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.

Pseudomonas stutzeri bacteremia associated with hemodialysis.

Pseudomonas stutzeri bacteremia developed in six patients undergoing hemodialysis. Fever, shaking chills, nausea, and vomiting were observed. All patients recovered, although only two received specific antibiotic therapy. The infections occurred sporadically over a period of nine months. Pseudomonas stutzeri was subsequently isolated from the dialysate that circulates within the hemodialysis machine. The ultimate source was the deionized water that is combined with the liquid concentrate to form the dialysate. Pseudomonas stutzeri could be localized to the top cannister of the dialysis machine but was also isolated throughout the machine, including the bottom reservoir and the recirculating pump. The emphasis on handwashing, strict compliance with disinfection procedures, and elimination of prolonged sitting times for the filled machine after disinfection resulted in no further cases of P stutzeri infection.

A pink-pigmented, oxidative, nonmotile bacterium as a cause of opportunistic infections.

We describe two cases of bacteremia due to a pink-pigmented, oxidative, nonmotile, gram-negative, rod-shaped organism. One case occurred in a febrile neutropenic patient and another in a chronically debilitated patient with pancreatic abscess. The first patient was cured with gentamicin and ticarcillin, but the second patient died while receiving cefamandole therapy. The organisms described here are similar to Methylobacterium mesophilicum (Pseudomonas mesophilica) and the "unnamed taxon" organisms. A major difference from M mesophilicum is the lack of methanol utilization. Further distinctions between our isolates and M mesophilicum are the lack of flagella in our organisms, growth at 42 degrees C, growth on MacConkey's agar, lack of acetamide assimilation, and citrate utilization. The lack of flagella is the principle difference between our isolates and those in the unnamed taxon. Both of the isolates were resistant to the cephalosporins, but susceptible to the aminoglycosides, ticarcillin-clavulanic acid, sulfamethoxazole and trimethoprim, and imipenem. With the growing population of immunocompromised and chronically ill patients, these organisms may emerge as important pathogens.

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.

Simultaneous infection with Legionella pneumophila and Pittsburgh pneumonia agent. Clinical features and epidemiologic implications.

Nosocomial pneumonia caused simultaneously by two organisms, Legionella pneumophila and the Pittsburgh pneumonia agent, was documented in seven patients in one institution. In all seven cases, both organisms were demonstrated by isolation from culture or visualization by direct immunofluorescence. Four patients died as a result of pneumonia, including two who received erythromycin therapy. The hospital water distribution system appeared to be the reservoir for both L. pneumophila and Pittsburgh pneumonia agent. These seven cases constituted 26.9 percent and 17.9 percent of the cases of Pittsburgh pneumonia agent and Legionnaires' disease, respectively, at one institution. Given this relatively high incidence of dual infection, it is likely that the mode of transmission for both organisms is identical. Dual infection may account for some cases of antibody response to more than one Legionella species. Historical parallels of the discovery of L. pneumophila and Pittsburgh pneumonia agent are reviewed.

Acute cardiac tamponade due to cardiac actinomycosis.

Cardiac actinomycosis occurs in less than 2 percent of the patients with infections due to Actinomyces israelii. We describe the findings in a patient with acute cardiac tamponade who survived through pericardial drainage and aggressive medical therapy. Although uncommon, this disorder is important to recognize because it is curable with current medical and surgical therapy.

A pseudo-outbreak of Streptomyces infections linked to laboratory construction.

Over a four-week period, nine isolates of Streptomyces species were identified from seven blood cultures, one infected gortex graft, and one cerebrospinal fluid (CSF) specimen. None of the patients were clinically infected or received antimicrobial therapy. Blood culture collecting tubes, iodoform prep solutions, BAC-TEC filters, and alcohol prep pads were cultured but did not reveal a source of contamination. Six additional clinical specimens became positive, and the occurrence of construction in the laboratory was considered as a source of the contaminating organism. Blood agar plates were placed for 20 minutes in an area undergoing construction. Streptomyces grew in heavy concentrations on nine of ten plates in the construction site, but only one colony on ten plates grew from a hallway not undergoing construction. Isolation of the organism ceased with cessation of the construction. The authors conclude that the Streptomyces pseudo-outbreak was related to construction around the laboratory, with probable airborne contamination of clinical specimens.

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.

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.

Antimicrobial susceptibility patterns of the Bacteroides fragilis group in the United States, 1989.

A national survey of Bacteroides fragilis group was continued in 1989 for the ninth consecutive year. Seven hundred thirty-nine isolates of B fragilis group from eight centers were tested for susceptibility to 14 antimicrobials. Sulbactam and clavulanic acid, beta-lactamase inhibitors, were tested at a constant concentration of 8 micrograms/ml and 2 micrograms/ml, respectively. Sulbactam was also tested in a fixed ratio of 1:2. Imipenem, ampicillin+sulbactam, and ticarcillin+clavulanic acid had resistance of less than 1% at breakpoints of 8 micrograms/ml, 16 micrograms/ml, and 64 micrograms/ml, respectively. At 32 micrograms/ml, resistance to cefoxitin, cefotetan, ceftizoxime, and ceftriaxone were 4%, 25%, 26%, and 46%, respectively. Clindamycin resistance was 10% at a breakpoint of 4 micrograms/ml. No isolates were resistant to chloramphenicol or metronidazole. Resistance for five B fragilis species to cefoxitin, ceftizoxime, and cefotetan varied greatly among both species and participating institutions. The addition of a beta-lactamase inhibitor increased the potency of the beta-lactam drugs tested as combinations. This finding suggests that beta-lactamase production is the major resistance factor in members of the B fragilis group.

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.

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.

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.

Invasive aspergillosis in AIDS.

Invasive pulmonary aspergillosis was diagnosed in two patients with HIV infection, one with prolonged neutropenia and another receiving corticosteroid therapy. We found 17 additional cases in the literature. A known predisposing risk factor for invasive aspergillosis, eg, neutropenia, corticosteroid use, or intravenous drug abuse, was present in 79% of the cases. That the known immunologic defect of AIDS is not a major host defense against Aspergillus is supported by the empiric observation of the relative rarity of aspergillosis in patients with AIDS. The lung was the most common site of Aspergillus infection (75%), and transbronchial biopsy is diagnostically useful. Central nervous system involvement was seen in 55% and appears to be more frequent in HIV-infected patients than in other immunosuppressed patients with invasive aspergillosis. Prognosis is grim. Despite early institution of amphotericin B therapy in a few cases, the disease was uniformly fatal. Efficacy of therapy with amphotericin B plus rifampin or itraconazole remains to be evaluated. We conclude that aspergillosis is not an AIDS-related opportunistic infection.

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.

Isolation of Legionella pneumophila from blood with the BACTEC system: a prospective study yielding positive results.

In a prospective study of 16 patients with Legionnaires disease confirmed by cultural isolation of Legionella pneumophila from the respiratory tract, 38% (6 of 16) had positive blood cultures. Daily subcultures were made onto buffered charcoal-yeast extract plates from 6B aerobic and 7C anaerobic BACTEC blood culture bottles (Johnston Laboratories, Inc., Towson, Md.). Isolation of L. pneumophila was achieved from both aerobic and anaerobic bottles. L. pneumophila growth indices failed to exceed the BACTEC threshold limits; thus, the organism would have been overlooked despite its presence in the blood culture bottles. Bacteremic patients had statistically significant higher quantities of L. pneumophila isolated from sputum and visualized on direct fluorescent antibody stains. Thus, the potential exists for improved diagnosis of Legionella infection by a relatively noninvasive procedure (blood culture) with an instrument already in use in many hospital laboratories.

Oerskovia xanthineolytica implicated in peritonitis associated with peritoneal dialysis: case report and review of Oerskovia infections in humans.

Oerskovia species are nocardialike bacteria that have been implicated as pathogens only rarely. These organisms are branched, gram-positive bacilli that are oxidase negative, catalase positive, and non-acid fast. Unlike Nocardia species, these organisms are motile, do not produce aerial mycelia, and possess a cell wall with large amounts of galactose. Colonies are bright yellow and produce branched vegetative hyphae on nutrient agar. A 70-year-old patient undergoing chronic ambulatory peritoneal dialysis for end-stage renal dysfunction developed recurrent peritonitis. Five peritoneal fluid cultures and one catheter specimen obtained over a period of two weeks yielded a gram-positive bacillus; Oerskovia xanthineolytica was isolated from all six cultures. Prolonged systemic therapy with gentamicin and vancomycin was unsuccessful in curing the peritonitis, but the infection resolved following removal of the peritoneal catheter. This is the first reported case of peritonitis associated with this microorganism. A review of previously described Oerskovia infections, most of which were associated with foreign bodies, showed that removal of infected foci was usually necessary for cure.

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.

A note on symbiosis of Legionella pneumophila and Tatlockia micdadei with human respiratory flora.

Sixteen micro-organisms, representing clinically important respiratory microflora, were tested for their ability to stimulate the growth of Legionella pneumophila and Tatlockia micdadei in nutritionally-deficient agar media. Nutritional symbiosis, indicated by the appearance of satellite colonies of L. pneumophila or T. micdadei, was observed for H. influenzae and N. meningitidis. This interaction between normal flora and pathogens of the respiratory tract may have clinical relevance in the pathogenesis of Legionnaires' disease and Pittsburgh pneumonia.