agar and Bacteremia

From 1 January to 31 December 2022, fifty-five institutions across Australia participated in the Australian Staphylococcus aureus Surveillance Outcome Program (ASSOP). The aim of ASSOP 2022 was to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on susceptibility to methicillin and on characterisation of the molecular epidemiology of the methicillin-resistant isolates. A total of 3,214 SAB episodes were reported, of which 77.5% were community-onset. Overall, 15.0% of S. aureus were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 21.4%, which was significantly different to the 16.8% all-cause mortality associated with methicillin-susceptible SAB (p = 0.02). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S. aureus was rare. However, in addition to the β-lactams, approximately 31% of methicillin-resistant S. aureus (MRSA) were resistant to ciprofloxacin; 30% to erythromycin; 13% to tetracycline; 11% to gentamicin; and 2% to co-trimoxazole. One MRSA isolate, with a daptomycin MIC of 1.5 mg/L, harboured the A302V mprF and A23V cls2 mutations. When applying the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, teicoplanin resistance was detected in one MRSA isolate. Resistance to vancomycin or linezolid was not detected. Resistance to non-β-lactam antimicrobials was largely attributable to the healthcare-associated MRSA (HA-MRSA) clone ST22-IV [2B] (EMRSA-15), and to the community-associated MRSA (CA-MRSA) clone ST45-V [5C2&5] which has acquired resistance to multiple antimicrobials including ciprofloxacin, clindamycin, erythromycin, gentamicin, and tetracycline. The ST22-IV [2B] (EMRSA-15) clone is the predominant HA-MRSA clone in Australia. Nonetheless, 86% of methicillin-resistant SAB episodes were due to CA-MRSA clones. Although polyclonal, approximately 72% of CA-MRSA clones were characterised as ST93-IV [2B] (Queensland clone); ST5-IV [2B]; ST45-V [5C2&5]; ST1-IV [2B]; ST30-IV [2B]; ST97-IV [2B]; ST953-IV [2B]; and ST8-IV [2B]. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S. aureus bacteraemia.

Topics: Agar; Anti-Bacterial Agents; Anti-Infective Agents; Australia; Bacteremia; Ciprofloxacin; Cross Infection; Drug Resistance, Bacterial; Erythromycin; Gentamicins; Humans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Staphylococcal Infections; Staphylococcus aureus; Tetracycline

From 1 January to 31 December 2020, forty-nine institutions around Australia participated in the Australian Enterococcal Sepsis Outcome Programme (AESOP). The aims of AESOP 2020 were to determine the proportion of enterococcal bacteraemia isolates in Australia that were antimicrobial-resistant, and to characterise the molecular epidemiology of the E. faecium isolates. Of the 1,230 unique episodes of enterococcal bacteraemia investigated, 93.9% were caused by either E. faecalis (54.2%) or E. faecium (39.7%). Ampicillin resistance was not detected in E. faecalis but was detected in 88.2% of E. faecium . Vancomycin non-susceptibility was detected in 0.2% of E. faecalis and 32.6% of E. faecium . Overall, 35.2% of E. faecium harboured vanA and/or vanB genes. For the vanA/B positive E. faecium isolates, 38.8% harboured the vanA gene, 60.6% the vanB gene, and 0.6% harboured both vanA and vanB . Although the percentage of E. faecium bacteraemia isolates was significantly lower than that detected in the 2019 AESOP (presumably due to the COVID-19 elective surgery restrictions placed on hospitals), it remains substantially higher than that recorded in most European countries. The E. faecium isolates detected consisted of 71 multilocus sequence types (STs), with 81.7% of these isolates classified into eight major STs each containing ten or more isolates. All major STs belonged to clonal cluster 17 (CC17), a major hospital-adapted polyclonal E. faecium cluster. The major STs (ST17, ST1424, ST80, ST796, ST78, ST1421, ST555 and ST117) were found across most regions of Australia. The predominant clone was ST17, which was identified in all regions except the Northern Territory. Overall, 40.9% of isolates belonging to the eight major STs harboured the vanA or vanB gene. The AESOP 2020 has shown enterococcal bacteraemia episodes in Australia are frequently caused by polyclonal ampicillin-resistant high-level gentamicin-resistant vanA - or vanB -positive E. faecium which have limited treatment options.

Topics: Agar; Anti-Bacterial Agents; Bacteremia; COVID-19; Drug Resistance, Bacterial; Enterococcus; Gram-Positive Bacterial Infections; Humans; Northern Territory; Sepsis

From 1 January to 31 December 2020, forty-nine institutions around Australia participated in the Australian Staphylococcus aureus Sepsis Outcome Programme (ASSOP). The aims of ASSOP 2020 were to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on susceptibility to methicillin; and to characterise the molecular epidemiology of the methicillin-resistant isolates. A total of 2,734 SAB episodes were reported, of which 79.7% were community-onset. Of S. aureus isolates, 17.6% were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 14.2%, which was not significantly different from the 13.3% mortality associated with methicillin-susceptible SAB (p = 0.6). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S. aureus was rare. However, in addition to the β-lactams, approximately 35% of methicillin-resistant S. aureus (MRSA) were resistant to erythromycin, 33% to ciprofloxacin, 13% to tetracycline, 13% to gentamicin and 4% to co-trimoxazole. When applying the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, teicoplanin resistance was detected in four S. aureus isolates. Resistance was not detected for vancomycin and linezolid. Resistance to non-beta-lactam antimicrobials was largely attributable to two healthcare-associated MRSA (HA-MRSA) clones: ST22-IV [2B] (EMRSA-15) and ST239-III [3A] (Aus-2/3 EMRSA). The ST22-IV [2B] (EMRSA-15) clone is the predominant HA-MRSA clone in Australia. However, 85% percent of methicillin-resistant SAB isolates were community-associated MRSA (CA-MRSA) clones. Although polyclonal, approximately 77% of CA-MRSA clones were characterised as: ST93-IV [2B] (Queensland CA-MRSA); ST5-IV [2B]; ST45-V [5C2&5]; ST1-IV [2B]; ST30-IV [2B]; ST8-IV [2B]; and ST97-IV [2B]. The CA-MRSA clones, in particular ST45-V [5C2&5], have acquired multiple antimicrobial resistance determinants including ciprofloxacin, erythromycin, clindamycin, gentamicin and tetracycline. The multi-resistant ST45-V [5C2&5] clone accounted for 11.0% of CA-MRSA. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S. aureus sepsis.

Topics: Agar; Anti-Bacterial Agents; Anti-Infective Agents; Australia; Bacteremia; Ciprofloxacin; Cross Infection; Drug Resistance, Bacterial; Erythromycin; Gentamicins; Humans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Sepsis; Staphylococcal Infections; Staphylococcus aureus; Tetracyclines

Since the introduction of pneumococcal conjugate vaccine, there have been warnings of an increase in infections caused by non-vaccine type of Streptococcus pneumoniae strains. Among them, nonencapsulated Streptococcus pneumoniae (NESp) has been reported to cause invasive infections, especially in children and the elderly. Due to low virulence, however, basic experimental reports on invasive infections are limited.. We applied a liquid-agar method to establish a mouse model of invasive NESp infection. Mice were intratracheally administered a bacterial suspension including agar. With this technique, we investigated the pathogenicity of NESp and the effect of Pneumococcal surface protein K (PspK), a specific surface protein antigen of NESp. NESp wild-type strain (MNZ11) and NESp pspK-deleted mutant strain (MNZ1131) were used in this study. The survival rate, number of bacteria, cytokine/chemokine levels in the bronchoalveolar lavage fluid, and histology of the lung tissue were evaluated.. Mice that were intratracheally administered MNZ11 developed lethal pneumonia with bacteremia within 48 h. Conversely, MNZ1131 showed predominantly low lethality without significant pro-inflammatory cytokine production. NESp was found to cause severe pneumonia and bacteremia upon reaching the lower respiratory tract, and PspK was a critical factor of NESp for developing invasive infections.. The current study demonstrated the ability of NESp to develop invasive diseases, especially in connection with PspK by use of a mouse pneumonia model.

Topics: Agar; Animals; Bacteremia; Cytokines; Mice; Pneumococcal Infections; Pneumonia, Pneumococcal; Streptococcus pneumoniae; Virulence

In 2019, the CLSI lowered the susceptible levofloxacin breakpoints for Enterobacterales from a MIC of ≤2 μg/mL to ≤0.5 μg/mL. The study evaluated the correlation between the susceptibility profiles obtained by the Vitek 2 and agar dilution (AD) methods in levofloxacin MIC ≤2 μg/mL isolates and its clinical impacts. Two hundred fifty-three Enterobacterales isolates and 222 patients treated with levofloxacin for Enterobacterales bacteremia were enrolled for analysis. There was 86.2% categorical agreement, 5 very major errors, and 30 minor errors based on the 2019 CLSI breakpoints. Higher levofloxacin MICs (1 or 2 μg/mL) determined using Vitek 2 or AD predicted early clinical failure (P < 0.001 for Vitek 2 and P = 0.001 for AD). In conclusion, Vitek 2 performance for levofloxacin susceptibility testing of Enterobacterales declined according to the 2019 CLSI criteria compared with the pre-2019 criteria. Although discrepant results were obtained, the MICs measured by Vitek 2 could still predict treatment outcomes.

Topics: Agar; Anti-Bacterial Agents; Bacteremia; Humans; Levofloxacin; Microbial Sensitivity Tests

From 1 January to 31 December 2021, forty-eight institutions around Australia participated in the Australian Enterococcal Surveillance Outcome Programme (AESOP). The aim of AESOP 2021 was to determine the proportion of enterococcal bacteraemia isolates in Australia that were antimicrobial resistant, and to characterise the molecular epidemiology of the Enterococcus faecium isolates. Of the 1,297 unique episodes of enterococcal bacteraemia investigated, 94.4% were caused by either E. faecalis (54.1%) or E. faecium (40.3%). Ampicillin resistance was detected in one E. faecalis isolate and in 89.3% of E. faecium isolates. Vancomycin non-susceptibility was not detected in E. faecalis but was detected in 37.9% of E. faecium. Overall, 39.6% of E. faecium harboured the vanA and/or vanB genes. For the vanA/vanB positive E. faecium isolates, 35.8% harboured the vanA gene and 64.2% the vanB gene. Although the percentage of vancomycin-resistant E. faecium bacteraemia isolates was significantly lower than that reported in the 2020 AESOP report (presumably due to the COVID-19 elective surgery restrictions placed on hospitals), it remains substantially higher than that recorded in most European countries. Isolates of E. faecium consisted of 73 multi-locus sequence types (STs); 77.2% of isolates were classified into seven major STs each containing more than ten isolates. All major STs belonged to clonal cluster (CC) 17, a major hospital-adapted polyclonal E. faecium cluster. The major STs (ST17, ST1424, ST796, ST78, ST80, ST1421 and ST555) were found across most regions of Australia. The predominant ST was ST17 which was identified in all regions except the Northern Territory. Overall, 46.5% of isolates belonging to the seven major STs harboured the vanA or vanB gene. The AESOP 2021 has shown that enterococcal bacteraemia episodes in Australia are frequently caused by polyclonal ampicillin-resistant high-level gentamicin resistant vanA- or vanB-positive E. faecium which have limited treatment options.

Topics: Agar; Anti-Bacterial Agents; Bacteremia; COVID-19; Drug Resistance, Bacterial; Enterococcus; Gram-Positive Bacterial Infections; Humans; Microbial Sensitivity Tests; Northern Territory; Vancomycin

From 1 January to 31 December 2021, forty-eight institutions around Australia participated in the Australian Staphylococcus aureus Surveillance Outcome Programme (ASSOP). The aim of ASSOP 2021 was to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on susceptibility to methicillin and on characterisation of the molecular epidemiology of the methicillin-resistant isolates. A total of 2,928 SAB episodes were reported, of which 78.4% were community-onset. Overall, 16.9% of S. aureus isolates were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 15.0%, which was not significantly different from the 14.4% all-cause mortality associated with methicillin-susceptible SAB (p = 0.7). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S. aureus was rare. However, in addition to the β-lactams, approximately 36% of methicillin-resistant S. aureus (MRSA) were resistant to ciprofloxacin; 30% to erythromycin; 15% to tetracycline; 16% to gentamicin; and 3% to co-trimoxazole. When applying the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, teicoplanin resistance was detected in three S. aureus isolates. Resistance to vancomycin or linezolid was not detected. Resistance to non-β-lactam antimicrobials was largely attributable to the healthcare-associated MRSA (HA-MRSA) clone ST22-IV [2B] (EMRSA-15), and the community-associated MRSA (CA-MRSA) clone ST45-V [5C2&5] which has acquired multiple antimicrobial resistance determinants including ciprofloxacin, erythromycin, clindamycin, gentamicin and tetracycline. The ST22-IV [2B] (EMRSA-15) clone is the predominant HA-MRSA clone in Australia. Nonetheless, 85% of methicillin-resistant SAB episodes were due to CA-MRSA clones. Although polyclonal, approximately 68% of CA-MRSA clones were characterised as ST93-IV [2B] (Queensland clone); ST45-V [5C2&5]; ST5-IV [2B]; ST1-IV [2B]; ST30-IV [2B]; and ST97-IV [2B]. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S. aureus bacteraemia.

Topics: Agar; Anti-Bacterial Agents; Anti-Infective Agents; Australia; Bacteremia; Ciprofloxacin; Cross Infection; Drug Resistance, Bacterial; Erythromycin; Gentamicins; Humans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Staphylococcal Infections; Staphylococcus aureus; Tetracycline

The main objective of the present study is to analyze different genotypic and phenotypic traits related to virulence in Enterococcus faecalis, as well as evaluated the agar invasion phenotype in a collection of isolates with different clinical origins.. Seventy-nine E. faecalis isolates, with invasive and non-invasive clinical origins, have been used in this work. Presence of cytolysin activator (cylA), gelatinase (gelE), surface protein (esp), aggregation substance (asa1), endocarditis antigen (efaA), and collagen-binding protein (ace) have been analyzed by PCR. Phenotypic characterization included gelatinase activity, haemolysin production, biofilm formation and agar invasion.. All the isolates tested harboured at least one of the virulence determinants. The 95.5% of isolates from haematologic samples were positive for agar invasion test, significantly higher than isolates from non-invasive diseases. A significant reduction in relative invasion area was observed in three selected agar-invasive strains after 15 serial passages.. It has been observed a significant high prevalence of agar-invasion positive isolates among strains belonged to haematological samples. Agar invasiveness is reduced after adaptation of clinical isolates to laboratory conditions, showing that agar invasion phenotype can be modulate by culture conditions as other virulence factors observed in different bacterial species.

Topics: Agar; Bacteremia; Bacterial Proteins; Biofilms; Enterococcus faecalis; Gelatinases; Genes, Bacterial; Genotype; Gram-Positive Bacterial Infections; Hemolysin Proteins; Humans; Microbial Sensitivity Tests; Phenotype; Virulence Factors

Listeria monocytogenes infection in pregnant women and newborns is a cause for serious concern, and invasive disease outcome strongly depends on prompt antibiotic therapy. To provide sooner identification from neonatal bacteremia we performed a CAMP test directly on positive blood aliquots and inoculated the Liofilchem(®) O.A. Listeria chromogenic agar as well, thus providing a 24-h turn-around time for response.

Topics: Agar; Bacteremia; Bacteriological Techniques; Chromogenic Compounds; Early Diagnosis; Humans; Infant, Newborn; Listeria monocytogenes; Listeriosis; Time Factors

Tigecycline is a semi-synthetic tetracycline with activity against most multidrug-resistant (MDR) bacteria.. We studied in vitro activity of tigecycline by agar dilution (AD) and Etest methods to evaluate their correlation. The study included 206 isolates of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli, Klebsiella pneumoniae and MDR Acinetobacter baumannii recovered from blood cultures of patients of Baskent University between 2008 and 2010.. ESBL-producing E. coli had MIC50/MIC90 values of 0.5/0.5 µg/ml by AD and 0.25/0.5 µg/ml by Etest. ESBL-producing K. pneumoniae had MIC50/MIC90 values of 1/2 µg/ml by AD and 0.75/2 µg/ml by Etest, whereas MDR A. baumannii had MIC50/MIC90 values of 4/4 µg/ml by AD and 2/4 µg/ml by Etest. The correlation between AD and Etest was weak for ESBL-producing E. coli and strong for ESBL-producing K. pneumoniae and MDR A. baumannii. Tigecycline MIC values for ESBL-producing E. coli were lower than the tigecycline concentration, while they were higher than the concentrations attainable by treatment doses for A. baumannii.. Tigecycline is an appropriate agent in the treatment of E. coli bacteremia, but it is not for treating A. baumannii bacteremia. Tigecycline could be used for K. pneumoniae bacteremia treatment after determining its MIC value. Determining the MIC value by gold-standard methods is more appropriate due to the correlation between Etest and AD at high MIC values.

Topics: Agar; Anti-Bacterial Agents; Bacteremia; Drug Resistance, Multiple, Bacterial; Gram-Negative Bacteria; Humans; Microbial Sensitivity Tests; Minocycline; Tigecycline

Staphylococcus is the most prevalent pathogen causing bacteremia and many of its isolates possess the ability to form biofilm. In this study Staphylococcus isolates from the blood of patients with bacteremia were analyzed by two biofilm detection phenotypic methods: Congo red agar (CRA) and microtiter-plate adherence (MPA) in relation to the presence of ica genes, detected by PCR. Their oxacillin susceptibility was also evaluated. Among 127 isolates evaluated, 47 were S. aureus and 80 were coagulase negative staphylococci (CNS). Seventy-four (58.3%) isolates were mecA gene positive (27.7%S. aureus and 76.3% CNS isolates). Among the 40 S. aureus isolates which were positive for the ica genes, 25 (62.5%) were positive in MPA and 27 (67.5%) in CRA, whereas both methods combined detected 34 (85%) isolates as biofilm producers. Among 12 S. epidermidis isolates carrying ica genes, 8 were positive in MPA and 5 in CRA. The combination of CRA and MPA methods provided a better prediction of the presence of ica genes in S. aureus isolates than did either method alone.

Topics: Agar; Bacteremia; Bacterial Adhesion; Bacteriological Techniques; Biofilms; Congo Red; Humans; Staphylococcus

The incidence of Bacillus cereus bacteremia is increasing, but the identification of Bacillus species remains difficult. Brilliance Bacillus cereus agar (BBC agar; Oxoid, UK) is a new CHROMagar medium that allows selective isolation and identification of B. cereus; however, its clinical usefulness is seldom studied. We evaluated the usefulness of BBC agar to identify B. cereus isolates recovered from blood cultures.. We analyzed a total of 53 blood isolates that showed a Bacillus-like morphology on Gram staining. All isolates were identified by using both the API Coryne (bioMérieux, France) and API 50CH/B (bioMérieux) systems. They were subsequently subcultured on BBC agar, incubated for 24 hr, and then examined for characteristic blue-green colonies. The clinical characteristics of patients whose isolates were identified as B. cereus were assessed.. Of the 53 isolates, 18 were identified as B. cereus by API 50CH/B. With the API 50CH/B system used as gold standard, the sensitivity and specificity for the identification of B. cereus were 100% (18/18) and 100% (35/35), respectively, using BBC agar, and 67% (12/18) and 100% (35/35), respectively, using the API Coryne system. Of the 18 patients with B. cereus bacteremia, 15 showed infectious signs, and 3 had more than 2 blood cultures positive for B. cereus on separate days.. Our study shows, for the first time, that BBC agar, with its good agreement and ease of use, is a valuable alternative to the API 50CH/B system for the presumptive identification of B. cereus isolates from blood cultures.

Topics: Adolescent; Adult; Agar; Aged; Bacillus cereus; Bacteremia; Child; Child, Preschool; Chromogenic Compounds; Culture Media; Female; Gram-Positive Bacterial Infections; Humans; Infant; Infant, Newborn; Male; Middle Aged; Reagent Kits, Diagnostic; Sensitivity and Specificity

Topics: Agar; Bacteremia; Bacterial Proteins; Bacteriological Techniques; beta-Lactamases; Blood; Cephalosporin Resistance; Cephalosporins; Chromogenic Compounds; Culture Media; Enterobacteriaceae; Enterobacteriaceae Infections; Humans; Microbial Sensitivity Tests; R Factors; Sensitivity and Specificity

We experienced a case of a 3-year-old boy who presented signs and symptoms of Kawasaki syndrome. Two blood culture sets were processed by the hospital microbiology laboratory using a standard blood culturing system. The anaerobic bottles gave a positive result at day 3 after inoculation. The biochemical profiles produced by the RapID ANA II System showed that the organism was Clostridium baratii with a probability of 99%. Our case highlights the importance of C. baratii as a potential human pathogen and reports the associations with manifestations, which, to our knowledge, have not been previously described concomitantly with a clostridial infection.

Topics: Agar; Anaerobiosis; Bacteremia; Child, Preschool; Clostridium; Clostridium Infections; Humans; Male; Mucocutaneous Lymph Node Syndrome; Reagent Kits, Diagnostic

We describe a novel mouse model of acute staphylococcal pneumonia induced by intravenous injection of Staphylococcus aureus enmeshed in agar beads. For comparison, we also used various strains of bacteria, including three strains of S. aureus, two strains of Staphylococcus epidermidis, one strain of Streptococcus pyogenes, three strains of Pseudomonas aeruginosa, and one strain of Klebsiella pneumoniae. All except two strains of S. aureus were cleared rapidly from the lungs. When S. aureus NUMR1 enmeshed in agar beads was injected intravenously, the organisms concentrated and remained in the lung for a period longer than several weeks. Multiple lung abscesses were evident macroscopically, and histological examination of the infected lung showed multiple lung abscesses around the pulmonary arterioles, consisting of bacterial colonies encircled with fibrin filaments and surrounded by inflammatory cells of neutrophils and macrophages. When 14 strains of clinically isolated S. aureus were injected intravenously, the number of bacteria recovered from the lung tissue 7 days after infection correlated with the titer of staphylocoagulase (P < 0.01) but not with the titer of clumping factor. Injection of coagulase-deficient mutant strain DU5843 was associated with a markedly reduced number of viable bacteria isolated from the lung, compared with its coagulase-positive parental strain DU5789. Our results suggest that coagulase may play a role in the development of blood-borne staphylococcal pneumonia in our model. Our animal model is simple and reproducible and resembles blood-borne staphylococcal pneumonia in humans, and it could be useful for investigating the pathogenicity or treatment of staphylococcal pulmonary infection, including infections with methicillin-resistant S. aureus.

Topics: Agar; Animals; Bacteremia; Coagulase; Colony Count, Microbial; Disease Models, Animal; Injections, Intravenous; Lung; Male; Mice; Mice, Inbred Strains; Microspheres; Pneumonia, Staphylococcal; Staphylococcal Infections; Staphylococcus aureus

From 1 February through 12 October 1990, 27 blood cultures processed at Shiprock Hospital were positive for Enterobacter cloacae; only 3 had been reported in the preceding 12 months. Twenty (74%) of the cultures were obtained from patients without clinical evidence of gram-negative septicemia. The increase in E. cloacae-positive blood cultures was temporally associated with the introduction of a new blood culturing system. To evaluate potential risk factors for an E. cloacae-positive blood culture (case-culture), we conducted a case-control study. Case-cultures were compared with 81 randomly selected cultures that were processed during the epidemic period and that were not positive for E. cloacae (controls). Because several factors suggested the possibility of pseudoinfection, we limited our analysis to the 20 blood cultures that appeared to be contaminants. Blood samples received in the laboratory during the midnight shift (5 of 20 [25%] versus 5 of 81 [6%]; odds ratio, 5.1; 95% confidence intervals, 1.01 to 24.6; P = 0.02) or present in the incubator with other E. cloacae-positive samples (17 of 20 [85%] versus 29 of 81 [36%]; odds ratio, 10.2, 95% confidence interval, 2.6 to 57.3; P < 0.001) were at increased risk for contamination. During mock experiments of the procedures for processing blood samples for culture, several breaks in aseptic technique and leakage from the blood culturing system were observed. Cultures of samples obtained from several environmental sites in the laboratory and the hand washings of two laboratory technicians grew E. cloacae. Plasmid and restriction enzyme analyses of E. cloacae isolates recovered from the patients' blood cultures, the two technicians' hand washings, and environmental sites in the laboratory indicated that all had identical plasmid profiles. Our findings suggest that the breaks in aseptic technique and the environmental contamination that occurred in association with the use of the new blood culturing system resulted in contamination of the blood cultures. This outbreak highlights the importance of routine environmental cleaning, periodic quality control assessments, and adherence to aseptic practices in clinical laboratories, particularly when new methods or equipment are introduced and/or new personnel are hired.

Topics: Agar; Bacteremia; Bacteriological Techniques; Case-Control Studies; Cluster Analysis; Disease Outbreaks; Enterobacter cloacae; Enterobacteriaceae Infections; Humans; Quality Control