Bacterial challenge study of a porous carbon percutaneous implant

Numerous percutaneous devices for power transmission and control to electrically powered, intracorporeal blood pumps have been used for periods ranging from 12 months to 4 yrs; however, consistent and reliable performance has not been achieved, due most frequently to the development of infection and sinus tracts at the percutaneous lead exit site. The present study showed that percutaneous devices fabricated from porous vitreous carbon can function satisfactorily in vivo over extended periods. The implant sites successfully resisted infection by normal flora bacteria for as long as 48 months, although superficial surface colonization and infection did occur after deliberate application of pathogens.     

Processing-structure-functional property relationship in organic-inorganic nanostructured scaffolds for bone-tissue engineering: The response of preosteoblasts

We elucidate here for the first time the structure-processing-functional property relationship in chitosan (CS)-based scaffolds, where molecular machinery governing proliferation and growth of osteoblasts is mediated by nanostructured carbon. The interconnected network structure of organic-inorganic scaffolds was obtained by covalent linkage of carboxyl group of functionalized single-walled carbon nanohorn with the amine group of CS. The molecular-scale dispersibility of functionalized nanostructured carbon was an important physicochemical factor influencing cellular interactions and biological response. Furthermore, it was beneficial in promoting the biocompatibility and the degradation product of the scaffolds. The hydrophilicity, good water retention ability, and interconnected porous structure of organic-inorganic scaffolds enabled pronounced cell attachment and proliferation and enhanced the stability toward enzymatic degradation. The infiltration of cells and colonization of the pores of the scaffolds and cellular interactions were promoted due to covalent linkage of nanostructured carbon with CS. Additionally, the interconnectivity of porous scaffolds facilitated cells to infiltrate inside the pores of CS-nanostructured scaffolds, implying that nanostructured carbon merits consideration in tissue engineering. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A:3080-3091, 2012.     

Modulation of colonization by black-pigmented Bacteroides species in squirrel monkeys by immunization with Bacteroides gingivalis.

Periodontal diseases are inflammatory responses thought to be triggered by specific microorganisms colonizing in the gingival crevice. Theoretically, periodontal diseases could be prevented if the etiologic organisms were not allowed to colonize the subgingival area. The humoral immune response is one mechanism which may modulate bacterial colonization in the gingival crevice. To test the effect of systemic humoral immunity on subgingival colonization by bacteria, squirrel monkeys (Saimiri sciureus) were immunized with Bacteroides gingivalis, a black-pigmented Bacteroides sp. and putative periodontal pathogen. Immunized and sham-immunized monkeys were orally inoculated with 10(10) viable B. gingivalis during ligation of five teeth in one quadrant with bacterium-soaked suture material and distribution over the entire dentogingival margin. Immunization resulted in an increased level of immunoglobulin G anti-B. gingivalis in serum and was associated with a strong trend toward a statistically significant reduction in colonization of the gingival crevice by black-pigmented bacteroides.     

Risk factors for colonization with extended-spectrum beta-lactamase-producing enterobacteriaceae on admission to rehabilitation centres

Patients newly admitted to rehabilitation centres are at high risk of colonization with multidrug-resistant bacteria because many of them have experienced prolonged stays in other healthcare settings and have had high exposure to antibiotics. We conducted a prospective study to determine the prevalence of and risk factors for colonization with extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) in this population. Subjects were screened by rectal swab for ESBL-PE within 2 days of admission. Swabs were plated on chromagar ESBL plates and the presence of ESBL was verified by a central laboratory. A multilevel mixed effects model was used to identify risk factors for ESBL-PE colonization. Of 2873 patients screened, 748 (26.0%) were positive for ESBL-PE. The variables identified as independently associated with ESBL-PE colonization were: recent stay in an acute-care hospital for over 2 weeks (OR = 1.34; 95% CI, 1.12, 1.6), history of colonization with ESBL-PE (OR = 2.97; 95% CI, 1.99, 4.43), unconsciousness on admission (OR = 2.59; 95% CI, 1.55, 4.34), surgery or invasive procedure in the past year (OR = 1.49; 95% CI, 1.2, 1.86) and antibiotic treatment in the past month (OR = 1.80; 95% CI, 1.45, 2.22). The predictive accuracy of the model was low (area under the ROC curve 0.656). These results indicate that ESBL-PE colonization is common upon admission to rehabilitation centres. Some risk factors for ESBL-PE colonization are similar to those described previously; however, newly identified factors may be specific to rehabilitation populations. The high prevalence and low ability to stratify by risk factors may guide infection control and empirical treatment strategies in rehabilitation settings.     

Sialic acid transport contributes to pneumococcal colonization

Streptococcus pneumoniae is a major cause of pneumonia and meningitis. Airway colonization is a necessary precursor to disease, but little is known about how the bacteria establish and maintain colonization. Carbohydrates are required as a carbon source for pneumococcal growth and, therefore, for colonization. Free carbohydrates are not readily available in the naso-oropharynx; however, N- and O-linked glycans are common in the airway. Sialic acid is the most common terminal modification on N- and O-linked glycans and is likely encountered frequently by S. pneumoniae in the airway. Here we demonstrate that sialic acid supports pneumococcal growth when provided as a sole carbon source. Growth on sialic acid requires import into the bacterium. Three genetic regions have been proposed to encode pneumococcal sialic acid transporters: one sodium solute symporter and two ATP binding cassette (ABC) transporters. Data demonstrate that one of these, satABC, is required for transport of sialic acid. A satABC mutant displayed significantly reduced growth on both sialic acid and the human glycoprotein alpha-1. The importance of satABC for growth on human glycoprotein suggests that sialic acid transport may be important in vivo. Indeed, the satABC mutant was significantly reduced in colonization of the murine upper respiratory tract. This work demonstrates that S. pneumoniae is able to use sialic acid as a sole carbon source and that utilization of sialic acid is likely important during pneumococcal colonization.     

Anaerobic bacteria commonly colonize the lower airways of intubated ICU patients

Objectives  To investigate respiratory tract colonization by aerobic and anaerobic bacteria in mechanically ventilated patients.
Methods  Bacterial colonization of the stomach and the respiratory tract was qualitatively and quantitatively analyzed over time in 41 consecutive mechanically ventilated patients in a Swedish intensive care unit (ICU), with special emphasis on elucidation of the role of anaerobic bacteria in the lower respiratory tract. Samples were taken from the oropharynx, gastric juice, subglottic space and trachea within 24 h (median 14 h) of intubation, and then every third day until day 18 and every fifth day until day 33.
Results  The patients were often heavily colonized with microorganisms not considered to belong to a healthy normal oropharyngeal and gastric flora on admission to the ICU. A majority harbored enterococci, coagulase-negative staphylococci and Candida spp. in at least one site on day 1. Anaerobic bacteria, mainly peptostreptococci and Prevotella spp., were isolated from subglottic and/or tracheal secretions in 59% of the patients. Different routes of tracheal colonization for different groups of microorganisms were found. Primary or concomitant colonization of the oropharynx with staphylococci, enterococci, enterobacteria and Candida was often seen, while Pseudomonas spp., other non-fermenting Gram-negative rods and several anaerobic species often primarily colonized the trachea, indicating exogenous or direct gastrointestinal routes of colonization.
Conclusions  Mechanically ventilated patients were heavily colonized in their lower airways by potential pathogenic microorganisms, including a high load of anaerobic bacteria. Different routes of colonization were shown for different species.     

Colonization resistance of the human intestinal microflora: Testing the hypothesis in normal volunteers

Colonization resistance is the mechanism whereby the intestinal microflora protects itself against incursion by new and often harmful microorganisms. Some authors have claimed that colonization resistance is related to the integrity of the anaerobic flora, but this point has not been established in humans. In previous studies in our laboratory cefoxitin, piperacillin, cefoperazone or aztreonam were administered intravenously to healthy volunteers in order to study changes in the intestinal flora and acquisition of new strains. Seven of 16 antibiotic-treated subjects were colonized with gram-negative bacilli, but no correlation was observed between this colonization and the suppression of either anaerobes or any other component of the fecal flora. Marked strains ofEscherichia coli andPseudomonas aeruginosa were also administered by mouth in order to test acquisition of new bacteria. The fed bacteria were found in the stools of both antibiotic-treated and control subjects; the antibiotics had no apparent influence on the ability of these strains to colonize the intestinal tract. Our work, along with findings of others, supports the concept that colonization resistance occurs in humans and is diminished by antibiotic administration. However, it does not support the hypothesis that colonization resistance is related to the anaerobic microflora.     

Role of anaerobic flora in the translocation of aerobic and facultatively anaerobic intestinal bacteria.

It is thought that the normal enteric microflora acts not only to prevent intestinal colonization but also to prevent subsequent systemic dissemination of ingested, potentially pathogenic bacteria. To determine the relative roles of specific components of the intestinal bacterial flora in bacterial translocation out of the gut, mice were given various antimicrobial agents to selectively eliminate specific groups of intestinal bacteria. The cecal flora and the translocating bacteria in mesenteric lymph nodes were monitored both before and after oral inoculation with antibiotic-resistant Escherichia coli C25. Orally administered streptomycin selectively eliminated cecal facultative gram-negative bacilli, orally administered bacitracin-streptomycin eliminated all cecal bacterial species except low numbers of aerobic sporeformers, and parenterally administered metronidazole selectively eliminated cecal anaerobic bacteria. Compared with control mice, only metronidazole-treated mice had significantly increased rates of dissemination of intestinal bacteria into mesenteric lymph nodes, indicating that the exclusive absence of anaerobic bacteria facilitated the translocation of the intestinal facultative bacteria. In a parallel experiment with streptomycin-resistant E. coli C25 as a marker, parallel results were obtained. Metronidazole increased the translocation of the marker strain and the indigenous strains of intestinal bacteria. Thus, anaerobes appeared to play a key role in confining indigenous bacteria to the gut. However, intestinal colonization and translocation of E. coli C25 occurred most readily after bacitracin-streptomycin treatment, suggesting that in addition to anaerobic bacteria, other bacterial groups may play a role in limiting the intestinal colonization and extraintestinal dissemination of E. coli C25.     

Bacterial colonization of disposable soft contact lenses is greater during corneal infiltrative events than during asymptomatic extended lens wear

Microorganisms, especially gram-negative bacteria, are considered to play a role in the etiology of certain corneal infiltrative events (CIEs) observed during soft contact lens wear. This study explored the possibility of microbial colonization of soft contact lenses as a risk factor leading to CIEs. In a clinical trial conducted from March 1993 to January 1996, 330 subjects wore disposable soft contact lenses on a 6-night extended-wear and disposal schedule. During this period, 4,321 lenses (118 during CIEs; 4,203 during asymptomatic lens wear) were recovered aseptically and analyzed for microbial colonization. A greater percentage of lenses were free from microbial colonization during asymptomatic wear than during CIEs (42 versus 23%; P < 0.0001). The incidence of gram-positive bacteria, gram-negative bacteria and fungi was greater during CIEs than during asymptomatic lens wear (P < 0.05). During asymptomatic lens wear, gram-positive bacteria were isolated most frequently and were usually normal external ocular microbiota. Of the gram-positive bacteria, the incidence of Streptococcus pneumoniae was greater during CIE than during asymptomatic wear (7.6 versus 0.6%; P < 0. 0001). While gram-negative bacteria were seen in few cases during asymptomatic wear, their incidence during CIE in comparison to asymptomatic wear was substantial and significant (23.7 versus 3.8%; P < 0.0001). Also, the level of colonization was high. Of CIEs, events of microbial keratitis, contact lens acute red eye, and asymptomatic infiltrative keratitis were associated with lens colonization with gram-negative bacteria or S. pneumoniae. Colonization of soft contact lenses with pathogenic bacteria, especially gram-negative bacteria and S. pneumoniae, appears to be a significant risk factor leading to CIE.     

Immunization with recombinant Streptococcus pneumoniae neuraminidase NanA protects chinchillas against nasopharyngeal colonization

Immunization with recombinant S. pneumoniae neuraminidase NanA (rNanA) resulted in a significant reduction in pneumococcal colonization in the chinchilla model. The bacteria were eliminated from the nasopharynx 1 week earlier than that from the control cohort. Our data suggest that rNanA affords protection against pneumococcal nasopharyngeal colonization.     

Modes of Helicobacter colonization and gastric epithelial damage

A total of 144 gastric biopsies colonized by Helicobacter-like organisms were studied under light and differential interference contrast microscopy for the modes of bacterial colonization. Biopsies were also graded for the degree of epithelial damage (epithelial-damage-grade: 0 to 6, in ascending order of severity) and density of Helicobacter-like organism (Helicobacter-grade: 0 to 6, in ascending order of bacterial density). Three modes of colonization were identified: free-in-mucus, surface-adhesion and intercellular colonization. Because light microscopy cannot definitely prove the presence of intracellular colonization, bacteria located between cells and below the apical cell border were counted together as intercellular colonization. Bacteria free-in-mucus were seen in all biopsies. Surface adhesion was seen in 50-87.9% of biopsies, without obvious correlation with the epithelial-damage- and Helicobacter-grades. The incidences of intercellular and intracellular colonization were directly proportional to the epithelial-damage- and Helicobacter-grades. Free-in-mucus as the predominant mode of colonization was mainly seen in biopsies with lower (1-3) epithelial-damage- and Helicobacter-grades. Conversely, biopsies with intercellular colonization as the predominant mode of colonization were mainly cases with higher (4-6) epithelial-damage- and Helicobacter-grades. In cases showing predominantly bacteria between cells, 69.2% had a gastric ulcer whereas only 38.8% of cases showing predominantly bacteria free-in-mucus showed ulceration (P < 0.01). These results indicate that Helicobacter-like organisms can invade and penetrate between epithelial cells. When free-in-mucus, Helicobacter-like organisms are less likely to induce epithelial damage. However, the more invasive modes of colonization (intercellular) were associated with severe epithelial damage and high Helicobacter density.(ABSTRACT TRUNCATED AT 250 WORDS)     

Proteomic analysis of Staphylococcus aureus biofilms grown in vitro on mechanical heart valve leaflets

The in vitro colonization of three commercial heart valve leaflets by Staphylococcus aureus was investigated. The leaflets, made of pyrolytic carbon alloyed with or without silicon, displayed similar surface properties (wettability, roughness) and were readily colonized by S. aureus that formed patchy biofilms on the three supports. A proteomic approach was used to assess the physiological status of biofilm populations by comparing their protein maps to those of bacteria cultured as free cells in the presence or absence of biofilm substratum. Principal component analysis (PCA) revealed, for each tested leaflet, statistical relationships between the protein maps of the biofilm and free-floating microbial populations. A spot-by-spot comparison of protein levels on two-dimensional electropherograms showed that many proteins were accumulated or underproduced by microbial populations grown in the presence of a leaflet compared with protein levels in control free populations. The number of accumulated proteins was noticeably higher than that of underproduced polypeptides. This protein overproduction was emphasized in biofilm populations. Several proteins, some of which were identified, were differentially produced by both surface-associated planktonic and biofilm-grown cell populations compared with control free-cell ones cultured in the absence of leaflet, whatever the leaflet tested. The potential of this proteomic approach for fighting against microbial adhesion and biofilm formation is discussed.     

Efficacy of titanium dioxide photocatalyst for inhibition of bacterial colonization on percutaneous implants

The purpose of this study was to evaluate the efficacy of titanium dioxide photocatalyst in inhibition of bacterial colonization on percutaneous implants. Titanium dioxide photocatalyst was prepared by direct oxidization of pure titanium substrate, and a comparative study with pure titanium was performed. The bactericidal ability of the photocatalyst was examined using methicillin-resistant Staphylococcus aureus (MRSA) suspensions in a colony-forming assay according to the Japanese Industrial Standards committee standard. After exposing the MRSA suspension on sample plates to ultraviolet A (UVA) light, the number of surviving bacteria was estimated. Next, an animal model for inhibition of colonization was examined in vivo. Pins were inserted into the femurs of rabbits, were infected with 10(8) colony-forming units of MRSA suspension, and were illuminated with UVA light for 60 min daily; the number of colonizing bacteria was estimated after 7 days. The bactericidal ability of the photocatalyst was apparent after 60 min, when the bacteria had almost disappeared. The number of colonizing bacteria on photocatalytic pins was decreased significantly in vivo. The photocatalyst was effective even against resistant bacterial colonization. Clinically, the incidence of percutaneous implant infection such as pin tract infection in external fixation could be reduced using the titanium photocatalyst.     

Accuracy of American Thoracic Society/Infectious Diseases Society of America criteria in predicting infection or colonization with multidrug-resistant bacteria at intensive-care unit admission

The aim of this prospective observational study was to determine the accuracy of American Thoracic Society (ATS)/Infectious Diseases Society of America (IDSA) criteria in predicting infection or colonization related to multidrug-resistant (MDR) bacteria at intensive-care unit (ICU) admission. MDR bacteria were defined as methicillin-resistant Staphylococcus aureus, ceftazidime-resistant or imipenem-resistant Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, and extended-spectrum β-lactamase-producing Gram-negative bacilli. Screening for MDR bacteria (using nasal and rectal swabs and tracheal aspirates from intubated patients) was performed at ICU admission. Risk factors for infection or colonization with MDR bacteria at ICU admission were determined using univariate and multivariate analyses. The accuracy of ATS/IDSA criteria in predicting infection or colonization with these bacteria at ICU admission was calculated. Eighty-three (13%) of 625 patients were infected or colonized with MDR bacteria at ICU admission. Multivariate analysis allowed identification of prior antimicrobial treatment (OR 2.3, 95% CI 1.2–4.3; p 0.008), residence in a nursing home (OR 2, 95% CI 1.1–3.7; p <0.001), and prior hospitalization (OR 3.9, 95% CI 1.7–8.8; p <0.001) as independent predictors of infection or colonization with MDR bacteria at ICU admission. Although sensitivity (89%) and negative predictive values (96%) were high, low specificity (39%) and a positive predictive value (18%) were found when ATS/IDSA criteria were used in predicting infection or colonization with MDR bacteria at ICU admission. In patients with pneumonia, adherence to guidelines was associated with increased rates of appropriate initial antibiotic treatment and de-escalation. ATS/IDSA criteria had an excellent negative predictive value and a low positive predictive value concerning infection or colonization with MDR bacteria at ICU admission.     

Microbial skin flora of selected cancer patients and hospital personnel.

The bacterial flora of the skin from five anatomical sites on 10 leukemia patients, 10 patients with malignant melanoma, and a control group of 10 medical personnel was examined quantitatively and qualitatively. This was done to determine whether malignant disease results in changes in skin flora and to establish carrier rates of gram-negative bacteria on the skin of personnel in hospital environments. Gram-negative bacteria were isolated more frequently (74 isolates from 100 cultures) from the skin of leukemia patients than from either patients with malignant melanoma (8 isolates from 100 cultures) or the medical personnel (9 isolates from 100 cultures). Klebsiella pneumoniae and Pseudomonas aeruginosa were isolated exclusively from leukemia patients. Relative proportions of gram-negative bacteria in total populations were determined. The axilla was the only site with a uniformly high proportion of gram-negative bacteria. From all other sites cultured, gram-negative populations were low (1 to 5 bacteria/cm2 of skin), although a high proportion of gram-negative populations occurred randomly throughout all subject groups. It was concluded that leukemia patients tend to carry gram-negative bacteria on the skin. The factors permitting colonization of skin by gram-negative bacteria are discussed.     

Bacterial motility is a colonization factor in experimental urinary tract infection.

In an experimental urinary tract infection of the mouse, colonization of the urinary bladder by isogenic strains of Salmonella enterica serovar Typhimurium was found to depend on the motility of the bacteria. Strains were obtained by genetic recombination between a highly motile O-6,7 and a poorly motile O-4,5,12 strain. The O antigen did not interfere with the colonization, whereas motility did; flagellated and motile O-6,7 and O-4,5,12 bacteria colonized the bladder equally well.     

Intestinal Microbiota Containing Barnesiella Species Cures Vancomycin-Resistant Enterococcus faecium Colonization

Bacteria causing infections in hospitalized patients are increasingly antibiotic resistant. Classical infection control practices are only partially effective at preventing spread of antibiotic-resistant bacteria within hospitals. Because the density of intestinal colonization by the highly antibiotic-resistant bacterium vancomycin-resistant Enterococcus (VRE) can exceed 10⁹ organisms per gram of feces, even optimally implemented hygiene protocols often fail. Decreasing the density of intestinal colonization, therefore, represents an important approach to limit VRE transmission. We demonstrate that reintroduction of a diverse intestinal microbiota to densely VRE-colonized mice eliminates VRE from the intestinal tract. While oxygen-tolerant members of the microbiota are ineffective at eliminating VRE, administration of obligate anaerobic commensal bacteria to mice results in a billionfold reduction in the density of intestinal VRE colonization. 16S rRNA gene sequence analysis of intestinal bacterial populations isolated from mice that cleared VRE following microbiota reconstitution revealed that recolonization with a microbiota that contains Barnesiella correlates with VRE elimination. Characterization of the fecal microbiota of patients undergoing allogeneic hematopoietic stem cell transplantation demonstrated that intestinal colonization with Barnesiella confers resistance to intestinal domination and bloodstream infection with VRE. Our studies indicate that obligate anaerobic bacteria belonging to the Barnesiella genus enable clearance of intestinal VRE colonization and may provide novel approaches to prevent the spread of highly antibiotic-resistant bacteria.     

Effects of intrapartum penicillin prophylaxis on intestinal bacterial colonization in infants

Early-onset group B streptococcal (GBS) infections remain a leading cause of morbidity and mortality in infants. To prevent the vertical transmission of GBS and neonatal GBS infection, guidelines recommend intrapartum penicillin or amoxicillin prophylaxis. This intrapartum antibiotic prophylaxis (IAP) is suspected to favor colonization by antibiotic-resistant bacteria. However, the effects of this prophylaxis on the patterns of acquisition of gastrointestinal bacterial flora in infants have never been studied. We collected stool samples from 3-day-old infants born to mothers who received intrapartum amoxicillin (antibiotic-exposed group; n = 25) and to untreated mothers (non-antibiotic-exposed group; n = 25). The groups were matched for factors known to affect intestinal microbial colonization: gestational age, type of delivery, and type of feeding. Qualitative and quantitative differential analyses of the bacterial flora in stool samples were performed. Similar numbers of infants in the non-antibiotic-exposed and antibiotic-exposed groups were colonized by aerobic bacteria and amoxicillin-resistant enterobacteria (75 and 77%, respectively) (P = 0.79). In contrast, significantly fewer infants in the antibiotic-exposed group than in the non-antibiotic-exposed group were colonized by anaerobic bacteria, especially Clostridium (12 and 40%, respectively) (P < 0.05). Regarding intestinal bacterial colonization, the differences between antibiotic-exposed and non-antibiotic-exposed infants were remarkably few. The only statistically significant effect was the reduced initial bacterial colonization by Clostridium in the antibiotic-exposed group. In our study, the use of IAP did not favor colonization by beta-lactam-resistant bacteria. However, further evaluations are required to highlight the potential risks of the widespread use of antibiotics to prevent early-onset GBS infection.     

An experimental intraarticular implantation of woven carbon fiber pad into osteochondral defect of the femoral condyle in rabbit

The defects of the articular cartilage structure are not replaced unless the subchondral plate has been breached. However, following the creation of a defect in the subchondral plate, the area is filled in with a fibrous tissue which gradually transforms to hyaline cartilage. The porous nontoxic materials of both biologic and synthetic origin have reportedly been used as matrices for repairing bone and cartilage. Following implantation, carbon fibre, chemically inert and well-tolerated by the body, induces a proliferation of ordered fibrous tissue. We implanted carbon fiber pads in osteochondral defects in rabbits. Those repairs were compared to control holes with no implants. The pads appeared to induce the gross appearance of a restored joint surface, mechanically strong to loading for periods from 2 to 6 weeks. Also, carbon fiber pads promoted the healing of the osteochondral defects in the rabbit femoral condyle, supplying well-organized cartilagenous tissue over repaired subchondral bone. The use of carbon fiber pads as implant material is suggested for the restoration of articular surface in osteoarthritis and osteochondritis dissecans.