Real-life experience of hepatitis C treatment in a Spanish prison.

OBJECTIVE: Hepatitis C virus (HCV) infection is a major public health problem that causes multiple comorbidities. People in prisons who inject intravenous drugs are at increased risk of HCV infection, and HCV infection is 15-fold more prevalent among prisoners compared with the community. The objective of this study was to analyse the clinical and epidemiological characteristics of residents of a Spanish prison with HCV infection who received antiviral treatment. METHODS: An observational, descriptive and retrospective study was performed. All patients with HCV infection diagnosed or followed up in an Infectious Diseases attached to a penitentiary were included in this study. RESULTS: Of 81 patients analysed, sixty-nine (83.1%) patients were male. The mean age was 50.1 (SD8.8) years, and 70% of the inmates had a history of injection drug use. Coinfection with HIV was detected in 30%. In up to 25% of the sample, there were data on chronic liver disease in the degree of liver cirrhosis. The diagnosis of HCV infection had been made more than 15 years earlier in 28% of those studied. Decompensations from liver disease, hepatocellular carcinoma, or hospital admissions were exceptional. Most of the inmates with HCV accepted treatment, and approximately 94% of the patients who completed treatment achieved a sustained virological response without interactions or complications of interest. CONCLUSIONS: The availability of direct-acting antivirals and their exceptional side effects constitute an opportunity to reduce the burden of HCV infection in Spain, particularly in these high-risk populations.

Mechanisms of resistance to daptomycin in Staphylococcus aureus.

Daptomycin is a cyclic lipopeptide active against multidrug-resistant Gram-positives, including methicillin-resistant Staphylococcus aureus (MRSA) and S. aureus with reduced susceptibility to vancomycin. It is 4-8 fold as active as vancomycin against methicillin-susceptible S. aureus (MSSA) and MRSA, and retains most of this activity against S. aureus with reduced susceptibility to vancomycin. The mechanism of action of daptomycin is not fully understood. Daptomycin binds to the bacterial cytoplasmic membrane, leading to depolarization due to the loss of potassium ions from the cytoplasm. Daptomycin non-susceptibility is unusual in the clinical setting. Different mechanisms have been proposed to explain daptomycin-resistance, most of them associated to changes in composition, charge and fluidity of the cell wall. The mprF mutations, which lead to an increase in the lysyl-phosphatidyl glycerol production, and rpoB and rpoC mutations (rpo genes encode for bacterial RNA polymerase subunits) have been proposed as associated to daptomycin-resistance, but a number of mutations in other genes ( walK, cls, ggrA…) have been proposed.

Full coding hepatitis E virus genotype 3 genome amplification method.

Hepatitis E virus (HEV) genotype 3 produces zoonotic infection associated with the consumption of infected animals. HEV infections can become chronic in immunocompromised (IC) patients. The viral genome has three well defined open reading frames (ORF1, ORF2 and ORF3) within which various domains and functions have been described. This paper (i) describes a new method of complete sequencing of the HEV coding region through overlapping PCR systems, (ii) establishes a consensus sequence and polymorphic positions (PP) for each domain, and (iii) analyzes the complete coding sequence of an IC patient. With regard to the consensus, a high percentage of PP was observed in protease (PP=19%) and the X domain (PP=22%) within ORF1, the N-terminal region of the S domain (PP=22%) in ORF2, and the P1 (PP=35%) and P2 (PP=25%) domains in ORF3. In contrast, the ORF1 Y, ORF2 S, ORF2 M and ORF3 D1 domains were conserved in the reference sequences (0.40, 1, 0.70 and 0% of PP, respectively). The sequence from the IC patient had more mutations in the RpRp (D1235G, Q1242R, S1454T, V1480I, I1502 V, K1511R, G1373 V, E1442D, V1693 M), the terminal ORF2 S- domain (F10L, S26T, G36S, S70P, A105 V, I113 V), the X domain (T938 M, T856 V, S898A) and the helicase (S1014N, S975T, Q1133 K).

Imported transmissible diseases in minors coming to Spain from low-income areas.

We prospectively studied the prevalence of imported transmissible diseases in 373 immigrant children and adolescents coming from Sub-Saharan Africa, North Africa and Latin America to Salamanca, Spain. The most frequent transmissible diseases in this group were latent tuberculosis (12.7%), chronic hepatitis B virus infection (4.2%), hepatitis C virus infection (2.3%), syphilis (1.5%) and human T-lymphotropic virus type 1 or 2 infections (1.4%). A total of 24.2% of patients had serologic profiles suggesting past hepatitis B virus infection. Anti-human immunodeficiency virus antibodies were not detected in any subject. Largely asymptomatic immigrant children show a high prevalence of communicable diseases. Thus, infectious disease screenings are highly advisable in immigrant children coming from low-income countries.

Pretreatment of urine samples with SDS improves direct identification of urinary tract pathogens with matrix-assisted laser desorption ionization-time of flight mass spectrometry.

We pretreated with SDS 71 urine samples with bacterial counts of >10(5) CFU/ml and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) identification scores of <2, in order to minimize failure rates. Identification improved in 46.5% of samples, remained unchanged in 49.3%, and worsened in 4.2%. The improvement was more evident for Gram-negative (54.3%) than for Gram-positive (32%) bacteria.

Rapid method for direct identification of bacteria in urine and blood culture samples by matrix-assisted laser desorption ionization time-of-flight mass spectrometry: intact cell vs. extraction method.

Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a fast and reliable technology for the identification of microorganisms with proteomics approaches. Here, we compare an intact cell method and a protein extraction method before application on the MALDI plate for the direct identification of microorganisms in both urine and blood culture samples from clinical microbiology laboratories. The results show that the intact cell method provides excellent results for urine and is a good initial method for blood cultures. The extraction method complements the intact cell method, improving microorganism identification from blood culture. Thus, we consider that MALDI-TOF MS performed directly on urine and blood culture samples, with the protocols that we propose, is a suitable technique for microorganism identification, as compared with the routine methods used in the clinical microbiology laboratory.

Microorganisms direct identification from blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows a fast and reliable bacterial identification from culture plates. Direct analysis of clinical samples may increase its usefulness in samples in which a fast identification of microorganisms can guide empirical treatment, such as blood cultures (BC). Three hundred and thirty BC, reported as positive by the automated BC incubation device, were processed by conventional methods for BC processing, and by a fast method based on direct MALDI-TOF MS. Three hundred and eighteen of them yield growth on culture plates, and 12 were false positive. The MALDI-TOF MS-based method reported that no peaks were found, or the absence of a reliable identification profile, in all these false positive BC. No mixed cultures were found. Among these 318 BC, we isolated 61 Gram-negatives (GN), 239 Gram-positives (GP) and 18 fungi. Microorganism identifications in GN were coincident with conventional identification, at the species level, in 83.3% of BC and, at the genus level, in 96.6%. In GP, identifications were coincident with conventional identification in 31.8% of BC at the species level, and in 64.8% at the genus level. Fungaemia was not reliably detected by MALDI-TOF. In 18 BC positive for Candida species (eight C. albicans, nine C. parapsilosis and one C. tropicalis), no microorganisms were identified at the species level, and only one (5.6%) was detected at the genus level. The results of the present study show that this fast, MALDI-TOF MS-based method allows bacterial identification directly from presumptively positive BC in a short time (<30 min), with a high accuracy, especially when GN bacteria are involved.

[Problematic bacteria]. / Bacterias problemáticas.

INTRODUCTION: Because of the special characteristics of the critical patients, infections are one of the primary complications they suffer so that multiresistant microorganisms take on a special importance in this type of patient. SOURCES: Search in Medline using the words ICU (Intensive Care Unit), multidrug resistant, critical patient. DEVELOPMENT: Glycopeptide resistant enterococci show a reduced prevalence in our setting and the VISA and hVISA are isolated sporadically. MRSA is, on the other hand, a major problem. In 2003-2005, it was already accounting for 28 %-38 % of the S. aureus isolated in the ICU, with a high percentage of fluoroquinolone (>90 %) and macrolide (>65 %) co-resistance. The extended-spectrum beta-lactamase producing enterobacteria (BLEE) also are a growing problem, worsened by their frequent co-resistance with fluoroquinolones, about 30 % according to some studies. Carbapenem resistance in A. baumannii has doubled in recent years, with values greater than 50 %, almost always associated to enzymes of the OXA group. P. aeruginosa also maintains high resistance values (25 %-30 % of resistance to imipenem, ceftazidime or ciprofloxacin), but more stable. However, high rates of multi-resistance are also observed, now about 50 % of the isolations of imipenem resistant P. aeruginosa are also to fluoroquinolones. As a whole, recent studies show that the multiresistance has multiplied in recently years by 5 in P. aeruginosa and by 7 in A. baumannii. CONCLUSIONS: Multiresistant bacteria infections are one of the greatest problems to combat in critical patients and control of their spreading and the development of active antimicrobials against them is one of the principal challenges at present.

In vitro susceptibility of community-acquired urinary tract pathogens to commonly used antimicrobial agents in Spain: a comparative multicenter study (2002-2004).

The susceptibility patterns of 2724 uropathogens isolated in 9 Spanish regions during 2002, and 3013 obtained in 2004 were determined. The antibiotics tested were fosfomycin trometamol, amoxicillin, co-amoxiclav, cefixime, cefuroxime-axetil, pipemidic, ceprofloxacin, trimethoprim plus sulphamethoxazole and nitrofurantoin. Escherichia coli was the main pathogen in both studies (73% vs. 68.3%) followed by Proteus mirabilis 7.2% vs. 6.4%) and Klebsiella pneumoniae (5.4% vs. 5.2%). Enteroccocus spp. (4.7% vs. 6.8%), Streptoccocus agalactiae (1.7% vs. 3.1%) and Staphyloccocus saprophyticus (0.7% vs. 1.3%)were the most frequent Gram-positive pathogens. 31.3% of E. coli in 2002 and 32% in 2004 were susceptible to all antibiotics tested. Around 40% of E. coli were resistant to a single agent. 21.6-24.1% were resistant to two antibiotics. 35.4% of first period isolates, and 37.6% of second period ones were resistant to two or more classes of antibiotics. Fosfomycin (2.1- 2.8%) and nitrofurantoin (3.5-5.7%) had the lowest resistance rates for E. coli. Amoxicillin (58.2-58.7%), co-trimoxazole (30.8-33.8%) and ciprofloxacin (22.6-22.7%) showed the highest resistance rates, and their suitability as empiric treatments for UTI should probably be re-evaluated.

Molecular epidemiology of drug-resistant Salmonella Typhimurium in Spain.

One hundred and forty-seven Salmonella serotype Typhimurium strains isolated in three provinces in the midwest of Spain were studied. Of these, 93.6% were drug resistant. There were two predominant resistance phenotypes: 43 isolates (29.3%) were resistant to amoxicillin, tetracyclines, chloramphenicol, streptomycin and sulphamethoxazole and 27 isolates (18.4%) to amoxicillin, amoxicillin/clavulanic acid, tetracyclines, chloramphenicol, streptomycin and sulphamethoxazole. Randomly amplified polymorphic DNA (RAPD) analysis and pulsed field gel electrophoresis (PFGE) were performed for molecular typing. Thirty-six DNA band profiles were differentiated by RAPD, and 38 by PFGE. We found a high level of clonality; 27% of strains were identical by both methods. There were additional smaller clonal lines within every area. The highest discriminatory power was obtained with PFGE, but the greatest degree of genetic diversity was observed among Salmonella Typhimurium using both RAPD and PFGE.

[Prevalence of anti-Chlamydophila pneumoniae antibodies in patients with intrinsic asthma]. / Prevalencia de anticuerpos frente a Chlamydophila pneumoniae en pacientes con asma intrinseca.

Chlamydophila pneumoniae is a respiratory pathogen which has been involved in the pathogenesis of a number of chronic diseases. We studied the association between IgG antibodies against C. pneumoniae and intrinsic asthma in adults. C. pneumoniae IgG serum titers were determined by enzymatic immunoassay in 55 adult patients and 87 healthy controls. A significant association was found between anti-C. pneumoniae antibodies and intrinsic asthma, as compared to the control group (23.6% vs. 10.3%, p <0.05). C. pneumoniae may therefore be involved in the origin or in exacerbations of intrinsic asthma.

[Antimicrobial drug resistance in non-typhi Salmonellae in Castilla y Leon]. / Resistencia a antimicrobianos en Salmonella no typhi en Castilla y León.

We studied the antibiotic susceptibility of 309 Salmonella isolates obtained from three hospitals serving the provinces of Salamanca, Avila and Zamora in the region of Castilla y Leon (mid-west Spain). The susceptibility to 18 antibiotics was studied using the agar dilution method, according to NCCLS guidelines, and the most common multiresistance phenotypes were determined for each province. We observed clear susceptibility differences between the two main serotypes found, S. enteritidis and S. typhimurium. Seventy percent of S. typhimurium were resistant to amoxicillin. In 44% of these isolates, amoxicillin resistance was associated with resistance to streptomycin, sulfonamides, tetracyclines and chloramphenicol. S. enteritidis was susceptible to most antibiotics tested; amoxicillin resistance was observed in 23.3%, and nalidixic acid resistance in 49.6%. Resistance to nalidixic acid was higher in S. enteritidis than in any other serotypes. According to NCCLS breakpoints, no strain was resistant to fluoroquinolones. However, according to MENSURA criteria, 9% of S. typhimurium isolates were resistant to ciprofloxacin. Resistance to cotrimazole and gentamicin was less than 10% for all the serotypes tested. The results indicate that S. typhimurium showed greater resistance and a high multidrug resistance rate. Conversely, S. enteritidis showed high resistance only to amoxicillin and nalidixic acid, though in most cases there was no correlation between this resistance and reduced susceptibility to fluoroquinolones.

In vitro activity of newer antibiotics against Corynebacterium jeikeium, Corynebacterium amycolatum and Corynebacterium urealyticum.

The in vitro activity of ciprofloxacin, erythromycin, telithromycin, teicoplanin, linezolid and quinupristin-dalfopristin was tested against human derived pathogenic corynebacteria. The MICs of these antibiotics were measured using the agar dilution method against 31 strains of Corynebacterium jeikeium, 58 Corynebacterium amycolatum (including 33 multidrug-resistant strains) and 64 Corynebacterium urealyticum clinical strains. A high resistance rate to ciprofloxacin and erythromycin was found in the three species. Telithromycin was much more active than erythromycin (MIC(90) of erythromycin >or=128 mg/l for all three species; MIC(90) of telithromycin: 4 mg/l for C. jeikeium, 64 mg/l for C. amycolatum and 1 mg/l for C. urealyticum). There were no teicoplanin-resistant (MIC(90) 1, 0.5 and 1 mg/l, respectively) or linezolid-resistant strains (MIC(90) 1, 0.2 and 0.5 mg/l, respectively). Quinupristin-dalfopristin was active against most strains with an activity similar to linezolid, but three C. jeikeium and one C. amycolatum showed MICs >or=4 mg/l. Telithromycin showed much better activity against corynebacteria than older macrolides. Synercid and linezolid were active against most isolates tested, including multidrug resistant strains.

In vitro activity of linezolid, synercid and telithromycin against genetically defined high level fluoroquinolone-resistant methicillin-resistant Staphylococcus aureus.

The in vitro activity of levofloxacin, moxifloxacin, gatifloxacin, erythromycin, telithromycin, linezolid, synercid and vancomycin was measured against 36 genetically defined, gyrA/grlA double mutant MRSA clinical strains with an MIC to ciprofloxacin > or = 8 mg/l. The three newer fluoroquinolones tested were more active than ciprofloxacin. Resistance rates for levofloxacin and gatifloxacin were high (44.5 and 36.1%, respectively). All the strains were moxifloxacin-susceptible, though most of them had MICs close to the break point. All the strains were intermediate or resistant to erythromycin and most were also resistant to telithromycin. No strains were resistant to linezolid, synercid or vancomycin (MIC(90): 2, 1 and 2 mg/l, respectively).