sputum preparations for elastin and no cavitation on chest x-ray film for whom tissue was available for study. All had pulmonary necrosis histologically. Our observations suggest that the KOH preparation of sputum for elastin fibers may be more sensitive than the chest roentgenogram in the detection of pulmonary necrosis and may be a useful adjunct in the diagnosis of necrotizing disease.

Introduction of a plasmid encoding the OHIO-1 beta-lactamase to an intermediate care ward by patient transfer.

A prospective study of 147 intermediate care ward (ICW) patients for acquisition of gentamicin-resistant Enterobacteriaceae (GRE) was carried out. Fifty (34%) were colonized or infected with one or more strains of GRE. Fifteen of these patients and one nurse were colonized with 22 strains (including ten species) of GRE bearing identical 60 kb plasmids encoding a novel beta-lactamase determinant, OHIO-1 and ANT(2"). Analysis of the time course of colonized patients on the ICW revealed one probable episode of cross-transmission. Five colonized patients had been residing in the ICW from one to four months prior to study initiation. Eight patients were admitted to the ICW from other hospital areas already colonized and one additional patient acquired colonization on the ICW from an unknown source. Thus, eight of ten patients admitted to the ICW during the prospective study were already colonized on admission to the ICW. To control this level of colonization it would therefore be necessary to direct efforts at limiting admission of colonized patients or attempting to eliminate the strain from persistently colonized patients, rather than trying to limit transmission within the ward.

Risk factors associated with permanent access-site infections in chronic hemodialysis patients.

OBJECTIVE: To determine the epidemiological risk factors associated with permanent access-site (PAS) infection in a population of chronic hemodialysis patients. DESIGN: Retrospective cohort analysis. SETTING: Hemodialysis unit of a 400-bed Department of Veterans' Affairs hospital. RESULTS: A cohort of 94 males (1,316 patient months) was studied. Fifty-one PAS infections in 31 patients were observed. Six patients had two PAS infections, four patients had three infections, and two patients had four infections. Twenty-nine of the 31 patients with PAS infections were bacteremic at least once. Univariate analysis identified seven factors significantly associated with PAS infection in this population: location of PAS other than forearm, type of vascular access (polytetrafluoroethylene [PTFE] versus endogenous arteriovenous [AV] fistula), limited ambulatory status, residence in a nursing home, bacterial infection at a distant site, number of access-site revisions, and number of hospitalizations. In a logistic regression analysis, only graft type and number of PTFE graft revisions were associated independently with PAS infection. The odds ratio for PAS infection in PTFE grafts compared to endogenous AV fistula was 7.8; the odds ratio for PAS infection with each PTFE graft revision was 1.5. CONCLUSIONS: PAS infections were associated independently with the type of graft and the number of PTFE graft surgical revisions.

Society for Healthcare Epidemiology of America and Infectious Diseases Society of America Joint Committee on the Prevention of Antimicrobial Resistance: guidelines for the prevention of antimicrobial resistance in hospitals.

Antimicrobial resistance results in increased morbidity, mortality, and costs of health care. Prevention of the emergence of resistance and the dissemination of resistant microorganisms will reduce these adverse effects and their attendant costs. Appropriate antimicrobial stewardship that includes optimal selection, dose, and duration of treatment, as well as control of antibiotic use, will prevent or slow the emergence of resistance among microorganisms. A comprehensively applied infection control program will interdict the dissemination of resistant strains.