First detection of plasmid-encoded blaOXY beta-lactamase.

Three Klebsiella oxytoca isolates and one Klebsiella pneumoniae isolate from three children admitted to the Hematology Unit of Hospital Vall d'Hebron (Barcelona, Spain) exhibited a susceptibility pattern suggesting OXY beta-lactamase hyperproduction. All the isolates contained a 95-kb plasmid that harbored bla(OXY-1), which was transferred by electrotransformation but could not be self-transferred by conjugation. A qnrS1 gene was also harbored in the bla(OXY-1)-carrying plasmid. This is the first report of a plasmid-encoded OXY beta-lactamase.

Prevalence of qnr genes among extended-spectrum beta-lactamase-producing enterobacterial isolates in Barcelona, Spain.

OBJECTIVES: To evaluate the presence of qnr genes among enterobacterial isolates carrying extended-spectrum beta-lactamases (ESBLs) in Barcelona, Spain. METHODS: Screening for the qnrA, qnrB and qnrS genes was carried out by PCR amplification with specific primers in 305 non-duplicate, clinically relevant ESBL-producing enterobacterial isolates obtained from February 2003 to August 2004. ESBLs from all qnr-positive isolates were characterized by isoelectric focusing, PCR amplification and DNA sequencing. Plasmid analysis was performed by S1 digestion and hybridization with specific probes for the qnr and bla genes. Plasmids containing qnr genes were transferred by conjugation or transformation. The genetic environment of qnrA1 in selected isolates was characterized by cloning experiments. RESULTS: Fifteen isolates, each from a different individual, carried qnr. Among them, 14 had qnrA1 (6 Klebsiella pneumoniae, 6 Enterobacter cloacae and 2 Escherichia coli isolates) and 1 had qnrS1 (K. pneumoniae). None of the isolates carried qnrB. Among the qnrA1-carrying isolates, 10 possessed both bla(CTX-M-9) and bla(SHV-12), 2 had both bla(CTX-M-9) and bla(SHV-92) and 2 had bla(CTX-M-9) alone. The isolate with qnrS1 possessed bla(SHV-12). The qnrA1 and ESBL genes were located together on plasmids ranging in size from 40 to 320 kb. qnrS1 and bla(SHV-12) were not located on the same plasmid. Transfer of quinolone resistance was successfully achieved from all but three isolates. The cloned region surrounding qnrA in two K. pneumoniae isolates revealed a novel genetic organization. CONCLUSIONS: The prevalence of qnr among enterobacterial clinical isolates carrying ESBLs between 2003 and 2004 in Barcelona was 4.9%. qnrA1 was the most prevalent, whereas only one qnrS and no qnrB were detected.

Dissemination of extended-spectrum beta-lactamase-producing bacteria: the food-borne outbreak lesson.

OBJECTIVES: Commensal and opportunistic bacteria producing extended-spectrum beta-lactamases (ESBL-PB) have undergone a broad and rapid spread within the general population; however, the routes of dissemination have not been totally elucidated. The aim of this study was to determine whether individuals involved in an outbreak of acute gastroenteritis, in addition to the enteropathogenic microorganism, share an ESBL-PB as indirect demonstration of its transmission from a common food source. METHODS: From 2003 to 2004 in Barcelona, Spain, stool samples from 905 people involved in 132 acute gastroenteritis outbreaks and 226 food handlers related to the outbreaks were investigated. RESULTS: In 31 outbreaks, 58 diners carrying one or more ESBL-PB were detected. In 10 outbreaks, two or more diners shared the same ESBL-PB, and in four of them, the strain was shared with the food handlers. CONCLUSIONS: This study provides circumstantial evidence that foods can be a transmission vector for ESBL-PB, probably from two reservoirs, food animals and food handlers.

Pathogenicity island markers in commensal and uropathogenic Escherichia coli isolates.

Uropathogenic isolates of Escherichia coli (UPEC) contain blocks of DNA, termed pathogenicity islands (PAIs), that contribute to their virulence. Two multiplex PCR assays were developed to detect eight PAI markers among 50 commensal E. coli and 100 UPEC isolates. In total, 40% of commensal isolates and 93% of UPEC carried PAIs. Despite this difference, the distribution of various PAIs showed the same pattern in both groups, with the most prevalent being PAI IV(536) (38% commensal vs. 89% UPEC), followed by PAI I(CFT073) (26% vs. 73%), PAI II(CFT073) (14% vs. 46%), PAI II(J96) (8% vs. 34%), PAI I(536) (8% vs. 33%) and PAI II(536) (4% vs. 20%). PAI III(536) was detected only in UPEC (2%), while PAI I(J96) was not detected in any isolate. Although the mean number of PAIs per isolate was higher among UPEC (2.97) than in commensal (0.98) isolates, there were no statistical differences among group B2 E. coli from the two origins; however, commensal isolates from groups D and B1 appeared to be less virulent than pathogenic isolates. Regardless of their phylogenetic group, nearly all the commensal and UPEC isolates with the same number of PAIs had the same PAI combinations. Although group B2 E. coli are uncommon among commensal intestinal flora, they are highly virulent when present, suggesting that the intestinal flora may act as a reservoir for bacteria that can cause urinary tract infection.

[Patterns of susceptibility to antibiotics of Enterobacteriaceae causing intra-abdominal infection in Spain: SMART 2003 study outcomes]. / Patrones de sensibilidad a antimicrobianos de Enterobacteriaceae causantes de infecciones intraabdominales en España: resultados del estudio SMART 2003.

SMART (Study for Monitoring Antimicrobial Resistance Trends) is an ongoing global antimicrobial surveillance program focused on clinical isolates from intra-abdominal infections. The objective of this subanalysis was to assess antimicrobial susceptibility patterns among Entero-bacteriaceae recovered at 13 participating Spanish sites during 2003. Antimicrobial susceptibility testing was performed using broth microdilution techniques according to the CLSI (formerly NCCLS) guidelines for MIC testing. The presence of extended-spectrum beta-lactamases (ESBL) was confirmed in isolates with a MIC of ceftriaxone, ceftazidime, or cefepime>or=2 mg/l by comparing cefepime MICs with and with-out clavulanate. A total of 981 Enterobacteriaceae recovered from 840 patients were tested, of which 398 (41%) were community-acquired. Escherichia coli was the most common isolate (571 isolates; 58%), followed by Klebsiella spp. (153; 16% Enterobacter spp. (97; 10%), and Proteus spp. (63; 6%). A total of 191 isolates (19%) from 176 patients produced inducible beta-lactamases. The carbapenems and amikacin were the most consistently active agents against the Enterobacteriaceae (susceptibility>or=99%). Resistance rates for ceftazidime, cipro-floxacin, and levofloxacin exceeded 10%. ESBLs were detected phenotypically in 61 (6%) isolates, being the most common E. coli (61%), Klebsiella spp. (20%), and Enterobacter spp. (8%). Antimicrobial resistance among Enterobacteriaceae isolated from intra-abdominal infections is a problem in Spain. A significant proportion of inducible beta-lactamase and ESBL-producing Enterobacteriaceae causing intra-abdominal infection were acquired in the community. The carbapenems ertapenem, imipenem and meropenem and the aminoglycoside amikacin were highly active in vitro against Enterobacteriaceae isolated from intra-abdominal sites, including ESBL-producing organisms.

Predictors of tuberculosis transmission in prisons: an analysis using conventional and molecular methods.

OBJECTIVE: To determine the tuberculosis (TB) transmission patterns within the prison system in Catalonia, conventional epidemiological techniques were combined with DNA fingerprinting of Mycobacterium tuberculosis. METHODS: IS6110- and polymorphic GC-rich repeat sequence (PGRS)-based restriction fragment length polymorphism (RFLP) were combined with epidemiological studies to assess the relatedness of isolates from all patients with confirmed TB at five prisons in the province of Barcelona (Catalonia, Spain), between 1 July 1994 and 31 December 1996. Risk factors for transmission were analysed to a logistic regression. The extent of drug-resistant TB was also assessed. RESULTS: The incidence of TB during the study period was 2775 cases per 100,000 inmate years. Of the 247 culture-positive cases, 126 (51%) appeared to have active TB as a result of recent transmission. Using conventional epidemiological methods, 14 active chains of transmission were identified in prison involving 65 isolates (52% of clustered patients). A lengthy history of imprisonment [odds ratio (OR) 2.8, 95% confidence interval (CI) 1.52-5.11] and pulmonary TB (OR 2.36, 95% CI 1.17-4.75) were independently associated with clustering. Low rates of both initial (2.9%) and acquired drug resistance (5.8%) were identified and there was no evidence of the transmission of drug-resistant TB. CONCLUSION: In the prison system studied, the recent transmission of TB contributes substantially to the overall incidence of the disease. Both lengthy incarcerations and delays in identifying inmates with pulmonary symptoms play a key role in this recent transmission. Directly observed therapy (DOT) is a critical control strategy for reducing the emergence of drug resistance and for avoiding the transmission of resistant organisms.

Antiviral susceptibility of Herpes simplex viruses and its clinical correlates: a single center's experience.

The in vitro susceptibility to acyclovir of 204 herpes simplex virus isolates from 165 immunocompromised patients treated at our hospital was determined by the cytopathic effect reduction assay. Approximately 95% of herpes simplex virus 1 and 73% of herpes simplex virus 2 isolates were inhibited by acyclovir at concentrations of <2 microgram/mL. From 8 patients (5%), an isolate with low susceptibility to acyclovir (50% inhibitory dose, >3 microgram/mL) was recovered. Medical records of 83 patients were reviewed. Lesions resolved in most of the patients, independent of treatment. Treatment failures were not always associated with isolation of an in vitro-resistant virus. On the contrary, when a virus with low susceptibility to acyclovir was isolated, resolution of the lesion was the rule. In 9 of 10 patients with subsequent recurrent episodes of disease, the susceptibility of the viruses isolated was similar to that of the first episode. Routine susceptibility testing in our geographic area is not encouraged because of the low incidence of acyclovir-resistant herpes simplex viruses.

Infection of pulmonary artery catheters. Epidemiologic characteristics and multivariate analysis of risk factors.

Sixty-nine consecutive pulmonary artery catheters (PACs) were prospectively studied in a medical-surgical intensive care unit. Fifteen (21.7 percent) and two (2.9 percent) of the PACs were associated with colonization and bacteremia, respectively. These data represent an incidence of 4.98 and 0.66 episodes per 100 catheterization-days, respectively. Coagulase-negative staphylococci were the most common isolates. The source of the colonizing microorganism was the skin in 56 percent of cases, hubs in 28 percent, and unknown in 16 percent. From multivariate analysis, only more than 5 days of catheterization was significantly associated with a greater risk of colonization. Antimicrobial use was associated with negative cultures. The most useful method to diagnose colonization was the examination of both tip and intradermal segments. In addition, we suggest associate hub cultures when catheter-related bacteremia is suspected. These data may be useful in improving efficacy in the diagnosis and prevention of PAC-related infection.

Impact of previous antimicrobial therapy on the etiology and outcome of ventilator-associated pneumonia.

OBJECTIVE: To define the influence of prior antibiotic use on the etiology and mortality of ventilator-associated pneumonia (VAP). SETTING: A university hospital medical-surgical ICU. DESIGN: Prospective clinical study. METHODS: Over a 35-month period, we prospectively studied 129 consecutive episodes of VAP. Etiologic diagnosis was established using a protected specimen brush and quantitative culture techniques. We examined prognostic factors by univariate and multivariate analyses using a statistical software package (SPSS). RESULTS: The rate of VAP caused by Gram-positive cocci or Haemophilus influenzae was statistically lower (p < 0.05) in the patients who had received antibiotics previously, while the rate of VAP caused by Pseudomonas aeruginosa was statistically higher (p < 0.01). Patients died of causes directly related to the infection in 18 (14.0 percent) episodes, P aeruginosa being isolated in 9 of these fatal cases. Indeed, we found that 27.7 percent (15/54) of patients who had received prior antimicrobial therapy before the onset of pneumonia died, compared with only 4.0 percent (3/75) of those who did not. In the univariate analysis, the variables significantly associated with attributable mortality were age older than 45 years, use of corticosteroids, presence of shock, hospital day of VAP over 9, antecedent COPD, and a prior antibiotic use. A step-forward logistic regression analysis defined only prior antibiotic use (p < 0.0001, OR = 9.2) as significantly influencing the risk of death from VAP. The same result was obtained when severity was included in the model. However, prior antibiotic use entirely dropped out as a significant risk factor when the etiologic agent was included in the regression equation. CONCLUSIONS: Distribution of infecting microorganisms responsible for VAP differs in patients who received prior antimicrobial therapy, and this factor determines a higher mortality rate. We suggest a restrictive antibiotic policy in mechanically ventilated patients with the purpose of reducing the risk of death from VAP.

A three-year study of severe community-acquired pneumonia with emphasis on outcome.

Fifty-eight consecutive patients with severe community-acquired pneumonia were studied prospectively during a three-year period. The group included 44 men and 14 women (mean age: 45.0 +/- 15.7 years). The cause of pneumonia was diagnosed in 35 (60.3 percent) cases, and the most common pathogens were Streptococcus pneumoniae (37.1 percent), Legionella pneumophila (22.8 percent) and Gram-negative bacilli (11.4 percent). The fact that Mycobacterium tuberculosis was present in four (11.4 percent) patients and Pneumocystis carinii in three (8.5 percent) is worthy of note. The overall death rate was 22.4 percent. More than 50 percent of deaths occurred within the first five days and were caused by septic shock, hemoptysis (tuberculosis) or hypoxia. However, hypoxia remains the main fatal complication and all late-occurring deaths (> 5 days) observed were due to this cause. These data could be important in planning strategies and protocols to improve prognosis.

Pneumonia due to Haemophilus influenzae among mechanically ventilated patients. Incidence, outcome, and risk factors.

Incidence and potential risk factors for pneumonia due to Haemophilus influenzae in adults treated with mechanical ventilation in a medical-surgical ICU were investigated. Diagnosis was established in 91 episodes and H influenzae was isolated in 20 of them. Mean onset of ventilator-associated pneumonia (VAP) due to H influenzae was 10.8 days after intubation. Six patients with H influenzae VAP died in the ICU. Of 13 risk factors for developing VAP due to H influenzae, an absence of prior antibiotic treatment was the only variable which had statistical significance (p < 0.001). In these mechanically ventilated patients, Haemophilus influenzae was a common causative agent for VAP, frequently associated with Gram-positive cocci. Episodes of H influenzae VAP were associated with a lower mortality compared with other etiologies. The epidemiologic and clinical findings indicate that patients without a prior antimicrobial treatment have increased susceptibility to infections of the airway by H influenzae.

Nosocomial respiratory tract infections in multiple trauma patients. Influence of level of consciousness with implications for therapy.

A prospective study of 161 multiple trauma patients was carried out to determine the incidence, the causative agents, and the outcome of nosocomial respiratory tract infections in this highly selected population. Thirty-eight (23.6 percent) patients developed a nosocomial pneumonia (NP). In addition, there were four superinfections in three patients, representing an incidence of 26 percent (42 of 161). Incidence of NP was significantly greater among comatose patients (42.2 vs 13.3 percent, p less than 0.05). Furthermore, purulent tracheobronchitis was diagnosed in six patients. The causative agent of NP was identified in 36 (85.7 percent) episodes by means of fiberoptic bronchoscopies with protected specimen brush sampling. Staphylococcus aureus (55.8 percent) was the predominant pathogen isolated in multiple trauma patients in coma (Glasgow coma score [GCS] below 9 during a period greater than 24 h), while aerobic Gram-negative bacilli were responsible for the majority of cases in the remaining population studied. The overall mortality rate was 19.8 percent, but only five deaths were related to NP. We conclude that nosocomial respiratory tract infections are a frequent problem in multiple trauma patients, especially in those with GCS below 9, although this complication is associated with a relatively low mortality. Among patients with GCS below 9, S aureus was a frequent finding; consequently, antimicrobial therapy in this population needs to be different than that for the remaining multiple trauma patients with NP.

Incidence, etiology, and outcome of nosocomial pneumonia in mechanically ventilated patients.

This study assessed the incidence, etiology, and consequences of ventilator-associated pneumonia in 1,000 consecutive patients admitted in a medical-surgical intensive care unit (ICU). A total of 264 patients were submitted to mechanical ventilation (MV) for more than 48 hours. Fifty-eight (21.9 percent) patients developed a bacterial pneumonia after a mean of 7.9 days (range, 2 to 40 days) of MV. In addition, they were ten superinfections in nine patients, raising the mean incidence to 25.7 percent. Five patients developed secondary bacteremia, and another five had septic shock. Identification of the causative agent of pneumonia was possible in 47 episodes by means of highly specific techniques (telescoping plugged catheter, blood cultures, and/or necropsy). Thirteen (27.6 percent) of these cases were polymicrobial. The predominant pathogens isolated in the first episode of pneumonia were Gram-negative bacilli (62.6 percent), but a high incidence of Staphylococcus aureus infection (23.2 percent) was detected. Gram-negative bacilli represented 66.6 percent of the total organisms isolated in superinfections. The mortality rate in the pneumonia group was 42 percent; this percentage is similar to mortality rate among MV patients without pneumonia (37 percent). We conclude that nosocomial pneumonia is a frequent complication of MV in the medical-surgical ICU. Ventilator-associated pneumonia does not appear to increase fatality in critically ill patients with a high mortality rate (38 percent); however, it significantly prolongs the length of stay in the ICU for survivors.

Epidemiologic evidence of transmission of donor-related bacterial infection through a transplanted heart.

This study describes a patient who had fulminant infectious myocarditis as a result of methicillin-resistant Staphylococcus aureus after receiving a heart transplant from an infected donor. There was complete concordance of typing results between donor and recipient strains that were different from the 20 isolates with which they were compared. Molecular epidemiologic study provided compelling evidence that a transplanted organ can transmit a bacterial infection from the donor to the recipient.

Polymicrobial bacteremia in critically ill patients.

OBJECTIVE: To characterize the epidemiology of polymicrobial bacteremia (PMB) among critically ill patients. DESIGN: Prospective clinical study. SETTING: University medical center. PATIENTS: All patients with positive blood cultures in a medical-surgical ICU. MEASUREMENTS: PMB represents 8.4% of all true bacteremia in our ICU. Most of these patients were postoperative but none had malignancies or significant immunodepression. Over three-quarters of the episodes were nosocomial. No significant differences in factors associated with PMB were found when they were compared with a cohort of 154 monomicrobial episodes. Enterobacteriaceae were the most common organisms. Intravascular devices (42.8%) were the most common source of PMB, followed by intra-abdominal origin (21.4%). The overall mortality was 7.1%, a lower rate than has previously been described. CONCLUSIONS: We suggest catheter replacement in patients who develop PMB and improving techniques of catheter maintenance in order to reduce its incidence.

Risk factors for infection by Pseudomonas aeruginosa in patients with ventilator-associated pneumonia.

OBJECTIVE: to investigate the epidemiology of infection by Pseudomonas aeruginosa in patients with ventilator-associated pneumonia (VAP). DESIGN: prospective clinical study. SETTING: a medical-surgical ICU in a university hospital. PATIENTS: we followed-up 568 mechanically ventilated patients and 83 episodes of VAP with etiologic diagnosis in 72 patients were retained for analysis. RESULTS: Ps. aeruginosa was isolated in 22 (26.5%) episodes in 18 patients. Of these episodes 7 were directly responsible for death. Using logistic regression analysis, the risk of VAP due to Ps. aeruginosa was increased in patients with chronic obstructive pulmonary disease (relative risk (RR) = 29.9, 95% confidence interval (CI) = 4.86-184.53), a mechanical ventilation period longer than 8 days (RR = 8.1, 95% CI = 1.01-65.40) and prior use of antibiotics (RR = 5.5, 95% CI = 0.88-35.01). CONCLUSIONS: patients with VAP and these factors have a greater risk of infection by Ps. aeruginosa and empirical therapy for these episodes should include anti-pseudomonal activity until etiologic diagnosis is established.

Nosocomial bacteremia in a medical-surgical intensive care unit: epidemiologic characteristics and factors influencing mortality in 111 episodes.

OBJECTIVE: To analyze the epidemiology and factors influencing mortality of ICU-acquired bacteremia. DESIGN: Prospective clinical study. SETTING: A medical-surgical ICU in an university hospital. PATIENTS: We recorded variables from 111 consecutive ICU-acquired episodes for a 3-year period. RESULTS: The attack rate was 1.9 episodes per 100 patient-days. The commonest isolates were coagulase-negative staphylococci, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. Intravascular catheters were the most frequent source of infection. Overall mortality was 31.5%, and 65.7% of all deaths were directly attributable to infection. Bacteremia from intra-abdominal, lower respiratory tract or unknown origin were associated with a poor prognosis. A logistic regression analysis defined intraabdominal origin (p = 0.01, OR = 15.7) and presence of shock (p = 0.04, OR = 3.3) as independently influencing the risk of death. No significant differences were found for the remaining variables studied. CONCLUSIONS: Epidemiology and etiology of ICU-acquired bacteremia does not differ seriously in respect to nosocomial bacteremia among unselected populations, although it is associated with a greater incidence and overall mortality. Presence of shock is the most important modificable variable affecting the outcome.

Increased resistance to quinolone in Catalonia, Spain.

From 1989 to 1991, the level of resistance to ciprofloxacin in our hospital increased from 0.47% to 6.7% in opportunistic Enterobacteriaceae, from 9.9% to 16% in Pseudomonas aeruginosa and from 8.27% to 31.8% in Campylobacter jejuni-coli. We also observed an increase in quinolone consumption from 1.1 in 1989 to 1.5 defined daily doses per 1000 inhabitants per day in 1991.

Comparison of conventional ribotyping and PCR-RFLP ribotyping for the analysis of endemic strains of Neisseria meningitidis isolated in a local community over 7 years.

Conventional ribotyping was compared with the PCR amplification of the intergenic spacer region between 16S and 23S rRNA genes (PCR-RFLP ribotyping) when applied to the subtyping of sporadic Neisseria meningitidis strains. Thirty isolates out of a total of 36 meningococcal disease cases, reported as having occurred in a particular community over a 7-year endemic period, were analyzed by each of the methods. Only ribotyping with three restriction enzymes (EcoRI, ClaI and XhoI) gave acceptable discriminatory power for short-term epidemiological purposes. We conclude that conventional ribotyping is a suitable method for typing sporadic meningococcal strains and that it cannot be replaced by the more straightforward PCR-RFLP ribotyping method.

Comparison of IS1245 restriction fragment length polymorphism and pulsed-field gel electrophoresis for typing clinical isolates of Mycobacterium avium subsp avium.

SETTING: Little is still known about the epidemiology and pathogenesis of Mycobacterium avium subsp avium (MASA) infection. OBJECTIVE: Examination of the reproducibility and the stability over time of pulsed-field gel electrophoresis (PFGE) and IS1245 restriction fragment length polymorphism (IS1245-RFLP) techniques. The ability of these typing systems for differentiating clinical isolates of MASA was also assessed. DESIGN: Clinical isolates recovered from 63 patients (59 human immunodeficiency virus [HIV] positive and four HIV-negative) were studied by insertion sequence IS1245 and PFGE. For the study of in vivo and in vitro stability, strains collected over time from four patients and five strains chosen at random, respectively, were used. RESULTS: The stability of PFGE and IS1245-RFLP in vitro was excellent. PFGE was also stable in vivo, but IS1245-RFLP patterns showed some variation. The discriminatory power of IS1245-RFLP and PFGE was 0.995 and 0.989, respectively. The cluster analysis did not reveal differences between strains recovered from HIV-negative and HIV-positive patients or between patients with colonisation, local infection or disseminated disease. CONCLUSION: IS1245-RFLP and PFGE are useful tools for typing MASA strains. However, IS1245 variations in vivo may complicate the analysis of epidemiological relationships.