Cross-reactivity of the IDEXX Legiolert method with other Gram-negative bacteria and waterborne pathogens leads to false-positive assay results.

Legionella species are the causative agent of Legionnaires' disease, a potentially fatal bacterial pneumonia. New regulations and standards have prioritized the development of water safety plans to minimize the growth and spread of Legionella species in buildings. To determine the presence and type of Legionella in a water system, microbiological culturing is the gold standard method. However, recently new methodologies have been developed that claim to be sensitive and specific for Legionella at the genus or L. pneumophila at the species level. Published and anecdotal reports suggest that one of these newer culture-based, enzyme-substrate methods, the IDEXX Legiolert test, may exhibit false positivity with other microbes common to water sources. We experimentally evaluated the IDEXX Legiolert method using these other waterborne bacteria including Elizabethkingia meningoseptica, Pseudomonas aeruginosa, Proteus mirabilis and Serratia marcescens at real-world environmental concentrations. We saw false-positive results for the Legiolert test with several of these organisms, at sample concentrations as low as 60 CFU per ml. False-positive Legionella results can trigger costly remediation and water-use restrictions, that may be implemented while waiting for additional, confirmatory microbiological testing that could, in this case, yield no L. pneumophila.

National survey on the susceptibility of Bacteroides fragilis group: report and analysis of trends in the United States from 1997 to 2004.

The susceptibility trends for the species of the Bacteroides fragilis group against various antibiotics from 1997 to 2004 were determined by using data for 5,225 isolates referred by 10 medical centers. The antibiotic test panel included ertapenem, imipenem, meropenem, ampicillin-sulbactam, piperacillin-tazobactam, cefoxitin, clindamycin, moxifloxacin, tigecycline, chloramphenicol, and metronidazole. From 1997 to 2004 there were decreases in the geometric mean (GM) MICs of imipenem, meropenem, piperacillin-tazobactam, and cefoxitin for many of the species within the group. B. distasonis showed the highest rates of resistance to most of the beta-lactams. B. fragilis, B. ovatus, and B. thetaiotaomicron showed significantly higher GM MICs and rates of resistance to clindamycin over time. The rate of resistance to moxifloxacin of B. vulgatus was very high (MIC range for the 8-year study period, 38% to 66%). B. fragilis, B. ovatus, and B. distasonis and other Bacteroides spp. exhibited significant increases in the rates of resistance to moxifloxacin over the 8 years. Resistance rates and GM MICs for tigecycline were low and stable during the 5-year period over which this agent was studied. All isolates were susceptible to chloramphenicol (MICs < 16 microg/ml). In 2002, one isolate resistant to metronidazole (MIC = 64 microg/ml) was noted. These data indicate changes in susceptibility over time; surprisingly, some antimicrobial agents are more active now than they were 5 years ago.

In vitro activities of newer quinolones against bacteroides group organisms.

The activities of BMS-284576, clinafloxacin, moxifloxacin, sitafloxacin, trovafloxacin, imipenem, cefoxitin, and clindamycin against 589 Bacteroides fragilis group isolates were determined. The activity of BMS-284576 was comparable to that of trovafloxacin. Sitafloxacin and clinafloxacin were the most active quinolones, and moxifloxacin was the least active. B. fragilis was the most susceptible of the species, and Bacteroides vulgatus was the most resistant. Association of specific antibiotic resistance with Bacteroides species was noted for all quinolones.

National survey on the susceptibility of Bacteroides Fragilis Group: report and analysis of trends for 1997-2000.

The results of a multicenter US survey using the National Committee for Clinical Laboratory Standards currently recommended methodology for measuring in vitro susceptibility of 2673 isolates of Bacteroides fragilis group species were compared from 1997 to 2000. The test panel consisted of 14 antibiotics: 3 carbapenems, 3 beta-lactam-beta-lactamase inhibitors, 3 cephamycins, 2 fluoroquinolones, clindamycin, chloramphenicol, and metronidazole. Declines in the geometric mean minimum inhibitory concentrations were seen with imipenem, meropenem, ampicillin-sulbactam, and the cephamycins. Increased geometric means were observed with the fluoroquinolones and were usually accompanied by an increase in resistance rates. Bacteroides distasonis shows the highest resistance rates among beta-lactam antibiotics, whereas Bacteroides vulgatus shows the highest resistance levels among fluoroquinolones. B. fragilis shows the lowest resistance rates for all antibiotics. All strains were susceptible to chloramphenicol and metronidazole concentrations <8 microgram/mL. The data underscore the need for species identification and continued surveillance to monitor resistance patterns.

Control of an outbreak of infection due to extended-spectrum beta-lactamase--producing Escherichia coli in a liver transplantation unit.

We report an outbreak of infection due to genotypically identical extended-spectrum beta-lactamase--producing Escherichia coli among patients in a liver transplantation unit. Control of the outbreak was achieved by a combination of contact isolation, feedback on hand hygiene, and gut decontamination with an orally administered fluoroquinolone. These interventions led to abrupt curtailment of the outbreak.

Evolving trends in multiple-antibiotic-resistant bacteria in liver transplant recipients: a longitudinal study of antimicrobial susceptibility patterns.

The incidence, sources, impact on outcome, and temporal trends in multiple-antibiotic-resistant bacteria in liver transplant recipients over the last decade (from 1990 through 1999) were assessed. Of 165 consecutive patients who underwent transplantation, 31% (51 of 165 patients) had at least 1 infection caused by multiple-antibiotic-resistant bacteria. Overall, 69% (66 of 96 infections) of all bacterial infections were multiple-antibiotic resistant. Ninety-one percent (45 of 49 isolates) of the Staphylococcus aureus isolates, 50% (6 of 12 isolates) of the enterococci, and 54% of the gram-negative bacteria (47%; 7 of 15 Pseudomonas aeruginosa, and 60%; 12 of 20 Enterobacteriaceae) were multiple-antibiotic resistant. A significant trend toward an increase in infections caused by multiple-antibiotic-resistant bacteria (P =.003), largely caused by an increase in gram-positive infections, was documented through the decade. There was a significant increase in infections caused by methicillin-resistant S aureus (P =.0001) and vancomycin-resistant enterococci (P =.04) over time. The proportion of gram-negative isolates that were multiple-antibiotic resistant (P =.447) did not increase significantly over time. However, a strikingly high frequency of resistance to piperacillin or ceftazidime suggests that extended-spectrum beta-lactamase production in our Enterobacteriaceae may have been more prevalent than realized. Mortality at 1 year was significantly greater in patients with multiple-antibiotic resistant bacteria compared with all other patients (P =.001). These longitudinal trends have implications not only for guiding therapeutic practices, but ultimately for devising strategies to curtail multiple-antibiotic resistance in liver transplant recipients.

Multicenter study of in vitro susceptibility of the Bacteroides fragilis group, 1995 to 1996, with comparison of resistance trends from 1990 to 1996.

Antimicrobial resistance, including plasmid-mediated resistance, among the species of the Bacteroides fragilis group is well documented. An analysis of the in vitro susceptibility of B. fragilis group species referred between 1995 and 1996 as well as during a 7-year (1990 to 1996), prospective, multicenter survey of over 4,000 clinical isolates of B. fragilis group species was undertaken to review trends in the percent resistance to and geometric mean MICs of the antibiotics tested. There was a trend toward a decrease in the geometric mean MICs of most beta-lactam antibiotics, while the percent resistance to most agents was less affected. Within the species B. fragilis, the geometric mean MICs showed significant (P < 0.05) decreases for piperacillin-tazobactam, ticarcillin-clavulanate, piperacillin, ticarcillin, ceftizoxime, cefotetan, and cefmetazole; a significant increase was observed for clindamycin and cefoxitin. For the non-B. fragilis species, a significant decrease in the geometric mean MICs was observed for meropenem, ampicillin-sulbactam, ticarcillin-clavulanate, piperacillin, ticarcillin, ceftizoxime, and cefmetazole; a significant increase was observed for cefoxitin. Significant increases in percent resistance were observed within the B. fragilis strains for ticarcillin and ceftizoxime and within the non-B. fragilis isolates for cefotetan. Significant increases in percent resistance among all B. fragilis group species were observed for clindamycin, while imipenem showed no significant change in resistance trends. The trend analysis for trovafloxacin was limited to 3 years, since the quinolone was tested only in 1994, 1995, and 1996. During the 7 years analyzed, there was no resistance to metronidazole or chloramphenicol observed. The data demonstrate that resistance among the B. fragilis group species has decreased in the past several years, the major exception being clindamycin. The majority of the resistance decrease has been for the beta-lactams in B. fragilis, compared to other species. The reasons for these changes are not readily apparent.

Infection and colonization with vancomycin-resistant Enterococcus faecium in an acute care Veterans Affairs Medical Center: a 2-year survey.

BACKGROUND: After our first known patient with vancomycin-resistant enterococci (VRE) infection was admitted in 1993, we observed a gradual increase in infections and colonization caused by this organism. Thus we initiated a prospective study to quantitate the incidence of VRE infection versus colonization, to identify risk factors for VRE, and to define the natural history of VRE colonization among our patients. METHODS: Stool/rectal cultures were performed for patients admitted to the intensive care units at the time of admission and weekly thereafter. Patients found to be carrying VRE were followed with cultures every 2 weeks, and this protocol was continued after transfer to the medical-surgical wards. A surveillance form was initiated on each VRE patient and included demographics, underlying diseases, and risk factors. Environmental cultures in the intensive care units were randomly performed. Patients with positive cultures were isolated. RESULTS: During a 27-month period, 210 patients were found to be colonized or infected with VRE. Ages ranged from 35 to 97 years; the mean age was 65 years. Fourteen percent (29 of 210) of the patients were VRE positive on admission. Nosocomial colonization or infection occurred at an average of 28 days after admission. Seventeen percent (25 of 216) of patients cleared VRE during their hospital stay; 19% (40 of 210) developed 47 infections. One third of infections involved the urinary tract. Liver transplantation, chemotherapy, and total parenteral nutrition were each associated with infection. CONCLUSION: Routine measures as advocated by the Centers for Disease Control and Prevention were not effective in controlling VRE in our patient population.

Analysis of trends in antimicrobial resistance patterns among clinical isolates of Bacteroides fragilis group species from 1990 to 1994.

Antimicrobial resistance, including plasmid-mediated resistance, among Bacteroides fragilis group species is well documented. A 5-year (1990-1994) prospective, eight-center survey of 3,177 clinical isolates of Bacteroides species was undertaken to review trends in resistance, using the breakpoints for full and intermediate susceptibility established by the National Committee for Clinical Laboratory Standards. No documented resistance to either metronidazole or chloramphenicol was found in this survey. Among B. fragilis isolates virtually no resistance was seen to imipenem, meropenem, ampicillin/sulbactam, piperacillin/tazobactam, or ticarcillin/clavulanate. Significant increases in resistance among B. fragilis isolates to cefotetan, ceftizoxime, and clindamycin (p < .01) were noted. Resistance to cefoxitin remained unchanged. Among the non-fragilis species of the B. fragilis group, there was virtually no resistance to imipenem, meropenem, chloramphenicol, or metronidazole. The three beta-lactamase inhibitors had increasing levels of resistance, although 95%-98% of strains were susceptible (p < .05). There was a significant decline in cefoxitin, cefmetazole, and clindamycin activity over time against these strains (p <.01). There was a significant (P < .001) increase in geometric mean minimum inhibitory concentration for most drugs and species tested from 1990 to 1994. Clusters in the eight institutions could not account for this rise in resistance. This survey demonstrates that rates of resistance of B. fragilis and non-fragilis species of B. fragilis group are increasing.

Brain abscess caused by Schizophyllum commune: an emerging basidiomycete pathogen.

Despite the worldwide distribution and prevalence of Schizophyllum commune, an emerging basidiomycetous pathogen, human infections occur only rarely. We describe the first well-documented pulmonary infection caused by S. commune which disseminated to the brain of a 58-year-old patient undergoing empiric corticosteroid therapy. Magnetic resonance imaging scans revealed ring-enhancing masses. Histologic examination of biopsy tissue from lungs and brain showed hyaline, septate, branched hyphae with clamp connections. Cultures of the lung tissue grew S. commune, which produced numerous, characteristic flabelliform and medusoid fruiting bodies on Czapek's agar. The isolate was susceptible to amphotericin B (MIC, < 0.03 microgram/ml) and fluconazole (MIC, 8 micrograms/ml). Despite treatment with antifungal and antibacterial agents, the patient developed progressive pulmonary failure and bacterial sepsis and died.

An outbreak of infusion-related Klebsiella pneumoniae bacteremia in a liver transplantation unit.

Over a 1-month period, there were five episodes of infusion-related Klebsiella pneumoniae bacteremia in four liver transplantation patients housed in the same ward. Investigation of nursing practices revealed that a common normal-saline bag, to which intravenous (iv) tubing and a stopcock were attached, was used to flush iv catheters. The iv tubing and stopcock were changed at sporadic intervals. Cultures of the normal saline and iv equipment yielded K. pneumoniae, which had the same susceptibility pattern as the patients' isolates. Isolates recovered during the outbreak from the patients and from the iv saline/equipment were of the same strain, as determined by pulsed-field electrophoresis of Xba I-digested genomic DNA. Termination of the practice of flushing iv catheters with a common normal-saline bag halted the outbreak.

Recurrent methicillin-resistant Staphylococcus aureus osteomyelitis: combination antibiotic therapy with evaluation by serum bactericidal titers.

OBJECTIVE: To report on a patient with recurrent methicillin-resistant Staphylococcus aureus (MRSA) osteomyelitis and bacteremia successfully treated with combination antibiotic therapy. CASE SUMMARY: Two sets of blood cultures from a 55-year-old man with fever, malaise, and low back pain grew MRSA. Radiologic studies of the spine showed bony changes consistent with osteomyelitis. Soon after completing 6 weeks of vancomycin, the patient experienced a recurrence of back pain. Laboratory values included an increase in the sedimentation rate to 53 mm/h and positive blood cultures for MRSA. Vancomycin, gentamicin, and rifampin were administered for 8 weeks. Serum inhibitory and bactericidal titers were more than 1:1024 for both the peak and trough concentrations. Radiologic studies of the spine showed healing osteomyelitis. Two years after completion of antibiotic therapy, the infection has not recurred. DISCUSSION: Antibiotic therapy alone was attempted because the patient was considered a risky surgical candidate. Serum inhibitory and bactericidal titers documented the high in vivo activity of the vancomycin, gentamicin, and rifampin combination. Initiation of vancomycin, gentamicin, and rifampin combination. Initiation of vancomycin therapy led to disappearance of the fever and back pain. Cure was documented by sustained normalization of the erythrocyte sedimentation rate and radiologic evidence of healing. CONCLUSIONS: Combination antibiotic therapy with vancomycin, rifampin, and low-dose gentamicin (1 mg/kg q12h) may be useful for deep-seated tissue infection caused by MRSA.

Efficacy of topical amoxicillin plus clavulanate/ticarcillin plus clavulanate and clindamycin in contaminated head and neck surgery: effect of antibiotic spectra and duration of therapy.

This prospective, randomized trial was designed to determine the efficacy and mechanism of action of topical mouthwash versus parenterally administered perioperative prophylactic antibiotics in contaminated head and neck surgery. Patients were randomly assigned to 1 of 4 treatment groups: 1 day of parenteral clindamycin (standard prophylaxis), 1 day of topical clindamycin, 5 days of topical clindamycin, or 1 day of topical amoxicillin plus clavulanate/ticarcillin plus clavulanate. Patients who received the latter regimen had fewer bacteria postoperatively compared with the other 3 treatment groups. The number of gram-negative aerobic bacilli on postoperative oral cavity cultures was increased in all 3 clindamycin groups but not in the amoxicillin plus clavulanate/ticarcillin plus clavulanate group. Parenteral clindamycin appears to exert its effect by being in the neck tissues at the time of surgery; however, all 3 topical regimens were more effective at reducing the number of bacteria in the neck viscera. Topical antibiotic prophylaxis was simple, safe, effective, and well tolerated.

Cutaneous cryptococcosis mimicking bacterial cellulitis in a liver transplant recipient: case report and review in solid organ transplant recipients.

Cutaneous cryptococcosis is an uncommon manifestation of disseminated cryptococcal disease. We report a liver transplant recipient presenting with onset over 3 days of progressive pain and swelling of the lower extremity and foot with erythema and heat. Cryptococcal cellulitis was documented, but in retrospect the clinical presentation was indistinguishable in presentation and appearance from acute bacterial cellulitis. The patient also proved to have concomitant cryptococcal septic arthritis; the presence of cryptococcal antigen in the synovial fluid allowed a rapid and definitive diagnosis. Subsequently, cultures of skin, synovial fluid, and blood were all positive for cryptococci. Cryptococcus should be considered in the differential diagnosis of bacterial cellulitis in a liver transplant recipient not responding to antibacterial therapy.

The efficacy of topical antibiotic prophylaxis for contaminated head and neck surgery.

This study was undertaken to determine the feasibility of using perioperative topical antibiotics in contaminated head and neck surgery and to standardize the culture methodology (both qualitative and quantitative) which could serve as bacteriologic endpoints for evaluation. Following preliminary studies to establish oral cavity indicator organisms and the impact of a single antibiotic mouthwash dose on oral microflora, 10 consecutive patients undergoing contaminated head and neck surgery were recruited into a clinical trial where clindamycin mouthwash and intraoperative irrigation containing clindamycin were used instead of traditional parenteral antibiotics. The bacteriologic efficacy of topical clindamycin was assessed by comparing the presence of four indicator microorganisms (two aerobic and two anaerobic) cultured from two oral cavity culture sites before and after antibiotic prophylaxis. The patients included in the study underwent total laryngectomy plus neck dissection(s) for laryngeal or hypopharyngeal carcinoma from 1991 to 1992 at a large university hospital specializing in head and neck cancer surgery. The main outcome measures used were the development of a postoperative wound infection and quantitative and qualitative bacteriology of the intraoperative neck wound and postoperative oral cavity. Two aerobic and two anaerobic organisms proved useful as a practical indicator for bacteriologic efficacy. Preoperative mouthwash resulted in a 99% reduction of both aerobic and anaerobic bacteria in intraoperatively cultured neck sites. Irrigation during surgery with the clindamycin solution further reduced the bacterial neck counts by an additional 90%. There was a consistent overgrowth of Hemophilus species on postoperative oral cavity cultures. No patient developed a postoperative wound infection. A topical prophylactic antibiotic alone was efficacious and safe for patients undergoing major contaminated head and neck surgery. Culture methods for assessment of bacteriologic efficacy were reproducible and cost-effective. This pilot study furnishes the ethical and scientific basis for large-scale prospective trials comparing topical versus parenteral antimicrobial agents.

Improved detection of spontaneous bacterial peritonitis with Bactec as compared with conventional culture methods. A prospective study.

Bacteriologic diagnosis of spontaneous bacterial peritonitis is difficult due to the low yield of isolating the bacteria from the ascitic fluid. We prospectively compared the conventional culture method with the nonradiometric Bactec culture system for the detection of bacteria in 20 episodes of spontaneous bacterial peritonitis. The ascitic fluid culture was positive by the conventional culture method in 25% and by Bactec in 79% of the episodes of spontaneous bacterial peritonitis (P = 0.004). Of culture-positive episodes, Gram-negative bacteria were detected by conventional cultures in 20% and by Bactec in 47%. Bactec culture system was significantly better than the conventional cultures for the detection of streptococci (viridans streptococci and Enterococcus fecalis), 33% versus 0 (P < 0.05). Conventional cultures did not detect bacteria that were not also detected by Bactec cultures. In conclusion, the Bactec nonradiometric culture method is superior to conventional cultures for the diagnosis of spontaneous bacterial peritonitis.

Roseomonas, a new genus associated with bacteremia and other human infections.

In the 1980s, a pink bacterium different from species of the genus Methylobacterium was implicated in human infection. Using biochemical tests and DNA hybridization, we examined 42 strains of pink-pigmented, gram-negative bacteria that were not members of the genus Methylobacterium. The isolates included 6 strains each of CDC "pink coccoid" groups I, II, III, and IV; 10 isolates from Gilardi's "unnamed taxon"; and 8 blood isolates from ill, debilitated, or immunosuppressed patients. The DNA hybridization studies supported the creation of six genomospecies encompassing the 42 strains. Reactions for esculin hydrolysis, glycerol oxidation, and D-mannose oxidation enabled separation of genomospecies 1 through 4. These tests, as well as motility, nitrate reduction, citrate utilization, and oxidation of L-arabinose, D-galactose, and D-xylose, differentiated genomospecies 5 and 6 from each other and from genomospecies 1 through 4. These organisms were susceptible in vitro to the aminoglycosides, tetracycline, and imipenem and generally susceptible to the quinolones. We propose the new genus, Roseomonas, for these bacteria to include three named species, Roseomonas gilardii sp. nov., Roseomonas cervicalis sp. nov., and Roseomonas fauriae sp. nov., and three unnamed genomospecies.

Staphylococcus epidermidis bacteremia from transfusion of contaminated platelets: application of bacterial DNA analysis.

Septicemia is a rare complication of platelet transfusion. A case is reported of transfusion-associated septicemia in a 66-year-old man who received a 10-unit pool of platelets. During transfusion, he experienced rigors, wheezing, dyspnea, and fever. A total of four blood cultures drawn 10 and 36 hours after discontinuation of the transfusion grew Staphylococcus epidermidis. Culture of the residual platelet pool yielded S. epidermidis with a colony count of 10(5) organisms per mL. Strain identity of all four blood isolates and the platelet pool isolate was confirmed by gel electrophoresis of EcoRI and HindIII restriction digests of whole-cell DNA. There have been 31 prior reported cases of platelet transfusion-associated septicemia, of which 9 have been caused by coagulase-negative staphylococci. Systemic reactions to platelet transfusions should prompt consideration of transfusion-associated bacteremia as the cause.

Clinical demonstration of isolation of Nocardia asteroides on buffered charcoal-yeast extract media.

Nocardia asteroides was isolated only from sputum samples, obtained from three patients with pulmonary nocardiosis, that had been cultured onto buffered charcoal-yeast extract (BCYE) and selective BCYE media as part of laboratory workups for Legionella species. A decontamination procedure with low-pH pretreatment (KCl-HCl solution) had been performed on the sputa prior to culture onto the BCYE media because direct cultures on the media were overgrown with commensal microflora. Chalky white colonies, 0.5 to 1.0 mm in diameter, that were subsequently identified as N. asteroides grew well on the BCYE media. Thus, the techniques and the selective media used for Legionella species were useful for isolating Nocardia species from sputum.