Abstract Clogging of endoscopic stents necessitates their replacement in many patients with malignant obstructive jaundice and limits their use in benign strictures. We studied the basic mechanism ...

You have accessJournal of UrologyStone Disease: Basic Research I1 Apr 2014MP20-01 AURICLOSENE IRRIGATION SOLUTION PREVENTS ENCRUSTATION BY CRYSTALLINE BIOFILM DUE TO PROTEUS MIRABILIS IN AN IN VITRO URINARY CATHETER PATENCY MODEL Suriani Abdul Rani, Ramin (Ron) Najafi, Keith Bley, William Costerton, David Stickler, Bernard Churchill, and Dmitri Debabov Suriani Abdul RaniSuriani Abdul Rani More articles by this author , Ramin (Ron) NajafiRamin (Ron) Najafi More articles by this author , Keith BleyKeith Bley More articles by this author , William CostertonWilliam Costerton More articles by this author , David SticklerDavid Stickler More articles by this author , Bernard ChurchillBernard Churchill More articles by this author , and Dmitri DebabovDmitri Debabov More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.723AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Long-term indwelling urinary catheters are susceptible to blockage due to formation of crystalline biofilms by urease-producing microorganisms such as Proteus mirabilis. An in vitro catheter biofilm model (CBM) was developed to compare current methods for maintaining urinary catheter patency. We compared various bladder irrigation solutions or antimicrobial-coated urinary catheters, versus a novel antimicrobial catheter irrigation solution containing auriclosene (N,N-dichloro-2,2-dimethyltaurine; formerly designated NVC-422). METHODS CBM units were fed artificial urine at 0.5 mL per min. The artificial bladder chamber was inoculated with 108 colony forming units (CFU) of P. mirabilis and biofilm was allowed to establish for 48 hours before daily treatments commenced. A single treatment consisted of two sequential 50 mL irrigations. Each irrigation was retained in the catheter for 15 min and then drained, with a 30 min washout period between the two irrigations. Experiments were conducted for up to 10 days or until catheter blockage. The pH of the effluent, CFU counts in the bladder chamber and the time to catheter blockage were recorded. The area of catheter encrustation was measured using Stereo Zoom imaging. RESULTS Inoculation of the CBM reactor with 108 CFU of P. mirabilis resulted in blockage of the urinary catheters within 5 days. The use of silver-hydrogel or nitrofurazone-coated catheters did not extend the period of catheter patency. Catheter irrigation with 0.25% acetic acid or isotonic saline blocked at the same rate as untreated catheters. Catheters irrigated with a citrate-buffered formulation of 0.2% auriclosene resulted in maintenance of catheter patency for 10 days and complete eradication of P. mirabilis and biofilm within one treatment day. In contrast, daily irrigations of infected catheters with 10 mM acetate-buffered saline (at pH 4) or Renacidin® Irrigation Solution had no effect on P. mirabilis colonization of the bladder chamber, even though catheter patency was maintained throughout 10-day studies. CONCLUSIONS Irrigation with the rapidly bactericidal antimicrobial auriclosene in a buffered acidic formulation - termed Auriclosene Irrigation Solution - significantly enhanced catheter patency in vitro versus other irrigation solutions and antimicrobial-coated urinary catheters. Clinical evaluation of Auriclosene Irrigation Solution is ongoing. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e196 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Suriani Abdul Rani More articles by this author Ramin (Ron) Najafi More articles by this author Keith Bley More articles by this author William Costerton More articles by this author David Stickler More articles by this author Bernard Churchill More articles by this author Dmitri Debabov More articles by this author Expand All Advertisement Advertisement PDF DownloadLoading ...

The realization at the beginning of the current millennium that there are very substantial differences in gene content among the component strains of bacterial species, together with the observation that there exists profound phenotypic heterogeneity amongst these strains, led directly to the development of the field of comparative bacterial genomics, and the concepts of the core and supra (pan) genomes. The core genome is composed of the set of genes shared by all members of a given species (or other taxonomic grouping), and the supragenome is the set of all genes contained by the same grouping. Those genes not present in the core genome, but present in the supragenome, are referred to as distributed (accessory) genes, and it is this last group that is responsible for much of the intraspecies heterogeneity. In the current work we provide comparative genomic evidence for both the core genome hypothesis and the distributed genome hypothesis, and describe a testable means to determine if a strain or particular group of strains belongs within an extant genomically defined species (or other taxonomic) grouping.

The social soil bacterium, Myxococcus xanthus, displays a variety of complex and highly coordinated behaviours, including social motility, predatory rippling and fruiting body formation. Here we show that M. xanthus cells produce a network of outer membrane extensions in the form of outer membrane vesicle chains and membrane tubes that interconnect cells. We observed peritrichous display of vesicles and vesicle chains, and increased abundance in biofilms compared with planktonic cultures. By applying a range of imaging techniques, including three-dimensional (3D) focused ion beam scanning electron microscopy, we determined these structures to range between 30 and 60 nm in width and up to 5 μm in length. Purified vesicle chains consist of typical M. xanthus lipids, fucose, mannose, N-acetylglucosamine and N-acetylgalactoseamine carbohydrates and a small set of cargo protein. The protein content includes CglB and Tgl outer membrane proteins known to be transferable between cells in a contact-dependent manner. Most significantly, the 3D organization of cells within biofilms indicates that cells are connected via an extensive network of membrane extensions that may connect cells at the level of the periplasmic space. Such a network would allow the transfer of membrane proteins and other molecules between cells, and therefore could provide a mechanism for the coordination of social activities.

We report a case of an 86-year-old woman with an atypical femoral fracture (AFF) who was treated with intramedullary nailing followed by lateral femoral plating. She developed a second femoral shaft fracture distal to the intramedullary nail which required a second operation. Biopsy of the periosteum overlying the site of the initial proximal AFF was sent for pathogen analysis. Using the Ibis T5000 platform and the BAC plate assay, a polymicrobial infection was diagnosed consisting of Bifidobacterium subtile and Pseudomonas mendocina. This raises the possibility that bacterial infections may play some role in atypical fractures of the femur.

PCR combined with mass spectrometry is providing a whole new way to identify bacteria, particularly clinically important pathogens. For years, unculturable bacteria troubled physicians observing signs in their patients that were characteristic of infections while reading “no growth” on lab reports. Indeed, culture-based microbiological techniques are severely limited in their ability to detect pathogens. Culture-negative infections result primarily from bacteria adopting the biofilm mode of growth or responding to bacteriostatic antibiotics.

Biofilms associated with the human body, particularly in typically sterile locations, are difficult to diagnose and treat effectively because of their recalcitrance to conventional antibiotic therapy and host immune responses. The study of biofilms in medicine today requires a translational approach, with examination of clinically relevant biofilms in the context of specific anatomic sites, host tissues, and diseases, focusing on what can be done to mitigate their pathologic consequences. This review, which grew out of a discussion session on clinical biofilms at the 5th ASM Biofilm Conference in Cancun, Mexico, is designed to give an overview of biofilm-associated infections (BAI) and to propose a platform for further discussion that includes clinicians, medical microbiologists, and biofilm researchers who are stakeholders in advancing the scientific pursuit of better diagnosis and treatment of BAI to mitigate their human and healthcare costs. It also highlights the need for better diagnostic markers, which exploit the difference between planktonic and biofilm cells.

One of the most practical strategies that has been undertaken to fight biofilm implant-related infections has been the development of coatings on biomaterial devices that can elute antimicrobials into regions of patients' tissues. To date, the majority of animal studies that have been developed to model infections that accompany the use of these materials have primarily involved an initial inoculum of planktonic bacterial cells from batch cultures. Although valuable, data that have been derived from these experiments may not provide important clinical insight into how bacteria in well-established, mature biofilms impact device-related and other clinical infections when they contaminate a patient site or implanted device. In this review, a discussion is presented on the impact that a shift in biofilm research may have if initial inocula of well-established, mature biofilms are used to model biomaterial device-related infections in animal models.

The hypothesis that a cold plasma jet has the antimicrobial effect againstEnterococcus faecalisbiofilms was testedin vitro. 27 hydroxyapatite discs were incubated withE. faecalisfor six days to form a monoculture biofilm on the disc surface. The prepared substrata were divided into three groups: the negative control, the positive control (5.25% NaOCl solution), and the plasma treatment group. Resultant colony-forming unit counts were associated with observations of bacterial cell morphology changes using scanning electron microscopy (SEM). Treatment ofE. faecalisbiofilm with the plasma and 5.25% NaOCl for 5 min resulted in 93.1% and 90.0% kill (P<0.0001), respectively. SEM detected that nearly no intact bacteria were discernible for the plasma-exposed HA disc surfaces. The demonstrated bactericidal effect of the plasma with direct surface contact may be due to the enhanced oxidation by the locally produced reactive plasma species.

BACKGROUND:Understanding nosocomial pathogen transmission is restricted by culture limitations. Novel platforms, such as PCR-based electron spray ionization-time-of-flight-mass spectrometry (ESI-TOF-MS), may be useful as investigational tools.METHODS:Traditional clinical microbiology (TCM) and PCR/ESI-TOF-MS were used to recover and detect microorganisms from the hands and personal protective equipment of 10 burn intensive care unit (ICU) healthcare workers providing clinical care at a tertiary care military referral hospital. High-use environmental surfaces were assessed in 9 burn ICU and 10 orthopedic patient rooms. Clinical cultures during the study period were reviewed for pathogen comparison with investigational molecular diagnostic methods.RESULTS:From 158 samples, 142 organisms were identified by TCM and 718 by PCR/ESI-TOF-MS. The molecular diagnostic method detected more organisms (4.5 ± 2.1 vs. 0.9 ± 0.8, p < 0.01) from 99% vs. 67% of samples (p < 0.01). TCM detected S. aureus in 13 samples vs. 21 by PCR/ESI-TOF-MS. Gram-negative organisms were less commonly identified than gram-positive by both methods; especially by TCM. Among all detected bacterial species, similar percentages were typical nosocomial pathogens (18-19%) for TCM vs. PCR/ESI-TOF-MS. PCR/ESI-TOF-MS also detected mecA in 112 samples, vanA in 13, and KPC-3 in 2. MecA was associated (p < 0.01) with codetection of coagulase negative staphylococci but not S. aureus. No vanA was codetected with enterococci; one KPC-3 was detected without Klebsiella spp.CONCLUSIONS:In this pilot study, PCR/ESI-TOF-MS detected more organisms, especially gram-negatives, compared to TCM, but the current assay format is limited by the number of antibiotic resistance determinants it covers. Further large-scale assessments of PCR/ESI-TOF-MS for hospital surveillance are warranted.

SUMMARY Microorganisms coexist in a complex milieu of bacteria, fungi, archaea, and viruses on or within the human body, often as multifaceted polymicrobial biofilm communities at mucosal sites and on abiotic surfaces. Only recently have we begun to appreciate the complicated biofilm phenotype during infection; moreover, even less is known about the interactions that occur between microorganisms during polymicrobial growth and their implications in human disease. Therefore, this review focuses on polymicrobial biofilm-mediated infections and examines the contribution of bacterial-bacterial, bacterial-fungal, and bacterial-viral interactions during human infection and potential strategies for protection against such diseases.

ABSTRACT Gardnerella vaginalis is associated with a spectrum of clinical conditions, suggesting high degrees of genetic heterogeneity among stains. Seventeen G. vaginalis isolates were subjected to a battery of comparative genomic analyses to determine their level of relatedness. For each measure, the degree of difference among the G. vaginalis strains was the highest observed among 23 pathogenic bacterial species for which at least eight genomes are available. Genome sizes ranged from 1.491 to 1.716 Mb; GC contents ranged from 41.18% to 43.40%; and the core genome, consisting of only 746 genes, makes up only 51.6% of each strain's genome on average and accounts for only 27% of the species supragenome. Neighbor-grouping analyses, using both distributed gene possession data and core gene allelic data, each identified two major sets of strains, each of which is composed of two groups. Each of the four groups has its own characteristic genome size, GC ratio, and greatly expanded core gene content, making the genomic diversity of each group within the range for other bacterial species. To test whether these 4 groups corresponded to genetically isolated clades, we inferred the phylogeny of each distributed gene that was present in at least two strains and absent in at least two strains; this analysis identified frequent homologous recombination within groups but not between groups or sets. G. vaginalis appears to include four nonrecombining groups/clades of organisms with distinct gene pools and genomic properties, which may confer distinct ecological properties. Consequently, it may be appropriate to treat these four groups as separate species.

During the recent transition between acute diseases caused by swarms of single planktonic bacteria, and chronic infections caused by bacteria growing in slime-enclosed biofilms, a general clinical con

Implant infections in orthopaedics, as well as in many other medical fields, are chiefly caused by staphylococci. The ability of growing within a biofilm enhances the chances of staphylococci to protect themselves from host defences, antibiotic therapies, and biocides. Advances in scientific knowledge on structural molecules (exopolysaccharide, proteins, teichoic acids, and the most recently described extracellular DNA), on the synthesis and genetics of staphylococcal biofilms, and on the complex network of signal factors that intervene in their control are here presented, also reporting on the emerging strategies to disrupt or inhibit them. The attitude of polymorphonuclear neutrophils and macrophages to infiltrate and phagocytise biofilms, as well as the ambiguous behaviour exhibited by these innate immune cells in biofilm-related implant infections, are here discussed. Research on anti-biofilm biomaterials is focused, reviewing materials loaded with antibacterial substances, or coated with anti-adhesive/anti-bacterial immobilized agents, or surfaced with nanostructures. Latter approaches appear promising, since they avoid the spread of antibacterial substances in the neighbouring tissues with the consequent risk of inducing bacterial resistance.

INTRODUCTION:The numbers of inflatable penile prosthesis (IPP) implanted has increased yearly due to the large numbers of patients treated for prostate cancer, patients becoming refractory to the five phosphodiesterase inhibitors and Peyronie's disease.AIM:Prosthesis implantation can be associated with a variety of complications with device infection being the most dreaded one.MAIN OUTCOME MEASURES:An understanding of the pathogenesis of these infections is necessary to allow the surgeon to plan treatment.METHODS:Infection begins with colonization of planktonic bacteria in the implant space. Biofilm forms around the bacterial mass within 48 hours. Bacteria in biofilm have reduced growth rates, may change phenotypically, and develop resistance to drugs. Antibiotics and the body's macrophages will kill the planktonic bacteria released from the biofilm but never eliminate the infecting organisms. This review will delineate present thinking on infection prevention and biofilm's role in device infection. IPP infection before and after the coated implants will be characterized. Future ideas for prevention and treatment of infection will be explored.RESULTS:The coated implants have reduced the incidence of IPP infections. The bacteria that cause the majority of infections in the era of the coated implant seem to have changed from predominantly nosocomial coagulase-negative Staphylococcus to more virulent organisms. Device infection requires new paradigms of prevention and treatment strategy because the infecting bacteria are different and the patients are sicker.CONCLUSIONS:The problem of infection is considerably decreased with coated IPP, yet those infections that do occur are systemic in nature and seem to be caused by more aggressive organisms. These infections are not usually amenable to salvage because the virulence of the bacteria. Future research to prevent these infections must be directed to magnifying the effective dosage of antibiotics to penetrate the biofilm or eliminating the bacteria's ability to secrete the slime.

Increasing attention has been focused on understanding bacterial biofilms and this growth modality's relation to human disease. In this review we explore the genetic regulation and molecular components involved in biofilm formation and maturation in the context of the Gram-positive cocci, Staphylococcus aureus. In addition, we discuss diseases and host immune responses, along with current therapies associated with S. aureus biofilm infections and prevention strategies.

ABSTRACT Introduction Infection is the worst complication seen with inflatable penile prosthesis (IPP). Both the American Medical Systems (AMS) and Coloplast IPP have infection retardant coatings. AMS is coated at the factory with rifampicin and minocycline (InhibiZone). The Coloplast IPP has a hydrophilic coating covalently bonded to its components that will absorb any aqueous solution before implantation and provides increased surface lubricity to decrease bacterial adherence. Aim We tested several antibiotic dips comparing zones of inhibition (ZOI) against five commonly infecting bacteria with coated Coloplast implants. Results were compared with those ZOI created with strips of an AMS IPP precoated with InhibiZone. Methods Pieces of sterile Coloplast Titan IPP were dipped in (i) trimethoprim/polymixin B ophthalmic solution; (ii) trimethoprim/sulfamethoxazole infusion solution; (iii) bacitracin; (iv) rifampicin/minocycline; and (v) rifampin/trimehtoprim/sulfamethoxazole. ZOI for the Titan strips and for AMS InhibiZone coated strips were tested against Staphylococcus epidermidis, Staphylococcus lugdunensis, Staphylococcus aureus, Pseudomonas, and Enterococcus. Main Outcome Measure ZOIs of the Coloplast Titan for each of the medicated solutions were compared with ZOI created by undipped strips of a sterile InhibiZone coated IPP placed on plates of the identical bacteria. Results All dips except bacitracin showed ZOI ≥ InhibiZone (P ≥ 0.005) for most organisms. Because of broad-spectrum effectiveness, ease of handling, and cost, infusion vial of trimehtoprim/sulfamethoxazole seemed optimal at this time. If trimehtoprim/sulfamethoxazole is unavailable; the ZOI with Polytrim ophthalmic solution zones were almost as good. Conclusions The Coloplast strips when dipped in several solutions showed equal or significantly larger ZOI against commonly infecting organisms than the InhibiZone coated strips. At the present time using off the shelf trimethoprim sulfamethoxazole infusion solution seems optimum. The flexibility of choosing the drug eluting from the Coloplast device seems promising in the changing bacterial environment.

The detection and identification of bacteria present in natural and industrial ecosystems is now entirely based on molecular systems that detect microbial RNA or DNA. Culture methods were abandoned, in the 1980s, because direct observations showed that <1% of the bacteria in these systems grew on laboratory media. Culture methods comprise the backbone of the Food and Drug Administration-approved diagnostic systems used in hospital laboratories, with some molecular methods being approved for the detection of specific pathogens that are difficult to grow in vitro. In several medical specialties, the reaction to negative cultures in cases in which overt signs of infection clearly exist has produced a spreading skepticism concerning the sensitivity and accuracy of traditional culture methods. We summarize evidence from the field of orthopedic surgery, and from other medical specialties, that support the contention that culture techniques are especially insensitive and inaccurate in the detection of chronic biofilm infections. We examine the plethora of molecular techniques that could replace cultures in the diagnosis of bacterial diseases, and we identify the new Ibis technique that is based on base ratios (not base sequences), as the molecular system most likely to fulfill the requirements of routine diagnosis in orthopedic surgery.

Gallo, Phillip H. PhD; Melton-Kreft, Rachael BS, RN; Nistico, Laura PhD; Sotereanos, Nicholas G. MD; Sewecke, Jeffrey J. DO; Stoodley, Paul PhD; Ehrlich, Garth D. PhD; Costerton, J. William PhD; Kathju, Sandeep MD, PhD Author Information