Outbreak of Pseudomonas fluorescens bloodstream infection in a coronary care unit.

An outbreak of Pseudomonas fluorescens infection in six patients in a coronary care unit was associated with a source not previously reported, namely the ice bath used for cardiac output determinations. Outbreaks of pseudobacteraemia caused by P. fluorescens and occasional blood transfusion-associated bloodstream infection (BSI) have been described. However, during the last two decades, two outbreaks of P. fluorescens BSI have been described and this article reports a third. Isolation of P. fluorescens in blood cultures must alert clinicians to the possibility of contamination of infusate, lock solutions or catheter flush.

Prevalence of acquired AmpC beta-lactamases in Enterobacteriaceae lacking inducible chromosomal ampC genes at a Spanish hospital from 1999 to 2007.

In 2007, a significant increase in acquired ampC genes in Enterobacteriaceae from 0.06% in 1999 to 1.3% was observed. Proteus mirabilis showed the highest prevalence (0.95%) and CMY-2 was the most prevalent AmpC enzyme (66.7%). Other enzymes such as CMY-4, DHA-1, ACC-1, and three new enzymes called CMY-25, CMY-27 and CMY-40 were detected. Seven out of the 117 isolates (6%) also produced an extended-spectrum beta-lactamase. As acquired AmpC enzymes are likely to become a serious public health issue worldwide, close surveillance is necessary to curb their spread.

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.

Increase in beta-lactam-resistant Proteus mirabilis strains due to CTX-M- and CMY-type as well as new VEB- and inhibitor-resistant TEM-type beta-lactamases.

OBJECTIVES: The aim of this study was to characterize the different inhibitor-resistant TEM beta-lactamases, extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC beta-lactamases implicated in beta-lactam resistance in Proteus mirabilis, which has increased over recent years. METHODS: From February 2000 to December 2005, 1423 clinical isolates of P. mirabilis were collected. The AmpC phenotype was checked by means of a double-disc synergy test using cloxacillin as an inhibitor of AmpC enzymes. The production of ESBL was assessed by the double-disc synergy method and by Etest ESBL. Analytical isoelectric focusing, determination of kinetic constants, conjugation, PCR and a sequencing strategy were used to characterize the enzymes. The possible relationships between isolates were analysed by PFGE. RESULTS AND CONCLUSIONS: Twenty-five of 1423 isolates were found to display intermediate or full resistance to co-amoxiclav, cefotaxime or ceftazidime. Seventeen isolates had reduced susceptibility to co-amoxiclav; of these, seven produced TEM-110, eight produced the new TEM-159, one the new TEM-160 and one TEM-1. Five isolates producing TEM-110, TEM-159 or TEM-160 enzymes shared the same PFGE profile. Three isolates produced an ESBL, CTX-M-1, CTX-M-32 and the new variant, VEB-4. Finally, five isolates with an AmpC phenotype produced CMY-2, two with the same PFGE profile. Our data emphasize the diversity of beta-lactamases found in this species.

Clinical and microbiological features of nocardiosis 1997-2003.

Nocardiosis has been believed to be caused by the members of the Nocardia asteroides complex and the Nocardia brasiliensis species. However, recent advances in genotypic identification have shown that the genus exhibits considerable taxonomic complexity and the phenotypic markers used in the past for its identification can be ambiguous. The aim of this study was to assess the species distribution of Nocardia isolates and to determine whether there are differences in pathogenicity or antimicrobial susceptibility between the different species identified. Nocardia isolates obtained over a 7 year period were retrospectively reviewed. The isolates were identified genotypically, their antibiotic susceptibility was tested and the clinical data of the 27 patients were retrieved. Eight different Nocardia species were identified: Nocardia farcinica (n=9), Nocardia abscessus (n=6), Nocardia cyriacigeorgica (n=6), Nocardia otitidiscaviarum (n=2), Nocardia nova (n=1), N. nova complex (n=1), Nocardia carnea (n=1) and Nocardia transvalensis complex (n=1). All species were susceptible to co-trimoxazole but different patterns of susceptibility to other agents were observed. All patients had active comorbidities at the time of infection. A total of 19 patients were immunosuppressed, due to human immunodeficiency virus infection, chronic corticosteroid therapy, immunosuppressive therapy or haematological malignancies. Six patients displayed a Charlson comorbidity index score above 4. Global mortality was 50 % while attributable mortality was 34.6 %. Patients infected with N. farcinica--the most resistant species--had the highest Charlson index score and the highest mortality rate. Accurate identification of the species and susceptibility testing of Nocardia isolates may play an important role in diagnosis and treatment.

Characterisation of fluoroquinolone-resistant clinical isolates of Streptococcus pyogenes in Barcelona, Spain.

Resistance mechanisms and clonal relationships were determined for six Streptococcus pyogenes isolates with low- or high-level ciprofloxacin resistance. Four isolates displayed reduced susceptibility to ciprofloxacin and levofloxacin and had alterations in ParC: Ser80-->Pro (isolate emm3.1); Ser79-->Ala (two isolates emm6.0); and a double substitution Ser79-->Phe and Ala121-->Val (isolate emm12.27). Two isolates (emm12.26) displayed high-level resistance to ciprofloxacin and levofloxacin, as well as to other quinolones. These isolates had the same double substitution in ParC as isolate emm12.27, and an additional substitution (Ser81-->Tyr) in GyrA. Resistance patterns, emm typing and sequencing of the quinolone resistance-determining regions defined two clusters containing three and two isolates, respectively.

CMY-2-producing Salmonella enterica, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis and Escherichia coli strains isolated in Spain (October 1999-December 2000).

CMY-2 plasmid-mediated AmpC beta-lactamase (CMY-2) was detected in 21 isolates from two hospitals located in different geographical regions of Spain between October 1999 and December 2000. The isolates comprised two Salmonella enterica serovars (Mikawasima and Montevideo), 16 Escherichia coli, one Klebsiella pneumoniae, one Klebsiella oxytoca and one Proteus mirabilis. In addition to the expected resistance to beta-lactams, including extended-spectrum cephalosporins and cefoxitin, all isolates showed a broad spectrum of associated resistance. All were resistant to sulfamethoxazole, chloramphenicol, tetracycline and streptomycin, and all but two were also resistant to gentamicin. Five isolates were studied in detail and all transferred CMY-2 and other resistance determinants by conjugation. Genomic DNA restriction pattern analysis of the E. coli isolates excluded the dissemination of a single clone. To the best of our knowledge this is the first time that CMY-2 has been detected in P. mirabilis, K. oxytoca and S. enterica serovars Mikawasima and Montevideo. It is also the first time that CMY-2 has been described in Spain.

Salmonella enterica serovar virchow with CTX-M-like beta-lactamase in Spain.

Four Salmonella enterica serovar Virchow strains resistant to broad-spectrum cephalosporins were isolated from patients with gastroenteritis in 1997 and 1998 in Murcia and Barcelona, Spain. The isolates expressed a beta-lactamase with a pI of about 8 and a positive PCR when specific primers for CTX-M-9 were used. These results suggest the presence of a CTX-M-9 beta-lactamase in these strains.

Antibiotic resistance trends in enteropathogenic bacteria isolated in 1985-1987 and 1995-1998 in Barcelona.

Trends in resistance to antimicrobial agents used for therapy have been evaluated with 3,797 enteropathogenic bacteria, Campylobacter, Salmonella, Shigella, and Yersinia, between 1985-1987 and 1995-1998. The greater increase in the rate of resistance was observed in Campylobacter jejuni for quinolones (from 1 to 82%) and tetracycline (from 23 to 72%) and in gastroenteric salmonellae for ampicillin (from 8 to 44%), chloramphenicol (from 1.7 to 26%), and trimethoprim-sulfamethoxazole and nalidixic acid (from less than 0.5 to 11%). Multidrug resistance was detected in several Salmonella serotypes. In the 1995-1998 period, 76% of Shigella strains were resistant to trimethoprim-sulfamethoxazole, 43% were resistant to ampicillin, and 39% were resistant to chloramphenicol. Seventy-two percent of Yersinia enterocolitica O3 strains were resistant to streptomycin, 45% were resistant to sulfonamides, 28% were resistant to trimethoprim-sulfamethoxazole, and 20% were resistant to chloramphenicol.

Escherichia coli serotype O15:K52:H1 as a uropathogenic clone.

To clarify the clinical and bacteriological correlates of urinary-tract infection (UTI) due to Escherichia coli O15:K52:H1, during a 1-year surveillance period we prospectively screened all 1, 871 significant E. coli urine isolates at the Hospital de la Santa Creu i Sant Pau, Barcelona, Spain, for this serotype and assessed the epidemiological features of community-acquired UTI due to E. coli O15:K52:H1 versus other E. coli serotypes. We also compared the 25 O15:K52:H1 UTI isolates from the present study with 22 O15:K52:H1 isolates from other, diverse geographic locales and with 23 standard control strains (8 strains from the ECOR reference collection and 15 strains of nonpathogenic O:K:H serotypes) with respect to multiple phenotypic and genotypic traits. Although E. coli O15:K52:H1 caused only 1.4% of community-acquired E. coli UTIs during the surveillance period, these UTIs were more likely to present as pyelonephritis and to occur in younger hosts, with similar risk factors, than were UTIs due to other E. coli serotypes. Irrespective of geographic origin, E. coli O15:K52:H1 strains exhibited a comparatively restricted repertoire of distinctive virulence factor profiles (typically, they were positive for papG allele II, papA allele F16, and aer and negative for sfa, afa, hly, and cnf1), biotypes, ribotypes, and amplotypes, consistent with a common clonal origin. In contrast, their antimicrobial resistance profiles were more extensive and more diverse than those of control strains. These findings indicate that E. coli O15:K52:H1 constitutes a broadly distributed and clinically significant uropathogenic clone with fluid antimicrobial resistance capabilities.

[Incidence of extended spectrum beta lactamase in Escherichia coli in a university hospital from 1994-1996]. / Incidencia de betalactamasas de espectro ampliado en Escherichia coli en un hospital universitario durante 1994-1996.

BACKGROUND: The aim of this study was to evaluate the incidence of TEM- and SHV-type extended spectrum betalactamases (ESBLs) in Escherichia coli in a 700-bed teaching hospital between 1994 and 1996. MATERIAL AND METHODS: Strains that presented reduced diameters to third-generation cephalosporins, as identified by disc diffusion techniques, were studied. The betalactamases involved were characterized by determination of the isoelectric point, hydrolysis profile, gene detection by polymerase chain reaction (PCR), and sequencing of the amplified products. RESULTS: 96 strains (1.4%) out of 7,054 strains of E. coli isolated between 1994 and 1996 showed decreased susceptibility to third-generation cephalosporins and only 4 strains (0.06%) produced ESBLs. Two strains produced SHV-2 and two produced TEM-12. CONCLUSIONS: The contribution of ESBL production to resistance to third-generation cephalosporins was low: 0.06% of all the E. coli strains isolated between 1994 and 1996.

Infections caused by Escherichia coli resistant to norfloxacin in hospitalized cirrhotic patients.

Selective intestinal decontamination with norfloxacin is useful to prevent bacterial infections in several groups of cirrhotic patients at high risk of infection. However, the emergence of infections caused by Escherichia coli resistant to quinolones has recently been observed in cirrhotic patients undergoing prophylactic norfloxacin. Our aim is to determine the characteristics of the infections caused by E. coli resistant to norfloxacin in hospitalized cirrhotic patients. One hundred and six infections caused by E. coli in 99 hospitalized cirrhotic patients were analyzed and distributed into two groups: group I (n = 67), infections caused by E. coli sensitive to norfloxacin, and group II (n = 39), infections caused by E. coli resistant to norfloxacin. The clinical and analytical characteristics at diagnosis of the infection were similar in both groups. Previous prophylaxis with norfloxacin was more frequent in group II (15/67, 22.4% vs. 32/39, 82%, P <.0001), as a result of a higher number of patients submitted to continuous long-term prophylaxis in this group, whereas previous short-term prophylaxis was similar in both groups. Infections were more frequently nosocomial-acquired in group II than in group I (17/67, 25.3% vs. 20/39, 51.2%, P =.01). The type of infections was similar in both groups: urinary tract infections 38 in group I and 24 in group II, spontaneous bacterial peritonitis 8 and 2, spontaneous bacteremia 4 and 4, and bacterascites 1 and 0, respectively (pNS). Mortality during hospitalization was similar in the two groups (4/67, 5.9% vs. 5/39, 12.8%, pNS). None of the E. coli resistant to norfloxacin were also resistant to cefotaxime and only one of them was resistant to amoxicillin-clavulanic acid. Prophylaxis with norfloxacin, usually continuous long-term prophylaxis, favors the development of infections caused by norfloxacin-resistant E. coli. Long-term antibiotic prophylaxis should therefore be restricted to highly selected groups of cirrhotic patients at high-risk of infection. Infections caused by E. coli resistant to norfloxacin show a severity similar to those caused by sensitive E. coli. No significant associated resistance between norfloxacin and the antibiotics most frequently used in the treatment of bacterial infections in cirrhotic patients has been observed.

[Haemophilus influenzae type b invasive disease in Catalonia (1996)]. / Enfermedad invasiva por Haemophilus influenzae tipo b en Cataluña (1996).

BACKGROUND: The purpose of this study was to find out the incidence and characteristics of H. influenzae type b invasive disease (HibID) in Catalonia, Spain. MATERIAL AND METHODS: An active surveillance of H. influenzae isolated from normally sterile sites was carried out during 1996. Microbiology laboratories of hospitals of Catalonia were periodically contacted by telephone. The serotype of all the strains was studied. RESULTS: The incidence of H. influenzae invasive disease (HIID) was 7.1 per 100,000 in children under 5 years and 1.0 per 100,000 in those over 5 years. The incidence of serotype b was 6.4 per 100,000 children under 5 years and 0.2 above this age. Only three strains belonged to types other than b (d, e and f). CONCLUSIONS: The incidence of HIbID is uncommon in Catalonia, lower than that registered in the prevaccine era in other countries and regions of the same geographical area.

Increase in quinolone resistance in a Haemophilus influenzae strain isolated from a patient with recurrent respiratory infections treated with ofloxacin.

The increase in the level of quinolone resistance of Haemophilus influenzae clinical isolates during ofloxacin therapy of a patient with recurrent respiratory infections was investigated. The first isolate (MIC of ciprofloxacin of 2 microg/ml) and the second isolate (MIC of 32 microg/ml) belonged to the same clone, as shown by pulsed-field gel electrophoresis, and the increase in the resistance level was associated with a substitution in Ser-84 to Arg in the ParC protein. These results emphasize the potential risk of development of quinolone-resistant H. influenzae during fluoroquinolone therapy in patients with recurrent respiratory infection.

[Haemophilus influenzae meningitis in adults: analysis of 12 cases]. / Meningitis por Haemophilus influenzae en adultos: análisis de 12 casos.

Haemophilus influenzae is an infrequent etiologic agent of bacterial meningitis in adult patients. In the last 12 years, it was the cause in 12 out of 238 cases (5.0%) of acute bacterial meningitis in adults. There were 5 men and 7 women with a mean age (SD) of 45.4 (16) years (range: 18-68 years). Seven patients (60%) had a communication between subarachnoid space and skin surface or mucosal cavities, and five (41.7%) had otitis or sinusitis. Most of the strains (9/12) were serotype b. Only one patient (8.3%) developed severe neurologic and extra-neurologic complications, and was the one who died. One of the survivors (9.1%) had partial deafness. H. influenzae is not a negligible cause of bacterial meningitis in adults. Moreover, its detection has been increasing in the last years. Patients with a cerebrospinal fluid leak, otitis or sinusitis are at high risk. The outcome is usually favorable if an early adequate therapy is given.

Emergence of clinical Escherichia coli isolates with decreased susceptibility to ceftazidime and synergic effect with co-amoxiclav due to SHV-1 hyperproduction.

Between 1994 and 1996 we detected 28 out of 7054 (0.4%) clinical isolates of Escherichia coli with abnormal or reduced inhibition diameters to co-amoxiclav and ceftazidime in a disc diffusion test. The increased MIC of ceftazidime (1-32 mg/L) and the effect of synergy between this antibiotic and co-amoxiclav according to the disc diffusion test suggest the presence of an extended-spectrum beta-lactamase. However, enzymatic characterization and the nucleotide sequence confirm the hyperproduction of the SHV-1 enzyme.

[Detection of pathogenicity factors in strains of classical enteropathogenic Escherichia coli]. / Detección de factores de patogenicidad en cepas de Escherichia coli enteropatógena clásica.

OBJECTIVE: To determine the number of strains of classic enteropathogenic E. coli (EPEC) that have the eae gene, that is considered a pathogenicity factor. MATERIAL AND METHODS: The presence of the eae gene has been evaluated on 62 EPEC strains of ten different serogroups, isolated from children with gastroenteritis. RESULTS: Amplification of the eae gene was positive in 10 out of 62 EPEC strains analyzed (16%) corresponding to seven different serogroups. DISCUSSION: The low frequency of the detection of the eae gene on EPEC strains shows the limited correlation between the pathogenicity and the serogroup of the strains and would corroborate the need to reexamine this subject prospectively in our country.