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Introduction The under-utilisation of taps is associated with the generation of reservoirs of non-fermenting gram-negative bacilli with the ability to disseminate. We describe the detection and approach of the problem in an ICU. Methods Observational descriptive study in an ICU with individual cubicles with their own sink. We collected clinical samples from patients and environmental samples from tap aerators and reviewed the unit's hygiene measures. Results We detected four cases due to Chryseobacterium indologenes, one to Elizabethkingia meningoseptica and another to Pseudomonas aeruginosa; they were identified both in clinical and the environmental samples. The healthcare professionals reported that almost every hand hygiene opportunity was performed with a hydroalcoholic solution. After considered the daily flushing of water outlets as inefficient, it was decided to remove them. Conclusions National recommendations were insufficient for preventing, detecting and controlling tap contamination in units with a high risk of infection. The management of taps in these units needs to be improved.

Resumen Introduccion La infrautilizacion de grifos se relaciona con reservorios de bacilos gramnegativos no fermentadores en su interior con capacidad de diseminacion. Se describe la deteccion del problema y abordaje en una UCI. Metodos Estudio descriptivo en una UCI con boxes individuales con lavabo propio. Se recogieron muestras clinicas de pacientes y ambientales de los aireadores de los grifos. Se revisaron medidas de higiene. Resultados Se identificaron 4 casos de Chryseobacterium indologenes , uno de Elizabethkingia meningoseptica y otro por Pseudomonas aeruginosa , tanto en muestras clinicas como ambientales. Los profesionales indicaron utilizar solucion hidroalcoholica casi siempre en la higiene de manos. Tras descartar la apertura controlada diaria de grifos por ineficiente, se decidio su retirada. Conclusiones Las recomendaciones nacionales resultaron insuficientes para la prevencion, deteccion y control de la contaminacion de los grifos de una unidad de alto riesgo de infeccion. Es necesario mejorar el manejo de los grifos en estas unidades.

Background The development of sonication protocols over the last few years has improved the sensitivity of conventional cultures for the diagnosis of prosthetic-joint infection (PJI). However, the development of a new, specifically designed kit for the molecular diagnosis of PJI could provide a major improvement in this field. Methods Prostheses retrieved from patients who underwent implant removal from May 2014 to May 2015 were sent for culture, and processed according to a previously defined protocol that included sonication. Furthermore, 180 microlitres of sonication fluid were used to carry out the multiplex PCR test (Unyvero i60 system®). A comparison of the sensitivity, specificity, positive (PPV) and negative (NPV) predictive value, was performed. The study was approved by the Clinical Research Ethics Committee. Results The analysis included 88 prostheses from 68 patients (1.29 prostheses/patient). The type of prostheses studied were knee (n = 55), total hip (n = 26), partial hip (n = 5), and shoulder (n = 2). Twenty-nine patients were diagnosed with a PJI (15 delayed, 12 acute, and 2 haematogenous infections). In 24 cases, the result of the PCR was positive, all but 1 corresponding to patients with clinical criteria of PJI. Nine resistance mechanisms were detected from 5 samples. The Unyvero i60 system® showed slightly better results than traditional culture in terms of specificity and PPV. Conclusions The Unyvero i60 system® may play a role in rapid diagnosis of PJI, due to its high specificity and PPV. However, despite these results, cultures have to be performed to detect organisms not detected by the system.

The physiological factors that contribute to Mycobacterium abscessus lung infections remain unclear. We determined whether antibiotic treatment targeting a major cystic fibrosis pathogen (i.e., Pseudomonas aeruginosa ) could provide the ideal conditions for the establishment of M. abscessus infection. Our data showed that P. aeruginosa inhibited M. abscessus biofilm formation under control conditions and that antimicrobial therapy selectively targeting P. aeruginosa diminished this competitive interaction, thereby increasing M. abscessus survival.

Implant-related infection is a devastating complication in orthopedic surgery. Aiming to minimize this problem, many material modifications have been developed. Here we report a study of a surface modification of Ti-6 Al-4 V alloy using a methodology that enables the study of interactions between bacteria and the material in the presence of eukaryotic cells. We mixed different concentrations of collection or clinical strains of staphylococci isolated from implant-related infections with preosteoblastic cells using a previously published methodology, analyzing the minimal concentration of bacteria able to colonize the surface of the material through image analysis. Ti-6 Al-4 V alloy was modified by anodization to obtain two F-doped nanostructured surfaces that have been previously described to have antibacterial properties.

Despite many advances, the management of prosthetic joint infection is still a complex issue. Moreover, in recent years the problem of antimicrobial resistance has emerged as an important challenge.We analysed recent advances in different aspects of prosthetic joint infections. The importance of biofilms needs to be considered for antibiotic selection because, when embedded in these structures, bacteria acquire resistant behaviour. Moreover, the presence of resistance mechanisms in some species of organisms increases the difficulty of management. In this sense, the growing importance of methicillin-resistant staphylococci, multidrug-resistant Enterobacteriaceae or Pseudomonas aeruginosa is of increasing concern. Together with these organisms, others with constitutive resistance against most antibiotics (like Enterococcus sp., mycobacteria or fungi) represent a similar problem for selection of therapy. Research into new materials that can be used as drug carriers opens a new field for management of these infections and will likely come to the front line in the coming years.Individualised therapies should carefully consider the aetiology, pathogenesis and antimicrobial susceptibility. Satisfactory clinical outcome could be further fostered by enhancing the multidisciplinary approach, with better collaboration in the antibiotic selection and the surgical management.

Lung disease in cystic fibrosis (CF) is characterized by the progressive colonization of the respiratory tract by different bacteria, which develop polymicrobial biofilms. In the past decades, there has been an increase in the number of CF patients infected with Non-Tuberculous Mycobacteria (NTM). Although Mycobacterium abscessus is the main NTM isolated globally, little is known about M. abscessus multispecies biofilm formation. In the present study we developed an in vitro model to study the phenotypic characteristics of biofilms formed by M. abscessus and Pseudomonas aeruginosa, a major pathogen in CF. For that purpose, dual species biofilms were grown on polycarbonate membranes with a fixed concentration of P. aeruginosa and different inoculums of M. abscessus. The biofilms were sampled at 24, 48, and 72 h and bacteria were quantified in specific media. The results revealed that the increasing initial concentration of M. abscessus in dual species biofilms had an effect on its population only at 24 and 48 h, whereas P. aeruginosa was not affected by the different concentrations used of M. abscessus. Time elapsed increased biofilm formation of both species, specially between 24 and 48 h. According to the results, the conditions to produce a mature dual species biofilm in which the relative species distribution remained stable were 72 h growth of the mixed microbial culture at a 1:1 ratio. A significant decrease in mycobacterial population in dual compared to single species biofilms was found, suggesting that P. aeruginosa has a negative influence on M. abscessus. Finally, in a proof of concept experiment, young and mature dual species biofilms were exposed to clarithromycin.

The aim of this study was to evaluate the proliferation and mineralization ability of mouse osteoblastic MC3T3-E1 cells on F-containing TiO2 films with different morphology and nanostructure that previously confirmed antibacterial properties. F-containing TiO2 films were fabricated by anodizing Ti-6Al-4V alloy ELI -grade 23. By using a mixture of H2SO4/HF acid at 20 V for 5 and 60 min, a TiO2 film grows with nanoporous (NP) and nanotubular (NT) features, characterized with a pore diameter of 20 and 100 nm, respectively. Fluoride-TiO2 barrier films (FBL) were produced in 1M NH4H2PO4/0.15M NH4F solution at constant voltage controlled at 20 V for 120 min. The amount of F incorporated in the nanostructured oxide films was 6 at % and of 4 at %, for the NP and NT, respectively, while for the FBL film was 12 at %. MC3T3-E1 cells exhibited different behavior when seeded and grown onto these surfaces. Thus, F-doped TiO2 films with NP structures increased proliferation as well as osteogenic gene expression and the mineralization capacity of these osteoblastic cells. These results confirm that anodizing process is suitable to fabricate multifunctional surfaces on Ti-6Al-4V alloy with improved not only antibacterial but also osteogenic properties useful for bone fixation of prosthetic devices

Orthopaedic device-related infections are closely linked to biofilm formation on the surfaces of these devices. Several modified titanium (Ti-6Al-4V) surfaces doped with fluorine were studied in order to evaluate the influence of these modifications on biofilm formation by Gram-positive and Gram-negative bacteria as well as a yeast. The biofilm studies were performed according to the standard test method approved by ASTM (Designation: E2196-12) using the Rotating Disk Reactor. Four types of Ti-6Al-4V samples were tested; chemically polished (CP), two types of nanostructures containing fluorine, nanoporous (NP) and nanotubular (NT), and non-nanostructured fluorine containing samples (fluoride barrier layers, FBL). Different species of Gram-positive cocci, (Staphylococcus aureus and epidermidis), Gram-negative rods (Escherichia coli, Pseudomonas aeruginosa), and a yeast (Candida albicans) were studied. For one of the Gram-positive (S. epidermidis) and one of the Gram-negative (E. coli) species a statistically-significant decrease in biofilm accumulation for NP and NT samples was found when compared with the biofilm accumulation on CP samples. The results suggest an effect of the modified materials on the biofilm formation. Graphical Abstract: [InlineMediaObject not available: see fulltext.], This work was funded by the following grants from the Spanish MINECO (MAT2013-48224-C2-2-R and MAT2013-48224-C2-1-R). Pérez-Jorge C wishes to thank J William Costerton and the Medical Biofilm Laboratory at the Center for Bio-film Engineering in Montana, USA, for the opportunity to be a visiting researcher and the National Spanish Society of Infection Diseases and Microbiology (SEIMC) for funding with a Mobility grant 2012. Hernández-López J.M. is receptor of the JAE-predoctoral grant funded by the CSIC.

Staphylococcus aureus and Staphylococcus epidermidis are human pathogens involved in implant-related infections. During those diseases, they are able to form biofilms showing resistance to the effect of many different antibiotics. Drug delivery systems allow a local and effective delivery of antibiotics at high concentrations in the infected tissue without causing the cytotoxic effects commonly linked to systemic administration. We report the use of a porous ceramic biomaterial, such as SBA-15 loaded with antibiotics, to deliver them directly to the infected tissue. SBA-15 discs were loaded with Vancomycin, Rifampin and a combination of both, introduced in a suspension of S. aureus 15981 and S. epidermidis ATCC 35984 and incubated during 6 and 24 h. A statistically significant decrease in the biofilm density and the number of viable bacteria was detected for all antibiotics at 6 h in both bacteria. Rifampin showed an increase in the biofilm density and the number of viable bacteria at 24 h. No differences were detected between Vancomycin and the combination of antibiotics. S. epidermidis was more sensitive to the effect of the antibiotics than S. aureus. Here we have demonstrated that SBA-15 is able to act as an effective drug delivery system not only from a pharmaceutical point of view, but also from a biological one.

The aim of this study was to compare the bacterial adhesion of Staphylococcus spp. on Ti-6Al-4V with respect to Ti-6Al-V modified alloys with a set of Cubic yttria stabilized zirconia (YSZ) and Ag-YSZ nanocomposite films. Silver is well known to have a natural biocidal character and its presence in the surface predicted to enhance the antimicrobial properties of biomedical surfaces. Microbial adhesion tests were performed using collection strains and twelve clinical strains of Staphylococcus aureus and Staphylococcus epidermidis. The adherence study was performed using a previously published protocol by Kinnari et al. Both collection strains and clinical isolates have shown lower bacterial adhesion to materials modified with respect to the alloy Ti-6Al-4V and the modification with silver reduced the bacterial adhesion for most of all the strains studied. Moreover the percentage of dead bacteria have been evaluated, demonstrating increased proportion of dead bacteria for the modified surfaces. Nanocrystalline silver dissolves releasing both Ag+ and Ag-0 whereas other silver sources release only Ag+. We can conclude that YSZ with nanocrystalline silver coating may lead to diminished postoperative infections and to increased corrosion and scratch resistance of YSZ incorporating alloys Ti-6Al-4V.

© 2015 Elsevier B.V. All rights reserved. The use of osteoarticular implants has improved the quality of life of millions of patients. In this work nanotubular structures tailored made on Ti6Al4 V substrates was used as drug delivery system of ibuprofen as a proof of concept. Three different nanotubular films with different sizes and forms (NT, NT+ and NTb) were analysed. Samples were soaked in a solution of 660 mg ibuprofen/20 mL n-pentane. The ibuprofen release in aqueous medium was evaluated by liquid chromatography reversed-phase (RP-HPLC). To calculate the observed constant k, the amount of ibuprofen released was plotted versus the time using linear regression according to the zero-order, first-order, second-order and Higuchi model. The release of ibuprofen was constant and independent of the concentration. The kinetic constant obtained was 0.021 (NT), 0.022 (NT+) and 0.013 (NTb) being the correlation factor of 0.98 (zero-order) where the maximum correlation factor was reached. These results indicate that the delivery process from NT and NT+ is similar and slower that NTb. In all the cases was inside the therapeutically range. These results showed the potential of these modifications in order to develop implants that can carry different molecules of medical importance., This work was funded by the grants from the Spanish MICINN (CS02010-11384-E, MAT2012-35556) and MINECO (MAT2013-48224-C2-2-R and MAT2013-48224-C2-1-R).

Bacterial colonization and biofilm formation on orthopedic implants is one of the worst scenarios in orthopedic surgery, in terms of both patient prognosis and healthcare costs. Tailoring the surfaces of implants at the nanoscale to actively promote bone bonding while avoiding bacterial colonization represents an interesting challenge to achieving better clinical outcomes. Herein, a Ti6Al4V alloy of medical grade has been coated with Ti nanostructures employing the glancing angle deposition technique by magnetron sputtering. The resulting surfaces have a high density of nanocolumnar structures, which exhibit strongly impaired bacterial adhesion that inhibits biofilm formation, while osteoblasts exhibit good cell response with similar behavior to the initial substrates. These results are discussed on the basis of a >lotus leaf effect> induced by the surface nanostructures and the different sizes and biological characteristics of osteoblasts and Staphylococcus aureus., This study was supported by research Grants from the Ministerio de Ciencia e Innovación (MICINN) – Spain through the projects MAT2012-35556 and CSO2010-11384-E (Agening Network of Excellence), the Spanish MINECO (Projects CONSOLIDER CSD2008-00023, MAT2011-29194-C02-01, MAT2013-40852-R and MAT2013-43299-R) and the Junta de Andalucía (projects P12-FQM-2265 and P10-FQM-6900).

Bacterial adherence is a complex process that involves different phases, which end with bacteria being firmly adhered to a material’s surface. Different mechanisms are involved in this process, including different molecules in the bacterial cell wall, and also different properties of the material. These latter properties can be modified to minimize bacterial adhesion, a strategy that is currently being developed. In this chapter we review the currently available data on bacterial adherence, and describe some of the strategies used to overcome this phenomenon.

The treatment of Chagas disease is limited by the frequent cutaneous side effects of benznidazole. We tested the use of steroids plus escalating doses of benznidazole to prevent this complication in 17 adult patients with chronic Trypanosoma cruzi infection and found extremely good tolerance. A randomized trial is warranted.

Ti-6Al-4V joint replacement implants foster uncemented fixation in orthopaedic surgery. However, bacterial colonization competes with host cells and ultimately may produce implant-related difficult-to-treat infections, justifying the efforts to obtain infection-resistant materials. In a previous work, the authors demonstrated the antibacterial properties of anodic fluoride-TiO2 nanostructured layers on Ti-6Al-4V alloy. In this work, the anodizing bath has been modified in order to grow fluoride-TiO2 barrier layers (FBL). A bacterial adherence protocol, run with reference and six different clinical strains of Staphylococcus aureus and Staphylococcus epidermidis, showed a statistically significant decrease in the percentage of covered surface (p

Diagnosis of Prosthetic Joint Infection is still a complex matter in clinical practice. Syndromic diagnosis includes the presence of several symptoms and signs together with the use of a combination of different techniques, including image analysis, biochemical markers and intraoperatory histopatology. Microbiological diagnosis is an essential part of this process, because it allows selection of the best possible antibiotic therapy, and even orients the surgical management and the potential outcome of the patient in some cases. Cultures of synovial fluids, periprosthetic tissues and implant sonication are the commonly recommended techniques, and a combination of these techniques allows reaching an etiologic diagnosis in most cases. However, rapid diagnosis is still a problem, because the lack of sensitivity and specificity of Gram stain of surgical products.

The aim of this study was to evaluate Staphylococcus sp. adhesion to modified surfaces of titanium alloy (Ti-6Al-4V). Specimens of Ti-6Al-4V alloy 6-4 ELI-grade 23 that meets the requirements of ASTM F136 2002A (AMS 2631B class A1) were anodized in a mixture of sulfuric/hydrofluoric acid at 20 V for 5 and 60 min to form nanoporous (NP) and nanotubular (NT) oxide layers with pore diameter of 20 and 100 nm, respectively. The amount of fluorine incorporated in the oxide films from the electrolyte was 6 and 4 wt %, respectively. Bacterial adherence was studied using laboratory strains and six clinical strains each of Staphylococcus aureus and Staphylococcus epidermidis. Lower adherence of laboratory strains was demonstrated on fluoride nanostructured surfaces in comparison with the fluoride-free surfaces. Significant differences between clinical strains and laboratory strains were also found (p < 0.0001, Kruskal-Wallis test) when NP and NT specimens were compared with chemically polished (CP) surfaces. The results of the tests using multiple clinical strains confirmed a decrease in bacterial adherence on F-containing titanium oxide surfaces, suggesting a potential applicability of this surface, with a confirmed added value of decreasing clinical staphylococci adherence, for medical prosthetic devices. © 2012 Wiley Periodicals, Inc.

Spread of cfr gene among staphylococci conferring resistance to linezolid in a patient under treatment

The aim of this study was to evaluate Staphylococcus sp. adhesion to modified surfaces of titanium alloy (Ti-6Al-4V). Specimens of Ti-6Al-4V alloy 6-4 ELI-grade 23 that meets the requirements of ASTM F136 2002A (AMS 2631B class A1) were anodized in a mixture of sulfuric/hydrofluoric acid at 20 V for 5 and 60 min to form nanoporous (NP) and nanotubular (NT) oxide layers with pore diameter of 20 and 100 nm, respectively. The amount of fluorine incorporated in the oxide films from the electrolyte was 6 and 4 wt %, respectively. Bacterial adherence was studied using laboratory strains and six clinical strains each of Staphylococcus aureus and Staphylococcus epidermidis. Lower adherence of laboratory strains was demonstrated on fluoride nanostructured surfaces in comparison with the fluoride-free surfaces. Significant differences between clinical strains and laboratory strains were also found (p0.0001, Kruskal-Wallis test) when NP and NT specimens were compared with chemically polished (CP) surfaces. The results of the tests using multiple clinical strains confirmed a decrease in bacterial adherence on F-containing titanium oxide surfaces, suggesting a potential applicability of this surface, with a confirmed added value of decreasing clinical staphylococci adherence, for medical prosthetic devices.

Background To analyze the incidence and clinical-microbiological characteristics of osteomyelitis (OM) in a tertiary Spanish hospital. Methods All cases diagnosed with OM between January 2007 and December 2010 were retrospectively reviewed. The variables examined include epidemiological characteristics, risk factors, affected bone, radiographic changes, histology, microbiological culture results, antibiotic treatment, and the need for surgery. Results Sixty-three cases of OM were diagnosed. Twenty-six patients (41.3%) had acute OM whereas 37 patients (58.7%) were classified as chronic OM. OM may result from haematogenous or contiguous microbial seeding. In this group, 49 patients (77.8%) presented with OM secondary to a contiguous source of infection and 14 patients had hematogenous OM (22.2%). Staphylococcus aureus was the most commonly found microorganism. Conclusions OM mainly affected patients with risk factors related to the presence of vascular diseases. Antibiotic treatment must be guided by susceptibility patterns of individual microorganisms, although it must be performed together with surgery in most of the cases.