Black-pigmented Bacteria Research Articles

Black-Pigmented Marine Pseudomonas aeruginosa Exhibiting Anti-Bacterial Activity against Multidrug-Resistant (MDR) Wound Infection Bacteria

The urgency of multidrug-resistant (MDR) bacterial infections in wounds is a significant concern due to the high prevalence of MDR in healthcare settings. Black pigmented marine bacteria, strain PS1C, were isolated from marine sediment on Awur Beach Jepara, Central Java, Indonesia, and cultured in the laboratory. The aim of this research includes molecular identification of strain PS1C, extracting black pigment from strain PS1C, isolation of MDR bacteria from wounds, and evaluating the antibacterial activity of black pigment from strain PS1C against MDR bacteria isolates of wound infections. We examined the 16S rRNA gene sequences strain PS1C to identify the species. Then, the black pigment from PS1C was extracted using methanol: acetone (7:3) solvent. Antibacterial activity was evaluated against MDR bacteria from wounds with the microdilution method. A black-pigmented bacterium was isolated and identified as Pseudomonas aeruginosa PS1C. We found that the black pigment from P. aeruginosa PS1C can be developed as an antibacterial agent against the MDR bacterial isolate of wounds with MIC and MBC values of 6.25-12.5 mg/ml and 6.25-25 mg/ml, respectively. In conclusion, the study's findings highlight the potential of the extract of black pigment from P. aeruginosa PS1C as an antibacterial agent against wound-causing MDR bacteria and reinforce previous research into P. aeruginosa can be isolated from marine sources. Additional in vivo investigations and the identification of the antibacterial activity's mode of action are required.

Presence of zoonotic black-pigmented periodontal pathogens in the oral microbiota of pet and stray cats

Black-pigmented bacteria are one of the neglected species to cause periodontal disease in cats, and they are also zoonotic agents that pose an infection risk to humans. In this study, we aimed to determine the presence of Porphyromonas gingivalis, Porphyromonas gulae and Prevotella nigrescens in the oral microbiota of pet and stray cats. Dental swab samples were taken from 25 pet cats and 25 stray cats with symptoms of periodontal disease and then investigated by multiplex polymerase chain reaction using 16S rRNA species-specific primers. As a result of the multiplex PCR analysis, P. gingivalis 3/25 (12%), P. nigrescens 1/25 (4%), P. gingivalis + P. gulae 7/25 (28%), P. gingivalis + P. nigrescens 1/25 (4%), P. gulae + P. nigrescens 1/25 (4%), and P. gingivalis + P. gulae + P. nigrescens 2/25 (8%) were molecularly typed in the pet cats. In addition, 1/25 (4%) of P. gulae and 21/25 (84%) of P. gingivalis + P. gulae were typed in the stray cats. In 10/25 (40%) pet and 3/25 (12%) stray cat samples, no bacteria were detected by molecular typing. In summary, the results provide strong evidence that black-pigmented zoonotic pathogens are associated with cat periodontal disease.

Evaluation of PCR primers to identify Porphyromonas endodontalis in apical periodontitis clinical samples.

We aimed to evaluate whether two commonly used PCR primers are effective to identify P. endodontalis and discriminate from other prevalent black-pigmented bacteria in apical periodontitis (AP). Endodontic canal samples from patients with asymptomatic AP (n = 20) were collected and cultured in anaerobiosis. Two primer sets to detect P. endodontalis were selected from the literature and first analyzed for their specificity in silico; and then tested on clinical isolates in vitro and finally, in apical exudates ex vivo. The identity of P. endodontalis was verified by PCR and Sanger sequencing with universal primers for bacterial V3-V6 regions 16S rDNA. Only one primer set showed specificity only for P. endodontalis clones in silico and also was specific for P. endodontalis in vitro and ex vivo.

Clinical and Microbiological Evaluation of Stabilised Periodontal Patients Undergoing Early Stage of Orthodontic Treatment

Orthodontic treatment may affect the equilibrium of oral microbiota which plays a major role in aetiology of periodontal disease. This prospective clinical study aimed to assess the periodontal health and microbiological profile of healthy (Group 1) and stabilised periodontal (Group 2) patients throughout three months of orthodontic treatment. Upper and lower fixed orthodontic appliances were bonded. Periodontal health was assessed using plaque score (PS), bleeding on probing (BOP) and pocket depth (PD). 29 sites were taken for subgingival plaque sampling. Plaque samples were inoculated on Trypticase Soy Blood Agar (TSBA) and Trypticase Soy Bacitracin Vancomycin (TSBV) agar for assessment of aerobe, anaerobe, black pigmented bacteria (BPB) and Aggregatibacter actinomycetemcomitans. All the measurements were taken before bonding (T0), 1 week (T1), 1 month (T2) and 3 months post-bonding (T3). Generally, periodontal health in both groups were almost similar. After 1 week, the number of aerobes was significantly higher in Group 1 (88%) while the anaerobes were significantly higher in Group 2 (45%). A. actinomycetemcomitans was higher in Group 1 at T0 and T1 but was significantly higher in Group 2 at T3. BPB was minimal at all time with no significant difference. Thus, during the first 3-month of orthodontic treatment, there were significant changes in the number of aerobes-anaerobes in both healthy and stabilised periodontal patients. Pathogenic bacteria would increase during early treatment of orthodontics.

Gestational diabetes mellitus increases the detection rate and the number of oral bacteria in pregnant women.

In the present study, we aimed to investigate the association between gestational diabetes mellitus (GDM) and the oral microbial imbalance in the second trimester of pregnancy.Three hundred thirtyone women in the second trimester of pregnancy who underwent prenatal examinations at the Third Affiliated Hospital of Soochow University from February 2018 to August 2018 were included in this study. Personal parameters including the age, education level, and body mass index (BMI) at 28 weeks of gestation were recorded. Gestational diabetes mellitus (GDM) was diagnosed according to the Standards of Medical Care in Diabetes-2011 recommended by American Diabetes Association (ADA). After the significant difference of each parameter was erased by a propensity-score matched (PSM) analysis at a 1:1 ratio, the oral health conditions and the oral flora in pregnant women with GDM were compared with those in nondiabetic pregnant women.Our data demonstrated that 65 of the 331 pregnant women (19.6%) were diagnosed with GDM. Results from the matched data including 59 matched pairs of pregnant women showed that the gingival index (GI), plaque index (PI), tooth mobility degree (TMD), probing depth (PD), and bleeding on probing (BOP) of pregnant women with GDM were higher or more severe than those of nondiabetic pregnant women (P < .05). The detection rate of tuberculosis bacilli, Black-pigmented bacteria, and Capnocytophaga in pregnant women with GDM was higher than those in nondiabetic pregnant women (P = .000, P = .026, and P = .030, respectively). In addition, pregnant women with GDM had fewer oral streptococci (P = .000) and lactobacilli (P = .000) and more oral anaerobic bacteria (P = .000), tuberculosis bacilli (P = .000), Black-pigmented bacteria (P = .007), Capnocytophaga (P = .000), and actinomycetes (P = .000).The detection rate and the number of oral bacteria in pregnant women with GDM were higher than those in nondiabetic pregnant women in the second trimester of pregnancy.

Antibacterial effect of binahong (Anredera cordifolia (Ten.) Steenis) leaf infusion against black pigmented bacteria

Periodontitis is a highly prevalent periodontal disease in Indonesia. Black pigmented bacteria, such as Porphyromonas gingivalis and Prevotella intermedia, are the major pathogens associated with chronic periodontitis. Binahong has been scientifically proven to have an antimicrobial effect. This study investigated the effectiveness of a binahong leave infusion as an antibacterial agent against black pigmented bacteria in vitro. The concentrations of the infusion used in the study were 50%, 65%, 80%, 95%, and 100%. The blank disc diffusion method was performed to measure the zone of inhibition in brucella agar. From the diffusion test on brucella agar, the inhibitory zones were 0.42 mm (50%), 1.21 mm (65%), 1.18 mm (80%), 1.19 mm (95%), and 1.36 (100%). In conclusion, a binahong leaf infusion inhibits the growth of black pigmented bacteria.

Alterations in oral microbial flora induced by waterpipe tobacco smoking.

BackgroundWaterpipe smoking is a global health problem and a serious public concern. Little is known about the effects of waterpipe smoking on oral health. In the current study, we examined the alterations of oral microbial flora by waterpipe smoking.MethodsOne hundred adult healthy subjects (59 waterpipe smokers and 41 non-smokers) were recruited into the study. Swabs were taken from the oral cavity and subgingival regions. Standard culturing techniques were used to identify types, frequency, and mean number of microorganisms in cultures obtained from the subjects.ResultsIt was notable that waterpipe smokers were significantly associated with a history of oral infections. In subgingiva, Acinetobacter and Moraxella species were present only in waterpipe smokers. In addition, the frequency of Candida albicans was higher in the subgingiva of waterpipe smokers (p = 0.023) while the frequency of Fusobacterium nucleatum was significantly lower in the subgingiva of waterpipe smokers (p = 0.036). However, no change was observed in other tested bacteria, such as Campylobacter species; Viridans group streptococci, Enterobacteriaceae, and Staphylococcus aureus. In oral cavity and when colony-forming units were considered, the only bacterial species that showed significant difference were the black-pigmented bacteria (p < 0.001).ConclusionThis study provides evidence indicating that some of the oral microflora is significantly altered by waterpipe smoking.

THE EFFICACY OF AN INFUSION OF BINAHONG LEAVES (ANREDERA CORDIFOLIA (TEN.) STEENIS) AGAINST WILD STRAIN BLACK-PIGMENTED BACTERIA

Objective: Black-pigmented bacteria represent one of the most prominent causes of chronic periodontitis. The objective of this study is to investigate the in vitro antibacterial effect of an infusion of binahong leaves (Anredera cordifolia (Ten.) Steenis) against black-pigmented bacteria.Methods: The black-pigmented bacteria were isolated from the periodontal pocket of a patient with chronic periodontitis. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by means of a dilution test using 105 cfu/ml of bacteria cultured on Brucella broth containing blood and kanamycin, which was exposed to a 20%, 30%, 40%, 50%, 65%, 80%, and 95% infusion of binahong leaves and then incubated for 24, 48, and 72 h. In the blank disc diffusion test, a 50%, 65%, 80%, 95%, and 100% infusion was dripped onto a paper disc placed at the center of a Brucella agar bacterial culture and then incubated for 24 h.Results: The MIC and MBC of the binahong leaf infusion were &gt;65% and 80%, respectively, while the antibacterial activity was not correlated with the duration of the exposure to the infusion. The diffusion test showed that an increased concentration of the infusion resulted in an increased diameter of the inhibition zone that ranged from 0.42 mm (50% infusion) to 1.42 mm (100% infusion).Conclusion: Binahong leaves infusion has potential efficacy against black-pigmented bacteria

Porphyromonas gingivalis: An Overview of Periodontopathic Pathogen below the Gum Line.

Periodontal disease represents a group of oral inflammatory infections initiated by oral pathogens which exist as a complex biofilms on the tooth surface and cause destruction to tooth supporting tissues. The severity of this disease ranges from mild and reversible inflammation of the gingiva (gingivitis) to chronic destruction of connective tissues, the formation of periodontal pocket and ultimately result in loss of teeth. While human subgingival plaque harbors more than 500 bacterial species, considerable research has shown that Porphyromonas gingivalis, a Gram-negative anaerobic bacterium, is the major etiologic agent which contributes to chronic periodontitis. This black-pigmented bacterium produces a myriad of virulence factors that cause destruction to periodontal tissues either directly or indirectly by modulating the host inflammatory response. Here, this review provides an overview of P. gingivalis and how its virulence factors contribute to the pathogenesis with other microbiome consortium in oral cavity.

Culture-based identification of pigmented Porphyromonas and Prevotella species in primary endodontic infections.

Background. The most common species isolated from primary endodontic infections are black-pigmented bacteria. These species are implicated in apical abscess formation due to their proteolytic activity and are fastidious in nature. Therefore, the present study was carried out to evaluate the presence and identification of various pigmented Porphyromonas and Prevotella species in the infected root canal through culture-based techniques.Methods. Thirty-one patients with primary endodontic infections were selected. Using sterile paper points, samples were collected from the root canals after access opening and prior to obturation, which were cultured using blood and kanamycin blood agar. Subsequently, biochemical test was used to identify the species and the results were analyzed using percentage comparison analysis, McNemar and chi-squared tests, Wilcoxon match pair test and paired t-test.Results. Out of 31 samples 26 were positive for black-pigmented organisms; the predominantly isolated species were Prevotella followed by Porphyromonas. In Porphyromonas only P. gingivalis was isolated. One of the interesting features was isolation of P. gingivalis through culture, which is otherwise very difficult to isolate through culture.Conclusion. The presence of Prevotella and Porphyromonas species suggests that a significant role is played by these organisms in the pathogenesis of endodontic infections.

Lethal effect of blue light-activated hydrogen peroxide, curcumin and erythrosine as potential oral photosensitizers on the viability of Porphyromonas gingivalis and Fusobacterium nucleatum.

Recently, photodynamic therapy (PDT) has been introduced as a new modality in oral bacterial decontamination. Current research aims to evaluate the effect of photodynamic killing of visible blue light in the presence of hydrogen peroxide, curcumin and erythrosine as potential oral photosensitizers on Porphyromonas gingivalis associated with periodontal bone loss and Fusobacterium nucleatum associated with soft tissue inflammation. Standard suspension of P. gingivalis and F. nucleatum were exposed to Light Emitting Diode (LED) (440-480 nm) in combination with erythrosine (22 µm), curcumin (60 µM) and hydrogen peroxide (0.3 mM) for 5 min. Bacterial samples from each treatment groups (radiation-only group, photosensitizer-only group and blue light-activated photosensitizer group) were subcultured onto the surface of agar plates. Survival of these bacteria was determined by counting the number of colony forming units (CFU) after incubation. RESULTS for antibacterial assays on P. gingivalis confirmed that curcumin, Hydrogen peroxide and erythrosine alone exerted a moderate bactericidal effect which enhanced noticeably in conjugation with visible light. The survival rate of P. gingivalis reached zero present when the suspension exposed to blue light-activated curcumin and hydrogen peroxide for 2 min. Besides, curcumin exerted a remarkable antibacterial activity against F. nucleatum in comparison with erythrosine and hydrogen peroxide (P=0.00). Furthermore, the bactericidal effect of visible light alone on P. gingivalis as black-pigmented bacteria was significant. Our result suggested that visible blue light in the presence of erythrosine, curcumin and hydrogen peroxide would be consider as a potential approach of PDT to kill the main gramnegative periodontal pathogens. From a clinical standpoint, this regimen could be established as an additional minimally invasive antibacterial treatment of plaque induced periodontal pathologies.

Black pigmentation in primary dentition: Case report and literature review

Abstract Black tooth staining is an extrinsic discoloration found in both primary and permanent dentition, and seen as dark pigmented lines extending to the gingival margin or an incomplete coalescence of dark dots that rarely extend beyond the cervical third of the crown. An association between black tooth staining and Actinomyces bacterial strains has been reported, while black-pigmented bacteria associated with such staining are known to be harbored in the oral cavity. Prevotella intermedia and Prevotella nigrescens are black-pigmented bacteria known to be dependent on the heme portion of hemoglobin as an iron source required for their growth. Recently, developments in molecular biological techniques have enabled rapid and easy detection of periodontopathic bacterial species using bacterial DNA extracted from oral specimens, such as plaque and saliva. Here, we report a case of black pigmentary deposition identified on all teeth of a 2-year-old girl, as well as the results of analysis of the distribution of oral bacteria in saliva and plaque specimens obtained from the patient using a molecular biological technique. In addition, a literature search found a case of disease related to the oral bacteria detected in our patient. We concluded that the bacteria detected in this case may have a strong relationship with black pigmentation, although the route of bacterial infection and cause of staining remain to be elucidated.

Bactericidal effect of visible light in the presence of erythrosine on Porphyromonas gingivalis and Fusobacterium nucleatum compared with diode laser, an in vitro study.

Recently, photodynamic therapy (PDT) has been introduced as a new modality in oral bacterial decontamination. Besides, the ability of laser irradiation in the presence of photosensitizing agent to lethal effect on oral bacteria is well documented. Current research aims to evaluate the effect of photodynamic killing of visible blue light in the presence of plaque disclosing agent erythrosine as photosensitizer on Porphyromonas gingivalis associated with periodontal bone loss and Fusobacterium nucleatum associated with soft tissue inflammation, comparing with the near-infrared diode laser. Standard suspension of P. gingivalis and F. nucleatum were exposed to Light Emitting Diode (LED) (440-480 nm) used to photopolymerize composite resine dental restoration in combination with erythrosine (22 µm) up to 5 minutes. Bacterial sample were also exposed to a near-infrared diode laser (wavelength, 830 nm), using identical irradiation parameters for comparison. Bacterial samples from each treatment groups (radiation-only group, erythrosine-only group and light or laser with erythrosine group) were subcultured onto the surface of agar plates. Survival of these bacteria was determined by counting the number of colony forming units (CFU) after incubation. Exposure to visible blue light and diode laser in conjugation with erythrosine significantly reduced both species examined viability, whereas erythrosine-treated samples exposed to visible light suggested a statically meaningful differences comparing to diode laser. In addition, bactericidal effect of visible light or diode laser alone on P. gingivalis as black-pigmented bacteria possess endogenous porphyrins was noticeably. Our result suggested that visible blue light source in the presence of plaque disclosing agent erythrosine could can be consider as potential approach of PDT to kill the main gram-negative periodontal pathogens. From a clinical standpoint, this regimen could be established as an additional minimally invasive antibacterial treatment of plaque induced periodontal pathologies.

Dental Caries and Microbiota in Children with Black Stain and Non-discoloured Dental Plaque

Aim: We aimed to assess caries experience and microbiota in systemically healthy children with black stain (BS) and non-discoloured plaque. Methods: Forty-six children with BS and 47 counterparts with non-discoloured plaque aged 7.9 ± 1.3 years were clinically examined. Dental caries was scored using WHO criteria. Samples of BS and non-discoloured dental plaque were collected from tooth surfaces. The DNA of the samples was extracted and real-time PCR was performed to determine the total number of bacteria and the species Streptococcus mutans, S. sobrinus, Lactobacillus sp., Actinomyces naeslundii, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum. Results: Children with BS had lower DMFT (p = 0.013), lower DT values (p = 0.005) and a tendency to lower caries prevalence (p = 0.061) than children with non-discoloured plaque. Plaque samples of the BS group contained higher numbers of A. naeslundii (p = 0.005) and lower numbers of F. nucleatum (p = 0.001) and Lactobacillus sp. (p = 0.001) compared to the non-discoloured plaque samples of the control group. Comparing the children with BS and non-discoloured plaque, higher counts for A. naeslundii (p = 0.013) were observed in caries-free children with BS while in caries-affected children with BS, lower counts of F. nucleatum (p = 0.007) were found. Counts of Lactobacillus sp. were higher in non-discoloured plaque samples than in BS of caries-free and caries-affected children. Conclusion: Results suggest that the different microbial composition of BS might be associated with the lower caries experience in affected subjects. The role of black-pigmented bacteria associated with periodontitis needs further studies.

Detection of Porphyromonas endodontalis, Porphyromonas gingivalis and Prevotella intermedia in primary endodontic infections in a Chinese population

To assess the prevalence of three black-pigmented bacterial species (Porphyromonas endodontalis, Porphyromonas gingivalis and Prevotella intermedia) using microarray technology in root canals of teeth associated with primary endodontic infections in a Chinese population. Microbial samples were taken from root canals of 80 teeth with pulp necrosis and primary endodontic infections in a Chinese population. DNA extracted from the samples was amplified by PCR with universal bacterial primers based on 16S rRNA gene sequences, and the products hybridized with the microarrays in which the specific oligonucleotide probes were added. The results of hybridization were screened by a confocal laser scanner. Pearson chi-square test and the two-sided Fisher exact test were used to analyse whether a significant association existed between the species and symptoms as well as in co-existence of two target organisms by a statistical software package (SAS 8.02). The 16S rRNA gene microarray detected at least one of the three test species in 76% of the study teeth. P. endodontalis, P. gingivalis and P. intermedia were found in 50%, 33% and 45%, respectively. A significant association was found in the presence of the pair P. endodontalis / P. gingivalis (P < 0.005). Both P. endodontalis (P <0.05) and P. gingivalis (P <0.005) had a statistically significant association with the presence of a sinus tract. The simultaneous presence of P. endodontalis and P. gingivalis was also associated with the presence of a sinus tract (P<0.005) and abscess formation (P<0.05). The three black-pigmented bacteria were prevalent in teeth with pulp necrosis and primary endodontic infections in a Chinese population. P. gingivalis and P. endodontalis were associated with the presence of sinus tract and abscess formation.

Clinical and microbiological evaluation of high intensity diode laser adjutant to non-surgical periodontal treatment: a 6-month clinical trial

This randomized split-mouth clinical trial was designed to evaluate the efficacy of scaling and root planing associated to the high-intensity diode laser on periodontal therapy by means of clinical parameters and microbial reduction. A total of 36 chronic periodontitis subjects, of both genders, were selected. One pair of contralateral single-rooted teeth with pocket depth >5mm was chosen from each subject. All patients received non-surgical periodontal treatment, after which the experimental teeth were designated to either test or control groups. Both teeth received scaling, root planing and coronal polishing (SRP) and teeth assigned to the test group (SRP + DL) were irradiated with the 808 ± 5nm diode laser, for 20s, in two isolated appointments, 1week apart. The laser was used in the continuous mode, with 1.5W and power density of 1,193.7W/cm(2). Clinical and microbiological data were collected at baseline, 6weeks and 6months after therapy. There was a significant improvement of all the clinical parameters-clinical attachment level (CAL), probing depth (PD), plaque index (PI) and Bleeding on Probing (BOP)-for both groups (P < 0.001), with no statistical difference between them at the 6weeks and the 6months examinations. As for microbiological analysis, a significant reduction after 6weeks (P > 0.05) was observed as far as colony forming units (CFU) is concerned, for both groups. As for black-pigmented bacteria, a significant reduction was observed in both groups after 6months. However, the difference between test and control groups was not significant. There was no association between group and presence of Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans at any time of the study. After 6months of evaluation, the high-intensity diode laser has not shown any additional benefits to the conventional periodontal treatment. The high intensity diode laser did not provide additional benefits to non-surgical periodontal treatment. More studies are necessary to prove the actual need of this type of laser in the periodontal clinical practice.

Purification and characterization of a novel secondary fimbrial protein from Porphyromonas gulae

Background:Porphyromonas gulae are black-pigmented anaerobic bacteria isolated from the gingival sulcus of various animal hosts and are distinct from Porphyromonas gingivalis originating in humans. We previously reported the antigenic similarities of 41-kDa fimbriae between P. gulae ATCC 51700 and P. gingivalis ATCC 33277. In this study, to clarify the presence of another type of fimbriae of P. gulae, we have purified and characterized the secondary fimbrial protein from P. gulae ATCC 51700.Methods: The secondary fimbrial protein was purified from P. gulae ATCC 51700 using an immunoaffinity column coupling with antibodies against the 41-kDa fimbrial protein. The expression of fimbriae on the cell surface of P. gulae ATCC 51700 was investigated by transmission electron microscopy. The N-terminal amino acid sequence was determined by an amino acid sequencer system.Results: The molecular mass of this protein was approximately 53-kDa, as estimated by SDS-PAGE. The polyclonal antibodies against the 53-kDa protein did not react with the 41-kDa fimbrial protein of P. gulae ATCC 51700. Immunogold electron microscopy revealed that anti-53-kDa fimbrial serum bound to fimbria on the cell surface of P. gulae ATCC 51700. The amino acid sequence of the N-terminal 15 residues of the 53-kDa fimbrial protein showed only 1 of 15 residues identical to the 41-kDa fimbrial protein.Conclusion: The 53-kDa fimbriae are different in molecular weight and antigenicity from the 41-kDa fimbrial protein of P. gulae ATCC 51700. These results clearly suggest that the 41-kDa and the 53-kDa fimbriae are distinct types of fimbriae expressed simultaneously by this organism.

Detection of bacterial species involved in perimplantitis concerned with cultural and RT-PCR

Dental implants offer new treatment options for edentulous either partially or completely, now represent a viable alternative to conventional fixed protheses. Dental implants are colonized by a flora dominated by Gram-positive facultative aerobic, while in patients with bone loss and formation of pockets peri-implant diseases was found a significant difference in the composition of microflora, bacteria, Gram-negative anaerobes in particular Fusobacterium spp., Treponema denticola (Spirochetes), Tannerella forsythensis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia as interim black-pigmented bacteria, Porphyromonas gingivalis, often in high concentrations. Aims. The purpose of this study was to identify those at risk of perimplantitis using 2 techniques: RT-PCR examination of trade and culture. The results were compared taking into consideration the advantages and disadvantages of both methods. Materials and methods.We studied 24 patients (14 women and 10 men), aged, women between 43 and 76 years, with an average of 63.8 + / - 10.9 years, men between 45 and 88 years with a average of 64.3 years + / - 12.5 years. Was performed a double levy of sub-gingival plaque at multiple sites that had an implant CAL (clinical attachment level)> 4mm in order to assess the microbiological identification with the two techniques: Examining culture and Real-Time PCR of Commerce ( Gum-Sunstar) that identifies 4 bacterial species: A. actinomycetemcomitans (A.a.), P.gingivalis (P.g.), T.forsythensis (T.f.), and T.denticola (T.d.). Results. All patients studied were positive to both tests with charger high: the consideration of tenure, with CFU / ml > 105, was positive in 66.6% of samples by:T.f., and P.g., in 12.5% for A.a., while T.d. not been sought by examining culture, the RT-PCR was positive, with high loads, in 95.8% of samples for T.f., in 79.1% for P.g., in 12.5% for A.a. and 20.8% for T.d.The test crop showed the presence of even P.intermedia in 91.6% of the samples, as well as Capnocytophaga spp. in 54.1%, F.nucleatum spp. in 50% and Campylobacter spp. 25%. Conclusions.The data show that RT-PCR has greater sensitivity than culture examination as well as response times are in favor of RT-PCR, but the kits Trade identify a limited number of species present no bacterial resistance if it were taking antibiotics.There are several factors (genetic, environmental and systemic diseases) of the subject that may affect the results of microbiological contamination. It is perhaps for this reason that many dentists often consider the microbiological examination of “unsafe” because there are not always matching the survey clinical microbiological examination then the same situation microbiological not always the same clinical situation. Being able to pinpoint those responsible for periodontal infection and inflammation, however, offers the opportunity to vary quite rightly, on the basis of virulence factors and evaluation of bacteria, the timing of recalls for the TPS (periodontal therapy support), leaving as a last choice antibiotic therapy targeted.

Microbial Adhesion on Different Bracket Types in vitro

To test the hypothesis that there are differences in total bacterial counts and capacity for biofilm formation between seven different bracket types. By means of an in vitro experiment, seven commercially available bracket systems (Damon [A], Clarity [B], Mystique [C], Speed [D], Victory MBT [E], Micro-loc [F], and Generus [G]) were compared. A total of 25 premolar brackets of each bracket system were incubated in brain heart infusion medium containing the saliva and bacteria of two orthodontic patients. After 72 hours, the amounts of aerobe and anaerobe bacteria were determined by counting the colony-forming units (CFU). The CFU ratio (aerobe/anaerobe) also was calculated, and the black pigmented bacteria were analyzed. Significant differences between the different bracket types in terms of biofilm formation were found. Bracket types can be arbitrarily divided into low, intermediate, and high plaque-retaining brackets. The group with low adhesion consists of bracket types E, F, and G; the group with high adhesion of bracket types A, B, and C; and type D exhibits intermediate adhesion. The group with high microbial adhesion (A, B, and C) did present significantly lower CFU ratios (aerobe/anaerobe) than were exhibited by the other bracket systems (P < .05). The hypothesis is accepted. Orthodontic brackets serve as different loci for biofilm formation; in this in vitro study, significant differences were noted between the different types of brackets.

The upregulation of receptor activator NF‐κB ligand expression by interleukin‐1α and Porphyromonas endodontalis in human osteoblastic cells

To investigate the receptor activator of nuclear factor-kappa B (NF-kappaB) ligand (RANKL) in osteoblastic cells stimulated with inflammatory mediators. The expression of RANKL in human osteoblastic cell line U2OS stimulated by pro-inflammatory cytokine interleukin (IL)-1alpha and black-pigmented bacteria Porphyromonas endodontalis was investigated by Western blot and enzyme-linked immunosorbent assay (ELISA). The significance of the results obtained from control and treated groups was statistically analysed by the paired Student's t-test. IL-1alpha was found to upregulate RANKL production in U2OS cells (P < 0.05). Investigations of the time dependence of RANKL expression in IL-1alpha-treated cells revealed a rapid accumulation of RANKL protein after 1 h of exposure; it remained elevated throughout the 24-h incubation period shown by Western blot and ELISA. In addition, P. endodontalis also increased RANKL expression in U2OS cells after 4-h incubation period demonstrated by Western blot and ELISA (P < 0.05). IL-1alpha and P. endodontalis may be involved in developing apical periodontitis through the stimulation of RANKL production.