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Osteomyelitis is an inflammatory process of bone and bone marrow that may even lead to patient death. Even though this disease is mainly caused by Gram-positive organisms, the proportion of bone infections caused by Gram-negative bacteria, such as Escherichia coli, has significantly increased in recent years. In this work, mesoporous silica nanoparticles have been employed as platform to engineer a nanomedicine able to eradicate E. coli- related bone infections. For that purpose, the nanoparticles have been loaded with moxifloxacin and further functionalized with Arabic gum and colistin (AG+CO-coated MX-loaded MSNs). The nanosystem demonstrated high affinity toward E. coli biofilm matrix, thanks to AG coating, and marked antibacterial effect because of the bactericidal effect of moxifloxacin and the disaggregating effect of colistin. AG+CO-coated MX-loaded MSNs were able to eradicate the infection developed on a trabecular bone in vitro and showed pronounced antibacterial efficacy in vivo against an osteomyelitis provoked by E. coli. Furthermore, AG+CO-coated MX-loaded MSNs were shown to be essentially non-cytotoxic with only slight effect on cell proliferation and mild hepatotoxicity, which might be attributed to the nature of both antibiotics. In view of these results, these nanoparticles may be considered as a promising treatment for bone infections caused by enterobacteria, such as E. coli, and introduce a general strategy against bone infections based on the implementation of antibiotics with different but complementary activity into a single nanocarrier. Statement of significance: In this work, we propose a methodology to address E.coli bone infections by using moxifloxacin-loaded mesoporous silica nanoparticles coated with Arabic gum containing colistin (AG+CO-coated MX-loaded MSNs). The in vitro evaluation of this nanosystem demonstrated high affinity toward E. coli biofilm matrix thanks to the Arabic gum coating, a disaggregating and antibacterial effect of colistin, and a remarkable antibiofilm action because of the bactericidal ability of moxifloxacin and colistin. This anti-E. coli capacity of AG+CO-coated MX-loaded MSNs was brought out in an in vivo rabbit model of osteomyelitis where the nanosystem was able to eradicate more than 90% of the bacterial load within the infected bone, Financial support from the Eu-ropean Research Council through ERC-2015-AdG-694160 (VERDI) grant. AM is funded by grants from Instituto de Salud Carlos III through the “Miguel Servet” program (CP15/00053)

This study was designed to propose alternative therapeutic compounds to fight against bacterial pathogens. Thus, a library of nitrogen-based compounds bis(triazolyl)methane (1T–7T) and bis(pyrazolyl)methane (1P–11P) was synthesised following previously reported methodologies and their antibacterial activity was tested using the collection strains of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Moreover, the novel compound 2P was fully characterized by IR, UV–Vis and NMR spectroscopy. To evaluate antibacterial activity, minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), minimum biofilm inhibitory concentrations (MBICs), and minimum biofilm eradication concentrations (MBECs) assays were carried out at different concentrations (2–2000 µg/mL). The MTT assay and Resazurin viability assays were performed in both human liver carcinoma HepG2 and human colorectal adenocarcinoma Caco-2 cell lines at 48 h. Of all the synthesised compounds, 2P had an inhibitory effect on Gram-positive strains, especially against S. aureus. The MIC and MBC of 2P were 62.5 and 2000 µg/mL against S. aureus, and 250 and 2000 µg/mL against E. faecalis, respectively. However, these values were > 2000 µg/mL against E. coli and P. aeruginosa. In addition, the MBICs and MBECs of 2P against S. aureus were 125 and > 2000 µg/mL, respectively, whereas these values were > 2000 µg/mL against E. faecalis, E. coli, and P. aeruginosa. On the other hand, concentrations up to 250 µg/mL of 2P were non-toxic doses for eukaryotic cell cultures. Thus, according to the obtained results, the 2P nitrogen-based compound showed a promising anti-Gram-positive effect (especially against S. aureus) both on planktonic state and biofilm, at non-toxic concentrations.

Prosthetic joint infections (PJIs) are typically caused by microorganisms that grow in biofilms. Traditional antimicrobial susceptibility tests are based on the study of planktonic bacteria that might lead to missing the biofilm behavior and to a treatment failure. This study was designed to analyze the antimicrobial susceptibility of clinical Gram-negative bacilli (GNB) isolates from PJIs in planktonic and sessile states and the possible relationship between antimicrobial resistance and biofilm formation. A total of 46 clinical isolates from patients with PJIs (mainly hip and knee prostheses) plus three GNB ATCC isolates were studied. The Minimal Inhibitory Concentration (MIC), minimal bactericidal concentration (MBC), minimal biofilm inhibitory concentration (MBIC), and minimal biofilm eradication concentration (MBEC) were assessed using a previously published methodology. Almost all of the GNB clinical isolates tested were biofilm forming. Pseudomonas aeruginosa was the largest biofilm-forming species. A comparison of MBIC

Non-tuberculous Mycobacteria (NTM) are a group of organisms whose importance in medicine seems to be increasing in recent times. The increasing number of patients susceptible to these diseases make it necessary to expand our knowledge of therapeutic options and to explore future possibilities for the development of a therapeutic arsenal.In this review, the authors provide a brief introduction about the present importance of NTM and describe the present recommendations of the available guidelines for their treatment. They include a description of the future options for the management of these patients, especially focusing on new antibiotics. The authors also look at possibilities for future therapeutic options, such as antibiofilm strategies.No actual changes have been made to the current recommendations for the management of most NTM infections (except perhaps the availability of nebulized amikacin). However, it is also true that we have increased the number of available antibiotic treatment options with good in vitro activity against NTM. The use of these drugs in selected cases could increase the therapeutic possibilities. However, some problems are still present, such as the knowledge of the actual meaning of a NTM isolate, and will probably be a key part of future research.

Background: Debridement, antibiotic agents, and implant retention (DAIR) is a currently accepted approach for the treatment of early prosthetic joint infections (PJI). The success of a DAIR procedure has shown variable results throughout the published literature. Scoring systems such as the Kidney, Liver, Index surgery, Cemented prosthesis, and C-reactive protein value (KLIC) score for the selection of patients that are likely to benefit from DAIR have proved to be helpful in decision making. Our study aims to further validate the KLIC score using a large external multicentric cohort and to evaluate other risk factors for failure. Patients and Methods: A retrospective analysis of patients with an early acute PJI who were treated with DAIR and recorded in a database of eight Spanish university hospitals was performed. According to pre-operative variables of the KLIC study, patients were categorized into five groups: group A, =2 points; group B, 2.5-3.5 points; group C, 4-5 points; group D, 5.5-6.5 points; and group E, =7 points. Failure rates were compared between groups at 60 days and after 60 days of DAIR. Further variables for risk of failure were also analyzed. Results: A total of 455 patients with early acute PJI were included in the analyses. At 60 days, patients presenting with pre-operative elevated C-reactive protein serum levels, Staphylococcus aureus, and polymicrobial infections were associated with failure. Failure rates recorded were 12% for group A (n = 210), 18% for group B (n = 83), 26% for group C (n = 89), 24% for group D (n = 66), and 0% for group E (n = 7). Univariable analysis between consecutive groups of the KLIC score showed no differences for failure before 60 days of the DAIR procedure. Scheduled surgery and having the procedure performed by a specialized unit were also identified as important factors for DAIR success. Conclusions: Our results suggest the KLIC score was not useful for predicting failure in our cohort. Furthermore, our results indicate a specialized unit should conduct DAIR procedures.

Methylobacterium sp. is isolated from water distribution systems and has been linked in the biofilms of the systems with a lower presence of Mycobacterium avium. In this study we aimed to determine the in vitro activity of Methylobacterium sp. in the development of rapidly growing mycobacteria (RGM) biofilms. Methylobacterium sp. CECT 7805 was added as a suspension of living bacteria (LB), an autoclaved suspension (AS), and an extract obtained after sonication (ES) at different times (24, 48, and 72 h), to preformed biofilms of Mycobacterium abscessus DSM 44196, Mycobacterium chelonae ATCC 19235, and Mycobacterium fortuitum ATCC 6841, using a 96 h control of each species. The biofilms were analyzed by confocal laser scanning microscopy and by the Calgary biofilm device using the plates MBECTM Biofilm Inoculator. A statistically significant reduction in the thickness and covered surface was observed in all mycobacterial biofilms with all forms of Methylobacterium sp. A statistically significant increase in the autofluorescence was observed in M. abscessus biofilms but not in other biofilms. The increased percentage of dead mycobacteria was statistically significant in all cases. The reduced log CFU (colony-forming units)/peg recount was statistically significant in M. chelonae biofilms after treatment with AS and ES, but in M. fortuitum biofilms the recount decreased only with AS. M. abscessus biofilms were always significantly reduced with AS at 72 h and with ES. Methylobacterium sp. could inhibit RGM biofilm formation. Living cells of Methylobacterium sp. were not necessary to inhibit the growth of a preformed biofilm. M. chelonae biofilms were the most greatly reduced.

Background Cutibacterium species are common pathogens in periprosthetic joint infections (PJI). These infections are often treated with β-lactams or clindamycin as monotherapy, or in combination with rifampin. Clinical evidence supporting the value of adding rifampin for treatment of Cutibacterium PJI is lacking. Methods In this multicenter retrospective study, we evaluated patients with Cutibacterium PJI and a minimal follow-up of 12 months. The primary endpoint was clinical success, defined by the absence of infection relapse or new infection. We used Fisher’s exact tests and Cox proportional hazards models to analyze the effect of rifampin and other factors on clinical success after PJI. Results We included 187 patients (72.2% male, median age 67 years) with a median follow-up of 36 months. The surgical intervention was a 2-stage exchange in 95 (50.8%), 1-stage exchange in 51 (27.3%), debridement and implant retention (DAIR) in 34 (18.2%), and explantation without reimplantation in 7 (3.7%) patients. Rifampin was included in the antibiotic regimen in 81 (43.3%) cases. Infection relapse occurred in 28 (15.0%), and new infection in 13 (7.0%) cases. In the time-to-event analysis, DAIR (adjusted hazard ratio [HR] = 2.15, P = .03) and antibiotic treatment over 6 weeks (adjusted HR = 0.29, P = .0002) significantly influenced treatment failure. We observed a tentative evidence for a beneficial effect of adding rifampin to the antibiotic treatment—though not statistically significant for treatment failure (adjusted HR = 0.5, P = .07) and not for relapses (adjusted HR = 0.5, P = .10). Conclusions We conclude that a rifampin combination is not markedly superior in Cutibacterium PJI, but a dedicated prospective multicenter study is needed.

Prosthetic joint infection (PJI) is one of the most devastating complications in orthopedic surgery. One approach used to prevent PJI is local antibiotic therapy. This study evaluates the antibiotic release, in vitro cytocompatibility and in vivo effectiveness in preventing PJI caused by Staphylococcus aureus (S. aureus) of the fluorine‐ and phosphorus‐doped, bottle‐shaped, nanostructured (bNT) Ti–6Al–4V alloy loaded with a mixture of gentamicin and vancomycin (GV). We evaluated bNT Ti–6Al–4V loading with a mixture of GV, measuring the release of these antibiotics using high‐performance liquid chromatography. Further, we describe bNT Ti–6Al–4V GV cytocompatibility and its efficacy against S. aureus using an in vivo rabbit model. GV was released from bNT Ti–6Al–4V following a Boltzmann non‐linear model and maximum release values were obtained at 240 min for both antibiotics. The cell proliferation of MCT3T3‐E1 osteoblastic cells significantly increased at 48 (28%) and 168 h (68%), as did the matrix mineralization (52%) of these cells and the gene expression of three of the most important markers related to bone differentiation (more than threefold for VEGF and BGLAP, and 65% for RunX) on bNT Ti–6Al–4V GV compared with control. In vivo study results show that bNT Ti–6Al–4V GV can prevent S. aureus PJI according to histopathological and microbiological results. According to our results, bNT Ti–6Al–4V loaded with a mixture of GV using the soaking method is a promising biomaterial with favorable cytocompatibility and osteointegration, demonstrating local bactericidal properties against S. aureus., Secretaría de Estado de Investigación, Desarrollo e Innovación. Grant Numbers: BES‐2014‐069007, MAT2013‐48224‐C2‐1‐R, MAT2013‐48224‐C2‐2‐R, MAT2017‐86163‐C2‐1‐R

Septic arthritis and prosthetic joint infection (PJI) are conditions commonly associated with Gram-positive cocci, however, a drastic increase in cases derived from enterobacterial species has recently been observed. Recently it has been reported by multiple groups that staphylococci rapidly form free-floating aggregates in the presence of synovial fluid. These aggregates are comparatively more resistant to antimicrobial challenge than their planktonic counterparts, and thus may play a role in the pathogenesis of joint infection. While staphylococcal aggregates have been the primary focus of interest in the field, it is unclear just how widespread synovial fluid mediated aggregation (SFMA) is in Gram negative enterobacteria (GNE). Through this work we have evaluated SFMA in clinical GNE isolated from PJIs. Two PJI clinical strains each of Enterobacter cloacae, Escherichia coli, Klebsiella pneumonia and Proteus mirabilis strains representing a range of antibiotic susceptibilities were exposed to 10% bovine synovial fluid supernatant (BSF) using a relatively simple, quick semi-quantitative method using an imaging plate reader. BSF stimulated aggregation within 0.5 hr both strains of E. cloacae and P. mirabilis and one strain of E.coli. In one strain of P. mirabilis and E.coli, the size of the aggregates significantly increased from 0.5 to 2 hr exposure. In contrast, neither K. pneumoniae strain aggregated in BSF. These preliminary findings show that aggregation can occur quickly in GNE, but the extent appears strain and species specific. Further work is required to assess the impact of SFMA on antibiotic tolerance, host innate immunity and the establishment of biofilms.

Catheter-related bloodstream infections (CRBSI) constitute an important cause of hospital-acquired infection associated with morbidity, mortality, and cost. The aim of these guidelines is to provide updated recommendations for the diagnosis and management of CRBSI in adults. Prevention of CRBSI is excluded. Experts in the field were designated by the two participating Societies (the Spanish Society of Infectious Diseases and Clinical Microbiology and [SEIMC] and the Spanish Society of Spanish Society of Intensive and Critical Care Medicine and Coronary Units [SEMICYUC]). Short-term peripheral venous catheters, non-tunneled and long-term central venous catheters, tunneled catheters and hemodialysis catheters are covered by these guidelines. The panel identified 39 key topics that were formulated in accordance with the PICO format. The strength of the recommendations and quality of the evidence were graded in accordance with ESCMID guidelines. Recommendations are made for the diagnosis of CRBSI with and without catheter removal and of tunnel infection. The document establishes the clinical situations in which a conservative diagnosis of CRBSI (diagnosis without catheter removal) is feasible. Recommendations are also made regarding empirical therapy, pathogen-specific treatment (coagulase-negative staphylococci, Staphylococcus aureus, Enterococcus spp., Gram-negative bacilli, and Candida spp.), antibiotic lock therapy, diagnosis and management of suppurative thrombophlebitis and local complications.

Background.: Streptococci are not an infrequent cause of periprosthetic joint infection (PJI). Management by debridement, antibiotics, and implant retention (DAIR) is thought to produce a good prognosis, but little is known about the real likelihood of success.Methods.: A retrospective, observational, multicenter, international study was performed during 2003-2012. Eligible patients had a streptococcal PJI that was managed with DAIR. The primary endpoint was failure, defined as death related to infection, relapse/persistence of infection, or the need for salvage therapy.Results.: Overall, 462 cases were included (median age 72 years, 50% men). The most frequent species was Streptococcus agalactiae (34%), and 52% of all cases were hematogenous. Antibiotic treatment was primarily using β-lactams, and 37% of patients received rifampin. Outcomes were evaluable in 444 patients: failure occurred in 187 (42.1%; 95% confidence interval, 37.5%-46.7%) after a median of 62 days from debridement; patients without failure were followed up for a median of 802 days. Independent predictors (hazard ratios) of failure were rheumatoid arthritis (2.36), late post-surgical infection (2.20), and bacteremia (1.69). Independent predictors of success were exchange of removable components (0.60), early use of rifampin (0.98 per day of treatment within the first 30 days), and long treatments (≥21 days) with β-lactams, either as monotherapy (0.48) or in combination with rifampin (0.34).Conclusions.: This is the largest series to our knowledge of streptococcal PJI managed by DAIR, showing a worse prognosis than previously reported. The beneficial effects of exchanging the removable components and of β-lactams are confirmed and maybe also a potential benefit from adding rifampin.

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.

Joint prosthesis failure is mainly related to aseptic loosening and prosthetic joint infections, both of which are associated with high morbidity and substantial costs for patients and health systems. The development of a biomaterial that is capable of stimulating bone growth while minimizing bacterial adhesion would reduce the incidence of prosthetic failure. We report antibacterial and osteostimulatory effects in a novel fluorine-phosphorus (F-P)-doped TiO2 oxide film grown on Ti-6Al-4V alloy with a nanostructure of bottle-shaped nanotubes (bNT) using five bacterial species (Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia) and MCT3T3-E1 osteoblastic cells. The interaction between the bacteria and bNT Ti-6Al-4V was complex, as the adhesion of four bacterial species decreased (two staphylococcus species, E. coli, and S. maltophilia), and the viability of staphylococci and S. maltophilia also decreased because of the aluminum (Al) released by bNT Ti-6Al-4V. This released Al can be recruited by the bacteria through siderophores and was retained only by the Gram-negative bacteria tested. P. aeruginosa showed higher adhesion on bNT Ti-6Al-4V than on chemically polished (CP) samples of Ti-6Al-4V alloy and an ability to mobilize Al from bNT Ti-6Al-4V. The cell adhesion and proliferation of MCT3T3-E1 osteoblastic cells significantly increased at 48 and 168 h, as did the matrix mineralization of these cells and the gene expression levels of three of the most important markers related to bone differentiation. According to our results, the bNT Ti-6Al-4V alloy could have clinical application, preventing infection and stimulating bone growth and thus preventing the two main causes of joint prosthesis failure.IMPORTANCE This work evaluates F-P-doped bNT Ti-6Al-4V from microbiological and cellular approaches. The bacterial results highlight that the antibacterial ability of bNT Ti-6Al-4V is the result of a combination of antiadhesive and bactericidal effects exerted by Al released from the alloy. The cell results highlight that F-P bNT Ti-6Al-4V alloy increases osseointegration due to modification of the chemical composition of the alloy resulting from P incorporation and not due to the nanostructure, as reported previously. A key finding was the detection of Al release from inside the bNT Ti-6Al-4V nanostructures, a result of the nanostructure growth during the anodizing process that is in part responsible for its bactericidal effect., This work was funded by a grant from the Spanish Ministry of Economics and Competitiveness (MAT2013-48224-C2-2-R and MAT2013-48224-C2-1-R). J.-J.A.-C. is funded by an FPI grant from the same institution (BES-2014-069007). We acknowledge Oliver Shaw for his help with the English language and Isabelle Schalk for providing the P. aeruginosa ATCC 15692 ΔpchA and ΔpvdF ΔpchA mutants.

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.

In artificial metallic joint implants, the failure is provoked by two effects in most of the cases: mass loss and wear debris removed due to tribological-corrosive effects on the implant alloy, and infections due to the presence of bacteria. In this work, several Ti-C-N corrosion and wear protective coatings were developed by Physical Vapour Deposition technology, and deposited on Ti6Al4V alloy. In order to provide the implant antibacterial properties, an additional silver top coating has been deposited. Tribological behavior was characterized through tribocorrosion and fretting tests. On the other hand, wettability tests were performed to study the grade of hydrophilicity/hydrophobia. Additionally, antibacterial properties were evaluated by means of bacterial adhesion tests. As a result of these characterization studies, the coating with the best performance was selected. The as-coated material includes excellent protection against tribocorrosion and fretting effects (in relation to the uncoated one) and the silver layer has been confirmed to exhibit antibacterial properties.

We analyze the effect of amikacin, ciprofloxacin, and clarithromycin, alone and associated with N-acetylcysteine (NAC) and Tween 80, at different times and concentrations in nonpigmented rapidly growing mycobacteria (NPRGM) biofilms. For this purpose, confocal laser scanning microscopy and image analysis were used to study the development and behavior of intrinsic autofluorescence, covered area, thickness, and cell viability in NPRGM biofilms after adding antibiotics alone and associated with antibiofilm agents. In this study, ciprofloxacin is the most active antibiotic against this type of biofilm and thickness is the most affected parameter. NAC and Tween 80 combined with antibiotics exert a synergistic effect in increasing the percentage of dead bacteria and also reducing the percentage of covered surface and thickness of NPRGM biofilms. Tween 80 seems to be an antibiofilm agent more effective than NAC due to its higher reduction in the percentage of cover surface and thickness. In conclusion, the results obtained in this work show that phenotypic parameters (thickness, percentage of covered surface, autofluorescence, percentage of live/dead bacteria) are affected by combining antibiotics and antibiofilm agents, ciprofloxacin and Tween 80 being the most active agents against NPRGM biofilms.

We aimed to evaluate the release of two antibiotics: gentamicin and vancomycin loaded into F-doped nanotubular anodic oxide layers, as well as their bactericide effect. F-doped nanotubular oxide layers fabricated on Ti-6Al-4V loaded with gentamicin (Gm), vancomycin (Vm) and their mixture (Gm + Vm) by a previously described loading method. Antibiotic release was studied by RP-HPLC and by a biological method. Bactericidal activity was evaluated by a bacterial adherence protocol described previously using on three clinically important bacterial species. The antibiotic release steady up to 120 and 180 min for Gm and Vm, respectively, and despite the antibiotic concentration decreased, their biological activity was maintained over time. The number of living bacteria of three species tested on NT-Gm specimens was significantly lower than on NT specimens without antibiotics (P < 0.01). There are significant differences among NT-Gm and NT-Gm + Vm specimens (P < 0.05) for S. aureus 15981, S. epidermidis ATCC 35984, and P. aeruginosa ATCC 27853 and no differences between NT-Vm and NT-Gm + Vm for staphylococci (P > 0.05). In conclusion, this Gm + Vm loading method added to the properties of F-doped nanotubular oxide layers fabricated on Ti-6Al-4V, and therefore surfaces with antibacterial, biocompatible, tissue integration stimulating and spread-spectrum bactericidal properties can be obtained. [Figure 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). JJAC is funded by a FPI grant from the same origin (BES2014-069007).

Recent studies showed that a positive microbiological result from sonication of the PMMA spacer was associated with poor outcome of patients, but no quantitative analysis has yet been performed. For this purpose, a prospective analysis of 50 spacers (46 patients) was performed. All spacers were processed according to a previously described protocol, including centrifugation and quantitative culture. Clinical data and outcome were also analysed. A statistical relationship between the results of the cultures and the outcome of the patient was assessed. Sixteen patients were diagnosed with spacer-associated infection. Thirteen out of 50 spacers gave a positive culture. Nine of 13 presented with growth of an organism not isolated in the first-stage cultures, and in 7 out of 13 the organisms count was high (>10,000 CFU/ml). We have detected a significant statistical relationship between poor outcome and positive cultures, high colony counts, isolation of different organisms, positive periprosthetic cultures and spacer-associated infection. The detection in a sonicated, antibiotic-loaded PMMA spacer of organisms other than those isolated in the first surgical samples or high colony counts of any organisms is diagnostic with regard to spacer-associated infection.

The risk of knee arthroplasty infection and appropriateness of antibiotic treatment are not clearly established in patients with preoperative asymptomatic bacteriuria. It has been the purpose to analyze the prevalence of preoperative asymptomatic bacteriuria in knee arthroplasty patients, as well as the incidence of prosthetic joint infection in those with asymptomatic bacteriuria treated and not with specific antibiotics. This prospective study included 215 consecutive knee arthroplasty patients (73 ± 6 years, 168 females) with neither urinary symptoms nor perioperative urethral catheterization. A “clean-catch” urinalysis was obtained from all patients before surgery and an urine culture if urinalysis was abnormal. Asymptomatic bacteriuria was diagnosed if >100,000 colony-forming units/ml were cultured. Patients were treated (Group A) or not (Group B) with additional specific antibiotics for urine bacteria according to surgeon criteria. Minimum follow-up reached 48 months. No patient was lost to follow-up. Asymptomatic bacteriuria was diagnosed in 11/215 patients (5.1 %) (11/11 females), and four of these 11 were treated with specific antibiotics (Group A). Only one patient in Group A suffered a prosthesis infection along the first 3 months (1/125, 0.5 %), but bacteria cultured from the wound were absolutely different to those in urine culture. No patient in Group B suffered a prosthesis infection. Asymptomatic bacteriuria presents a low prevalence. We have not found any case of arthroplasty infection from urinary focus in patients with asymptomatic bacteriuria whether they received or not specific antibiotics.

Mycobacterium abscessus lung infection is a major health problem for cystic fibrosis (CF) patients. Understanding the in vivo factors that influence the outcome of therapy may help addressing the poor correlation between in vitro and in vivo antibiotic efficacy. We evaluated the influence of interspecies interactions and lung epithelial cells on antibiotic efficacy. Therefore, single and dual-species biofilms of M. abscessus and a major CF pathogen (Pseudomonas aeruginosa) were cultured on a plastic surface or on in vivo-like three-dimensional (3-D) lung epithelial cells, and the activity of antibiotics (colistin, amikacin, clarithromycin, ceftazidime) in inhibiting biofilm formation was evaluated. Using the most physiologically relevant model (dual-species biofilms on 3-D cells), we observed that treatment with antibiotics during biofilm development inhibited P. aeruginosa but not M. abscessus biofilms, resulting in a competitive advantage for the latter. Clarithromycin efficacy against P. aeruginosa was inhibited by 3-D lung cells. In addition, biofilm induction of M. abscessus was observed by certain antibiotics on plastic but not on 3-D cells. Pseudomonas aeruginosa influenced the efficacy of certain antibiotics against M. abscessus, but not vice versa. In conclusion, these results suggest a role of host cells and interspecies interactions in bacterial responses to antimicrobials.

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.

Catheter-related bloodstream infections (CRBSI) constitute an important cause of hospital-acquired infection associated with morbidity, mortality, and cost. The aim of these guidelines is to provide updated recommendations for the diagnosis and management of CRBSI in adults. Prevention of CRBSI is excluded. Experts in the field were designated by the two participating Societies (Sociedad Espanola de Enfermedades Infecciosas y Microbiologia Clinica and the Sociedad Espanola de Medicina Intensiva, Crftica y Unidades Coronarias). Short-term peripheral venous catheters, non-tunneled and long-term central venous catheters, tunneled catheters and hemodialysis catheters are covered by these guidelines. The panel identified 39 key topics that were formulated in accordance with the PICO format. The strength of the recommendations and quality of the evidence were graded in accordance with ESCMID guidelines. Recommendations are made for the diagnosis of CRBSI with and without catheter removal and of tunnel infection. The document establishes the clinical situations in which a conservative diagnosis of CRBSI (diagnosis without catheter removal) is feasible. Recommendations are also made regarding empirical therapy, pathogen-specific treatment (coagulase-negative staphylococci, Sthaphylococcus aureus, Enterococcus spp, Gram-negative bacilli, and Candida spp), antibiotic lock therapy, diagnosis and management of suppurative thrombophlebitis and local complications. (C) 2017 Elsevier Espana, S.L.U. and Sociedad Espanola de Enfermedades Infecciosas y Microbiologia Clinica. All rights reserved.

The Ti–6Al–4V alloy is one of the most commonly used in orthopedic surgery. Despite its advantages, there is an increasing need to use new titanium alloys with no toxic elements and improved biomechanical properties, such as Ti–13Nb–13Zr. Prosthetic joint infections (PJI) are mainly caused by Gram-positive bacteria; however, Gram-negative bacteria are a growing problem due to associated multidrug resistance. In this study, the bacterial adherence and viability on the Ti–13Nb–13Zr alloy have been compared to that of the Ti–6Al–4V alloy using 16 collection and clinical strains of bacterial species related to PJI: Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa. When compared with the Ti–6Al–4V alloy, bacterial adherence on the Ti–13Nb–13Zr alloy was significantly higher in most staphylococcal and P. aeruginosa strains and lower for E. coli strains. The proportion of live bacteria was significantly lower for both Gram-negative species on the Ti–13Nb–13Zr alloy than on the Ti–6Al–4V alloy pointing to some bactericidal effect of the Ti–13Nb–13Zr alloy. This bactericidal effect appears to be a consequence of the formation of hydroxyl radicals, since this effect is neutralized when dimethylsulfoxide was added to both the saline solution and water used to wash the stain. The antibacterial effect of the Ti–13Nb–13Zr alloy against Gram-negative bacteria is an interesting property useful for the prevention of PJI caused by these bacteria on this potential alternative to the Ti–6Al–4V alloy for orthopedic surgery.

Objective This NMA compared the efficacy and safety between IV antibiotics that are used in the current standard of care for managing adult patients (≥18 years of age) with ABSSSI. Methods Comparators were chosen on the basis that both direct and indirect comparisons between the interventions of interest could be performed. Outcomes of the analysis were selected on the basis that they are frequently measured and reported in trials involving ABSSSI patients, and only published randomised control trials of any size and duration and with any blinding status were eligible for inclusion in the analysis. The NMA was performed using both a fixed-effect and random-effect model. Efficacy-related endpoints were (1) clinical treatment success and (2) microbiological success at TOC visit. Safety-related endpoints were (1) number of discontinuations due to AEs/SAEs, (2) patients experiencing AEs, (3) patients experiencing SAEs and (4) all-cause mortality. Results Study interventions included daptomycin, dalbavancin, linezolid and tigecycline. Vancomycin was the comparator in all studies, except in two where it was linezolid and teicoplanin. The NMA showed that irrespective of patient subgroup, the likelihood of clinical and microbiological success with dalbavancin was statistically similar to the comparators studied. No statistically significant differences were observed between dalbavancin and any of the comparators in the discontinuation rate due to AEs/SAEs. In contrast, dalbavancin was associated with a significantly lower likelihood of experiencing an AE than linezolid, a significantly lower likelihood of experiencing a SAE than vancomycin and daptomycin, and a significantly lower risk of all-cause mortality than vancomycin, linezolid and tigecycline. Conclusion Dalbavancin affords a promising, new alternative IV antimicrobial agent which is as effective as traditional therapies, but with the added benefit of enabling clinicians to treat patients with ABSSSI in different organisational settings. Notwithstanding, any introduction of an effective treatment with a differential mode of administration into healthcare systems must be followed by a change in clinical practice and patient management in order to fully achieve desirable economic outcomes.

Purpose Fluoroquinolones are recommended for the treatment of pneumonia. The recognition of risk factors for invasive levofloxacin-resistant Streptococcus pneumoniae is important for the design of treatment. Methods A retrospective review of cases of invasive pneumococcal infections in adults was undertaken. Epidemiologic data, predisposing factors, clinical variables, and outcome were recorded from previously established protocols. Antimicrobial susceptibility was determined by disk diffusion and the Etest method. Serotyping was performed by latex agglutination and Quellung reaction. Results Twenty patients with infection caused by levofloxacin-resistant pneumococci [minimum inhibitory concentration (MIC) ≥2 μg/ml] were compared with 102 patients harboring levofloxacin-susceptible strains; 80 % of levofloxacin-resistant pneumococci were resistant to ≥3 antibiotics but susceptible to penicillin. Most levofloxacin-resistant strains (80 %) belonged to serotype 8. In comparison, only 8 % of levofloxacin-susceptible pneumococci belonged to serotype 8. In the multivariate analysis, residence in public shelters [odds ratio (OR) 26.13; p 0.002], previous hospitalization (OR 61.77; p

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.

Mycobacteria are a large group of microorganisms, multiple species of which are major causes of morbidity and mortality, such as tuberculosis and leprosy. At present, the emergence and spread of multidrug-resistant strains of Mycobacterium tuberculosis complex are one of the most serious health problems worldwide. Furthermore, in contrast to M. tuberculosis and Mycobacterium leprae, non-tuberculous mycobacteria (NTM) are more frequently isolated and, in many cases, treatment is based on drug susceptibility testing. This article is a review of the different methods to determine the in vitro drug susceptibility of M. tuberculosis complex and the most relevant NTM isolates. The molecular techniques currently used for rapid detection of resistance of clinical specimens are also analysed.

Levofloxacin plus rifampicin (L+R) is the treatment of choice for acute staphylococcal prosthetic joint infection (PJI) managed with debridement and implant retention (DAIR). Long courses have been empirically recommended, but some studies have suggested that shorter treatments could be as effective. Our aim was to prove that a short treatment schedule was non-inferior to the standard long schedule. An open-label, multicentre, randomised clinical trial (RCT) was performed. Patients with an early post-surgical or haematogenous staphylococcal PJI, managed with DAIR and initiated on L+R were randomised to receive 8 weeks of treatment (short schedule) versus a long schedule (3 months or 6 months for hip or knee prostheses, respectively). The primary endpoint was cure rate. From 175 eligible patients, 63 were included (52% women; median age, 72 years): 33 patients (52%) received the long schedule and 30 (48%) received the short schedule. There were no differences between the two groups except for a higher rate of polymicrobial infection in the long-schedule group (27% vs. 7%; P = 0.031). Median follow-up was 540 days. In the intention-to-treat analysis, cure rates were 58% and 73% in patients receiving the long and short schedules, respectively (difference −15.7%, 95% CI −39.2% to 7.8%). Forty-four patients (70%) were evaluable per-protocol: cure rates were 95.0% and 91.7% for the long and short schedules, respectively (difference 3.3%, 95% CI −11.7% to 18.3%). This is the first RCT suggesting that 8 weeks of L+R could be non-inferior to longer standard treatments for acute staphylococcal PJI managed with DAIR.

Prosthetic joint infections (PJI) are severe complications in Orthopedics, with Staphylococcus aureus and Staphylococcus epidermidis being the most commonly isolated pathogens. The variable antimicrobial susceptibility found in these microorganisms, along with the increasing number of methicillin-resistant strains, increases the difficulty of antibiotic selection and makes it necessary to perform individual susceptibility studies to select the optimal antibiotic treatment. The aim of this study was to evaluate the in vitro susceptibility pattern of 35 clinical strains isolated from PJI (17 S. aureus and 18 S. epidermidis) against rifampin, vancomycin, tygecicline, clindamycin, cotrimoxazole, cloxacillin, ciprofloxacin, daptomycin and fosfomycin. In vitro susceptibility assays were performed using the broth microdilution method and agar dilution for fosfomycin. MBC was also determined. Tygecicline and daptomycin showed the highest antimicrobial activity with low MIC(90) values, and no resistant strains were detected. On the other hand, ciprofloxacin and cloxacillin exhibited a poor antimicrobial effect with a high percentage of nonsusceptible strains in both species. Bactericidal activity rates revealed the bacteriostatic behavior of rifampin, tygecicline, cotrimoxazole, fosfomycin and clindamycin, whereas vancomycin and cloxacillin showed species- and strain-dependent behavior. Daptomycin and ciprofloxacin were observed to be efficient bactericidal agents against the tested strains. According to our data, rifampin, tigecycline, daptomycin and fosfomycin showed high in vitro activity against most staphylococcal strains isolated from the PJIs tested, although daptomycin seems to be the best alternative to vancomycin therapy.

The genus Corynebacterium includes a high number of species that are usually isolated from human skin as saprophytes. However, these microorganisms have also been reported as infectious agents in a broad group of patients and have showed broad-spectrum resistance. We studied the susceptibility profiles against macrolides, clindamycin, and streptogramins of 254 clinical strains belonging to the species Corynebacterium urealyticum (120), Corynebacterium amycolatum (66), Corynebacterium jeikeium (17), Corynebacterium striatum (20), Corynebacterium coyleae (12), Corynebacterium aurimucosum (11), and Corynebacterium afermentans subsp. afermentans (8). The MLS(B) phenotype was detected in 186 strains and was associated with the presence of methylase enzymes codified by the erm(X) gene in 171 strains. The erm(B) gene was only detected in two C. urealyticum strains. Fourteen strains showed macrolide resistance, but they did not carry erm genes. mef genes were not detected despite eight C. amycolatum strains showed the M phenotype. Also, the presence of hydrolytic enzymes codified by ere(B) was evaluated, but all results were negative. Resistance to macrolide in Corynebacterium sp. is mainly due to the presence of erm(X) methylase, although other resistance mechanisms could be involved.

Atypical mycobacteria are a heterogeneous group of organisms that are of increasing importance because of the growing number of infections they cause. This rising rate of infection is due mainly to the increase in the number of susceptible (and especially immunosuppressed) patients.To revise the currently used treatment schemes of the most commonly isolated atypical mycobacteria.Literature review using reference books and PubMed with specific keywords for each mycobacteria.The first important step in the management of atypical mycobacteria is to recognize the true infections caused by these organisms. The treatment required varies according to species. Well-characterized combinations exist for most common isolates, with the use of first-line antituberculous drugs (isoniazid, rifampin, ethambutol), clarithromycin, aminoglycosides and/or quinolones for slowly growing species (Mycobacterium avium complex, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium ulcerans, Mycobacterium marinum, Mycobacterium lentiflavum, Mycobacterium malmoense) and macrolides, quinolones, amikacin and other antibiotics for rapidly growing mycobacteria (Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium fortuitum). Surgical therapy is also important for some species (Mycobacterium ulcerans, Mycobacterium scrofulaceum) and for localized infections. The treatment of uncommon species is not well defined and is determined by the results of in vitro tests of individual strains. Because of the increasing number of resistant strains, new antibiotics need to be used for the treatment of these strains.

An in vitro study aimed to evaluate the effect of N-acetyl cysteine (NAC) or sub-ICs of erythromycin on antimicrobial susceptibility of staphylococcal biofilms was performed. Staphylococcus aureus and Staphylococcus epidermidis strains were isolated from patients with prosthetic joint infections using a previously published sonication procedure. Conventional susceptibility studies were performed using microdilution according to the CLSI procedures. Biofilm susceptibility was performed using the Calgary methodology. The addition of NAC showed no effect with the S. aureus strains, and a strain-dependent effect with the S. epidermidis strains. No effect was detected with erythromycin for almost all tested strains.

Prosthetic joint infections (PJIs) are related to the formation of biofilms, mainly by Staphylococcus aureus and Staphylococcus epidermidis. Therapy is usually selected according conventional susceptibility testing, but these data may be insufficient to detect the true antibiotic susceptibility in the biofilm. In total, 32 clinical strains (17 S. aureus and 15 S. epidermidis) isolated from patients with PJIs as well as 2 collection strains (S. aureus 15981 and S. epidermidis ATCC 35984) were tested against nine antibiotics commonly used in the treatment of PJIs (rifampicin, vancomycin, tigecycline, clindamycin, trimethoprim/sulfamethoxazole, ciprofloxacin, cloxacillin, daptomycin and fosfomycin) using the Calgary Biofilm Device. None of the antibiotics proved to be totally effective against biofilms in both species, with minimum biofilm eradication concentrations (MBECs) highly above the minimum inhibitory concentrations for most of the antibiotics (>1024 mg/L). Rifampicin and tigecycline showed MBECs slightly lower, mainly against S. epidermidis biofilms, and only two strains of this staphylococcal species were susceptible to almost all of the antibiotics tested. These results show that the search for new compounds with antimicrobial and antibiofilm properties is mandatory as well as the development of other strategies that could lead to the prevention and treatment of PJIs. In addition, more studies are necessary to obtain a better understanding of the mechanisms involved in antimicrobial resistance of biofilms.

We studied the in vitro activity of tigecycline and 10 other commonly used antibiotics against 135 clinical isolates of non-diphtheria Corynebacterium spp. using the Etest system. Tigecycline minimum inhibitory concentrations for 50% and 90% of the organisms (MIC(50) and MIC(90) values, respectively, in mg/L) were: Corynebacterium urealyticum, 0.094 and 0.125; Corynebacterium amycolatum, 0.125 and 2; Corynebacterium jeikeium, 0.094 and 0.75; Corynebacterium coyleae, 0.064 and 0.064; Corynebacterium striatum, 0.064 and 1; Corynebacterium aurimucosum, 0.094 and 0.125; and Corynebacterium afermentans, 0.064 and 0.094. The activities of all other antimicrobials were variable, with good activity of glycopeptides, linezolid, quinupristin/dalfopristin and daptomycin and with resistance to macrolides in a high number of isolates. Tigecycline is a good alternative for the therapy of infections caused by non-diphtheria corynebacteria.

The bactericidal activities of two new ketolides, HMR 3004 and HMR 3647, and the potential to develop resistance to these two antibiotics were studied in Gram-positive bacilli. As judged by time-kill kinetics both ketolides were mostly bacteriostatic, being bactericidal against only highly susceptible isolates of Corynebacterium striatum (two isolates) and Corynebacterium minutissimum (one isolate). Spontaneous resistant mutants were detected in seven of 30 strains tested, mainly in Rhodococcus equi, C. minutissimum and C. striatum, with a very low frequency of mutation (10(-12)-10(-15)).

The genus Capnocytophaga comprises a group of gramnegative rods that are part of the normal human oral flora. They are opportunistic pathogens associated with localized (periodontal, abscess, sinusitis, keratitis, endophthalmitis and osteomyelitis) and systemic (sepsis, peritonitis and endocarditis) infections, and they affect both the immunocompetent and the immunocompromised [1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14]. Most of the cases of Capnocytophaga sputigena bacteremia reported previously in the literature resulted from a breakthrough in the oral mucosa from which the organisms could have spread to the bloodstream [4, 5]. This kind of infection is usually associated with hematological malignancy and happens during the neutropenic period that follows chemotherapy. In most cases, the organism is recovered in pure culture, but cases of mixed bacteremia due to this organism are rare [2, 9]. We report a case of mixed bacteremia due to Capnocytophaga sputigena and Escherichia coli that occurred in a patient who had undergone bone marrow transplantation (BMT), and we review previously reported similar infections. An 18-year-old male who had been diagnosed with stage III-B bulky-disease Hodgkin’s lymphoma was referred to our hospital for rescue with high-dose therapy and autologous stem cell transplantation. The patient received a transplantation conditioning regimen of combined cyclophosphamide-carmustine-etoposide, and prophylaxis against infection with cotrimoxazole, acyclovir and fluconazole. On day 0 his leukocyte count came down to 0.670 103/mm3, and between days 5 and 11 it fell to numbers lower than 0.4 103/mm3. On day 1 the patient complained of mouth pain due to oral mucositis and ulceration. On day 2 he presented with low-grade fever (37.4 C). Urine and blood cultures (from the white and red lumen of the catheter and from the peripheral vein) were taken, and he was started on amoxicillin/clavulanate. On day 3 the blood culture taken through the red lumen yielded a gram-negative rod further identified as Escherichia coli resistant to ampicillin and cotrimoxazole. Twenty-four hours later, a gram-negative rod, subsequently identified as Capnocytophaga sputigena resistant to aminoglycosides, grew from the sample taken through the white lumen. The patient was then given empiric antibiotic therapy with cefepime and amikacin up to day 13. A strain of Escherichia coli with a susceptibility pattern identical to that of the Escherichia coli strain isolated from blood culture and a strain of Proteus mirabilis were isolated from the urine culture, the latter being resistant to ampicillin, cotrimoxazole and gentamicin. The patient’s oral lesions healed gradually during the following 7 days. However, he remained febrile up to day 9, when fever finally resolved and his neutrophil level returned to normal. Blood cultures taken from the same four sites after the start of treatment were all negative. We searched the Medline database for cases of Capnocytophaga bacteremia in BMT patients using the search terms “Capnocytophaga”, “bacteremia” and “bone marrow”. We included all cases that met all three search criteria in our review (Table 1). To the best of our knowledge, the present case is the first in which Capnocytophaga sputigena and Escherichia coli were isolated concurrently from a BMT patient. Although Parenti and Snydman [2] identified six patients with polymicrobial infection in their series of patients infected with Capnocytophaga spp., none of these infections followed BMT. In two of their cases, monomicrobial Capnocytophaga bacteremia followed BMT, but it was not specified whether BMT and bacteremia were concomitant, and the organisms were not identified to the J. I. Garc a-C a · J. Esteban ()) · F. Santos-O’Connor · F. Soriano Department of Microbiology, Fundaci n Jim nez D az, Avenida Reyes Cat licos 2, 28040 Madrid, Spain e-mail: jesteban@fjd.es Tel.: +34-91-5504800 Fax: +34-91-5494764

It is important to know the spectrum of the microbial aetiology of prosthetic joint infections (PJIs) to guide empiric treatment and establish antimicrobial prophylaxis in joint replacements. There are no available data based on large contemporary patient cohorts. We sought to characterize the causative pathogens of PJIs and to evaluate trends in the microbial aetiology. We hypothesized that the frequency of antimicrobial-resistant organisms in PJIs has increased in the recent years. We performed a cohort study in 19 hospitals in Spain, from 2003 to 2012. For each 2-year period (2003-2004 to 2011-2012), the incidence of microorganisms causing PJIs and multidrug-resistant bacteria was assessed. Temporal trends over the study period were evaluated. We included 2524 consecutive adult patients with a diagnosis of PJI. A microbiological diagnosis was obtained for 2288 cases (90.6%). Staphylococci were the most common cause of infection (1492, 65.2%). However, a statistically significant rising linear trend was observed for the proportion of infections caused by Gram-negative bacilli, mainly due to the increase in the last 2-year period (25% in 2003-2004, 33.3% in 2011-2012; p 0.024 for trend). No particular species contributed disproportionally to this overall increase. The percentage of multidrug-resistant bacteria PJIs increased from 9.3% in 2003-2004 to 15.8% in 2011-2012 (p 0.008), mainly because of the significant rise in multidrug-resistant Gram-negative bacilli (from 5.3% in 2003-2004 to 8.2% in 2011-2012; p 0.032). The observed trends have important implications for the management of PJIs and prophylaxis in joint replacements. N. Benito, (C) 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Nontuberculous mycobacteria (NTM) are becoming increasingly important. A growing number of patients with underlying conditions that make them prone to diseases caused by NTM. These diseases include the appearance of new syndromes, such as mesotherapy and other cosmetic-related infections, or diseases that affect patients who are being treated with tumor necrosis factor.A literature search has been performed for each mycobacterium species. An introduction to the different aspects of the species and the diseases is provided, along with a review of the current therapeutic options; special emphasis is put on new research and discoveries.Recognition of the current role of NTM isolates remains the key step in the management of NTM infections. After recognition, treatment must be guided by attending to the isolated species, the specific syndromes, clinical experience and - for some species - the results of in-vitro susceptibility tests. Surgical therapy is also important for some species (Mycobacterium ulcerans, Mycobacterium scrofulaceum) and for localized infections. The treatment of uncommon species is not yet well defined and recent research on resistance mechanisms has described their importance. The role of biofilms is currently of special concern for various specific infections.

Despite being relatively infrequent, prosthetic joint infections are a devastating medical complication. However, recent advances in surgical techniques, new antibiotics, and knowledge about pathogenic mechanisms have improved the outcome for affected patients.We have analyzed recent advances in pathogenesis, medical and surgical therapy of prosthetic joint infections, with special focus on new antibiotics useful for this disease. Recent studies focused on the important role of biofilms and intracellular bacteria in the pathogenesis of biomaterial-related infections. These advances must guide the management of the patients. Together with more classical antibiotics, linezolid and daptomycin have shown their usefulness for the treatment of these infections. Recently developed lipoglycopeptides have the potential to be used for these infections. In this sense, the possibility of treating patients with oral antibiotics without lack of efficacy is of great interest.Individualized therapies that take into account the microbial etiology, pathogenesis of the disease, antimicrobial susceptibility, and efficacy of antibiotics against biofilms and intracellular organisms make it possible to treat even those infections caused by multidrug-resistant organisms. A multidisciplinary approach (including a surgeon, infectious diseases specialist and microbiologist) provides the best possible management of patients.

The frequency of isolation as well as the number of species of non-tuberculous mycobacteria (NTM) has increased in the last years. Nearly every pathogenic species of NTM may cause skin and soft tissue infections, but rapidly growing mycobacteria (Mycobacterium fortuitum, Mycobacterium chelonae and Mycobacterium abscessus), Mycobacterium marinum and Mycobacterium ulcerans are the most commonly involved. Many of these cutaneous mycobacteriosis, such as rapidly growing mycobacteria, M. marinum, Mycobacterium avium complex, Mycobacterium kansasii or Mycobacterium xenopi are world-wide distributed. In contrast, some others have a specific geographical distribution. This is the case of M. ulcerans, which causes a cutaneous diseases endemic of Central and West Africa (Buruli ulcer) and Australia (Bairnsdale ulcer), being the third mycobacterial infection after tuberculosis and leprosy. Cutaneous mycobacteriosis usually appear either after contact of traumatic or surgical wounds with water or other contaminated products, or, secondarily, as a consequence of a disseminated mycobacterial disease, especially among immunosuppressed patients. For an early diagnosis, it is necessary to maintain a high degree of suspicion in patients with chronic cutaneous diseases and a history of trauma, risk exposure and negative results of conventional microbiological studies. In general, individualized susceptibility testing is not recommended for most NTM infections, except for some species, and in case of therapeutic failure. Treatment includes a combination of different antimicrobial agents, but it must be taken into account that NTM are resistant to conventional antituberculous drugs. Severe cases or those with deep tissues involvement could also be tributary of surgical resection.

Previous studies of knee arthroplasty infections caused by high-virulence organisms suggest poor outcomes. Polymicrobial and Gram-negative infections are less studied.This study compared the results of treatment of knee arthroplasty infections by single versus polymicrobial isolates, Gram-positive versus Gram-negative, and methicillin-resistant versus -sensitive Staphylococci.We prospectively followed 47 patients with late knee arthroplasty infections. The mean age was 72 years (range, 20-87 years). The treatment protocol included two-stage exchange and a combination of two oral antibiotics given for 6 months. Minimum followup was 1 year (average, 4.8 +/- 3 years; range, 1-12 years). Control of the infection was judged by absence of clinical, serologic, and radiologic signs of infection. The functional outcome was evaluated by Knee Society score at the last followup.Infection was controlled in all 15 patients with polymicrobial and in 28 of 32 (88%) with monomicrobial infections, in eight of nine patients with Gram-negative and in 35 of 38 (92%) with Gram-positive isolates. Control was also achieved in 22 of 25 patients (88%) infected by methicillin-resistant Staphylococci and in 14 of 14 by methicillin-sensitive Staphylococci. The Knee Society scores averaged 81-63 in patients with polymicrobial infections and were higher than in monomicrobial infections (75-52). The mean KSS was 85-59 in Gram-negative infections compared to 75-55 in Gram-positive infections. The mean KSS was similar in methicillin-resistant (78-54) and methicillin-sensitive Staphylococci (73-56) infections.Polymicrobial and Gram-negative infections can be controlled in late knee arthroplasty infections. On the other hand, infections by methicillin-resistant Staphylococci are less likely to be controlled by the regimens we used.Level II, prognostic study. See Guidelines for Authors for a complete description of levels of evidence.

Infected arthroplasties reportedly have a lower eradication rate when caused by highly resistant and/or polymicrobial isolates and in these patients most authors recommend intravenous antibiotics. We asked whether two-stage revision with interim oral antibiotics could eradicate these infections. We prospectively followed 36 patients (mean age, 71.8 years) with late hip arthroplasty infections. Combinations of oral antibiotics were prescribed according to cultures, biofilm, and intracellular effectiveness. The minimum followup was 1 year (mean, 4.4 years; range, 1-12 years). We presumed eradication in the absence of clinical, serologic, and radiographic signs of infection. Infection was eradicated in all 13 patients with highly resistant bacteria who completed a two-stage protocol (10 with methicillin-resistant Staphylococci) and in eight of 11 patients treated with only the first stage (and six of nine with methicillin-resistant Staphylococci). Infection was eradicated in six of six patients with polymicrobial isolates (of sensitive and/or resistant bacteria) who completed a two-stage protocol and in five of seven with polymicrobial isolates treated with only the first surgery. The Harris hip score averaged 88.1 (range, 70-98) in patients who underwent reimplantation and 56.8 (range, 32-76) in patients who underwent resection arthroplasty. Long cycles of combined oral antibiotics plus a two-stage surgical exchange appear a promising alternative for infections by highly resistant bacteria, methicillin-resistant Staphylococci, and polymicrobial infections.

We evaluated the in vitro activities of tigecycline and 10 other antibiotics against clinical isolates of nonpigmented rapidly growing mycobacteria. Fifteen collection strains and 165 clinical isolates were included in the study. Tigecycline showed the highest activity among all antibiotics studied: all the strains were inhibited by 1 mg/liter.

Exchange surgery in late arthroplasty infection is directed against bacteria adhering to implants. Therapies based on antibiotics that are effective intracellularly have been proposed recently. We have combined both strategies to improve the cure rate.40 consecutive patients (16 hips, 24 knees) were diagnosed with late arthroplasty infection. The organisms isolated were 35 Staphylococcus, 19 of which were methicillin-resistant, 4 Enterococcus, 6 Gram-negative bacilli, and 4 Corynebacterium. The infections were managed by a combined therapy consisting of two-stage exchange surgery and two oral intracellularly-effective antibiotics. The antibiotics were selected according to bacterial sensitivity and intracellular and biofilm effectiveness. Second re-implantation surgery was delayed until clinical and analytical normalization. Patients were in hospital for only 1 week after each surgery, and were followed up prospectively on an outpatient basis (2-9 years). Cure of the infection was defined as absence of clinical, serological, and radiographic signs of infection during the whole follow-up.The infection was resolved in 38/40 patients (15/16 hips and 23/24 knees).Oral antibiotics that are effective intracellularly in combination with two-stage exchange surgery is a promising alternative for treating late arthroplasty infections. Oral antibiotics shorten hospitalization and reduce patient discomfort.

To investigate the potential implications (especially the implications in clinical significance and antimicrobial susceptibility) of polyclonality among rapidly growing mycobacteria, we performed random amplified polymorphic DNA analysis in 64 clinical isolates of which the clinical significance was established. Phenotypic characteristics (antimicrobial susceptibility test, colony morphology and growth rate) of each clone were studied. Polyclonality was detected in 13 of the isolates (20.3%). There was a relationship between monoclonality and clinical significance (p 0.0096). Monoclonal and polyclonal isolates showed different behaviour in antimicrobial susceptibility. There was a strong relationship between monoclonality and those species that are more pathogenic for humans, and also with clinical significance of the isolates.