Sensibilidad a azitromicina y otros antibióticos en aislados recientes de Salmonella,Shigella y Yersinia

Introduction

Azithromycin represents an alternative option to treat bacterial diarrhea when the antibiotic therapy is indicated. Little is known regarding the susceptibility to azithromycin in enteropathogens in Spain.

Methods

The MICs of azithromycin were determined by E-test against Salmonella non-typhi (SNT), Shigella and Yersinia isolates collected over the last three years (2010-2012). In addition, the susceptibility to other antibiotics usually used to treat gastrointestinal diseases was determined in these isolates by using a microdilution method.

Results

A total of 139 strains of SNT, Shigella and Yersinia were studied. All of them, except one strain, had a MIC ≤ 16 mg/L of azithromycin. In the adult population, 14.7% and 40.6% of SNT and Shigella isolates, respectively, were resistant to at least 2 of following antibiotics: amoxicillin, trimethoprim-sulfamethoxazole and ciprofloxacin. In the pediatric population, 10% of SNT clinical isolates and 28.6% (2/7) of Shigella isolates were resistant to amoxicillin and trimethoprim-sulfamethoxazole.

Conclusions

In our experience, azithromycin would be a useful antibiotic alternative to treat bacterial diarrhea.     

Staphylococcus coagulasa negativos resistentes al linezolid: características fenotípicas,genotípicas y sensibilidad a combinaciones de antibióticos

Objective

We recovered 22 coagulase-negative staphylococci isolates in our hospital to study their identity, susceptibility, epidemiological profile, linezolid resistance mechanisms, and the possibilities of different antibiotic combinations.

Methods

Isolate identification was performed using mass spectrometry (Vitek-MS, bioMérieux). Susceptibility testing was carried out with the Vitek-2 system and the broth microdilution method according to CLSI guidelines. Pulsed-field gel electrophoresis (PFGE) was performed to analyze the genetic relationship between isolates. Linezolid resistance mechanisms were evaluated by PCR/sequencing: presence of cfr gene, point mutations in domain V of 23S ribosomal RNA and additional ribosomal mutations (in the rplC, rplD and rplV genes).The in vitro activity of linezolid was investigated alone and in combination with another three antibiotics acting on different cellular targets, using E-test strips.

Results

Twenty isolates were identified as Staphylococcus epidermidis, and 2 as Staphylococcus hominis. PFGE showed that isolates belonged to diverse clones, 21 of them presented mutations in the domain V region of 23S rRNA and the cfr gene was found in 54.5%.Prior administration of linezolid was documented in most of cases.Linezolid in combination with gentamicin showed a synergistic activity in 45.5% of isolates.

Conclusions

Staphylococcus epidermidis was the most prevalent linezolid-resistant coagulase-negative staphylococci. All isolates showed increased MIC values compared to other anti-staphylococcal drugs and several linezolid resistance mechanisms. Our data suggest that linezolid plus gentamicin could be a synergistic combination against linezolid-resistant coagulase-negative staphylococci.     

Aplicación del análisis de los polimorfismos de los tamaños de los fragmentos de restricción del gen flaA amplificado por reacción en cadena de la polimerasa y resistotipo para identificar potenciales brotes de campilobacteriosis no diagnosticados en España

Introduction

Outbreaks of campylobacteriosis are infrequent and usually involve a low number of patients, although it is estimated that many more remain undiagnosed. The most successful techniques for outbreak investigation in Campylobacter spp. (PFGE, MLST) have the drawback of being laborious and not available in many laboratories.

Methods

During the year 2008, 352 isolates of C. jejuni and C. coli from 16 hospitals were received in our laboratory. All strains were genotyped by RFLP-PCR-flaA (flaA type) and phenotyped with their resistotype. It was established that the strains of the same species from the same hospital, isolated over a period of up to 11 days, with MIC values of ± 1 dilution with the same flaA type could belong to an outbreak. Strains that met these criteria would be later subtyped by KpnI-PFGE and MLST.

Results

A total of 23 out of 352 isolates, distributed in 10 groups, met the criteria for being associated with putative undiagnosed outbreaks. The similarity of the PFGE-profiles in 8 groups was greater than 95% among the isolates from each group. In 7 of the groups, the sequence types (MLST) were coincident.

Conclusions

The use of 2 easy markers (resistotype and RFLP-PCR-flaA) may detect isolates probably belonging to an undiagnosed outbreak of campylobacteriosis. Accurate diagnosis requires other molecular markers and epidemiological data of each isolate. The study suggests that, as in other countries, the number of outbreaks of campylobacteriosis in Spain is probably underestimated.     

Utilización de diferentes técnicas de biología molecular integradas en un algoritmo de identificación de micobacterias no tuberculosas

Introduction

The aim of the present work was to demonstrate the utility of a non-tuberculous mycobacteria (NTM) identification algorithm, which integrates different PCR-based techniques and basic phenotypic features. Moreover, the algorithm for pattern restriction analysis of hsp65 (hsp65 PRA) interpretation has been updated.

Methods

The workflow chosen consisted of the identification by a DNA hybridization probe method, followed by PCR-restriction enzyme analysis of hsp65 (hsp65 PRA) in those isolates that cannot be identified by hybridization probes. If necessary, 16S rRNA gene and hsp65 gene sequencing were used for speciation.

Results

A total of 236 NTM were collected, in which 102 (43.2%) isolates were identified by DNA specific probes and 76 (32.2%) isolates were identified with hsp65 PRA. Partial sequencing of the 16S rRNA gene was used for species identification of the remaining 58 (24.5%) isolates. Fifty-three (22.4%) were identified using this method. Five isolates (2.1%) were submitted for partial sequencing of hsp65 gene and one isolate was identified with this method. Four strains (1.7%) could not be identified at species level. Three new PRA patterns were found. Seven isolates tested positive with the AccuProbe Mycobacterium avium complex identification test but did not test positive with the M. avium or Mycobacterium intracellulare specific probes. Five and two of these isolates were identified as M. intracellulare and Mycobacterium colombiense, respectively.

Conclusion

This approach allowed us to identify almost all NTM isolates found in this study, including some recently described species.     

Análisis de la variación intraespecífica de Conidiobolus coronatus usando RAPD y secuencias ITS

Introduction

The fungus Conidiobolus coronatus (C. coronatus) has an extensive distribution of habitats and hosts. It is found saprophytically, and attacks insects and mammals, including humans. Although there are few reports on humans, and they are restricted to tropical areas. The aim of this work was to determine whether genetic variation exists between C. coronatus isolates coming from human lesions and other sources.

Methods

A total of 11 C. coronatus isolates obtained from soil, insects and humans were analyzed with the random amplification of polymorphic ADN (RAPD) and internal transcribed spacer (ITS) techniques; the maximum parsimony, neighbour-joining and minimum evolution methods were used for the ITS analysis.

Results

The analysis of the C. coronatus polymorphisms showed high intra-species variation levels between the evaluated isolates. The isolates coming from human lesions showed the greatest genetic divergence compared with the remaining isolates. The greatest genetic distance between isolate groups was found between those coming from humans and those taken from the insect Lycoriella ingenua.

Conclusions

This is the very first work evaluating and demonstrating that within species variation exists at molecular level in C. coronatus, and is related to the source where the isolates were taken from.     

Enterococcus faecium resistente a glucopéptidos. Análisis del genotipo de resistencia,virulencia y líneas genéticas

Objective

The objective of this study was to analyse the genotypic characteristics of all Enterococcus isolates with acquired vancomycin resistance (VRE) recovered in the Hospital Clinic (Barcelona, Spain) in a period of three years and two months.

Methods

All VRE isolated in the referred Hospital in the period January 2004-March 2007 were included in the study. The vancomycin resistance mechanism was investigated, as well as other antibiotic resistance mechanisms. Isolates were also typed by pulsed-field-gel-electrophoresis (PFGE) and multi-locus-sequence-typing (MLST).

Results

Thirty-nine VRE were recovered, all being identified as E. faecium, representing 2% of total enterococci obtained in that period. Thirty-eight of them carried the vanA gene, and one isolate the vanB2 gene. The 39 VRE were classified into 13 different pulsotypes (A-M), with one main pulsotype, A, which included 13 isolates. The sequence type was identified by MLST in 24 VRE (with unrelated or closely-related PFGE patterns), and they were ascribed to the clonal complex CC17, but two classified as CC9. All VRE showed a multiresistance phenotype, including, in most cases ampicillin, ciprofloxacin, erythromycin, streptomycin, gentamicin, kanamycin and chloramphenicol, harbouring multiple antibiotic resistance genes. The presence of esp and/or hyl genes was identified in 37 VRE.

Conclusion

All VRE, but one, showed the vanA genotype and they were mostly ascribed to the high-risk clonal complex CC17.     

Susceptibilidad frente a sarampión y varicela en el personal sanitario de un hospital de tercer nivel en Cataluña

Introduction

To estimate the susceptibility to measles and varicella (chickenpox) in healthcare workers in a public tertiary level teaching hospital, in Catalonia.

Methods

A prevalence study was conducted from January 2006 to December 2008 on 2,752 workers who had serology performed for the determination of measles or varicella by ELISA test during a health examination. Data were analysed by, sex, age, professional category and work unit.

Results

A total of 153 healthcare workers were susceptible to measles and 187 to varicella. The susceptibility of healthcare workers to measles was 6.04% (95% CI: 5.78 to 6.30), and to varicella it was 7.45% (95% CI: 7.14 to 7.75). The highest susceptibility to measles was in resident physicians with 14% (95% CI: 10.8 to 18.5). In high-risk services, where highly immunocompromised patients are attended, the susceptibility of workers was slightly higher than the rest to measles (6.32% vs 5.93%) and varicella (8.34% vs 7.09%). Healthcare workers born after 1980 were 20 times (95% CI: 11.0 to 37.2) more likely to be susceptible to measles, and 2 times (95% CI: 1.2 to 3.2) more likely to be susceptible to varicella than those those born before 1965.

Conclusions

The susceptibility to measles in healthcare workers in our centre is higher in younger cohorts, with values higher than expected in a community with high vaccination coverage against measles, mumps, rubella vaccine (MMR) in the paediatric population for many years.     

Colonización nasal por Staphylococcus aureus en estudiantes de medicina: importancia en la transmisión hospitalaria

Background

Staphylococcus aureus is the main pathogen causing nosocomial infections. Health professionals, including medical students, could be a source of transmission. The aims of the study were to determine the rate of nasal carriage of S. aureus susceptible and resistant to methicillin (MRSA) and evaluate the knowledge and adherence that students had about hand hygiene.

Methods

The study included medical students attached to the Hospital Universitario 12 de Octubre (Madrid, Spain). We collected samples from both nasal vestibules, and the antimicrobial susceptibility was determined on all isolates. Data collection was performed using a self-administered questionnaire that included risk factors for colonization, hygiene habits and knowledge of hand hygiene protocols.

Results

Of the 140 students included, 55 (39.3%) were colonized by S. aureus, and 3 (2,1%) by MRSA. The exposure to antibiotics in the last 3 months was lower in colonized students (12.3% vs. 25.9%, P = .03). Self-assessment showed that 56.4% of students almost never washed their hands before to attending to the first patient, and only 38.6% always washed after examining patients. More than a third (35.7%) ignored the hand hygiene protocol, and 38.6% had not received specific formation.

Conclusions

Medical students should be included in hospital infection control programs. Hand hygiene training should be given to students before they begin their practices in the hospital.     

Aumento significativo de la colonización por Staphylococcus aureus entre los estudiantes de medicina durante la realización de las prácticas en el hospital

Introduction

Staphylococcus aureus is a pathogen of major concern. The emergence of methicillin-resistant S. aureus (MRSA) has increasingly complicated the therapeutic approach of hospital-acquired infections. Surveillance of MRSA and control measures must be implemented in different healthcare settings, including screening programs for carriers. Our first aim was to determine the prevalence of methicillin-susceptible S. aureus (MSSA) and MRSA nasal carriage in medical students from the Clínico San Carlos Hospital (Madrid). As the MRSA carrier rate in healthcare workers is higher than in the general population, we hypothesised that carrier rate could be increased during their clinical practice in their last three years.

Methods

We performed an epidemiologic al study of the prevalence of S. aureus colonisation among a group of medical students, who were sampled in 2008 in their third-year, and in 2012 when this class was in its sixth year.

Results

We have found a significant increase in MSSA carriage, from 27% to 46%. There were no MRSA colonisations in the third-year, but one was found in the sixth-year group. The large majority of strains (89%) of strains were resistant to penicillin, and 27% to erythromycin and clindamycin. As 19 coagulase-negative Staphylococcus MR were also identified, a horizontal transfer of genes, such as mecA gene to S. aureus, could have occurred.

Conclusions

Medical students are both, at risk for acquiring, and a potential source of nosocomial pathogens, mainly MSSA. Therefore, they should take special care for hygienic precautions, such as frequent and proper hand washing, while working in the hospital.     

Resistencia a antibióticos y factores de virulencia en aislados clínicos de Salmonella enterica

Introduction

The increase of Salmonella enterica isolates multi-resistant to different antibiotics, including β-lactams and fluoroquinolones, is a problem of clinical importance. The dissemination of Salmonella Typhimurium resistant to ampicillin (AMP)-chloramphenicol (CHL)-streptomycin (STR)-sulphonamides and (SUL)-tetracycline (TET), that harbour the Salmonella Genomic Island type 1 (SGI1), and the acquisition of transferable genetic material have favoured the multi-resistance in this genus.

Methods

A total of 114 clinical S. enterica isolates were studied (period 2009-2010). The susceptibility to 20 antibiotics was determined by disc diffusion and microdilution. The antimicrobial resistance mechanisms and the integrons were analysed by PCR, and sequencing in the AMP^R isolates. In all the bla_PSE-1-positive isolates, the clonal relationship was determined by PFGE, as well as the presence of SGI1 and 29 virulence genes by PCR.

Results

Eighteen different serotypes were found among the 114 isolates studied, Typhimurium (61%) and Enteritidis (16%) being the most prevalent. High percentages of resistance to SUL (68%), TET (58%), AMP (55%) and STR (46%) were observed. The great majority (92%) of 63 AMP^R isolates were multi-resistant, with the AMP-STR-TET-SUL phenotype (19 isolates) being the most frequent one and associated with the bla_TEM-1b + strA-strB + tet(B) + sul2 genotype. Class 1 integrons (7 different structures) were observed in 48% AMP^R isolates, highlighting the bla_OXA-1 + aadA1 structure (8 isolates), one empty integron and non-classical integrons (5 isolates). The bla_PSE-1 gene was detected inside the classical SGI1 structure in 13 clonally-related isolates that showed the same virulence profile.

Conclusions

The high percentage of multi-resistant S. enterica isolates, especially associated to S. Typhimurium, to the AMP, STR, TET and SUL phenotype, and to the bla_TEM-1b + strA-strB + tet(B) + sul2 genotype, shows an important risk of possible failures in the treatment of serious infections caused by this serotype.     

Implantación electiva de stent recubierto en la coartación y recoartación en el joven y en el adulto

Introduction and objectives

Stent implantation is an effective therapy for aortic coarctation and recoarctation. However, in adolescents and adults, aortic wall rupture and dissection can occur, as well as aneurysms during follow-up. In order to reduce these complications, we electively implant covered stents.

Methods

Since 2005, we have performed the procedure using femoral access in 17 patients (2 adolescents and 15 adults), 16 electively and 1 as a rescue procedure. We used the Mullins technique in all cases, implanting a NuMED^® covered stent.

Results

Good stent apposition was achieved in all 17 procedures; 8 patients required a distal flare. Gradient was reduced from 40 (16) mmHg to 2 (2) mmHg (P<.001) and lumen diameter increased from 4 (2) mm to 19 (3) mm (P<.001). Two exceptional cases are discussed: one patient with aortic wall rupture who underwent a rescue procedure using a stent within a covered stent and another patient with total obstruction and intercostal aneurysm in whom the outcome was fatal at 48 h postprocedure (autopsy is shown). Four-year clinical follow-up included Doppler echocardiography; an additional imaging technique was required in 13 patients. All patients recovered well and there were no complications.

Conclusions

Covered stents are effective in treating coarctation and recoarctation in adolescents and adults, are the treatment of choice in patients with complex anatomy, and must be available in the operating room as a rescue device when implanting a conventional stent.Full English text available from:www.revespcardiol.org/en     

Caracterización molecular de cepas de Staphylococcus aureus resistentes a meticilina ST398 en pacientes con infecciones cutáneas y sus familiares

Introduction

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) of sequence type ST398 is a genetic lineage also described in human infections.

Methods

Cutaneous infections related with MRSA ST398 are described in 3 patients, two of them pig farmers. The MRSA nasal carriage by patients and their relatives was also studied. MRSA ST398 strains were typed (SCCmec, spa, agr and MLST) and the antimicrobial resistance pattern and virulence genes were determined.

Results

Twenty MRSA ST398 isolates were recovered in lesions of three patients, and in nasal samples of two patients and five relatives. Isolates were typed: spa-type t011 or t108; agr-type I and SCCmec IVa or V. MRSA strains were tetracycline-resistant and 15 of them showed a phenotype and genotype of multi-resistance, but they were free of tested virulence genes.

Conclusions

LA-MRSA ST398 is an emergent problem in our country, mainly associated with skin and soft tissue infections in people with professional relationships with pig farms. Tetracycline resistance is an important marker for MRSA ST398 detection.     

La infección neonatal tardía por Streptococcus agalactiae en el área de Barcelona (1996-2010)

Objetive

To study the characteristics and evolution of group B Streptococcus (GBS) late-onset diseases, over a period of 15 years in 8 hospitals the Barcelona area and analyze the possible impact of prophylactic measures for the prevention of early-onset neonatal infections.

Methods

Retrospective review of all patients diagnosed with late-onset neonatal disease due to GBS from 1996 to 2010.

Results

A total of 143 patients were diagnosed. Of these, 51 were born in others hospitals. The overalll incidence was 0.42 per 1000 live births, varying between 0.14‰ in the year 2000 and 0.80‰ in 2009. A slight but sustained tendency of increased risk was observed over the years, 6.9% in the overall disease (with no statistical significance). Sepsis/bacteremia was detected in 63.6% of the newborns, meningitis in 32.8%, and arthritis/osteomyelitis in 3.5%. In cases with known obstetrics dates, 53% of mothers had been colonized by GBS during pregnancy, 53.8% received intrapartum antibiotic prophylaxis, and 41.2% had some obstetric risk factors, particularly premature birth in 35.9%. There was a 2.8% mortality rate in the neonates, and predominant serotypes were III and Ia.

Conclusions

The incidence of GBS late-onset disease has not decreased despite the control practices of early-onset disease, and possibility of this appearing must be taken into account.     

Caractéristiques clinicobiologiques des méningites à Enterovirus de l’adulte : étude de 59 cas

Purpose

To describe the epidemiological and clinical features of enteroviral meningitis as well as the biological profile of the cerebrospinal fluid (CSF).

Methods

A retrospective study conducted in a single centre between 2004 and 2008. All aseptic meningitis due to Enterovirus were included.

Results

Fifty-nine patients were included. The triad including fever, headache and neck stiffness was reported in 62% of patients. Twelve patients (20%) had a neutrophilic leukocytosis and 23 (39%) an elevated CRP level. Twenty-eight patients (47%) had a prominent neutrophilic reaction in the CSF and nine (15.2%) had a low glucose concentration. A presumptive anti-bacterial treatment was initiated in 47 patients (80%) for an average of 3.2 days.

Conclusion

Due to the lack of specificity of clinical features and biological manifestations of Enterovirus meningitis, the widespread use of real-time Enterovirus PCR is a priority for reducing the number of unnecessary anti-bacterial treatment. Guidelines based on clinical and biological features may be associated to help physicians in the differential diagnosis between bacterial and viral meningitides.     

Candida: epidemiología y factores de riesgo para especies no albicans

Introduction

Nosocomial fungal infections have increased significantly in the last decade. Candida detection in clinical specimens can mean either colonization or an infection which can be local (muguet) or invasive. Knowledge of the species helps in choosing the best treatment. The aims of this study were to determine the frequency and distribution of Candida species detected in clinical samples, to analyze the clinical characteristics of the involved population and to determine the risk factors for Candida non-albicans species.

Methods

Retrospective, observational. Period: 2006-2010. Inclusion criteria: all isolates of Candida in clinical specimens from patients hospitalized —at least 48 hours in a neurological center. We analyzed epidemiological characteristics, co morbidities, risk factors, factors associated with Candida non-albicans detection, antifungal treatment, development of adverse events and mortality.

Results

Candida spp. was isolated from 321 clinical specimens: 139 (43.3%) were C. albicans and 182 (56.7%) Candida non-albicans. The distribution of the sample was: urine 122 (Candida non-albicans 67.2%), airway 81, oropharynx 45 (C. albicans) and candidemia 40 (Candida non-albicans 75%). The most frequent co-morbidity was solid tumor (35.5%). The main risk factors were antibiotic therapy (85.5%), steroid therapy (61.7%) and in ICU at diagnosis (61.6%). The analysis of risk factors and the isolation of Candida non-albicans shows that chemotherapy, previous surgery, treatment with aminopenicillins, carbapenems and glycopeptides were statistically significant (P < .05). There is a trend in neutropenic patients (P = .055) and in ICU at diagnosis (P = .076). Overall survival was 71%.

Conclusions

Candida species distribution varies with the type of sample analyzed. Non-albicans species make up the majority of the isolates. The identification of the species involved per sample helps to optimize treatment. The high frequency of isolation of Candida in patients on steroids and antibiotics and admitted to ICU, is worth pointing out. Patients with previous surgery, treated with the aforementioned antibiotics or chemotherapy, could receive non-azole antifungals in the initial empirical treatment strategy.     

Validez y aplicabilidad de un nuevo método de registro para la hipertensión arterial

Introduction and objectives

Blood pressure measurement methods and conditions are determinants of hypertension diagnosis. A recent British guideline recommends systematic 24-h ambulatory blood pressure monitoring. However, these devices are not available at all health centers and they can only be used by 1 patient per day. The aim of this study was to test a new blood pressure recording method to see if it gave the same diagnostic results as 24-h blood pressure monitoring.

Methods

One-hour blood pressure monitoring under routine clinical practice conditions was compared with standard method of day time recording by analyzing the coefficient of correlation and Bland-Altman plots. The Kappa index was used to calculate degree of agreement. Method sensitivity and specificity were also analyzed.

Results

Of the 102 participants, 89 (87.3%) obtained the same diagnosis regardless of method, with high between-method agreement (κ= 0.81; 95% confidence interval, 0.71-0.91). We observed robust correlations between diastolic (r = 0.85) and systolic blood pressure (r = 0.76) readings. Sensitivity and specificity for the new method for diagnosing white coat hypertension were 85.2% (95% confidence interval 67.5%-94.1%) and 92% (95% confidence interval, 83.6%-96.3%), respectively.

Conclusions

One-hour blood pressure monitoring is a valid and reliable method for diagnosing hypertension and for classifying hypertension subpopulations, especially in white coat hypertension and refractory hypertension. This also leads to a more productive use of monitoring instruments.Full English text available from:www.revespcardiol.org/en     

Molecular epidemiology and antimicrobial susceptibility of Campylobacter coli clinical isolates

Introduction

Campylobacter spp. is a major cause of acute bacterial diarrhea in humans worldwide, and C. coli is responsible for 10% of the cases.

Materials and methods

A study was made of the antimicrobial susceptibility using the E-test^®, and the clonal relationship using PCR-RFLP, of the flaA gene, as well as PFGE techniques on 43 C. coli clinical isolates.

Results

Only 49% and 2% of the isolates were susceptible to erythromycin and ciprofloxacin, respectively. Imipenem and clindamyicn, with 100% and 84% of the strains, respectively, being susceptible, were the most active antimicrobials. The PCR-RFLP of flaA gene technique grouped fourteen isolates into six clusters, while the PFGE technique grouped eleven isolates into five clusters.

Conclusion

Ciprofloxacin and erythromycin are not suitable for the treatment of C. coli infections. Clindamycin could be considered as a therapeutic alternative in cases of enteritis, while imipenem is the best alternative for extra-intestinal infections. Both PFGE and PCR-RFLP can be useful to detect clones.