EM evaluation of bacterial biofilm and microorganisms on the apical external root surface of human teeth

The aim of this study was to evaluate the presence of bacterial biofilm on the external surface of the root apex in teeth with pulp necrosis, with and without radiographically visible periapical lesions, and in teeth with a vital pulp. Twenty-one teeth were extracted, eight with pulp necrosis and periapical lesions, eight with pulp necrosis without radiographically visible periapical lesions, and five with a vital pulp. The roots were sectioned, and the root apexes (+/- 3 mm) were processed for scanning electron microscope evaluation. The surface of the apical root was evaluated for the presence of microorganisms, root resorption, and biofilm. There were no microorganisms on the apical root surface of either teeth with pulp vitality or with pulp necrosis with no radiographically visible periapical lesions. Microorganisms were always present in teeth with pulp necrosis and radiographically visible periapical lesions. These included cocci, bacilli, and filaments and the presence of an apical biofilm. Apical biofilm is clinically important because microbial biofilms are inherently resistant to antimicrobial agents and cannot be removed by biomechanical preparation alone. This may cause failure of endodontic treatment as a consequence of persistent infection.

The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability

Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) ≈500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) wide-spread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C. violaceum infection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications.

Analysis of susceptibility profile of Pseudomonas spp. and prevalence of bacterial samples from the surfaces of dental consulting-rooms

The aim of this research was to evaluate the susceptibility profile of Pseudomonas spp. and the prevalence of bacterial samples isolated from horizontal surfaces surrounding wash-basins used by dentists in several adjoined consulting-rooms, at points next to and at a distance from the basin, before and after surgical procedures. Our results showed a high percentage of Gram-positive cocci and Gram-negative bacilli; 34.66% were Staphylococcus spp. and 30.12% were non-fermentative Gram-negative bacilli among which Pseudomonas spp. (40.90%) was the commonest genus. Analysis of the susceptibility profile of Pseudomonas spp. isolates by determining the minimal inhibitory concentration (MIC) of 14 antibiotics showed a great variation among the strains and high rates of resistance to cefazolin, ceftazidime and aztreonan. Of the 14 antibiotics tested, 59.03% were found to be active against all the environmental isolates. Strains were resistant to aztreonan (62.82%), while susceptibility to third generation cephalosporins was variable.

Bacterial contamination of the catfish (Cathorops agassizii - Valenciennes, 1839 and Genidens genidens - Agassiz, 1829) sting

This work aimed to study the bacterial contamination in stings of the catfish Genidens genidens and Cathorops agassizii found in the São Vicente estuarine system (São Paulo State, Brazil). For bacteriological analyses, we used fish samples distributed into a group of 50 specimens (25 C. agassizii and 25 G. genidens) and a group of 14 specimens (7 C. agassizii and 7 G. genidens). Results showed contamination of 13 different bacterial species of Enterobacteriaceae, being Klebsiella pneumoniae the most frequent bacteria (26.80%) followed by Enterobacter sp and Escherichia coli (16.27%), and Serratia marcescens, Serratia sp. and Proteus mirabilis (1.16%). Gram-positive bacteria as well as fungi were not detected in the samples. According to the Gram-negative species characterized and with regard to the environmental conditions, it can also be considered that accidents with these catfish stings may develop significant acute secondary infections in humans.

Molecular modeling databases: A new way in the search of protein targets for drug development

DBMODELING is a relational database of annotated comparative protein structure models and their metabolic, pathway characterization. It is focused on enzymes identified in the genomes of Mycobacterium tuberculosis and Xylella fastidiosa. The main goal of the present database is to provide structural models to be used in docking simulations and drug design. However, since the accuracy of structural models is highly dependent on sequence identity between template and target, it is necessary to make clear to the user that only models which show high structural quality should be used in such efforts. Molecular modeling of these genomes generated a database, in which all structural models were built using alignments presenting more than 30% of sequence identity, generating models with medium and high accuracy. All models in the database are publicly accessible at http://www.biocristalografia.df.ibilce.unesp.br/tools. DBMODELING user interface provides users friendly menus, so that all information can be printed in one stop from any web browser. Furthermore, DBMODELING also provides a docking interface, which allows the user to carry out geometric docking simulation, against the molecular models available in the database. There are three other important homology model databases: MODBASE, SWISSMODEL, and GTOP. The main applications of these databases are described in the present article. © 2007 Bentham Science Publishers Ltd.

SEM evaluation of in situ early bacterial colonization on a Y-TZP ceramic: A pilot study

This study aimed to evaluate the effect of surface glazing and polishing of yttrium-stabilized tetragonal zirconia polycrystal ceramic on early dental biofilm formation, as well as the effect of brushing on the removal of adhered bacteria. Two subjects used oral appliances with polished and glazed samples fixed to the right and left sides. After 20 minutes, 1 hour, and 6 hours, the subjects manually brushed the samples on the right side. The samples were analyzed using scanning electron microscopy. Granular material was verified on the samples, especially on irregular surfaces. After 1 hour, there was no significant difference between glazed and polished surfaces in terms of bacterial presence. However, glazed surfaces tended to accumulate more biofilm, and brushing did not completely remove the biofilm. Polished surfaces seem to present a lower tendency for biofilm formation. Int J Prosthodont 2007;20:419-422.

Molecular typing of bacterial strains isolated from the Clinics of Surgery and Face Traumatology at Ribeirão Preto University

The number of infectious illnesses and cross infection is spreading drastically among the professionals of the dentistry area. Controlling infections in dental offices is one of the greatest challenges for dentists and researchers of this area. In practice, contacts between professionals and infected patients are relatively common. The transmission of infectious illnesses from the health professionals to their patients is also possible, either by direct contact or due to lack of cares in relation to biosafety, increasing the cycle of cross infection. Molecular typing is necessary since these methods are an important tool to investigate the epidemiology of bacterial infections. Moreover, they are important for supplying information and precedents through the analysis of the infectious agents eletrophoretic profile. The aim of the present work was to analyze by molecular typing the genomic profile of aerobic bacteria isolated from the Clinics of Surgery and Face Traumatology, Ribeirão Preto University, through the technique of Random Amplified Polymorphic DNA (RAPD) and grouped based on similarity coefficients. Of two carried out collections, 55 strains were isolates belonging to the following groups: 12 Staphylococcus aureus; 13 Klebsiella oxytoca; 7 Klebsiella pneumoniae; 8 Pseudomonas aeruginosa; 5 Hafnia alvei; 5 Proteus vulgaris; 4 Escherichia coli; and 1 Proteus mirabilis. The adopted molecular typing strategy allowed the determination of the persistence of definitive strains at the collection environment, besides the identification of strains proceeding from the hands and gloves of the surgeon dentists, which could have been found in distant places as sinks and reflectors.

SEM analysis of the in situ early bacterial colonization on two novel feldspathic ceramics submitted to different types of glazing

Aim: The aim of this study was to evaluate in situ, the early bacterial colonization on feldspar-ceramics submitted to different glazing. Methods and Materials: Fourteen standardized disc specimens (diameter: 5 mm, thickness: 1.5 mm) of each of two micro-particulate feldspathic ceramics (VM7 and VM13, Vita) were produced according to manufacturers' specifications for a total of 28 specimens (24 for the analysis of biofilm and 4 for topographic analysis analyzing the ceramic surfaces). Specimens from each type of ceramic were submitted to two different glazing methods composing four groups: VM7 glazed using glazing liquid Vita Akzent® 25 (G1) and glaze firing (G2), VM13 glazed using glazing liquid (G3) and glaze firing (G4). Six individuals (n=6) wore oral appliances with four ceramic specimens, fixed on the buccal face of the appliances. After 8 hours, each sample was evaluated for the presence (1) or absence (0) of bacterial colonization under a scanning electron microscope (SEM) on five randomly selected fields. The value for each sample was cumulative of the results observed in the fields. One sample from each group was evaluated under a SEM to verify the topographic pattern. Results: There was no difference with regard to bacterial colonization between the feldspar-ceramics and between the glazing types (Kruskal-Wallis non-parametric test). Conclusion: Feldspar-ceramics submitted to firing or glaze firing with Vita Akzent® 25 present a similar condition for in situ bacterial colonization. The similar topographic pattern of the ceramic surfaces seems to have influenced the bacterial colonization.

Bacterial cellulose from glucanacetobacter xylinus: Preparation, properties and applications

This chapter deals with the cellulose produced by the Glucanacetobacter xylinus strain, called bacterial cellulose, which is a remarkably versatile biomaterial usable in wide variety of domains, such as papermaking, optics, electronics, acoustics, and biomedical devices. Its unique structure shows entangled ultrafine fibers, which provide excellent mechanical strength, besides biodegradability, biocompatibility, high water-holding capacity, and high crystallinity. Some of its applications are described, such as complementary nutrition (. nata de coco), artificial temporary skin for wounds and burns, dental aid, artificial blood vessels and micronerve surgery, DNA separation, composite reinforcement, electronic paper, light emitting diodes, and fuel cell membranes. © 2007 Elsevier Ltd. All rights reserved.

Development and validation of a Xanthomonas axonopodis pv. citri DNA microarray platform (XACarray) generated from the shotgun libraries previously used in the sequencing of this bacterial genome

Background. From shotgun libraries used for the genomic sequencing of the phytopathogenic bacterium Xanthomonas axonopodis pv. citri (XAC), clones that were representative of the largest possible number of coding sequences (CDSs) were selected to create a DNA microarray platform on glass slides (XACarray). The creation of the XACarray allowed for the establishment of a tool that is capable of providing data for the analysis of global genome expression in this organism. Findings. The inserts from the selected clones were amplified by PCR with the universal oligonucleotide primers M13R and M13F. The obtained products were purified and fixed in duplicate on glass slides specific for use in DNA microarrays. The number of spots on the microarray totaled 6,144 and included 768 positive controls and 624 negative controls per slide. Validation of the platform was performed through hybridization of total DNA probes from XAC labeled with different fluorophores, Cy3 and Cy5. In this validation assay, 86% of all PCR products fixed on the glass slides were confirmed to present a hybridization signal greater than twice the standard deviation of the deviation of the global median signal-to-noise ration. Conclusions. Our validation of the XACArray platform using DNA-DNA hybridization revealed that it can be used to evaluate the expression of 2,365 individual CDSs from all major functional categories, which corresponds to 52.7% of the annotated CDSs of the XAC genome. As a proof of concept, we used this platform in a previously work to verify the absence of genomic regions that could not be detected by sequencing in related strains of Xanthomonas. © 2010 Moreira et al; licensee BioMed Central Ltd.

Easiness of use and validity testing of VS-SENSE device for detection of abnormal vaginal flora and bacterial vaginosis

Accessing vaginal pH is fundamental during gynaecological visit for the detection of abnormal vaginal flora (AVF), but use of pH strips may be time-consuming and difficult to interpret. The aim of this study was to evaluate the VS-SENSE test (Common Sense Ltd, Caesarea, Israel) as a tool for the diagnosis of AVF and its correlation with abnormal pH and bacterial vaginosis (BV). The study population consisted of 45 women with vaginal pH ≥ 4.5 and 45 women with normal pH. Vaginal samples were evaluated by VS-SENSE test, microscopy and microbiologic cultures. Comparing with pH strips results, VS-SENSE test specificity was 97.8% and sensitivity of 91%. All severe cases of BV and aerobic vaginitis (AV) were detected by the test. Only one case with normal pH had an unclear result. Concluding, VS-SENSE test is easy to perform, and it correlates with increased pH, AVF, and the severe cases of BV and AV. Copyright © 2010 Gilbert G. G. Donders et al.

Bacterial cellulose-hydroxyapatite nanocomposites for bone regeneration

The aim of this study was to develop and to evaluate the biological properties of bacterial cellulose-hydroxyapatite (BC-HA) nanocomposite membranes for bone regeneration. Nanocomposites were prepared from bacterial cellulose membranes sequentially incubated in solutions of CaCl2 followed by Na2HPO4. BC-HA membranes were evaluated in noncritical bone defects in rat tibiae at 1, 4, and 16 weeks. Thermogravimetric analyses showed that the amount of the mineral phase was 40-50 of the total weight. Spectroscopy, electronic microscopy/energy dispersive X-ray analyses, and X-ray diffraction showed formation of HA crystals on BC nanofibres. Low crystallinity HA crystals presented Ca/P a molar ratio of 1.5 (calcium-deficient HA), similar to physiological bone. Fourier transformed infrared spectroscopy analysis showed bands assigned to phosphate and carbonate ions. In vivo tests showed no inflammatory reaction after 1 week. After 4 weeks, defects were observed to be completely filled in by new bone tissue. The BC-HA membranes were effective for bone regeneration. © 2011 S. Saska et al.

Reduction of bacterial adhesion to biocompatible polymer surfaces via plasma processing

Plasma processing of the surfaces of biomaterials is interesting because it enables modification of the characteristics of a surface without affecting bulk properties. In addition, the results are strongly influenced by the conditions of the treatment. Therefore, by adjusting the plasma parameters it is possible to tailor the surface properties to best fulfill the requirements of a given application. In this work, polyurethane substrates have been subjected to sulfur hexafluoride glow discharge plasmas. The influences of different SF 6 plasma exposure times and pressures on the adhesion of Staphylococcus aureus and Pseudomonas aeruginosa to the polymer have been investigated. The wettability and surface free energy have been evaluated via contact angle measurements. At low pressure (6.7 Pa) the contact angle decreases with increasing exposure time in the 180 s to 540 s interval, but at higher pressure (13.3 Pa) it increases as a function of the same variable. Bacterial adhesion has been quantified from in vitro experiments by determining the growth of colonies on Petri dishes treated with agar nutrient. It has been observed that the surface properties play an important role in microbe adhesion. For instance, the density of adhered P. aeruginosa decreased as the surface contact angle increased. S. aureus preferred to adhere to hydrophobic surfaces. © 2011 by Begell House, Inc.

Preparation and characterization of the bacterial cellulose/polyurethane nanocomposites

New nanocomposites based on bacterial cellulose nanofibers (BCN) and polyurethane (PU) prepolymer were prepared and characterized by SEM, FT-IR, XRD, and TG/DTG analyses. An improvement of the interface reaction between the BCN and the PU prepolymer was obtained by a solvent exchange process. FT-IR results showed the main urethane band at 2,270 cm-1 to PU prepolymer; however, in nanocomposites new bands appear as disubstituted urea at 1,650 and 1,550 cm-1. In addition, the observed decrease in the intensity of the hydroxyl band (3,500 cm-1) suggests an interaction between BCN hydroxyls and NCO-free groups. The nanocomposites presented a non-crystalline character, significant thermal stability (up to 230 °C) and low water absorption when compared to pristine BCN. © 2013 Akadémiai Kiadó, Budapest, Hungary.

Bacterial nanocellulose for medical implants

Bacterial cellulose (BC) has established to be a remarkably versatile biomaterial and can be used in wide variety of applied scientific endeavours, especially for medical devices. In fact, biomedical devices recently have gained a significant amount of attention because of an increased interest in tissue-engineered products for both wound care and the regeneration of damaged or diseased organs. Due to its unique nanostructure and properties, microbial cellulose is a natural candidate for numerous medical and tissue-engineered applications. Hydrophilic bacterial cellulose fibers of an average diameter of 50 nm are produced by the bacterium Acetobacter xylinum, using a fermentation process. The microbial cellulose fiber has a high degree of crystallinity. Using direct nanomechanical measurement, determined that these fibers are very strong and when used in combination with other biocompatible materials, produce nanocomposites particularly suitable for use in human and veterinary medicine. Moreover, the nanostructure and morphological similarities with collagen make BC attractive for cell immobilization and cell support. The architecture of BC materials can be engineered over length scales ranging from nano to macro by controlling the biofabrication process. The chapter describes the fundamentals, purification and morphological investigation of bacterial cellulose. This chapter deals with the modification of microbial cellulose and how to increase the compatibility between cellulosic surfaces and a variety of plastic materials. Furthermore, provides deep knowledge of fascinating current and future applications of bacterial cellulose and their nanocomposites especially in the medical field, materials with properties closely mimic that of biological organs and tissues were described. © Springer-Verlag Berlin Heidelberg 2013.