Clinical, microbiological, and molecular characterization of pediatric invasive infections by Streptococcus pyogenes in Spain in a context of global outbreak.

In December 2022, an alert was published in the UK and other European countries reporting an unusual increase in the incidence of Streptococcus pyogenes infections. Our aim was to describe the clinical, microbiological, and molecular characteristics of group A Streptococcus invasive infections (iGAS) in children prospectively recruited in Spain (September 2022-March 2023), and compare invasive strains with strains causing mild infections. One hundred thirty isolates of S. pyogenes causing infection (102 iGAS and 28 mild infections) were included in the microbiological study: emm typing, antimicrobial susceptibility testing, and sequencing for core genome multilocus sequence typing (cgMLST), resistome, and virulome analysis. Clinical data were available from 93 cases and 21 controls. Pneumonia was the most frequent clinical syndrome (41/93; 44.1%), followed by deep tissue abscesses (23/93; 24.7%), and osteoarticular infections (11/93; 11.8%). Forty-six of 93 cases (49.5%) required admission to the pediatric intensive care unit. iGAS isolates mainly belonged to emm1 and emm12; emm12 predominated in 2022 but was surpassed by emm1 in 2023. Spread of M1UK sublineage (28/64 M1 isolates) was communicated for the first time in Spain, but it did not replace the still predominant sublineage M1global (36/64). Furthermore, a difference in emm types compared with the mild cases was observed with predominance of emm1, but also important representativeness of emm12 and emm89 isolates. Pneumonia, the most frequent and severe iGAS diagnosed, was associated with the speA gene, while the ssa superantigen was associated with milder cases. iGAS isolates were mainly susceptible to antimicrobials. cgMLST showed five major clusters: ST28-ST1357/emm1, ST36-ST425/emm12, ST242/emm12.37, ST39/emm4, and ST101-ST1295/emm89 isolates. IMPORTANCE: Group A Streptococcus (GAS) is a common bacterial pathogen in the pediatric population. In the last months of 2022, an unusual increase in GAS infections was detected in various countries. Certain strains were overrepresented, although the cause of this raise is not clear. In Spain, a significant increase in mild and severe cases was also observed; this study evaluates the clinical characteristics and the strains involved in both scenarios. Our study showed that the increase in incidence did not correlate with an increase in resistance or with an emm types shift. However, there seemed to be a rise in severity, partly related to a greater rate of pneumonia cases. These findings suggest a general increase in iGAS that highlights the need for surveillance. The introduction of whole genome sequencing in the diagnosis and surveillance of iGAS may improve the understanding of antibiotic resistance, virulence, and clones, facilitating its control and personalized treatment.

Lack of Functional Trehalase Activity in Candida parapsilosis Increases Susceptibility to Itraconazole.

Central metabolic pathways may play a major role in the virulence of pathogenic fungi. Here, we have investigated the susceptibility of a Candida parapsilosis mutant deficient in trehalase activity (atc1Δ/ntc1Δ strain) to the azolic compounds fluconazole and itraconazole. A time-course exposure to itraconazole but not fluconazole induced a significant degree of cell killing in mutant cells compared to the parental strain. Flow cytometry determinations indicated that itraconazole was able to induce a marked production of endogenous ROS together with a simultaneous increase in membrane potential, these effects being irrelevant after fluconazole addition. Furthermore, only itraconazole induced a significant synthesis of endogenous trehalose. The recorded impaired capacity of mutant cells to produce structured biofilms was further increased in the presence of both azoles, with itraconazole being more effective than fluconazole. Our results in the opportunistic pathogen yeast C. parapsilosis reinforce the study of trehalose metabolism as an attractive therapeutic target and allow extending the hypothesis that the generation of internal oxidative stress may be a component of the antifungal action exerted by the compounds currently available in medical practice.

The Fungicidal Action of Micafungin is Independent on Both Oxidative Stress Generation and HOG Pathway Signaling in Candida albicans.

In fungi, the Mitogen-Activated Protein kinase (MAPK) pathways sense a wide variety of environmental stimuli, leading to cell adaptation and survival. The HOG pathway plays an essential role in the pathobiology of Candida albicans, including the colonization of the gastrointestinal tract in a mouse model, virulence, and response to stress. Here, we examined the role of Hog1 in the C. albicans response to the clinically relevant antifungal Micafungin (MF), whose minimum inhibitory concentration (MIC) was identical in the parental strain (RM100) and in the isogenic homozygous mutant hog1 (0.016 mg/L). The cell viability was impaired without significant differences between the parental strain, the isogenic hog1 mutant, and the Hog1+ reintegrant. This phenotype was quite similar in a collection of hog1 mutants constructed in a different C. albicans background. MF-treated cells failed to induce a relevant increase of both reactive oxygen species (ROS) formation and activation of the mitochondrial membrane potential in parental and hog1 cells. MF was also unable to trigger any significant activation of the genes coding for the antioxidant activities catalase (CAT1) and superoxide dismutase (SOD2), as well as on the corresponding enzymatic activities, whereas a clear induction was observed in the presence of Amphotericin B (AMB), introduced as a positive control of Hog1 signaling. Furthermore, Hog1 was not phosphorylated by the addition of MF, but, notably, this echinocandin caused Mkc1 phosphorylation. Our results strongly suggest that the toxic effect of MF on C. albicans cells is not mediated by the Hog1 MAPK and is independent of the generation of an internal oxidative stress in C. albicans.

Sensitivity of the Candida albicans trehalose-deficient mutants tps1Δ and tps2Δ to amphotericin B and micafungin.

Purpose. Fungal infections have increased in recent decades, with Candida albicans being the fourth most common aetiological agent of nosocomial infections. Disaccharide trehalose has been proposed as a target for the development of new antifungals. In C. albicans we have examined the susceptibility shown by two mutants deficient in trehalose biosynthesis, namely tps1Δ and tps2Δ, to amphotericin B (AmB) and micafungin (MF).Methodology. Minimum inhibitory concentrations (MICs) were calculated according to the Clinical and Laboratory Standards Institute (CLSI) criteria. Cell viability was assessed by cell counting. Intracellular reactive oxygen species (ROS) and the mitochondrial membrane potential were measured by flow cytometry, while the trehalose content and biofilm formation were determined by enzymatic assays.Results. While the tps1Δ mutant was highly sensitive to AmB exposure, its resistance to MF was similar to that of the wild-type. Notably, the opposite phenotype was recorded in the tps2Δ mutant. In turn, MF induced a significant level of endogenous ROS production in the parental SC5314 and tps2Δ cells, whereas the ROS formation in tps1Δ cells was virtually undetectable. The level of endogenous ROS correlated positively with the rise in mitochondrial activity. Only AmB was able to promote intracellular synthesis of trehalose in the parental strain; it was absent from tps1Δ cells and showed low levels in tps2Δ, confirming the unspecific dephosphorylation of trehalose-6P in C. albicans. Furthermore, the capacity of both tps1Δ and tps2Δ mutants to form biofilms was drastically reduced after AmB exposure, whereas it increased in tps1Δ cells treated with MF.Conclusion. Our data lend weight to the idea of using trehalose biosynthesis as a potential target for antifungal therapy.

Infecciones nosocomiales por Pseudomonas aeruginosa multiresistente incluido carbapenémicos: factores predictivos y pronósticos. Estudio prospectivo 2016-2017 / Nosocomial infections caused by multiresistant Pseudomonas aeruginosa (carbapenems included): predictive and prognostic factors. A prospective study (2016-2017)

Introducción. Pseudomonas aeruginosa es uno de los principales microorganismos causante de infecciones nosocomiales. En los últimos años están aumentando las resistencias a antimicrobianos y las infecciones producidas por estas cepas se han asociado a un aumento de la mortalidad. El objetivo de este trabajo es valorar su significación clínica y analizar los valores predictivos y pronósticos. Métodos. Estudio prospectivo de casos y controles no emparejados realizado en 64 pacientes diagnosticados de infección nosocomial por P. aeruginosa, 32 de ellos por cepas sensibles y 32 por cepas multiresistentes incluido los carbapenémicos (MDR/XDR-C), ingresados en un hospital de tercer nivel. Se realizó un seguimiento hospitalario hasta el alta o fallecimiento y un control a los 30 días. Se analizaron variables clínico-epidemiológicas y microbiológicas. Resultados. La incidencia de cepas MDR/XDR-C fue de 2,3 por 1000 ingresos. Diez de las cuales fueron productoras de metalo-β-lactamasa tipo VIM. Los factores predictivos asociados de forma independiente con MDR/XDR-C fueron: la estancia previa en UCI o Reanimación (OR 14,01; IC95% 2,105-93,297), la aparición tras >20 días de estancia (OR 29,826; IC 95% 4,783-185,997) y la leucocitosis (OR 10,0190; IC95% 1,842-56,369). En cambio, no se detectaron diferencias estadísticamente significativas en cuanto a los factores de gravedad clínica y la mortalidad en ambos grupos. Conclusión. Los principales factores de riesgo asociados a infecciones por cepas MDR/XDR-C fueron la estancia previa en UCI o Reanimación, la aparición tras >20 días y la leucocitosis. La infección por cepas MDR/XDR-C no se asocia a un aumento de la mortalidad

Introduction. Pseudomonas aeruginosa is one of the major pathogens causing hospital-acquired infections. In recent years, antimicrobial resistance is increasing and multidrug resistant (MDR) and extremely drug resistant (XDR) isolates have been associated with an increase of mortality. The aim of this study is to assess the clinical significance and analyze predictors and prognostic factors. Methods. Prospective case-control non-paired study involving 64 patients with P. aeruginosa nosocomial infection, 32 caused by susceptible P. aeruginosa and 32 by MDR/XDR including to carbapenems (XDR-C) strains, admitted at a third level hospital. The follow-up period was till hospital discharge or death and at 30 days after discharge. For all patients, clinical epidemiology and microbiological data were analyzed. Results: The incidence of MDR/XDR-C strains was 2.3 per 1000 admissions. Ten of which were VIM metallo-β-lactamase-producing. Independent predictor factors associated with MDR/XDR-C infections were: previous ICU or Resuscitation unit admission (OR 14.01; IC 95% 2.105-93.297) appearance >20 days after admission (OR 29.826; IC 95% 4.783-185.997) and leukocytosis (OR 10.0190; IC 95% 1.842-56.369). However, there were not statistically significant differences in clinical severity or mortality between both groups. Conclusion: the major risk factors associated with MDR/XDR-C infections were previous ICU or Resuscitation unit admission, appearance >20 days after admission and leukocytosis. MDR/XDR-C infections were not associated to increased mortality

Emergence of cfr-Mediated Linezolid Resistance in a Methicillin-Resistant Staphylococcus aureus Epidemic Clone Isolated from Patients with Cystic Fibrosis.

Resistance to linezolid (LZD) in methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients with cystic fibrosis (CF) is due mainly to ribosomal mutations. We report on four CF patients with LZD-resistant MRSA bronchopulmonary infections by strains carrying the cfr gene. Strains from one patient also harbored the G2576U mutation (23S rRNA) and the G139R substitution (L3 protein). All strains belonged to the epidemic clone ST125 MRSA IVc. Our results support the monitoring of LZD resistance emergence in CF and non-CF MRSA isolates.

[Study of a cohort of patients with Enterococcus spp. Bacteraemia. Risk factors associated to high-level resistance to aminoglycosides]. / Estudio de una cohorte de pacientes con bacteriemias por Enterococcus spp. Factores de riesgo para resistencia de alto nivel a aminoglicósidos.

OBJECTIVES: To analyze a cohort of patients with Enterococcus sp. bacteraemia. PATIENTS AND METHODS: Retrospective and observational study of a cohort of non-pediatric in-patients with Enterococcus spp. bacteraemia (June 2007-September 2009). Data collection from clinical records was done according to a standard protocol. We analyzed epidemiological, clinical and microbiological data. Treatment with glycopeptides in non allergic patients or in case of betalactam susceptibility (ampicillin) was considered "optimizable". RESULTS: Three were 106 cases of bacteraemia (2.2/1000 admitted patients; 84% E. faecalis); 83% had an underlying condition; 88% nosocomial or health related cases. Urinary infection was present in 20% and primary bacteraemia in 47%. High level resistance to gentamicin (HLRG) was present in 60%; there was no vancomycin or linezolid resistance. Most frequent empiric treatments were penicillin-betalactamase inhibitor (25%) and glycopeptides (22%). Most frequent definitive treatment was glycopeptides (34%), being "optimized" 21% and 44% of empiric and definitive treatments, respectively. Mortality was 23% (related, 14%). In the multivariate analysis, risk factors associated with HLRG were nosocomial acquired infection (OR 6.083; 95CI% 1.428-25.915) and no-abdominal origin (OR 6.006; 95CI%1.398-25.805). In multivariate analysis, independent risk factors for mortality were: Pitt > 3 (OR 14.405; 95CI%2.236-92.808) and active empiric treatment (OR 8.849; 95CI% 1.101-71.429). Incidence in previous cohort was similar but HLRG rate has increased. CONCLUSIONS: Risk factors associated with HLRG were nosocomial acquired infection and no-abdominal origin. Risk factors for mortality were initial clinical severity and having received active empiric treatment. HLRG rate has increased.

Estudio de una cohorte de pacientes con bacteriemias por Enterococcus spp. Factores de riesgo para resistencia de alto nivel a aminoglicósidos / Study of a cohort of patients with Enterococcus spp. Bacteraemia. Risk factors associated to high-level resistance to aminoglycosides

Objetivos. Analizar una cohorte de pacientes con bacteriemia enterocócica. Pacientes y métodos. Estudio retrospectivo-observacional de adultos ingresados con aislamiento de Enterococcus spp en hemocultivos (Junio 2007-Septiembre 2009). Se revisaron las historias clínicas siguiendo un protocolo que consideraba variables epidemiológicas, clínicas y microbiológicas. El tratamiento con glicopéptidos en pacientes no alérgicos o cepas sin resistencia a ampicilina se consideró “optimizable”. Resultados. Se detectaron 106 episodios (2/1000 pacientes ingresados; 84% E. faecalis); 83% presentaban comorbilidad de base; 88% de adquisición nosocomial/asociada a cuidados sanitarios. El foco fue urinario en 20% y desconocido en 47%; 60% tenían resistencia de alto nivel a gentamicina (RANG); no hubo cepas resistentes a vancomicina o linezolid. Los tratamientos empíricos más usados fueron penicilina-inhibidor de betalactamasas (25%) y glicopéptidos (22%). En el tratamiento definitivo el antibiótico más usado fue glicopéptido (34%), considerándose “optimizado” 21% de tratamientos empíricos y 44% de definitivos. La mortalidad global fue 23% (relacionada 14%). Se asociaron a RANG: adquisición nosocomial (OR 6,083; IC95% 1,428-25,915) y no tener foco abdominal (OR 6,006; IC95%1,398-25.805). Se asociaron a mayor mortalidad la gravedad clínica inicial (Pitt > 3) (OR 14,405; IC95%2,236-92,808) y haber recibido un tratamiento empírico activo (OR 8,849; IC95% 1,101-71,429). La incidencia de la serie histórica fue similar y aumentó el porcentaje de RANG en la cohorte más reciente. Conclusiones. La adquisición nosocomial y el no tener foco abdominal se asociaron a RANG; la gravedad clínica inicial y recibir tratamiento empírico activo (que no optimizado) se asociaron a mayor mortalidad. Apreciamos un aumento en el porcentaje de RANG (AU)

Objectives. To analyze a cohort of patients with Enterococcus sp. bacteraemia. Patients and methods. Retrospective and observational study of a cohort of non-pediatric in-patients with Enterococcus spp. bacteraemia (June 2007-September 2009). Data collection from clinical records was done according to a standard protocol. We analyzed epidemiological, clinical and microbiological data. Treatment with glycopeptides in non allergic patients or in case of betalactam susceptibility (ampicillin) was considered “optimizable”. Results. Three were 106 cases of bacteraemia (2.2/1000 admitted patients; 84% E. faecalis); 83% had an underlying condition; 88% nosocomial or health related cases. Urinary infection was present in 20% and primary bacteraemia in 47%. High level resistance to gentamicin (HLRG) was present in 60%; there was no vancomycin or linezolid resistance. Most frequent empiric treatments were penicillin-betalactamase inhibitor (25%) and glycopeptides (22%). Most frequent definitive treatment was glycopeptides (34%), being “optimized” 21% and 44% of empiric and definitive treatments, respectively. Mortality was 23% (related, 14%). In the multivariate analysis, risk factors associated with HLRG were nosocomial acquired infection (OR 6.083; 95CI% 1.428-25.915) and no-abdominal origin (OR 6.006; 95CI%1.398-25.805). In multivariate analysis, independent risk factors for mortality were: Pitt > 3 (OR 14.405; 95CI%2.236-92.808) and active empiric treatment (OR 8.849; 95CI% 1.101-71.429).). Incidence in previous cohort was similar but HLRG rate has increased. Conclusions. Risk factors associated with HLRG were nosocomial acquired infection and no-abdominal origin. Risk factors for mortality were initial clinical severity and having received active empiric treatment. HLRG rate has increased (AU)

Infecciones por Acinetobacter baumannnii multirresistente y resistente a carbapenem: factores asociados a la mortalidad / Multidrug and carbapenem-resistant Acinetobacter baumannii infections: Factors associated with mortality

Background and objective: To analyse factors related to mortality and influence of antibiotic treatment on outcome in patients with nosocomial infection due to multidrug and carbapenem-resistant Acinetobacter baumannii (MDR-C AB). Patients and methods: Observational and prospective study of a cohort of adult patients with MDR-C AB infection. Data collection from clinical records was done according to a standard protocol (January 2007 through June 2008). Patients with MDR-C AB infection were identified by review of results of microbiology cultures from the hospital microbiology laboratory. Epidemiological and clinical variables and predictors of mortality were analysed. Results: 24 out of 101 cases were considered colonizations and 77 infections (27 bacteraemia); global mortality in infected patients was 49% (18 cases with bacteraemia and 20 with no bacteraemia). In the multivariate analysis, including the 77 cases of infection, the prognosis factors associated with mortality were age (OR 1.09; 95% CI 1.02-1.2), McCabe 1 (OR 33.98; 95% CI 4.33-266.85), bacteraemia (OR 9.89; 95% CI 1.13-86.13), inadequate empiric treatment (OR 16.7; 95% CI 2.15-129.79), and inadequate definitive treatment (OR 26.29; 95% CI 1.45-478.19). In the multivariate analysis including the 57 cases of infection with adequate definitive treatment, the prognosis factors associated with mortality were McCabe 1 (OR 24.08; 95% CI 3.67-157.96) and monotherapy versus combined treatment (OR 7.11; 95% CI 1.63-30.99). Conclusions: Our cohort of patients with MDR-C AB infection is characterised by a very high mortality (49%); the severity of patients and inadequate treatment or monotherapy are statistically associated with mortality (AU)

Fundamento y objetivo: Analizar los factores asociados a mortalidad y la influencia del tratamiento antibiótico en la evolución de los pacientes con infección por Acinetobacter baumannii multiresistente y resistente a carbapenem (ABMDR-C). Pacientes y métodos: Estudio observacional y prospectivo de una cohorte de pacientes adultos con infección por ABMDR-C (Enero 2007 a junio 2008). Los casos de infección se identificaron a partir de los resultados de los cultivos microbiológicos. Analizamos las variables epidemiológicas y clínicas asociadas a mortalidad. Resultados: Veinticuatro de los 101 aislamientos de ABMDR-C se consideraron colonizaciones y 77 fueron definidos como infecciones (27 bacteriemias); la mortalidad global de los casos de infección fue del 49% (18 pacientes con bacteriemia y 20 sin bacteriemia). En el análisis multivariado que incluía a los 77 pacientes con infección, los factores predictores de mortalidad fueron: edad (OR 1,09; IC95% 1,02-1,2), McCabe 1 (OR 33,98; IC95% 4,33-266,85), bacteriemia (OR 9,89; IC95% 1,13-86,13), tratamiento empírico inadecuado (OR 16,7; IC95% 2,15-129,79), tratamiento definitivo inadecuado(OR 26,29; IC95% 1,45-478,19). En el análisis multivariado que incluía los 57 pacientes que recibieron tratamiento definitivo adecuado, los factores predictores de mortalidad fueron McCabe 1 (OR 24,08; IC95% 3,67-157,96) y tratamiento en monoterapia versus combinaciones (OR 7,11; IC95% 1,63-30,99). Conclusiones: En nuestra cohorte de pacientes con infección por ABMDR-C la mortalidad es elevada (49%); los pacientes graves y el tratamiento antibiótico inadecuado o el tratamiento definitivo adecuado en monoterapia se asocian de forma estadísticamente significativa a mayor mortalidad (AU)

Epidemiology, species distribution and in vitro antifungal susceptibility of fungaemia in a Spanish multicentre prospective survey.

OBJECTIVES: To update the knowledge of the epidemiology of fungaemia episodes in Spain, the species implicated and their in vitro antifungal susceptibilities. METHODS: Episodes were identified prospectively over 13 months at 44 hospitals. Molecular methods were used to determine the cryptic species inside the Candida parapsilosis and Candida glabrata complexes. Susceptibility to amphotericin B, anidulafungin, caspofungin, fluconazole, flucytosine, itraconazole, micafungin, posaconazole and voriconazole was determined by a microdilution colorimetric method. New species-specific clinical breakpoints (SSCBPs) for echinocandins, fluconazole and voriconazole were applied. RESULTS: The incidence of the 1357 fungaemia episodes evaluated was 0.92 per 1000 admissions. The incidence of Candida albicans fungaemia was the highest (0.41 episodes/1000 admissions), followed by Candida parapsilosis sensu stricto (0.22). Candida orthopsilosis was the fifth cause of fungaemia (0.02), outnumbered by Candida glabrata and Candida tropicalis. Interestingly, the incidence of fungaemia by C. parapsilosis was 11 and 74 times higher than that by C. orthopsilosis and Candida metapsilosis, respectively. Neither Candida nivariensis nor Candida bracarensis was isolated. Fungaemia was more common in non-intensive care unit settings (65.2%) and among elderly patients (46.4%), mixed fungaemia being incidental (1.5%). Overall susceptibility rates were 77.6% for itraconazole, 91.9% for fluconazole and 96.5%-99.8% for the other agents. Important resistance rates were only observed in C. glabrata for itraconazole (24.1%) and posaconazole (14.5%), and in Candida krusei for itraconazole (81.5%). CONCLUSIONS: Fungaemia is more common in non-critical patients. C. albicans is the most common species, followed by C. parapsilosis and C. glabrata. Nearly 90% of yeasts are susceptible to all antifungal agents tested. Resistance rates change moderately when applying the new SSCBPs.

Multidrug and carbapenem-resistant Acinetobacter baumannii infections: Factors associated with mortality.

BACKGROUND AND OBJECTIVE: To analyse factors related to mortality and influence of antibiotic treatment on outcome in patients with nosocomial infection due to multidrug and carbapenem-resistant Acinetobacter baumannii (MDR-C AB). PATIENTS AND METHODS: Observational and prospective study of a cohort of adult patients with MDR-C AB infection. Data collection from clinical records was done according to a standard protocol (January 2007 through June 2008). Patients with MDR-C AB infection were identified by review of results of microbiology cultures from the hospital microbiology laboratory. Epidemiological and clinical variables and predictors of mortality were analysed. RESULTS: 24 out of 101 cases were considered colonizations and 77 infections (27 bacteraemia); global mortality in infected patients was 49% (18 cases with bacteraemia and 20 with no bacteraemia). In the multivariate analysis, including the 77 cases of infection, the prognosis factors associated with mortality were age (OR 1.09; 95% CI 1.02-1.2), McCabe 1 (OR 33.98; 95% CI 4.33-266.85), bacteraemia (OR 9.89; 95% CI 1.13-86.13), inadequate empiric treatment (OR 16.7; 95% CI 2.15-129.79), and inadequate definitive treatment (OR 26.29; 95% CI 1.45-478.19). In the multivariate analysis including the 57 cases of infection with adequate definitive treatment, the prognosis factors associated with mortality were McCabe 1 (OR 24.08; 95% CI 3.67-157.96) and monotherapy versus combined treatment (OR 7.11; 95% CI 1.63-30.99). CONCLUSIONS: Our cohort of patients with MDR-C AB infection is characterised by a very high mortality (49%); the severity of patients and inadequate treatment or monotherapy are statistically associated with mortality.

Prospective multicenter study of the epidemiology, molecular identification, and antifungal susceptibility of Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis isolated from patients with candidemia.

A 13-month prospective multicenter study including 44 hospitals was carried out to evaluate the epidemiology of Candida parapsilosis complex candidemia in Spain. Susceptibility to amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, caspofungin, and micafungin was tested by the microdilution colorimetric method. A total of 364 C. parapsilosis complex isolates were identified by molecular methods: C. parapsilosis (90.7%), Candida orthopsilosis (8.2%), and Candida metapsilosis (1.1%). Most candidemias (C. parapsilosis, 76.4%; C. orthopsilosis, 70.0%; C. metapsilosis, 100%) were observed in adults. No C. orthopsilosis or C. metapsilosis candidemias occurred in neonates. C. parapsilosis was most frequent in adult intensive care unit (28.8%), surgery (20.9%), and internal medicine (19.7%) departments; and C. orthopsilosis was most frequent in hematology (28.6%), pediatrics (12.0%), and neonatology (11.5%) departments. The geographic distribution of C. orthopsilosis and C. metapsilosis was not uniform. According to CLSI clinical breakpoints, all C. orthopsilosis and C. metapsilosis isolates were susceptible to the nine agents tested. Resistance (MICs > 1 mg/liter) was observed only in C. parapsilosis: amphotericin B, posaconazole, itraconazole, and caspofungin (0.3% each), anidulafungin (1.9%), and micafungin (2.5%). Applying the new species-specific fluconazole and echinocandin breakpoints, the rates of resistance to fluconazole for C. parapsilosis and C. orthopsilosis increased to 4.8% and 0.3%, respectively; conversely, for C. parapsilosis they shifted from 1.9 to 0.6% (anidulafungin) and from 2.5 to 0.6% (micafungin). Our study confirms the different prevalence of C. parapsilosis complex candidemia among age groups: neither C. orthopsilosis nor C. metapsilosis was isolated from neonates; interestingly, C. metapsilosis was isolated only from adults and the elderly. The disparity in antifungal susceptibility among species could be important for therapy.

[Bacteraemia due to Escherichia coli producing extended-spectrum beta-lactamases (ESBL): clinical relevance and today's insights]. / Bacteriemias por Escherichia coli productor de betalactamasas de espectro extendido (BLEE): significación clínica y perspectivas actuales.

Antibiotic resistance is an old problem with new face as the rate of infections due to multidrug resistant bacteria is higher everyday and the number of new antibiotics to overwhelm the problem is becoming smaller. E. coli is the most frequent agent causing nosocomial or community-acquired bacteraemia being in our country 10% of them extended-spectrum beta-lactamases (ESBL) producing E. coli isolates. Nowadays the number of community- acquired or health-related infections caused by these ESBL producing E. coli is increasing. CTX-M has also become the most frequent ESBL compared to other enzymes. The role of these enzymes as a virulence factor increasing mortality in patients with bacteraemia due to E. coli is not well defined. The relevance of ESBL-E. coli seems to be related with the higher frequency of inadequate treatment and therefore the importance of identifying factors or features that might predict that the patient's infection is due to one of these isolates. In terms of prevention and control of infection measures, the role of patient's isolation is not clear but a proper prescription of antibiotics and antibiotic control policies are probably important to reduce the problem.

Bacteriemias por Escherichia coli productor de betalactamasas de espectro extendido (BLEE): significación clínica y perspectivas actuales / Bacteraemia due to Escherichia coli producing extended-spectrum beta-lactamases (ESBL): clinical relevance and today’s’ insights

La resistencia bacteriana es un problema antiguo pero de gran actualidad, ya que en un marco de “austeridad” en cuanto al número de nuevas moléculas de antibiótico disponibles en el mercado, la presencia de microorganismos multirresistentes es cada vez más frecuente. E. coli es el microorganismo más frecuentemente implicado en bacteriemias nosocomiales y comunitarias, y el aislamiento de cepas productoras de BLEE se sitúa en torno al 10% en nuestro país. Las infecciones por E. coli con BLEE han experimentado importantes cambios epidemiológicos en los últimos tiempos y actualmente la atención se centra en el aumento de infecciones y colonizaciones en pacientes procedentes de la comunidad, sobre todo en relación con instituciones sanitarias, y la mayor incidencia de las CTX-M frente a otros tipos de BLEE. El papel de estas enzimas como factor de virulencia que aumente por sí mismo la mortalidad en los pacientes con bacteriemia por E. coli no queda claro. La principal repercusión clínica de las BLEE parece ser la mayor frecuencia con la que estos pacientes con infecciones graves reciben un tratamiento empírico inadecuado, de ahí la importancia de identificar qué factores predicen la presencia de una cepa con BLEE para poder ofrecer un tratamiento adecuado lo antes posible. En cuanto a las medidas de control de la diseminación de BLEE, la eficacia del aislamiento de contacto y la actuación frente a pacientes colonizados por E. coli con BLEE no están claras, pero es incuestionable la necesidad de implementar un uso correcto y responsable de los antibióticos para evitar la expansión de cepas resistentes(AU)

Antibiotic resistance is an old problem with new face as the rate of infections due to multidrug resistant bacteria is higher everyday and the number of new antibiotics to overwhelm the problem is becoming smaller. E. coli is the most frequent agent causing nosocomial or community-acquired bacteraemia being in our country 10% of them extended-spectrum betalactamases (ESBL) producing E. coli isolates. Nowadays the number of community- acquired or health-related infections caused by these ESBL producing E. coli is increasing. CTX-M has also become the most frequent ESBL compared to other enzymes. The role of these enzymes as a virulence factor increasing mortality in patients with bacteraemia due to E. coli is not well defined. The relevance of ESBL- E. coli seems to be related with the higher frequency of inadequate treatment and therefore the importance of identifying factors or features that might predict that the patient’s infection is due to one of these isolates. In terms of prevention and control of infection measures, the role of patient’s isolation is not clear but a proper prescription of antibiotics and antibiotic control policies are probably important to reduce the problem(AU)

Colonización/infección por Acinetobacter baumannii multirresistente y resistente a carbapenémicos: epidemiología y factores predictivos de infección / Carbapenem and multidrug-resistant Acinetobacter baumannii colonisation/infection: Epidemiology and factors associated with infection

Fundamento y objetivo: Estudio de un brote de colonización/infección nosocomial por Acinetobacter baumannii multirresistente y resistente a carbapenémicos (ABMDR-C).Pacientes y método: Estudio prospectivo de pacientes con aislamiento de ABMDR-C entre enero de 2007 y junio de 2008. Se analizaron las características epidemiológicas y clínicas de los pacientes y los factores predictivos de infección frente a colonización.Resultados: De 101 casos, 24 presentaban colonización y 77 presentaban infección (27 bacteriemia); la mortalidad global (colonizaciones e infecciones) fue del 42% (4 colonizaciones y 38 infecciones [18 bacteriemias]). La incidencia de colonización/infección fue de 3,2/1.000 ingresos/día; un 29% tenía antecedente de ingreso previo y un 79% había recibido antibióticos previos (carbapenémicos [29%], piperacilina-tazobactam [34%] y ambos [12,5%]). El 78% tenía enfermedades de base, un 81% ingresó en la unidad de cuidados intensivos/reanimación, un 90% tuvo algún tipo de manipulación y un 65% presentaba coinfección por otros microorganismos. En el análisis multivariante fueron factores predictivos de infección frente a colonización: aislamiento de ABMDR-C en las muestras respiratorias (odds ratio [OR]: 5,406; intervalo de confianza [IC] del 95%: 1,419–20,599); sexo masculino (OR: 8,842; IC del 95%: 1,988–39,325); ingreso previo (OR: 9,720; IC del 95%: 1,383–68,291) y gravedad clínica inicial (OR: 30,897; IC del 95%: 5,533–172,543).Conclusiones: Nuestra cohorte de pacientes con olonizaciones/infecciones por ABMDR-C está constituida por pacientes con enfermedad de base crónica, a los que se les realizó distintas manipulaciones instrumentales, con ingresos hospitalarios previos y uso previo de betalactámicos de amplio espectro, especialmente carbapenémicos. Los pacientes tienen una alta gravedad clínica al comienzo y el aislamiento respiratorio es un importante factor predictor de infección frente a colonización (AU)

Background and objective: To study an outbreak of nosocomial colonisation/infection due to multidrug and carbapenem resistant A. baumannii (ABMDR-C). Patients and methods: Prospective study of patients with ABMDR-C colonisation/infection (January 2007–June 2008). Epidemiological and clinical variables and predictors of infection versus colonization were analysed. Results: 24 out of 101 cases were considered colonisations and 77 infections (27 bacteraemia); global mortality (colonisations and infections) was 42% (4 colonisations and 38 infections -18 bacteraemia). All together, the incidence was 3.2/1000 admissions/day; 29% had been previously admitted and 79% had received previous antibiotic treatment (29% carbapenem; 34% piperacillin-tazobactam; 12.5% boths); 78% had an underlying condition; 81% were UCI patients; 90% had gone through invasive procedures; 65% had another microorganism isolated. In multivariate analysis, infection predictor factors were isolation of ABMDR-C in respiratory samples (OR 5.406; 95% CI 1.419–20.599); male patients (OR 8.842; 95% CI 1.988–39.325); previous hospitalization (OR 9.720; 95% CI 1.383–68.291) and initial clinical severity (OR 30.897; 95% CI 5.533–172.543). Conclusions: Our cohort of patients with ABMDR-C colonisation/infection is characterised by their underlying comorbidity, the high rate of previous invasive procedures, previous hospitalisation and previous broad-spectrum betalactam treatments (especially carbapenem). Initial severity and respiratory samples with ABMDR-C isolates were predictors of infection versus colonisation (AU)

[Carbapenem and multidrug-resistant Acinetobacter baumannii colonisation/infection: epidemiology and factors associated with infection]. / Colonización/infección por Acinetobacter baumannii multirresistente y resistente a carbapenémicos: epidemiología y factores predictivos de infección.

BACKGROUND AND OBJECTIVE: To study an outbreak of nosocomial colonisation/infection due to multidrug and carbapenem resistant A. baumannii (ABMDR-C). PATIENTS AND METHODS: Prospective study of patients with ABMDR-C colonisation/infection (January 2007-June 2008). Epidemiological and clinical variables and predictors of infection versus colonization were analysed. RESULTS: 24 out of 101 cases were considered colonisations and 77 infections (27 bacteraemia); global mortality (colonisations and infections) was 42% (4 colonisations and 38 infections -18 bacteraemia). All together, the incidence was 3.2/1000 admissions/day; 29% had been previously admitted and 79% had received previous antibiotic treatment (29% carbapenem; 34% piperacillin-tazobactam; 12.5% both); 78% had an underlying condition; 81% were UCI patients; 90% had gone through invasive procedures; 65% had another microorganism isolated. In multivariate analysis, infection predictor factors were isolation of ABMDR-C in respiratory samples (OR 5.406; 95% CI 1.419-20.599); male patients (OR 8.842; 95% CI 1.988-39.325); previous hospitalization (OR 9.720; 95% CI 1.383-68.291) and initial clinical severity (OR 30.897; 95% CI 5.533-172.543). CONCLUSIONS: Our cohort of patients with ABMDR-C colonisation/infection is characterised by their underlying comorbidity, the high rate of previous invasive procedures, previous hospitalisation and previous broad-spectrum betalactam treatments (especially carbapenem). Initial severity and respiratory samples with ABMDR-C isolates were predictors of infection versus colonisation.

A study of air microbe levels in different areas of a hospital.

Airborne transmission is an important route for many microbial pathogens in outdoor and indoor environments, including hospitals. A 2-year-long survey of bioaerosol quality in operating theatres (OT), hospital rooms (HR) and maternity wards (MW) at a hospital in Murcia, Spain, was performed. Total aerobic counts (TAC) and fungal load (FL) were assessed using a microbiological air sampler (MAS-100 single-stage impactor). While fungal levels were below 1 cfu/m(3) (0-7.33 cfu/m(3)) in OT, they were higher in MW (mean, 6.9 cfu/m(3); range 0.44-44.67 cfu/m(3)) and in HR (mean, 10.6 cfu/m(3); range, 0-266 cfu/m(3)). In OT the aerobic counts were considerably higher, with a mean of 25.6 cfu/m(3) (range, 1.67-157 cfu/m(3)). MW and HR also showed higher means for total aerobic counts compared to OT. Seasonal changes were not detected in mould and bacteria levels in OT. Hospital renovation occurred during this study and OT adjacent to renovated areas were closed. A survey of TAC and FL in OT resumed when renovation was completed. We observed an outstanding increase in FL (more than 100 cfu/m(3)), particularly Aspergillus spp., during this period, but no significant changes in TAC were observed after renovation.

Characterization of acyl-phosphatidylinositol from the opportunistic pathogen Corynebacterium amycolatum.

The aim of the present study was to characterize a new lipid detected in the opportunistic pathogen Corynebacterium amycolatum. It was identified as acyl-phosphatidylinositol (acyl-PI), and revealed as a mixture of homologues compounds by electrospray ionization mass spectrometry, with pseudomolecular ions, (M-H)-, observed at 1099 (the major one) 1113, and 1127. Acyl-PI exclusively contained octadecenoyl on the inositol moiety (as 3-O-acyl), an unsaturated fatty acyl (mostly octadecenoyl) at sn-1 position of the glycerol and a saturated fatty acyl (mainly hexadecanoyl) at the sn-2 position. Acyl-PI constitutes a new natural substance and seems to be unique among the phospholipids of C. amycolatum. Other more complex molecules, previously undetected, and assigned in this work to several acyl forms of phosphatidylinositol trimannosides, lacked octadecenoyl in their polar heads. The present study reveals the existence of acyl-PI in C. amycolatum as rather unexpected finding and, additionally, gives evidence for the ability of this species to synthesize a great variety of inositol-containing phospholipids.

Comparison of diagnostic sensitivities of three assays (Bartels enzyme immunoassay [EIA], Biotest EIA, and Binax NOW immunochromatographic test) for detection of Legionella pneumophila serogroup 1 antigen in urine.

The Bartels enzyme immunoassay (EIA), Biotest EIA, and Binax NOW immunochromatographic test (ICT) urinary antigen kits for the detection of Legionella pneumophila serogroup 1 were compared using 178 frozen urine samples. When nonconcentrated urine samples were used, the sensitivity levels of both enzyme EIAs were significantly higher than the sensitivity level of the ICT (Bartels EIA, 71.3%; Biotest EIA, 65.1%; Binax NOW ICT, 37% [P < 0.001]). After concentration of the urine samples, no significant differences in sensitivity were found among the three tests.

Phospholipid composition of several clinically relevant Corynebacterium species as determined by mass spectrometry: an unusual fatty acyl moiety is present in inositol-containing phospholipids of Corynebacterium urealyticum.

A comparative study on phospholipids of Corynebacterium amycolatum, Corynebacterium jeikeium and Corynebacterium urealyticum was carried out using fast-atom bombardment (FAB) and electrospray ionization (ESI) mass spectrometry. Data obtained indicate the presence of acylphosphatidylglycerol (APG), diphosphatidylglycerol, phosphatidylglycerol (PG), phosphatidylinositol (PI) and triacylphosphatidylinositol dimannosides (Ac(3)PIM(2)) in these bacteria. In general, octadecenoyl and hexadecanoyl fatty acyl moieties predominated in phospholipids of C. amycolatum, whereas high levels of hexadecenoyl were found in C. jeikeium and C. urealyticum. Mass spectra from purified APG and PG indicated that the sn-1 position of the glycerol was occupied by octadecenoyl in the three species studied. Notably, several major molecular species of PI and Ac(3)PIM(2) from C. urealyticum contained significant amounts of a moiety identified as 10-methyleneoctadecanoyl, located at the sn-1 position of these molecules. On the other hand, multiantibiotic resistant and susceptible strains of C. amycolatum differed in several minor phospholipid fatty acids of 19 carbon atoms, identified as 10-methyloctadecenoic, 10-methyloctadecanoic (tuberculostearic acid) and 10-methyleneoctadecanoic. The results demonstrate an overall similarity among the phospholipids of the different species studied but also significant differences related to the acyl chains of the glycerol moiety of these compounds, notably the high levels of an unusual fatty acyl moiety in inositol-containing phospholipids of C. urealyticum.