Adherence Of Streptococci Research Articles

Is microbial adhesion affected by the build orientation of a 3-dimensionally printed denture base resin?

Statement of problemHow the build orientation of a 3-dimensionally (3D) printed denture affects microbial adhesion is unclear. PurposeThe purpose of this in vitro study was to compare the adherence of Streptococcus spp. and Candida spp. on 3D-printed denture bases prepared at different build orientations with conventional heat-polymerized resin. Material and methodsResin specimens (n=5) with standardized 28.3 mm2 surface area were 3D printed at 0 and 60 degrees, and heat-polymerized (3DP-0, 3DP-60, and HP, respectively). The specimens were placed in a Nordini artificial mouth (NAM) model and exposed to 2 mL of clarified whole saliva to create a pellicle-coated substratum. Suspensions of Streptococcus mitis and Streptococcus sanguinis, Candida albicans and Candida glabrata, and a mixed species, each at 108 cfu/mL were pumped separately into the model for 24 hours to promote microbial adhesion. The resin specimens were then removed, placed in fresh media, and sonicated to dislodge attached microbes. Each suspension (100 μL) was aliquoted and spread on agar plates for colony counting. The resin specimens were also examined under a scanning electron microscope. The interaction between types of specimen and groups of microbes was examined with 2-way ANOVA and then further analysis with Tukey honest significant test and Kruskal-Wallis post hoc tests (α=.05). ResultsA significant interaction was observed between the 3DP-0, 3DP-60, and HP specimen types and the groups of microbes adhering to the corresponding denture resin specimens (P<.05). The difference was statistically significant among 3DP-0, 3DP-60, and HP specimens (P<.05). The adherence of candida was 3.98-times lower on the 3DP-0 than that of HP (P<.05). However, adherence of the mixed-species microbes and streptococci on the 3DP-60 were 1.75 times and 2-fold higher, respectively (P<.05). The scanning electron micrographs showed that 3DP-0 exhibited the lowest microbial adherence compared with HP and 3DP-60. ConclusionsAdherence affinity of denture base resin is affected by the build orientation rather than by the group of different microbes. Three-dimensionally printed denture base resin fabricated at a 0-degree build orientation exhibited low affinity for microbial adhesion. Three-dimensionally printed dentures may reduce microbial adhesion when printed at a 0-degree build orientation.

Construction and protective efficacy of a novel Streptococcus pneumoniae fusion protein vaccine NanAT1-TufT1-PlyD4.

During the past decades, with the implementation of pneumococcal polysaccharide vaccine (PPV) and pneumococcal conjugate vaccines (PCVs), a dramatic reduction in vaccine type diseases and transmissions has occurred. However, it is necessary to develop a less expensive, serotype-independent pneumococcal vaccine due to the emergence of nonvaccine-type pneumococcal diseases and the limited effect of vaccines on colonization. As next-generation vaccines, conserved proteins, such as neuraminidase A (NanA), elongation factor Tu (Tuf), and pneumolysin (Ply), are promising targets against pneumococcal infections. Here, we designed and constructed a novel fusion protein, NanAT1-TufT1-PlyD4, using the structural and functional domains of full-length NanA, Tuf and Ply proteins with suitable linkers based on bioinformatics analysis and molecular cloning technology. Then, we tested whether the protein protected against focal and lethal pneumococcal infections and examined its potential protective mechanisms. The fusion protein NanAT1-TufT1-PlyD4 consists of 627 amino acids, which exhibits a relatively high level of thermostability, high stability, solubility and a high antigenic index without allergenicity. The purified fusion protein was used to subcutaneously immunize C57BL/6 mice, and NanAT1-TufT1-PlyD4 induced a strong and significant humoral immune response. The anti-NanAT1-TufT1-PlyD4 specific IgG antibody assays increased after the first immunization and reached the highest value at the 35th day. The results from in vitro experiments showed that anti-NanAT1-TufT1-PlyD4 antisera could inhibit the adhesion of Streptococcus pneumoniae (S. pneumoniae) to A549 cells. In addition, immunization with NanAT1-TufT1-PlyD4 significantly reduced S. pneumoniae colonization in the lung and decreased the damage to the lung tissues induced by S. pneumoniae infection. After challenge with a lethal dose of serotype 3 (NC_WCSUH32403), a better protection effect was observed with NanAT1-TufT1-PlyD4-immunized mice than with the separate full-length proteins and the adjuvant control; the survival rate was 50%, which met the standard of the marketed vaccine. Moreover, we showed that the humoral immune response and the Th1, Th2 and Th17-cellular immune pathways are involved in the immune protection of NanAT1-TufT1-PlyD4 to the host. Collectively, our results support that the novel fusion protein NanAT1-TufT1-PlyD4 exhibits extensive immune stimulation and is effective against pneumococcal challenges, and these properties are partially attributed to humoral and cellular-mediated immune responses.

Secondary streptococcal infection following influenza.

Secondary bacterial infection following influenza type A virus (IAV) infection is a major cause of morbidity and mortality during influenza epidemics. Streptococcus pneumoniae has been identified as a predominant pathogen in secondary pneumonia cases that develop following influenza. Although IAV has been shown to enhance susceptibility to the secondary bacterial infection, the underlying mechanism of the viral-bacterial synergy leading to disease progression is complex and remains elusive. In this review, cooperative interactions of viruses and streptococci during co- or secondary infection with IAV are described. IAV infects the upper respiratory tract, therefore, streptococci that inhabit or infect the respiratory tract are of special interest. As many excellent reviews on the co-infection of IAV and S. pneumoniae have already been published, this review is intended to describe the unique interactions between other streptococci and IAV. Both streptococcal and IAV infections modulate the host epithelial barrier of the respiratory tract in various ways. IAV infection directly disrupts epithelial barriers, though at the same time the virus modifies the properties of infected cells to enhance streptococcal adherence and invasion. Mitis group streptococci produce neuraminidases, which promote IAV infection in a unique manner. The studies reviewed here have revealed intriguing mechanisms underlying secondary streptococcal infection following influenza.

Effects of 2% sodium fluoride solution on the prevention of streptococcal adhesion to titanium and zirconia surfaces

Streptococci are associated with dental plaque formation as the early-colonizing bacteria that adhere to titanium (CpTi) and zirconia (TZP) implant abutment surfaces. Effective prevention of peri-implantitis may be possible by removing streptococci as target. This study aimed to evaluate the effects of 2% NaF on the prevention of streptococcal adhesion to CpTi and TZP. After immersion in 2% NaF for 90 min, surface characterization of mirror-polished CpTi and TZP disks were assesed using XPS, EPMA, and SEM. S. sanguinis, S. gordonii, and S. oralis were used as the streptococcal bacterial strains. After 24 h culture, bacterial adhesion was evaluated using an ATP-bioluminescent assay and SEM. In XPS, EPMA, and SEM analyses, fluoride was detected on the CpTi and TZP surfaces after 2% NaF immersion with no signs of localization, and no corrosion on the CpTi disks. Based on the adhesion assay, the adherences of S. sanguinis, S. gordonii, and S. oralis were significantly lower with NaF than without NaF in CpTi (p = 0.005, 0.001, and 0.001, respectively) and TZP (p = 0.003, 0.002, and 0.001). This was also confirmed by SEM. In conclusion, 2% NaF reduced the adhesion of streptococci to the CpTi and TZP surfaces.

Inhibitory effects of 2-methacryloyloxyethyl phosphorylcholine polymer on the adherence of bacteria causing upper respiratory tract infection

ABSTRACT Objective We aimed to investigate the inhibitory effect of 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer on the adherence of Streptococcus pneumoniae (Spn) and nontypeable Haemophilus influenzae (NTHi) in vitro and in vivo. Materials and Methods Phosphorylcholine (PC) expression of 21 strains each of Spn and NTHi was evaluated using fluorescence-activated cell sorting; the adherence of bacteria to Detroit 562 cells and to the nasal mucosa of BALB/c mice was determined. MPC polymer-mediated inhibitory effects were compared with PC-keyhole limpet hemocyanin (PC-KLH)-mediated inhibitory effects. Results In vitro experiments showed that pretreatment with MPC polymer markedly inhibited the adherence of Spn and NTHi in a concentration dose–dependent manner independently of PC expression. No correlation was observed between PC expression and MPC polymer-mediated inhibitory effects. Contrarily, there was a significant negative correlation between PC-KLH-mediated inhibitory effects and PC expression in Spn and NTHi. The same results were obtained via in vivo experiments. The MPC polymer did not affect the histology of the nasal mucosa. Conclusions MPC polymer might be effective to reduce the occurrence of upper respiratory tract infection caused by Spn and NTHi and could be applied for the development of local treatments, such as topical gargles and nebulizer medications.

Microbial changes in biofilms on composite resins with different surface roughness: An in vitro study with a multispecies biofilm model

Statement of problemThe single-species biofilm method cannot represent the interaction and complex functions of microorganisms associated with oral biofilms. PurposeThe purpose of this in vitro study was to investigate microbial changes in biofilms on composite resins of varying surface roughness by using a multispecies biofilm model with early-colonizing streptococci, middle colonizer, and late-colonizing gram-negative anaerobes. Material and methodsComposite resin disks were prepared with different roughness: SR180, SR400, SR1500, and SRGlass roughened with 180-, 400-, and 1500-grit silicon carbide paper and glass (control surface without surface roughening). Surface roughness was analyzed by confocal laser scanning and scanning electron microscopy. After multispecies biofilms had been grown on the composite resin surfaces, the adhesion of Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and of total bacteria was determined after 1 (T1) and 4 (T2) days. Differences in surface roughness among the 4 groups were tested with 1-way ANOVA. Multifactorial analysis of variance was used to determine the time-related differences in the bacterial composition with respect to surface roughness (α=.05). ResultsThe order of SR, from highest to lowest, was SR180 (1.45 ±0.11 μm), SR400 (0.62 ±0.05 μm), SR1500 (0.35 ±0.02 μm), and SRGlass (0.15 ±0.01 μm) (SR180>SR400>SR1500>SRGlass, P<.001). Increased surface roughness was not proportional to bacterial adhesion. Significant differences in the adhesion of total bacteria was only found between SRGlass and SR180 (SR180>SRGlass, P=.029). The adhesion of S. mutans and S. sobrinus to SR180 and SR400 was higher than that to SRGlass (SR180=SR400>SRGlass; S. mutans, P=.003; S. sobrinus, P=.002). However, the adhesion of A. actinomycetemcomitans and P. gingivalis to composite resin was not significantly influenced by surface roughness. Adhesion of total bacteria, S. mutans, and S. sobrinus increased from T1 to T2 (T1<T2, P<.001), whereas the adhesion of periodontal pathogens decreased from T1 to T2 (T1>T2; A. actinomycetemcomitans, P<.001; P. gingivalis, P=.013). ConclusionsDecreased adhesion of cariogenic streptococci and total bacteria was observed at surface roughness values of around 0.15 μm. Periodic finishing of surface roughness should be considered to minimize the adhesion of cariogenic streptococci to composite resin surfaces.

Vitamin D attenuates rhinovirus-induced expression of intercellular adhesion molecule-1 (ICAM-1) and platelet-activating factor receptor (PAFR) in respiratory epithelial cells

Human rhinoviruses commonly cause upper respiratory infections, which may be complicated by secondary bacterial infection. Vitamin D replacement reduces risk of acute respiratory infections in vitamin D-deficient individuals, but the mechanisms by which such protection is mediated are incompletely understood. We therefore conducted experiments to characterise the influence of the major circulating metabolite 25-hydroxyvitamin D (25[OH]D) and the active metabolite 1,25-dihydroxyvitamin D (1,25[OH]2D) on responses of a respiratory epithelial cell line (A549 cells) to infection with a major group human rhinovirus (RV-16). Pre-treatment of A549 respiratory epithelial cells with a physiological concentration (10−7M) of 25(OH)D induced transient resistance to infection with RV-16 and attenuated RV-16-induced expression of the genes encoding intercellular adhesion molecule 1 (ICAM-1, a cell surface glycoprotein that acts as the cellular receptor for major group rhinoviruses) and platelet-activating factor receptor (PAFR, a G-protein coupled receptor implicated in adhesion of Streptococcus pneumoniae to respiratory epithelial cells). These effects were associated with enhanced expression of the genes encoding the NF-κB inhibitor IκBα and the antimicrobial peptide cathelicidin LL-37. Our findings suggest possible mechanisms by which vitamin D may enhance resistance to rhinovirus infection and reduce risk of secondary bacterial infection in vitamin D-deficient individuals.

Streptococcus pneumoniae galU gene mutation has a direct effect on biofilm growth, adherence and phagocytosis in vitro and pathogenicity in vivo.

Streptococcus pneumoniae, the most common cause of bacterial pneumonia, has developed a wide range of virulence factors to evade the immune system of which the polysaccharide capsule is the most important one. Formation of this capsule is dependent on the cps gene locus, but also involves other genes-like galU. The pyrophosphorylase encoded by galU plays a role in the UDP-glucose metabolism of prokaryotes and is required for the biosynthesis of capsular polysaccharides. In this paper, the effect of a galU mutation leading to a dysfunctional UDP-glucose pyrophosphorylase (UDPG:PP) on in vitro biofilm biomass, adherence to lung epithelial cells and macrophage phagocytosis is studied. Last, in vivo virulence using a Galleria mellonella model has been studied. We show that the mutation improves streptococcal adherence to epithelial cells and macrophage phagocytosis in vitro, while there is no definitive correlation on biofilm formation between parent and mutant strains. Moreover, in vivo virulence is attenuated for all mutated strains. Together, these results demonstrate that a galU mutation in S. pneumoniae influences host cell interactions in vitro and in vivo and can strongly influence the outcome of a streptococcal infection. As such, UDPG:PP is worth investigating further as a potential drug target.

The role of cell wall surface proteins and adherence of Streptococcus pneumoniae strains in host pathogen interaction

The role of cell wall surface proteins and adherence of Streptococcus pneumoniae strains in host pathogen interaction

Effects of Manufacturing and Finishing Techniques of Feldspathic Ceramics on Surface Topography, Biofilm Formation, and Cell Viability for Human Gingival Fibroblasts.

Feldspathic ceramic restorations can be obtained by different techniques (stratification or computer-aided design/computer-aided manufacturing [CAD/CAM] blocks) and finishing procedures (polishing or glaze application). This study evaluated the effects of techniques and finishing procedures on surface properties, biofilm formation, and viability of human gingival fibroblasts (FMM-1) in contact with these materials. Ceramic specimens were obtained through a stratification technique (Vita VM9) and from CAD/CAM blocks (Vita Blocs Mark II; both Vita Zahnfabrik) and their surfaces were finished by polishing (ceramisté diamond rubbers + polishing paste; "p" subgroups) or glaze spray application + sintering ("g" subgroups). Roughness (Ra and RSm parameters) and surface free energy (SFE) were measured. Early biofilm formation of Streptococcus mutans, Streptococcus sanguinis, and Candida albicans was evaluated by counting colony-forming units (CFU). MTT (3-[4.5-dimethyl-thiazol-2-yl-]-2.5-diphenyl tetrazolium bromide) cytotoxicity test evaluated cellular viability for the growth of FMM-1 after 24 hours and seven days of contact. Scanning electron microscopy (SEM) and three-dimensional optical profilometry were performed to qualitatively analyze the surface. Data were analyzed by analysis of variance, Tukey test, and t-test (all α=0.05). Polished samples presented lower roughness (Ra, p=0.015; RSm, p=0.049) and higher SFE ( p=0.00). Streptococci had higher CFU in all groups, but the CFU of C albicans was lower for polished samples. Biofilm formation was influenced by the interaction of all factors ( p=0.018), and the materials showed no cytotoxicity to FMM-1 growth. Polishing resulted in the lowest values for surface roughness and higher SFE values. Polished ceramics showed less C albicans adherence while the adherence of Streptococci was greater than C albicans in all conditions.

Adherence of Streptococcus pneumoniae and expression analysis of neuraminidase gene (NanA and NanB) after interaction of A549 human lung epithelial cells with pneumococcal strains of various serotypes

Adherence of Streptococcus pneumoniae and expression analysis of neuraminidase gene (NanA and NanB) after interaction of A549 human lung epithelial cells with pneumococcal strains of various serotypes

The Effect of Vinegar, Rose Water and Ethanolic Extract Green Tea Against Oral Streptococci, an In Vitro Study

Objective: Dental caries is the most common infectious diseases that are involved all groups, ages and classes of society people. Among the oral bacteria, streptococci especially Streptococcus mutans and Streptococcus salivarius are known as the most important microbial agents in dental caries and dental plaque. Dental caries treatment imposes heavy costs in all countries. Although there are different chemical antimicrobial agents for the prevention of of dental caries, but their important side effects have reported. Therefore, many attempts have done for finding alternative safe medications specially natural ingredients. The aim of this research is determination of antimicrobial effects of vinegar, rose water and green tea against four cariogenic bacteria.Methods: The antimicrobial effects of different concentrations of vinegar, rose water and green tea against Streptococcus mutans, Streptococcus salivarius, Streptococcus sanguis and Streptococcus salivarius are evaluated with disc diffusion, well plate and microtitre plate methods, also the effects of them against biofilm formation were studied.Results: The results showed vinegar, rose water and green tea significantly reduced biofilm formation of the streptococci. Vinegar and green tea decreased more then 70% the adhesion of streptococci but rose water decreased more then 70% of S. salivarius, more then 60% of S. sobrinus, S. sanguis and more then 50% of S. mutans adhesion. The diameter of highest inhibition zone was 24.2 mm for Streptococcus salivarius and 22 mm for Streptococcus salivarius against vinegar.The MIC of vinegar was for S. salivarius 0.0312 and MBC was 0.0625 (P<0.05), but the MIC of green tea for Streptococcus mutans and Streptococcus salivarius was 7.81 mg/ml and its MBC was 31.25 mg/ml which was significantly less than that for Streptococcus sanguis and Streptococcus salivarius which was 15.625 and 62.5 mg/ml, respectively (P<0.05). The MIC of rose water for Streptococcus salivarius, Streptococcus sanguis, Streptococcus salivarius was 1.Conclusions: This results indicated the potential capacity of vinegar, rose water and green tea for prevention or control of cariogenic streptococci proliferation.

Dental materials and their performance for the management of screw access channels in implant-supported restorations.

Unsuccessfully sealed screw access channels of prosthetic implant abutments may lead to malodor or peri-implant diseases in gingival tissues adjacent to implant-supported restorations. Therefore, 72 sets of screw channel analogs with six different materials incorporated (Polytetrafluoroethylene (PTFE), wax, gutta-percha, cavit, endofrost-pellets and cotton pellets) were exposed (2.5 h, 37°C) to Streptococcus mutans, oralis and Candida albicans suspensions. Bacterial adherence was quantified by using the fluorescence dye, Alamar Blue/resazurin, and an automated multifunctional reader. For quantification of fungal adherence the ATP-based bioluminescence approach was used. High relative fluorescence and luminescence intensities (>10,000), indicating high adhesion of streptococci and fungi were found for cotton and endofrost-pellets and low intensities (<5,000) for wax, gutta-percha, cavit and PTFE. The quantity of bacterial and fungal adhesion differed significantly between the assessed various sealing materials. In conclusion and within the limitations of this study, wax, gutta-percha, cavit and PTFE should be preferred as sealing materials.

Host-derived extracellular RNA promotes adhesion of Streptococcus pneumoniae to endothelial and epithelial cells.

Streptococcus pneumoniae is the most frequent cause of community-acquired pneumonia. The infection process involves bacterial cell surface receptors, which interact with host extracellular matrix components to facilitate colonization and dissemination of bacteria. Here, we investigated the role of host-derived extracellular RNA (eRNA) in the process of pneumococcal alveolar epithelial cell infection. Our study demonstrates that eRNA dose-dependently increased S. pneumoniae invasion of alveolar epithelial cells. Extracellular enolase (Eno), a plasminogen (Plg) receptor, was identified as a novel eRNA-binding protein on S. pneumoniae surface, and six Eno eRNA-binding sites including a C-terminal 15 amino acid motif containing lysine residue 434 were characterized. Although the substitution of lysine 434 for glycine (K434G) markedly diminished the binding of eRNA to Eno, the adherence to and internalization into alveolar epithelial cells of S. pneumoniae strain carrying the C-terminal lysine deletion and the mutation of internal Plg-binding motif were only marginally impaired. Accordingly, using a mass spectrometric approach, we identified seven novel eRNA-binding proteins in pneumococcal cell wall. Given the high number of eRNA-interacting proteins on pneumococci, treatment with RNase1 completely inhibited eRNA-mediated pneumococcal alveolar epithelial cell infection. Our data support further efforts to employ RNAse1 as an antimicrobial agent to combat pneumococcal infectious diseases.

Xylitol for preventing acute otitis media in children up to 12 years of age.

Acute otitis media (AOM) is the most common bacterial infection among young children in the United States. There are limitations and concerns over its treatment with antibiotics and surgery and so effective preventative measures are attractive. A potential preventative measure is xylitol, a natural sugar substitute that reduces the risk of dental decay. Xylitol can reduce the adherence of Streptococcus pneumoniae (S pneumoniae)andHaemophilus influenzae (H influenzae) to nasopharyngeal cells in vitro. This is an update of a review first published in 2011. To assess the efficacy and safety of xylitol to prevent AOM in children aged up to 12 years. We searched CENTRAL (to Issue 12, 2015), MEDLINE (1950 to January 2016), Embase (1974 to January 2016), CINAHL (1981 to January 2016), LILACS (1982 to January 2016), Web of Science (2011 to January 2016) and International Pharmaceutical Abstracts (2000 to January 2016). Randomised controlled trials (RCTs) or quasi-RCTs of children aged 12 years or younger where xylitol supplementation was compared with placebo or no treatment to prevent AOM. Two review authors independently selected trials from search results, assessed and rated study quality and extracted relevant data for inclusion in the review. We contacted trial authors to request missing data. We noted data on any adverse events of xylitol. We extracted data on relevant outcomes and estimated the effect size by calculating risk ratio (RR), risk difference (RD) and associated 95% confidence intervals (CI). We identified five clinical trials that involved 3405 children for inclusion. For this 2016 update, we identified one new trial for inclusion. This trial was systematically reviewed but due to several sources of heterogeneity, was not included in the meta-analysis. The remaining four trials were of adequate methodological quality. In three RCTs that involved a total of 1826 healthy Finnish children attending daycare, there is moderate quality evidence that xylitol (in any form) can reduce the risk of AOM from 30% to around 22% compared with the control group (RR 0.75, 95% CI 0.65 to 0.88). Among the reasons for dropouts, there were no significant differences in abdominal discomfort and rash between the xylitol and the control groups. Xylitol was not effective in reducing AOM among healthy children during a respiratory infection (RR 1.13, 95% CI 0.83 to 1.53; moderate quality evidence) or among otitis-prone healthy children (RR 0.90, 95% CI 0.67 to 1.21; low-quality evidence). There is moderate quality evidence showing that the prophylactic administration of xylitol among healthy children attending daycare centres can reduce the occurrence of AOM. There is inconclusive evidence with regard to the efficacy of xylitol in preventing AOM among children with respiratory infection, or among otitis-prone children. The meta-analysis was limited because data came from a small number of studies, and most were from the same research group.

Identification of a Monoclonal Antibody Against Pneumococcal Pilus 1 Ancillary Protein Impairing Bacterial Adhesion to Human Epithelial Cells.

The adhesion of Streptococcus pneumoniae is a key step during colonization of human respiratory tract mucosae. Here we demonstrate that pneumococcal type I pilus significantly increases the adhesiveness of poorly adhering highly capsulated strains in vitro. Interestingly, preincubation of bacteria with antibodies against the major pilus backbone subunit (RrgB) or the adhesin component (RrgA) impaired pneumococcal association to human epithelial cells. Screening for anti-RrgA monoclonal antibodies specifically affecting the adhesive capacity of S. pneumoniae led to the identification of the monoclonal 11B9/61 antibody, which greatly reduced pilus-dependent cell contact. Proteomic-based epitope mapping of 11B9/61 monoclonal antibody revealed a well-exposed epitope on the D2 domain of RrgA as the target of this functional antibody. The data presented here confirm the importance of pilus I for S. pneumoniae pathogenesis and the potential use of antipilus antibodies to prevent bacterial colonization.

Quantitative assessment of Mutans Streptococci adhesion to coated and uncoated orthodontic archwires(In vitro study)

Background: The development of orthodontic biomaterials that attract less biofilm has been a goal for decades. Adhesion and colonization of cariogenic streptococci are considered to play key roles in the development of enamel demineralization related to orthodontic materials. The aim of this study was to quantitatively evaluate the Mutans streptococci adhesion to coated orthodontic archwires (Epoxy and Teflon) and uncoated archwires (stainless steel and nickel-titanium) with respect to incubation time in the presence and absence of saliva. Material and Method: Six types of archwires stainless steel and nickel titanium with two type of coating (Epoxy, Teflon) were used in this study. Twelve specimens of each archwire were incubated in sterilized unstimulated whole saliva (for the study group) and phosphate-buffered saline (for control group) for 2 hours, then incubated with suspension of Mutans streptococci allowed to adhere for (5,90,180 minutes). Adhesion was quantitated by a microbial culture technique by treating the archwires with adhering bacteria with trypsin and enumerating the colony forming unit (CFU) counts of bacteria recovered after cultivation by using Dentocult SM kit. Results: There was significant difference among the tested archwire types in each time interval with the highest bacterial adhesion on the NiTi archwires in the absence of saliva. In the presence of saliva, the results revealed non-significant difference at 5 min. while there was significant difference at 90 min and highly significant difference at 180 min. Conclusion: The adherence of Mutans streptococci was decreased in the presence of saliva on different archwires and the extended incubation time was significantly related to increase colony forming unit of Mutans Streptococci.

Preceding human metapneumovirus infection increases adherence of Streptococcus pneumoniae and severity of murine pneumococcal pneumonia.

Coinfection with respiratory virus and Streptococcus pneumoniae has been frequently reported in several epidemiologic studies. The aim of this study was to explore the effect of preceding human metapneumovirus (hMPV) inoculation on subsequent pneumococcal infection. Hep-2 and A549 cells were infected with hMPV then inoculated with S. pneumoniae. Bacterial adhesion was measured using colony forming unit and cytometric-fluorescence assays. In vivo bacterial adhesion was examined in hMPV-infected mice after inoculation of fluorescence-conjugated S. pneumoniae. Pulmonary inflammation (bacterial titers, cytokine levels, and histopathology) of hMPV-infected mice was investigated after inoculation with S. pneumoniae. In vitro results of bacterial infection with S. pneumoniae on A549 and Hep-2 monolayer cells showed that even though cellular adherence was variable among different serotypes, there was significantly enhanced bacterial adherence in A549 cells with preceding hMPV infection. In addition, in vivo study of hMPV-infected mice showed increased adhesion of S. pneumoniae on the bronchial epithelium with delayed bacterial clearance and exacerbated histopathology. Furthermore, mice with preceding hMPV infection showed repressed recruitment of airway neutrophils with decreased expression of neutrophil chemoattractants during pneumococcal infection. These results suggest that hMPV-infected airway cells, especially the lower airway epithelium, express increased adherence with S. pneumoniae. Furthermore, hMPV-infected mice showed impaired recruitment of airway neutrophils, possibly leading to delayed bacterial clearance and exacerbated pulmonary inflammation, after secondary infection with pneumococcal isolates.

Investigating the Effects of Probiotics on Pneumococcal Colonization Using an &lt;em&gt;In Vitro&lt;/em&gt; Adherence Assay

Adherence of Streptococcus pneumoniae (the pneumococcus) to the epithelial lining of the nasopharynx can result in colonization and is considered a prerequisite for pneumococcal infections such as pneumonia and otitis media. In vitro adherence assays can be used to study the attachment of pneumococci to epithelial cell monolayers and to investigate potential interventions, such as the use of probiotics, to inhibit pneumococcalcolonization.The protocol described here is used to investigate the effects of the probiotic Streptococcus salivarius on the adherence of pneumococci to the human epithelial cell line CCL-23 (sometimes referred to as HEp-2 cells). The assay involves three main steps: 1) preparation of epithelial and bacterial cells, 2) addition of bacteria to epithelial cell monolayers, and 3) detection of adherent pneumococci by viable counts (serial dilution and plating) or quantitative real-time PCR (qPCR). This technique is relatively straightforward and does not require specialized equipment other than a tissue culture setup. The assay can be used to test other probiotic species and/or potential inhibitors of pneumococcal colonization and can be easily modified to address other scientific questions regarding pneumococcal adherence and invasion.

Adhesion of &lt;i&gt;Streptococcus mutans&lt;/i&gt; on Different Types of Orthodontic Brackets

Objective: The aim of this study was to evaluate the adhesion of Streptococcus mutans (S. mutans) on different types of orthodontic brackets commonly used in Thailand. Materials and Methods: The adhesion of S. mutans on 7 types of orthodontic brackets (Clarity, Spirit MB, Inspire Ice, 3M, Tomy, Ormco and W&amp;H) was investigated in vitro. The saliva-coated brackets were immersed in a flat-bottomed 96-well plate containing S. mutans suspension (108 CFU/ml) and incubated at 37 ̇C in 10% CO2 and 100 rpm for 2 hours. The adhered bacterial cells were fixed by 99% methanol and stained with 0.5% crystal violet. The quantity of adhesion was determined by measuring optical density (OD) at 590 nm. The data were analyzed with Kolmogorov-Smirnov test, Levene test, one way ANOVA and Games-Howell post hoc test. Results: Significantly higher bacteria adhesion was found on Clarity, Spirit MB, Tomy, Ormco and W&amp;H compared to Inspire Ice and 3M Gemini. However, there were no significant differences among the different types of bracket materials (ceramic, plastics and metal). Conclusion: The amount of bacteria adhesion was independent of the type of bracket materials. The size and shape of brackets also played an important role in the amount of adhesion of microorganism.