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.     

Asociación en un mismo plásmido de blaVIM-1 y qnrS2 en Klebsiella pneumoniae y Klebsiella oxytoca aisladas en Sevilla

Background

Combined resistance to quinolones and β-lactams is common in Enterobacteriaceae. The appearance in enterobacteria coding for metallo-β-lactamases and determinants of plasmid-mediated quinolone resistance are an emerging problem in our country.

Methods

The susceptibility was determined by E-test. The resistance genes were detected by PCR and the corresponding plasmids were characterised.

Results

This study describes 2 strains (1 Klebsiella oxytoca, 1 Klebsiella pneumoniae) carrying the genes qnrS2 and blaVIM-1 in a transferable plasmid of 70-Kb isolated in surveillance cultures at the University Hospital Virgen Macarena in Seville.

Conclusion

This is the first combination of qnrS2 and bla_VIM-1 on the same non-typeable plasmid isolated in our centre.     

Caracterización molecular de aislados humanos de Cryptosporidium spp. procedentes de 2 diferentes localizaciones de España

Molecular PCR based diagnostic techniques have enabled us to distinguish between the different, morphologically identical, Cryptosporidium species that can infect humans. Of the 23 recognized species in the genus, at least 9 are able to infect humans. As the intensity of the clinical manifestations, pathogenicity, excretion of oocysts, and incidence, are different between this species, molecular studies are crucial for a better understanding of the epidemiology of human cryptosporidiosis.     

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.     

Caracterización de cepas de Staphylococcus epidermidis y S. haemolyticus resistentes a meticilina y linezolid en un hospital español

Introduction

Linezolid resistance is mainly due to mutations in the 23S rRNA target. The aim of this study was to characterize linezolid and methicillin resistant Staphylococcus epidermidis (SE-LM^R) and S. haemolyticus (SH-LM^R) strains detected in a Spanish hospital.

Methods

SE-LM^R and SH-LM^R strains obtained in the period June 2009-August 2011 in a second level hospital were recorded along with the epidemiological characteristics of the patients. These strains were typed, and their resistance, phenotype, genotype and the factors determining their virulence were analysed.

Results

Linezolid resistance was explained by the presence of G2603T mutation (23S rRNA) and aminoacid changes in L3 and L4 ribosomal proteins. The 25 SE-LM^R strains belonged to sequence type ST2, presented SCCmec type III, and two different PFGE patterns. The two SH-LM^R strains showed non-typeable SCCmec. SE-LM^R strains harboured the resistance genes aac(6’)-aph(2”), and dfrS1. SH-LM^R strains contained these genes and the gene erm(C). No lincomycin resistance mechanism was identified in SE-LM^R strains regardless of showing lincomycin resistance and diminished susceptibility to clindamycin.

Conclusions

Linezolid resistance is of concern in hospitals, and requires continued vigilance. Several linezolid resistance mechanisms (mutation in 23S RNAr and amino acid changes in L3 and L4) were identified in this study.     

Prevalencia del determinante de resistencia plasmídica a quinolonas aac(6’)-Ib-cr en cepas de Escherichia coli y Klebsiella pneumoniae productoras de BLEE aisladas en diez hospitales de Chile

Introduction

The frequency of aac(6′)-Ib-cr gene in ESBL-producing strains of Klebsiella pneumoniae and Escherichia coli is unknown, in Chile.

Methodology

The aac(6′)-Ib and aac(6’)-Ib-cr genes were investigated using polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and sequencing, in strains isolated from 10 Chilean hospitals between 2008-2009.

Results

The aac(6’)-Ib-cr gene was detected in 54% of K. pneumoniae and 74% of E. coli strains. The CIM₅₀ of CIP was higher among strains harboring aac(6’)-Ib-cr, 8 times higher in K. pneumoniae and 4 times higher in E. coli. Moreover, both aac(6’)-Ib and aac(6’)-Ib-cr were simultaneously found in 13 K. pneumoniae and 3 E. coli isolates.

Conclusion

This is the first report of aac(6’)-Ib-cr in ESBL-producing strains of K. pneumoniae and E. coli isolated from in-patients in Chilean hospitals located along an area of more than 2,800 Km.     

Perfil serológico en gestantes extranjeras frente a VIH,VHB, VHC,virus de la rubéola, Toxoplasma gondii,Treponema pallidum, y Trypanosoma cruzi

Introduction

The increase in immigration is changing the prevalence of mother to child infectious diseases. Our aim is to determine the serological profile of foreign pregnant women against these infections.

Methods

A retrospective cross sectional study was performed in a tertiary hospital from Madrid between August 2007 and October 2008. The seroprevalence against HIV, HBV, HCV, rubeola, T. gondii, T. pallidum and T. cruzi was determined in every pregnant immigrant, as well as in a representative group of Spanish pregnant women.

Results

A total of 2526 immigrant and 157 Spanish pregnant women were studied. None of the Spanish and 0.5% of the foreigners showed antibodies against HIV; 18.9% of them were Sub-Saharan women. Antigen HBs was detected in 2% of the immigrant women and in 1.1% of the Spanish women. Asian women had the highest rate of type B Hepatitis (10.9%). There was 0.9% of type C Hepatitis among the immigrants and 1% among the Spanish. Within the cases with RPR ≥ 1/8, 1.6% were immigrants, most of whom were Latin American. Thirty-one per cent of the immigrants showed antibodies against T. gondii (37.5% from Central America, 2.5% from the Far East). More than 95% of the Spanish women had antibodies against Rubella, this being lower in the rest of the areas (75.5% in Sub-Saharan Africa). T. cruzi infection was detected in 12.1% of the Bolivian women studied.

Conclusion

The prevalence of mother-to-child transmitted infections depends on the origin of pregnant women. Knowledge of these differences may lead to improved control these diseases.     

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.     

Significado clínico del aislamiento de Staphylococcus aureus en muestras de orina

Introduction

To evaluate the clinical significance of the isolation of Staphylococcus aureus in urine samples.

Methods

A retrospective study was performed on adult patients identified from a microbiology database in a 200-bed general hospital between the years 2000 and 2009. The demographic data, comorbidities, and risk factors, were reviewed, particularly those associated with the concomitant isolation of S. aureus in blood cultures.

Results

The frequency of S. aureus found in urine samples was 0.63%. A total of 43 patients (mean age 68.7 years [SD ± 16], and 58.1% males) were identified in the database. A Charlson comorbidity index > 3 was observed in 20.9%. Concurrent bacteremia was seen in 48.8%. Two groups of patients were distinguished: with concomitant bacteremia (n = 21) or without (n = 22). Intervention in the urinary tract significantly predicted (P = .00004) bacteriuria without bacteremia (81.8%), compared to bacteremia cases (19%). The attributable mortality was 47.6% in patients with bacteremia compared to non-bacteremia (no deaths), even though the appropriate antibiotic treatment was more frequent among patients with bacteremia (92% and 60%, respectively).

Conclusion

The presence of S. aureus in urine was accompanied by bacteremia in half of the cases, but in patients without previous urinary tract intervention such a possibility increased to 81%. Concomitant bacteremia predicts a worse prognosis even with appropriate treatment.     

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     

Actualización de la eficacia de la terapia triple para la infección por Helicobacter pylori y de la resistencia a claritromicina en España (2007-2012)

Introduction

Triple therapy, which remains the standard treatment for Helicobacter pylori infection, should be discouraged when its efficacy is lower than 80% or when clarithromycin resistance rates are above 15-20%.

Aim

To update the available evidence on the effectiveness of triple therapy and clarithromycin resistance rates in adults in Spain over the last 6 years.

Methods

A literature search (2007-2012) was conducted in Medline and the abstracts books of the annual meetings of several Spanish gastroenterological and microbiological congresses. The search terms were «Helicobacter pylori», «Spain» and «clarithromycin». Studies were selected if they included triple therapy consisting of a proton pump inhibitor with clarithromycin and amoxicillin or if they analyzed H. pylori clarithromycin susceptibility in treatment-naïve patients.

Results

There were five articles and nine abstracts (3147 patients) on triple therapy, which showed a mean cure rate of 70.8% (95% CI = 66-76%). When stratified by the duration of therapy, the mean cure rates were 68.8% (60-76%) for 7-day regimens and 71.76% (68-78%) for 10-day regimens. For clarithromycin resistance rates, four articles and five abstracts (1709 patients) revealed a mean resistance rate of 18.3% (13-22%).

Conclusions

The efficacy of triple therapy seems to be unacceptable in recent studies conducted in Spain, possibly associated with clarithromcyin resistance rates higher than previously reported.     

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.     

Epidemiología y características clínicas de los episodios de bacteriemia por Streptococcus pyogenes en Cartagena (Murcia)

Introduction

A gradual increase in severe cases due to Streptococcus pyogenes or Streptococcus beta-hemolytic group A (SGA), has been detected in the last few decades.

Methods

Retrospective study of bacteremia due to S. pyogenes detected between January 2009 and January 2013 in Cartagena. The annual incidence for severe bacteremia has been estimated.

Results

Thirteen cases of SGA bacteremia were recorded. The incidence increased from 0.37 in 2009 to 2.5 cases/100,000 inhabitants in 2012. The predominant focus was skin and soft tissue infections (53%). Early mortality was 20%.

Conclusion

Severe streptococcal disease is rare, but affects individuals with good functional status, and is associated with a high mortality.     

Neisseria gonorrhoeae: resistencias antimicrobianas y estudio de la dinámica poblacional. Situación en 2011 en Barcelona

Background

Due to the high rates of antimicrobial resistance to certain antibiotics, together with the emergence of Neisseria gonorrhoeae (NG) with reduced susceptibility and resistance to third-generation cephalosporins, gonococcal infection is becoming a public health problem. The objectives of the study were: To keep track of the antimicrobial susceptibility of NG strains obtained from January to August 2011. To study the population dynamics.

Methods

The antimicrobial susceptibility was studied by disk-diffusion and E-test. The genotyping was performed by NG-MAST method.

Results

Of a total of 100 strains studied, 59% showed intermediate sensitivity to penicillin and 9% were resistant. According to EUCAST, we detected 3 gonococci with reduced susceptibility to ceftriaxone, 10 to cefixime and one with high-level resistance to both antibiotics (MIC 1.5 μg/ml). MIC₅₀ and MIC₉₀ to cefixime were 0.016 and 0.125 μg/ml, respectively, whereas to ceftriaxone they were < 0.016 and 0.064 μg/ml, respectively. Almost all (99%) of the strains were resistant to doxycycline, 53% to ciprofloxacin, 3% to azithromycin, and 1% to spectinomycin. The most prevalent ST was ST1407, predominantly associated to resistance or reduced sensitivity to cephalosporins or macrolides.

Conclusions

NG has developed significant rates of resistance to various antibiotics. One strain has been detected with high level resistance to third generation cephalosporins, and several strains with reduced susceptibility. An increase in MIC₅₀ and MIC₉₀ to these antibiotics has also been observed. NG population structure remains stable and common to the rest of Europe, although two new ST (ST7226 and ST7227) have been identified that could be selected and acquire high levels of resistance to cephalosporins.     

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.