Influence of periodontal treatment on subgingival and salivary microbiotas

DEFF Research Database (Denmark)

Belstrøm, Daniel; Grande, Maria Anastasia; Sembler-Møller, Maria Lynn

2018-01-01

BACKGROUND: The purpose of this study was to characterize and compare subgingival and salivary microbiotas before and after periodontal treatment to learn if any changes of the subgingival microbiota were reflected in saliva. We tested the hypothesis that salivary levels of specific periopathogens...... correlate with corresponding subgingival levels before and after periodontal treatment. METHODS: Twenty-five patients with generalized chronic periodontitis completed the study. Stimulated saliva samples and subgingival plaque samples were collected at baseline and 2, 6 and 12 weeks after non......-surgical periodontal therapy. Subgingival and salivary microbiotas were processed by means of the Human Oral Microbe Next Generation Sequencing (HOMINGS) technique, and characterized based on relative abundance. Spearman signed rank test was used to test correlation of periopathogens in subgingival and saliva samples...

Antimicrobial effect of chlorhexidine on Aggregatibacter actinomycetemcomitans biofilms associated with peri-implantitis

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Zeinab Kadkhoda

2016-08-01

Full Text Available Background. This study aimed to assessthe antimicrobial effect of chlorhexidine (CHX on Aggregatibacter actinomyce-temcomitans biofilms isolated from subgingival plaque of peri-implantitis lesions. Methods. Thirteen patients requiring peri-implantitis treatment were consecutively selected and their subgingival biofilm was collected by inserting fine sterile paper points into peri-implant pockets for 15 seconds. A. actinomycetemcomitans was isolated from the subgingival biofilm and cultured. In this study, the standard strain of A. actinomycetemcomitans served as the positive control group and a blank disc impregnated with water served as the negative control; 0.1 mL of the bacterial suspension was cultured on specific culture medium and blank discs (6 mm in diameter impregnated with 0.2%CHX mouthrinse (Behsa Pharmaceutical Co. and negative control discs were placed on two sides of the bacterial culture plate. The size of growth inhibition zone was measured by a blinded independent observer in millimetres. Results. According to the results of disc diffusion test, the mean diameter of growth inhibition zone of A. actinomycetem-comitans around discs impregnated with CHX was larger in both standard (positive control and biofilm samples of A. acti-nomycetemcomitans compared to the negative control group (blank disc (P<0.001. Conclusion. Use of0.2% CHX mouthwash had antibacterial effects on A. actinomycetemcomitans species isolated from peri-implantitis sites.

The Biofilm Community-Rebels with a Cause.

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Aruni, A Wilson; Dou, Yuetan; Mishra, Arunima; Fletcher, Hansel M

2015-03-01

Oral Biofilms are one of the most complex and diverse ecosystem developed by successive colonization of more than 600 bacterial taxa. Development starts with the attachment of early colonizers such as Actinomyces species and oral streptococci on the acquired pellicle and tooth enamel. These bacteria not only adhere to tooth surface but also interact with each other and lay foundation for attachment of bridging colonizer such as Fusobacterium nucleatum followed by late colonizers including the red complex species: Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola -the founders of periodontal disease. As the biofilm progresses from supragingival sites to subgingival sites, the environment changes from aerobic to anaerobic thus favoring the growth of mainly Gram-negative obligate anaerobes while restricting the growth of the early Gram-positive facultative aerobes. Microbes present at supragingival level are mainly related to gingivitis and root-caries whereas subgingival species advance the destruction of teeth supporting tissues and thus causing periodontitis. This review summarizes our present understanding and recent developments on the characteristic features of supra- and subgingival biofilms, interaction between different genera and species of bacteria constituting these biofilms and draws our attention to the role of some of the recently discovered members of the oral community.

Quantification of periodontal pathogens in vascular, blood, and subgingival samples from patients with peripheral arterial disease or abdominal aortic aneurysms.

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Figuero, Elena; Lindahl, Christeel; Marín, María José; Renvert, Stefan; Herrera, David; Ohlsson, Ola; Wetterling, Thomas; Sanz, Mariano

2014-09-01

The aim of this investigation is to quantify periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Campylobacter rectus, and Tannerella forsythia) in vascular, blood, and subgingival samples. As a secondary objective, two molecular bacterial identification methods (nested polymerase chain reaction [PCR] and quantitative PCR [qPCR]) are compared. Seventy consecutive patients provided a vascular lesion, a blood sample, and 36 subgingival samples. Bacterial DNA was extracted, and qPCR was used to determine the prevalence and amounts of the target pathogens in each sample. Nested PCR was performed only in the samples from vascular lesions. Periodontal examination was performed in 42 patients. Mann-Whitney U or χ(2) tests were used to compare microbiologic results according to periodontal diagnosis. All targeted periodontal pathogens (A. actinomycetemcomitans, P. gingivalis, T. forsythia, or C. rectus) were detected in subgingival samples, with a prevalence rate of 72.2%, 47.2%, 74.3%, and 82.9%, respectively. In 7.1% and 11.4% of vascular and blood samples, bacterial DNA was detected. One patient was positive for A. actinomycetemcomitans in the three types of samples. No differences were found in the levels of targeted bacteria when comparing patients with and without periodontitis. Prevalence rates obtained with nested PCR were significantly higher than those obtained with qPCR. The presence of A. actinomycetemcomitans was demonstrated in vascular, blood, and subgingival samples in one of 36 patients. These results, although with a very low frequency, may support the hypothesis of a translocation of periodontal pathogens from subgingival microbiota to the bloodstream and then to atheromatous plaques in carotid or other peripheral arteries. Nested PCR is not an adequate method for identifying DNA of periodontal pathogens in low quantities because of the high number of false-negative results.

Subgingival microbial profile of obese women with periodontal disease.

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Silva-Boghossian, Carina M; Cesário, Paola C; Leão, Anna Thereza T; Colombo, Ana Paula V

2018-02-01

This study compared the composition of subgingival microbiota between obese and non-obese women with or without periodontal disease. Full-mouth periodontal clinical assessments were carried out in 76 obese women (17 periodontally healthy and 59 with periodontal disease), and 34 non-obese women (12 periodontally healthy, 22 with periodontal disease). Subgingival biofilm samples were individually obtained from seven sites of each individual, and the prevalence and counts of 40 bacterial taxa were determined by the checkerboard method. The frequency and counts of each species were computed for each individual and across the groups. Differences among and between groups were sought by the Kruskal-Wallis and Mann-Whitney tests, respectively. Possible correlations between obesity and clinical and microbiologic parameters were tested with Spearman correlation coefficient. Streptococcus sanguinis, Streptococcus oralis, and Capnocytophaga ochracea were found in significantly higher levels in obese compared with non-obese women (P periodontal health, Porphyromonas gingivalis and Leptotrichia buccalis were detected in higher mean frequency and/or counts in obese women than in non-obese women, whereas in patients with periodontal disease, obese women harbored greater levels of C. ochracea than non-obese women (P periodontal disease presented significantly greater mean counts of P. gingivalis and Tannerella forsythia than non-obese women with periodontal health (P periodontal disease are present at the same time, significant positive correlations were detected with C. ocharcea, P. gingivalis, S. sanguinis, and T. forsythia. Few differences in the composition of the subgingival microbiota of obese and non-obese women with periodontal health or disease were found. However, a high prevalence of P. gingivalis in obese women with periodontal health was observed. © 2018 American Academy of Periodontology.

Subgingival bacterial recolonization after scaling and root planing in smokers with chronic periodontitis.

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Feres, M; Bernal, Mac; Matarazzo, F; Faveri, M; Duarte, P M; Figueiredo, L C

2015-06-01

The aim of this study was to compare subgingival bacterial recolonization patterns after scaling and root planing in current smokers and non-smokers. 15 smokers and 15 non-smokers with chronic periodontitis received scaling and root planing in six visits lasting one hour each, over a period of 21 days. Clinical monitoring was performed at baseline and 180 days, and microbiological monitoring was performed at baseline, immediately after scaling and root planing (Day 0) and at 42, 63 and 180 days post-therapy. Subgingival plaque samples were analysed by checkerboard DNA-DNA hybridization. An improvement in clinical condition was observed for smokers and non-smokers; however, non-smokers showed a greater reduction in mean clinical attachment level in intermediate sites in comparison with smokers (p < 0.05). At Day 0, there was a significant reduction in the mean counts of the three pathogens from the red complex, Eubacterium nodatum and Parvimonas micra only in non-smokers (p < 0.05). There was a significant increase in the proportion of host-compatible species in non-smokers and smokers from baseline to 180 days post-therapy (p < 0.05). However, a significant decrease in the pathogenic species was observed only in non-smokers. Smokers were more susceptible to the re-establishment of a pathogenic subgingival biofilm than non-smokers. © 2015 Australian Dental Association.

Profiling of subgingival plaque biofilm microbiota in female adult patients with clear aligners: a three-month prospective study.

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Guo, Runzhi; Zheng, Yunfei; Liu, Hao; Li, Xiaobei; Jia, Lingfei; Li, Weiran

2018-01-01

Clear aligners are well known for facilitating oral hygiene maintenance and decreasing susceptibility to periodontal diseases as compared to conventional fixed appliances. However, few research studies focus on the subgingival microbial community during clear aligner treatment (CAT). Hence, this study investigates changes of the subgingival microbial community and its association with clinical characteristics during the first three months of CAT. Ten female patients with clear aligners were enrolled in this study. Subgingival plaque samples were obtained at three time points: before orthodontic treatment (T0), one month after orthodontic treatment (T1) and three months after orthodontic treatment (T2). DNA was then extracted from plaque samples and analyzed by 16S rRNA gene sequencing. Periodontal examinations, including plaque index (PI) and gingival bleeding index (GBI) measurements were also recorded. The plaque indices (PIs) and gingival bleeding indices (GBIs) were slightly increased at T1 and T2, but no statistically significant difference was found. The alpha diversity indices, including the ACE, Chao1, Shannon indices, all showed a declining trend without significance, and a rising trend in the Simpson diversity index was observed. The weighted UniFrac distance was significantly higher at T1 and T2 compared with T0. Principal Coordinates Analysis (PCoA) demonstrated that the communities at T0 tended to cluster apart from the communities at T1 and T2. The relative abundance of the phylum Firmicutes and genus Mycoplasma was significantly increased at T0 compared with T2. There was no significant difference in the relative abundance of periodontal pathogens at the genus and species levels or core microorganisms at the genus level. A slightly decreasing microbial diversity with a significant change of microbial structure was found during the first three-month clear aligner treatment (CAT). However, subjects receiving clear aligner treatment were free from

Dental biofilm infections

DEFF Research Database (Denmark)

Larsen, Tove; Fiehn, Nils-Erik

2017-01-01

and cause gingival inflammation and breakdown of supporting periodontal fibers and bone and ultimately tooth loss, i.e., gingivitis, chronic or aggressive periodontitis, and around dental implants, peri-implantitis. Furthermore, bacteria from the dental biofilm may spread to other parts of the body......-fermenting bacteria causing demineralization of teeth, dental caries, which may further lead to inflammation and necrosis in the pulp and periapical region, i.e., pulpitis and periapical periodontitis. In supra- and subgingival biofilms, predominantly gram-negative, anaerobic proteolytic bacteria will colonize...

Profiling of subgingival plaque biofilm microbiota in female adult patients with clear aligners: a three-month prospective study

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Runzhi Guo

2018-01-01

Full Text Available Background Clear aligners are well known for facilitating oral hygiene maintenance and decreasing susceptibility to periodontal diseases as compared to conventional fixed appliances. However, few research studies focus on the subgingival microbial community during clear aligner treatment (CAT. Hence, this study investigates changes of the subgingival microbial community and its association with clinical characteristics during the first three months of CAT. Methods Ten female patients with clear aligners were enrolled in this study. Subgingival plaque samples were obtained at three time points: before orthodontic treatment (T0, one month after orthodontic treatment (T1 and three months after orthodontic treatment (T2. DNA was then extracted from plaque samples and analyzed by 16S rRNA gene sequencing. Periodontal examinations, including plaque index (PI and gingival bleeding index (GBI measurements were also recorded. Results The plaque indices (PIs and gingival bleeding indices (GBIs were slightly increased at T1 and T2, but no statistically significant difference was found. The alpha diversity indices, including the ACE, Chao1, Shannon indices, all showed a declining trend without significance, and a rising trend in the Simpson diversity index was observed. The weighted UniFrac distance was significantly higher at T1 and T2 compared with T0. Principal Coordinates Analysis (PCoA demonstrated that the communities at T0 tended to cluster apart from the communities at T1 and T2. The relative abundance of the phylum Firmicutes and genus Mycoplasma was significantly increased at T0 compared with T2. There was no significant difference in the relative abundance of periodontal pathogens at the genus and species levels or core microorganisms at the genus level. Conclusion A slightly decreasing microbial diversity with a significant change of microbial structure was found during the first three-month clear aligner treatment (CAT. However, subjects

A Biofilm Pocket Model to Evaluate Different Non-Surgical Periodontal Treatment Modalities in Terms of Biofilm Removal and Reformation, Surface Alterations and Attachment of Periodontal Ligament Fibroblasts.

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Tobias T Hägi

Full Text Available There is a lack of suitable in vitro models to evaluate various treatment modalities intending to remove subgingival bacterial biofilm. Consequently, the aims of this in vitro-study were: a to establish a pocket model enabling mechanical removal of biofilm and b to evaluate repeated non-surgical periodontal treatment with respect to biofilm removal and reformation, surface alterations, tooth hard-substance-loss, and attachment of periodontal ligament (PDL fibroblasts.Standardized human dentin specimens were colonized by multi-species biofilms for 3.5 days and subsequently placed into artificially created pockets. Non-surgical periodontal treatment was performed as follows: a hand-instrumentation with curettes (CUR, b ultrasonication (US, c subgingival air-polishing using erythritol (EAP and d subgingival air-polishing using erythritol combined with chlorhexidine digluconate (EAP-CHX. The reduction and recolonization of bacterial counts, surface roughness (Ra and Rz, the caused tooth substance-loss (thickness as well as the attachment of PDL fibroblasts were evaluated and statistically analyzed by means of ANOVA with Post-Hoc LSD.After 5 treatments, bacterial reduction in biofilms was highest when applying EAP-CHX (4 log10. The lowest reduction was found after CUR (2 log10. Additionally, substance-loss was the highest when using CUR (128±40 µm in comparison with US (14±12 µm, EAP (6±7 µm and EAP-CHX (11±10 µm. Surface was roughened when using CUR and US. Surfaces exposed to US and to EAP attracted the highest numbers of PDL fibroblasts.The established biofilm model simulating a periodontal pocket combined with interchangeable placements of test specimens with multi-species biofilms enables the evaluation of different non-surgical treatment modalities on biofilm removal and surface alterations. Compared to hand instrumentation the application of ultrasonication and of air-polishing with erythritol prevents from substance-loss and results

Detection and comparison of Selenomonas sputigena in subgingival biofilms in chronic and aggressive periodontitis patients

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Disha Nagpal

2016-01-01

Full Text Available Background: With the advent of DNA-based culture-independent techniques, a constantly growing number of Selenomonas phylotypes have been detected in patients with destructive periodontal diseases. However, the prevalence levels that have been determined in different studies vary considerably. Aim: The present study was undertaken to detect and compare the presence of Selenomonas sputigena in the subgingival plaque samples from generalized aggressive periodontitis (GAP, chronic generalized periodontitis, and periodontally healthy patients using conventional polymerase chain reaction (PCR technique. Materials and Methods: A total of 90 patients were categorized as periodontally healthy individuals (Group I, n = 30, chronic generalized periodontitis (Group II, n = 30, and GAP (Group III, n = 30. The clinical parameters were recorded and subgingival plaque samples were collected. These were then subjected to conventional PCR analysis.Statistical Analysis Used: Kruskal–Wallis ANOVA test was used for multiple group comparisons followed by Mann–Whitney U-test for pairwise comparison. Results: On comparison between three groups, all the clinical parameters were found to be statistically highly significant. Comparing Groups I-II and I-III, the difference in detection was found to be statistically highly significant whereas in Groups II-III, it was statistically nonsignificant. On comparison of S. sputigena detected and undetected patients to clinical parameters in various study groups, the difference was found to be nonsignificant. Conclusion:S. sputigena was found to be significantly associated with chronic and aggressive periodontitis. Although the difference in its detection frequency in both groups was statistically nonsignificant when compared clinically, S. sputigena was more closely associated with the GAP.

Reproducibility of subgingival bacterial samples from patients with peri-implant mucositis

DEFF Research Database (Denmark)

Hallström, Hadar; Persson, G Rutger; Strömberg, Ulf

2015-01-01

collected with paper points and analyzed using the checkerboard DNA-DNA hybridization technique. Whole genomic probes of 74 preselected bacterial species were used. Based on the bacterial scores, Cohen's kappa coefficient was used to calculate the inter-annotator agreement for categorical data......OBJECTIVE: The aim of the present study was to investigate the reproducibility of bacterial enumeration from subsequent subgingival samples collected from patients with peri-implant mucositis. MATERIAL AND METHODS: Duplicate microbial samples from 222 unique implant sites in 45 adult subjects were....... The percentage agreement was considered as "good" when the two samples showed the same score or differed by 1 to the power of 10. RESULTS: Moderate to fair kappa values were displayed for all bacterial species in the test panel (range 0.21-0.58). There were no significant differences between Gram...

Comparison of Subgingival and Peri-implant Microbiome in Chronic Periodontitis.

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Zhang, Qian; Qin, Xue Yan; Jiang, Wei Peng; Zheng, Hui; Xu, Xin Li; Chen, Feng

2015-09-01

To analyse the microbia composition of 10 healthy dental implants and 10 chronic periodontitis patients. Subgingival plaque and peri-implant biofilm were sampled at the first molar site before and after implant restoration. The analysis was conducted by 454-prosequencing of bacterial V1 to V3 regions of 16S rDNA. Chronic periodontitis subjects showed greater bacterial diversity compared with implant subjects. The relative abundance of sixteen genera and twelve species differed significantly between implant and chronic periodontitis subjects. The genera Catonella, Desulfovibrio, Mogibacterium, Peptostreptococcus and Propionibacterium were present in higher abundance in chronic periodontitis subjects, while implant subjects had higher proportions of Brevundimonas and Pseudomonas species. Our results demonstrate that implant restoration changes the oral microbiota. The analysis suggests that periodontal bacteria can remain for a prolonged period of time at non-dental sites, from where they can colonise the peri-implant.

Chlorhexidine with or without alcohol against biofilm formation: efficacy, adverse events and taste preference

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Gabriela Otero dos SANTOS

2017-05-01

Full Text Available Abstract In recent years, different chlorhexidine formulations have been tested, including an alcohol-free alternative, but the effect of this solution on early biofilm formation is not clear. A crossover, randomized, double-blind clinical trial was conducted to evaluate the effect of two chlorhexidine solutions against supra- and subgingival biofilm formation (NCT#02656251. Thirty-five participants were randomized and asked to rinse twice daily with 15 ml of an alcohol-containing 0.12% chlorhexidine solution, an alcohol-free 0.12% chlorhexidine solution, or placebo. The study was conducted in three experimental periods of 4 days each, with a 10-day washout between the periods. All the experimental periods followed the same protocol, except that the solutions were switched. Biofilm distribution was evaluated every 24 hours by the Plaque-Free Zone Index, during 96 hours. Adverse events were self-reported and sensory evaluation was performed using a hedonic scale. Compared to the placebo, the chlorhexidine solutions resulted in a significantly higher number of surfaces free of plaque over 96 hours (p < 0.01, and were able to prevent subgingival biofilm formation (p < 0.01. The alcohol-free chlorhexidine solution was associated with a lower incidence of adverse events, compared with alcohol-containing chlorhexidine (p < 0.05; it also received better sensory evaluation and acceptance by trial participants, compared with the alcohol-containing chlorhexidine (p = 0.007, and had a similar inhibitory effect on the formation of supra- and subgingival biofilms.

Identification of subgingival periodontal pathogens and association with the severity of periodontitis in patients with chronic kidney diseases: a cross-sectional study.

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Ismail, Fidan Bahtiar; Ismail, Gener; Dumitriu, Anca Silvia; Baston, Catalin; Berbecar, Vlad; Jurubita, Roxana; Andronesi, Andreea; Dumitriu, Horia Traian; Sinescu, Ioanel

2015-01-01

The aim of our study was to assess the subgingival profile of 9 periodontal pathogens, by means of real-time PCR, in a group of predialysis chronic kidney disease patients with and without periodontal disease and to identify the risk factors associated with periodontal disease in these patients. This is a single centre cross-sectional cohort study performed on 70 CKD patients. Patients received a full-mouth periodontal examination and the following parameters were assessed: periodontal pocket depth (PPD), clinical attachment level, bleeding on probing, and plaque index; subgingival biofilm samples were collected from the deepest periodontal pocket of each quadrant and were pooled in one transporting unit. Clinical data were drawn from the medical file of the patients. T. denticola (P = 0.001), T. forsythia (P < 0.001), and P. micros (P = 0.003) are significantly associated with periodontal disease in CKD subjects but in a multivariate model only age and T. forsythia remain independent risk factors for periodontal disease in patients with CKD. In our cohort, age and T. forsythia are independently associated with periodontitis in CKD patients. Within the limits of this study, CKD was not significantly associated with a particular subgingival periodontal pathogens profile in periodontitis patients.

Identification of Subgingival Periodontal Pathogens and Association with the Severity of Periodontitis in Patients with Chronic Kidney Diseases: A Cross-Sectional Study

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Fidan Bahtiar Ismail

2015-01-01

Full Text Available Background. The aim of our study was to assess the subgingival profile of 9 periodontal pathogens, by means of real-time PCR, in a group of predialysis chronic kidney disease patients with and without periodontal disease and to identify the risk factors associated with periodontal disease in these patients. Material and Methods. This is a single centre cross-sectional cohort study performed on 70 CKD patients. Patients received a full-mouth periodontal examination and the following parameters were assessed: periodontal pocket depth (PPD, clinical attachment level, bleeding on probing, and plaque index; subgingival biofilm samples were collected from the deepest periodontal pocket of each quadrant and were pooled in one transporting unit. Clinical data were drawn from the medical file of the patients. Results. T. denticola (P=0.001, T. forsythia (P<0.001, and P. micros (P=0.003 are significantly associated with periodontal disease in CKD subjects but in a multivariate model only age and T. forsythia remain independent risk factors for periodontal disease in patients with CKD. Conclusions. In our cohort, age and T. forsythia are independently associated with periodontitis in CKD patients. Within the limits of this study, CKD was not significantly associated with a particular subgingival periodontal pathogens profile in periodontitis patients.

Levels of Candidate Periodontal Pathogens in Subgingival Biofilm.

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Oliveira, R R D S; Fermiano, D; Feres, M; Figueiredo, L C; Teles, F R F; Soares, G M S; Faveri, M

2016-06-01

In recent years, several new periodontal taxa have been associated with the etiology of periodontitis. A recent systematic review provides further support for the pathogenic role of 17 species/phylotypes. Thus, the aim of this study was to assess the prevalence and levels of these species in subjects with generalized chronic periodontitis (GChP; n = 30), generalized aggressive periodontitis (GAgP; n = 30), and periodontal health (PH; n = 30). All subjects underwent clinical and microbiological assessment. Nine subgingival plaque samples were collected from each subject and analyzed for their content of 20 bacterial species/phylotypes through the RNA-oligonucleotide quantification technique. Subjects from the GChP and GAgP groups presented the highest mean values for all clinical parameters in comparison with the PH group (P GChP and GAgP showed significantly higher mean levels of Bacteroidetes sp. human oral taxon (HOT) 274, Fretibacterium sp. HOT 360, and TM7 sp. HOT 356 phylotypes, as well as higher mean levels of Filifactor alocis, Fretibacterium fastidiosum, Porphyromonas gingivalis, Tannerella forsythia, and Selenomonas sputigena species than PH subjects (P GChP subjects (P GChP and GAgP than in PH subjects. Mean levels of P. gingivalis (r = 0.68), T. forsythia (r = 0.62), F. alocis (r = 0.51, P = 0.001), and Fretibacterium sp. HOT 360 (r = 0.41) were correlated with pocket depth (P < 0.001). In conclusion, Bacteroidales sp. HOT 274, Desulfobulbus sp. HOT 041, Fretibacterium sp. HOT 360, Fretibacterium sp. HOT 362, and TM7 sp. HOT 356 phylotypes, in addition to F. alocis, F. fastidiosum, and S. sputigena, seem to be associated with periodontitis, and their role in periodontal pathogenesis should be further investigated. © International & American Associations for Dental Research 2016.

Impact of supragingival therapy on subgingival microbial profile in smokers versus non-smokers with severe chronic periodontitis

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Tatiana Meulman

2012-01-01

Full Text Available The aim of this study was to assess subgingival microbiological changes in smokers versus non-smokers presenting severe chronic periodontitis after supragingival periodontal therapy (ST.Non-smokers (n=10 and smokers (n=10 presenting at least nine teeth with probing pocket depth (PPD (≥5 mm, bleeding on probing (BoP, and no history of periodontal treatment in the last 6 months were selected. Clinical parameters assessed were plaque index (PI, BoP, PPD, relative gingival margin position (rGMP and relative clinical attachment level (rCAL. Subgingival biofilm was collected before and 21 days after ST. DNA was extracted and the 16S rRNA gene was amplified with the universal primer pair, 27F and 1492R. Amplified genes were cloned, sequenced, and identified by comparison with known 16S rRNA sequences. Statistical analysis was performed by Student's t and Chi-Square tests (α=5%.Clinically, ST promoted a significant reduction in PI and PPD, and gain of rCAL for both groups, with no significant intergroup difference. Microbiologically, at baseline, data analysis demonstrated that smokers harbored a higher proportion of Porphyromonas endodontalis, Bacteroidetes sp., Fusobacterium sp. and Tannerella forsythia and a lower number of cultivated phylotypes (p<0.05. Furthermore, non-smokers featured significant reductions in key phylotypes associated with periodontitis, whereas smokers presented more modest changes.Within the limits of the present study, ST promoted comparable clinical improvements in smokers and non-smokers with severe chronic periodontitis. However, in smokers, ST only slightly affected the subgingival biofilm biodiversity, as compared with non-smokers.

Comparative evaluation of subgingival plaque microflora in pregnant and non-pregnant women: A clinical and microbiologic study

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Rishi Emmatty

2013-01-01

Full Text Available Context: Gingival changes in pregnancy have been attributed to changes in the subgingival biofilm related to hormonal variations. Aims: To evaluate the subgingival plaque microflora in pregnant and nonpregnant women to determine if pregnancy induces any alterations in the subgingival plaque and to associate these changes with changes in periodontal status. Settings and Design: Thirty pregnant and 10 nonpregnant women within the age group of 20-35 years having a probing pocket depth (PPD of 3-4 mm were included in the study. The pregnant women were equally divided into 3 groups of 10, each belonging to I, II, and III trimester. Materials and Methods: Plaque index, gingival index, PPD, and microbiologic evaluation for specific bacterial counts for Prevotella intermedia, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Parvimonas micra, and Fusobacterium nucleatum were carried out for all subjects. Statistical Analysis: Mann-Whitney U test. Results: Increase in gingival inflammation was observed in II and III trimester as compared with I trimester and control. Plaque scores did not show any significant difference between pregnant and nonpregnant women. Specific bacterial examination revealed an increase in proportion of P. intermedia in pregnant women of both II and III trimester as compared with I trimester and nonpregnant women. Conclusions: A definite increase in proportions of P. intermedia occurs in subgingival plaque microflora in pregnancy that may be responsible for the exaggerated gingival response.

Subgingival Microbiome Colonization and Cytokine Production during Early Dental Implant Healing.

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Payne, Jeffrey B; Johnson, Paul G; Kok, Car Reen; Gomes-Neto, João C; Ramer-Tait, Amanda E; Schmid, Marian J; Hutkins, Robert W

2017-01-01

Little is known about longitudinal development of the peri-implant subgingival microbiome and cytokine production as a new sulcus forms after dental implant placement. Therefore, the purpose of this observational study was to evaluate simultaneous longitudinal changes in the oral microbiome and cytokine production in the developing peri-implant sulcus compared to control natural teeth. Four and 12 weeks after implant placement and abutment connection, a dental implant and a natural tooth were sampled in 25 patients for subgingival plaque and gingival crevicular fluid (GCF [around teeth] and peri-implant crevicular fluid [PICF] around implants). DNA from plaque samples was extracted and sequenced using Illumina-based 16S rRNA sequencing. GCF and PICF samples were analyzed using a customized Milliplex human cytokine and chemokine magnetic bead panel. Beta diversity analysis revealed that natural teeth and implants had similar subgingival microbiomes, while teeth had greater alpha diversity than implants. At the genus level, however, few differences were noted between teeth and dental implants over 12 weeks. Specifically, Actinomyces and Selenomonas were significantly elevated around teeth versus dental implants at both 4 weeks and 12 weeks, while Corynebacterium and Campylobacter were significantly elevated only at 4 weeks around teeth. The only difference between PICF and GCF biomarkers was significantly elevated granulocyte-macrophage colony-stimulating factor levels around teeth versus dental implants at the 4-week visit. The subgingival microbiome and cytokine production were similar between teeth and implants during early healing, suggesting that these profiles are driven by the patient following dental implant placement and are not determined by anatomical niche. IMPORTANCE Dental implants are a common treatment option offered to patients for tooth replacement. However, little is known regarding initial colonization of the subgingival microbiome and

Chlorhexidine with or without alcohol against biofilm formation: efficacy, adverse events and taste preference.

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Santos, Gabriela Otero Dos; Milanesi, Fernanda Carpes; Greggianin, Bruna Frizon; Fernandes, Marilene Issa; Oppermann, Rui Vicente; Weidlich, Patricia

2017-05-04

In recent years, different chlorhexidine formulations have been tested, including an alcohol-free alternative, but the effect of this solution on early biofilm formation is not clear. A crossover, randomized, double-blind clinical trial was conducted to evaluate the effect of two chlorhexidine solutions against supra- and subgingival biofilm formation (NCT#02656251). Thirty-five participants were randomized and asked to rinse twice daily with 15 ml of an alcohol-containing 0.12% chlorhexidine solution, an alcohol-free 0.12% chlorhexidine solution, or placebo. The study was conducted in three experimental periods of 4 days each, with a 10-day washout between the periods. All the experimental periods followed the same protocol, except that the solutions were switched. Biofilm distribution was evaluated every 24 hours by the Plaque-Free Zone Index, during 96 hours. Adverse events were self-reported and sensory evaluation was performed using a hedonic scale. Compared to the placebo, the chlorhexidine solutions resulted in a significantly higher number of surfaces free of plaque over 96 hours (p alcohol-free chlorhexidine solution was associated with a lower incidence of adverse events, compared with alcohol-containing chlorhexidine (p alcohol-containing chlorhexidine (p = 0.007), and had a similar inhibitory effect on the formation of supra- and subgingival biofilms.

Biofilm-mediated Antibiotic-resistant Oral Bacterial Infections: Mechanism and Combat Strategies.

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Kanwar, Indulata; Sah, Abhishek K; Suresh, Preeti K

2017-01-01

Oral diseases like dental caries and periodontal disease are directly associated with the capability of bacteria to form biofilm. Periodontal diseases have been associated to anaerobic Gram-negative bacteria forming a subgingival plaque (Porphyromonas gingivalis, Actinobacillus, Prevotella and Fusobacterium). Biofilm is a complex bacterial community that is highly resistant to antibiotics and human immunity. Biofilm communities are the causative agents of biological developments such as dental caries, periodontitis, peri-implantitis and causing periodontal tissue breakdown. The review recapitulates the latest advancements in treatment of clinical biofilm infections and scientific investigations, while these novel anti-biofilm strategies are still in nascent phases of development, efforts dedicated to these technologies could ultimately lead to anti-biofilm therapies that are superior to the current antibiotic treatment. This paper provides a review of the literature focusing on the studies on biofilm in the oral cavity, formation of dental plaque biofilm, drug resistance of bacterial biofilm and the antibiofilm approaches as biofilm preventive agents in dentistry, and their mechanism of biofilm inhibition. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Characteristics of subgingival calculus detection by multiphoton fluorescence microscopy

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Tung, Oi-Hong; Lee, Shyh-Yuan; Lai, Yu-Lin; Chen, How-Foo

2011-06-01

Subgingival calculus has been recognized as a major cause of periodontitis, which is one of the main chronic infectious diseases of oral cavities and a principal cause of tooth loss in humans. Bacteria deposited in subgingival calculus or plaque cause gingival inflammation, function deterioration, and then periodontitis. However, subgingival calculus within the periodontal pocket is a complicated and potentially delicate structure to be detected with current dental armamentaria, namely dental x-rays and dental probes. Consequently, complete removal of subgingival calculus remains a challenge to periodontal therapies. In this study, the detection of subgingival calculus employing a multiphoton autofluorescence imaging method was characterized in comparison with a one-photon confocal fluorescence imaging technique. Feasibility of such a system was studied based on fluorescence response of gingiva, healthy teeth, and calculus with and without gingiva covered. The multiphoton fluorescence technology perceived the tissue-covered subgingival calculus that cannot be observed by the one-photon confocal fluorescence method.

Comparison of subgingival bacterial sampling with oral lavage for detection and quantification of periodontal pathogens by real-time polymerase chain reaction

NARCIS (Netherlands)

Boutaga, Khalil; Savelkoul, Paul H. M.; Winkel, Edwin G.; van Winkelhoff, Arie J.

Background: Saliva has been studied for the presence of subgingival pathogens in periodontitis patients. With the anaerobic culture technique, the discrepancy between salivary recovery and subgingival presence has been significant, which makes this approach not suitable for practical use in the

[Study on Microbial Diversity of Peri-implantitis Subgingival by High-throughput Sequencing].

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Li, Zhi-jie; Wang, Shao-guo; Li, Yue-hong; Tu, Dong-xiang; Liu, Shi-yun; Nie, Hong-bing; Li, Zhi-qiang; Zhang, Ju-mei

2015-07-01

To study microbial diversity of peri-implantitis subgingival with high-throughput sequencing, and investigate microbiological etiology of peri-implantitis. Subgingival plaques were sampled from the patients with peri-implantitis (D group) and non-peri-implantitis subjects (N group). The microbiological diversity of the subgingival plaques was detected by sequencing V4 region of 16S rRNA with Illumina Miseq platform. The diversity of the community structure was analyzed using Mothur software. A total of 156 507 gene sequences were detected in nine samples and 4 402 operational taxonomic units (OTUs) were found. Selenomonas, Pseudomonas, and Fusobacterium were dominant bacteria in D group, while Fusobacterium, Veillonella and Streptococcus were dominant bacteria in N group. Differences between peri-implantitis and non-peri-implantitis bacterial communities were observed at all phylogenetic levels by LEfSe, which was also found in PcoA test. The occurrence of peri-implantitis is not only related to periodontitis pathogenic microbe, but also related with the changes of oral microbial community structure. Treponema, Herbaspirillum, Butyricimonas and Phaeobacte may be closely related to the occurrence and development of peri-implantitis.

Prevalence of "non-oral" pathogenic bacteria in subgingival biofilm of subjects with chronic periodontitis Prevalência de bactérias patogênicas "não-orais" no biofilme dental subgengival de pacientes com periodontite crônica

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Renata Souto

2006-09-01

Full Text Available The oral cavity may act as a reservoir for several pathogens related to systemic infections. Therefore, the purpose of this study was to determine the prevalence and levels of pathogenic bacteria in the subgingival biofilm of chronic periodontitis lesions and healthy periodontal sites using the checkerboard DNA-DNA hybridization technique. 200 samples of subgingival biofilm from sites with periodontitis (probing pocket depth > 4 mm and /or clinical attachment level > 4 mm and 200 samples from healthy sites of 14 patients with chronic periodontitis, as well as 200 samples from 3 periodontally healthy patients were obtained. The presence and levels of 11 pathogenic bacteria were determined using whole genomic DNA probes and the checkerboard method, computed for each site and then across sites within each subject group. Significance of differences in clinical and microbiological parameters among groups were examinated using the Mann-Whitney and Wilcoxon sign tests. The predominant species in all 600 samples included Corynebacterium diphtheriae, Enterococcus faecalis, Staphylococcus aureus,Acinetobacter baumannii and Escherichia coli. In general, most of the species were detected in greater prevalence and levels in sites with and without disease from patients with periodontitis in comparison to the periodontally healthy group. In particular, C. diphtheriae, E. coli, E. faecalis, P. aeruginosa and S. aureus were significantly more prevalent and detected in higher counts in diseased sites of patients with periodontal disease compared to healthy subjects (p Apesar da extensa literatura sobre a associação de bactérias orais e doenças sistêmicas, tem se dado pouca atenção à cavidade oral como um reservatório de bactérias patogênicas "não-orais". A microbiota oral é constituída de mais de 300 espécies bacterianas já caracterizadas, além de organismos não cultiváveis que vêm sendo descobertos através de técnicas moleculares. O

Near-ultraviolet removal rates for subgingival dental calculus at different irradiation angles

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Schoenly, Joshua E.; Seka, Wolf D.; Rechmann, Peter

2011-07-01

The laser ablation rate of subgingival dental calculus irradiated at a 400-nm-wavelength, 7.4-mJ pulse energy, and 85- and 20-deg irradiation angles is measured using laser triangulation. Three-dimensional images taken before and after irradiation create a removal map with 6-μm axial resolution. Fifteen human teeth with subgingival calculus are irradiated in vitro under a cooling water spray with an ~300-μm-diam, tenth-order super-Gaussian beam. The average subgingival calculus removal rates for irradiation at 85 and 20 deg are 11.1+/-3.6 and 11.5+/-5.9 μm/pulse, respectively, for depth removal and 4.5+/-1.7×105 and 4.8+/-2.3×105 μm3/pulse, respectively, for volume removal. The ablation rate is constant at each irradiation site but varies between sites because of the large differences in the physical and optical properties of calculus. Comparison of the average depth- and volume-removal rates does not reveal any dependence on the irradiation angle and is likely due to the surface topology of subgingival calculus samples that overshadows any expected angular dependence.

Reliability of recordings of subgingival calculus detected using an ultrasonic device.

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Corraini, Priscila; López, Rodrigo

2015-04-01

To assess the intra-examiner reliability of recordings of subgingival calculus detected using an ultrasonic device, and to investigate the influence of subject-, tooth- and site-level factors on the reliability of these subgingival calculus recordings. On two occasions, within a 1-week interval, 147 adult periodontitis patients received a full-mouth clinical periodontal examination by a single trained examiner. Duplicate subgingival calculus recordings, in six sites per tooth, were obtained using an ultrasonic device for calculus detection and removal. Agreement was observed in 65 % of the 22,584 duplicate subgingival calculus recordings, ranging 45 % to 83 % according to subject. Using hierarchical modeling, disagreements in the subgingival calculus duplicate recordings were more likely in all other sites than the mid-buccal, and in sites harboring supragingival calculus. Disagreements were less likely in sites with PD ≥  4 mm and with furcation involvement  ≥  degree 2. Bleeding on probing or suppuration did not influence the reliability of subgingival calculus. At the subject-level, disagreements were less likely in patients presenting with the highest and lowest extent categories of the covariate subgingival calculus. The reliability of subgingival calculus recordings using the ultrasound technology is reasonable. The results of the present study suggest that the reliability of subgingival calculus recordings is not influenced by the presence of inflammation. Moreover, subgingival calculus can be more reliably detected using the ultrasound device at sites with higher need for periodontal therapy, i.e., sites presenting with deep pockets and premolars and molars with furcation involvement.

Comparative evaluation of two subgingival irrigating solutions in the management of periodontal disease: A clinicomicrobial study

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Dhara Jayesh Pandya

2016-01-01

Full Text Available Context: Local administration of antimicrobial agents offer a “site-specific” approach to the periodontal therapy and it has several benefits. Aim: The present study was aimed to assess the clinical and microbial changes by subgingival irrigation using different subgingival irrigants in periodontitis patients and also to assess the mechanical effect of different local irrigation devices; if any. Settings and Design: Split-mouth design was employed on ten individuals. Materials and Methods: The study sample consisted of 10 individuals in whom full-mouth scaling and root planing was performed and subgingival irrigation therapy was instituted for an experimental period of 30 days. The clinical as well as microbiological parameters were evaluated. Statistical Analysis Used: To calculate baseline data with day thirty data,paired t-test was used. Intergroup comparison was carried out using one-way ANOVA. Multiple comparisons among groups were carried out using post hoc Tamhane's T2 test. Results: Among the different subgingival irrigants used, 0.2% chlorhexidine gluconate is most effective followed by ozonated water, whereas saline was found to be ineffective when compared to the other two subgingival irrigants. Subgingival irrigation using pulsated device may not have any additive effect in alteration of the subgingival microflora. Conclusion: Within the limits and scope of the study, it can be safely concluded that 0.2% chlorhexidine may be used as an adjunct to mechanical therapy for achieving a significant reduction in inflammatory periodontal changes and also reduction in periodontopathogenic microflora.

Changes in subgingival microflora after placement and removal of fixed orthodontic appliances

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Živković-Sandić Marija

2014-01-01

Full Text Available Introduction. The placement of fixed orthodontic appliances may lead to increased plaque accumulation and changes in subgingival microflora. Objective. The aim of this study was to examine the changes in frequency of subgingival microflora that occur after placement and removal of fixed orthodontic appliance using polymerase chain reaction (PCR. Methods. This study included 33 orthodontic patients, who were divided into two groups. Subgingival plaque samples were collected from the right upper incisor (U1 and right upper first molar (U6. In group A, the samples were taken three times: before placement appliance (T1, after one month (T2, and after 3 months (T3. In group B the samples were also taken three times: before appliance removal (T1, after one month (T2, and after three months (T3. PCR method was used to determine the presence of P. gingivalis, A. actinomycetemcomitans, T. forsythia, and P. intermedia. Results. In group A the frequency of P. gingivalis showed statistically significant decrease at U1 (p=0.049 and U6 (p=0.008, from T1 to T2, and at U1 (p=0.048 from T1 to T3. In group B only the frequency of T. forsythia showed a statistically significant decrease, at U6 (T1 vs. T2, p=0.004; T1 vs. T3, p=0.0003. Regarding other analyzed bacteria, changes in the presence were noticed but no statistical significance was found. Conclusion. Placement of fixed appliances may have an impact on subgingival microflora, but in the first months after the placement and removal of the appliance changes were not significant, probably due to good oral hygiene. [Projekat Ministarstva nauke Republike Srbije, br. 175075

Simultaneous detection of periodontal pathogens in subgingival plaque and placenta of women with hypertension in pregnancy.

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Swati, P; Thomas, Betsy; Vahab, Saadi Abdul; Kapaettu, Satyamoorthy; Kushtagi, Pralhad

2012-03-01

There are many studies documenting increased prevalence of periodontal infection in women with preeclampsia. But, very few studies have attempted to establish causal relationship between the two. To find out causal circumstantial evidence by isolating specific periodontal pathogens in oral and placental samples. Antenatal periodontal screening and subgingival plaque collection was carried out in ten women with hypertension in pregnancy and ten normotensive controls on their hospital admission at term for cesarean delivery. Placental biopsy was obtained after aseptic placental collection at the time of elective cesarean delivery. Subgingival plaque and placental biopsy were studied for Porphyromonas gingivalis, Fusobacterium nucleatum, Treponema denticola, Prevotella intermedia and Aggregatibacter actinomycetemcomitans using quantitative polymerase chain reaction technique. Periodontist and laboratory personnel were unaware of case or control status. Periodontal status was not informed to the obstetrician recruiting the cases and laboratory. Microbiology report was not revealed till end of the study. Periodontal pathogens were found to be high in the group with hypertension than the controls. P gingivalis was found in all the samples from subgingival plaque and placenta, irrespective of the periodontal disease status. In cases with hypertension, periodontal pathogens are present in higher proportion in subgingival plaque and placenta.

Effect of non-surgical periodontal treatment on the subgingival microbiota of patients with chronic kidney disease

OpenAIRE

Artese,Hilana Paula Carillo; Sousa,Celso Oliveira de; Torres,Maria Cynésia Medeiros de Barros; Silva-Boghossian,Carina Maciel; Colombo,Ana Paula Vieira

2012-01-01

This study investigated the effect of non-surgical periodontal therapy on the composition of subgingival microbiota of patients with chronic kidney disease (CKD). Sixteen CKD pre-dialysis individuals (CKD) and 14 individuals without clinical evidence of kidney disease (C) presenting chronic periodontitis were treated by scaling and root planing. Subgingival samples were collected from each patient and analyzed for their composition by checkerboard at baseline and 3 months post-therapy. Signif...

Pyrosequencing Analysis of Subgingival Microbiota in Distinct Periodontal Conditions.

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Park, O-J; Yi, H; Jeon, J H; Kang, S-S; Koo, K-T; Kum, K-Y; Chun, J; Yun, C-H; Han, S H

2015-07-01

Subgingival microorganisms are potentially associated with periodontal diseases. However, changes in the subgingival microbiota during the progress of periodontal diseases are poorly understood. In this study, we analyzed bacterial communities in the subgingival paper point samples from 32 Korean individuals with no sign of disease, gingivitis, or periodontitis using 454 FLX Titanium pyrosequencing. A total of 256,113 reads representing 26 phyla, 433 genera, and 1,016 species were detected. Bacteroidetes, Fusobacteria, Synergistetes, and Spirochaetes were the abundant phyla in periodontitis subjects, whereas Firmicutes and Proteobacteria were identified as the dominant phyla in the gingivitis and healthy subjects, respectively. Although high levels of Porphyromonas, Fusobacterium, Fretibacterium, Rothia, Filifactor, and Treponema genera were observed in the periodontitis subjects, Streptococcus, Capnocytophaga, Leptotrichia, and Haemophilus genera were found at high frequency in the gingivitis subjects. Species including Porphyromonas gingivalis, Fusobacterium nucleatum, and Fretibacterium fastidiosum were significantly increased in periodontitis subjects. On the other hand, Streptococcus pseudopneumoniae, Haemophilus parainfluenzae, and Leptotrichia hongkongensis were preferentially observed in the gingivitis subjects. Intriguingly, the halophile Halomonas hamiltonii was revealed as a predominant species in the healthy subjects. Based on Fast UniFrac analysis, distinctive bacterial clusters were classified for the healthy, gingivitis, and periodontitis state. The current findings might be useful for understanding the pathogenesis, diagnosis, and treatment of periodontal diseases. © International & American Associations for Dental Research 2015.

Subgingival Microbiota in White Patients With Desquamative Gingivitis: A Cross-Sectional Study.

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Arduino, Paolo G; Romano, Federica; Sasia, Danilo; Broccoletti, Roberto; Ricceri, Fulvio; Barbui, Anna Maria; Brossa, Silvia; Cipriani, Raffaella; Cricenti, Luca; Cabras, Marco; Aimetti, Mario

2017-07-01

Presence of epithelial desquamation, erythema, and erosions on gingival tissue is usually described in the literature as desquamative gingivitis (DG). A wide range of autoimmune/dermatologic disorders can manifest as DG, although the two more common are oral lichen planus and mucous membrane pemphigoid. The aim of this study is to investigate prevalence of 11 periodontopathogenic microorganisms in patients with DG and to compare it with the microbiologic status of individuals affected by plaque-induced gingivitis (pGI). Cross-sectional clinical and microbiologic data were obtained from 66 patients (33 in each group). Subgingival plaque samples were analyzed using semiquantitative polymerase chain reaction analysis. Statistically significant difference, but with little clinical significance, was observed in gingival conditions between the two groups, probably due to the worse home control hygiene of patients with DG. Prevalence and levels of Aggregatibacter actinomycetemcomitans, Eikenella corrodens, and Fusobacterium nucleatum/periodonticum were statistically higher in samples from patients with DG than in those with pGI. In multivariate regression models, subgingival colonization of A. actinomycetemcomitans and F. nucleatum/periodonticum was not statistically associated with DG, whereas, high levels of E. corrodens were associated with 13-fold increased odds for DG. Microbiologic differences were found in subgingival plaque for patients with DG and pGI. This may suggest possible association between periodontal pathogens and DG.

Evaluation of Subgingival Dental Plaque Microbiota Changes In Fixed Orthodontic Patients with Syber Green Real Time PCR

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Naser Sargolzaie

2014-06-01

Full Text Available Introduction: One of the most common problems we confront in orthodontic therapy is periodontal diseases. Initial factor which causes these diseases is colonization of anaerobic microorganisms in subgingival plaque. Technically, local environmental changes related to orthodontic band and brackets may influence the bacterial species in periodontal plaque. However, it seems necessary to assess variations in subgingival plaque caused by orthodontic appliances. The aim of this study was to investigate changes in subgingival microbiota and clinical parameters before and after bracket placement. Methods: Clinical parameters including probing depth (PD, plaque index (PI, clinical attachment level (CAL, bleeding on probing (BOP and gingival index (GI were recorded and subgingival microbial samples were collected in 30 people aged between 13 and 25. As a control group, 15 persons getting matched as to their age and sex with no need to orthodontic treatment were opted using specific primers, SYBER Green Real-Time PCR was carried out in order to determine bacterial flora in stored samples. All mentioned procedures were reassessed in experimental group and in control group three months after band and bracket bonding. A descriptive analysis was conducted, and paired t test and Wilcoxon test were used for differences between groups (P

Sampling natural biofilms: a new route to build efficient microbial anodes.

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Erable, Benjamin; Roncato, Marie-Anne; Achouak, Wafa; Bergel, Alain

2009-05-01

Electrochemically active biofilms were constructed on graphite anodes under constant polarization at -0.1V vs saturated calomel reference (SCE) with 10 mM acetate as substrate. The reactors were inoculated with three different microbial samples that were drawn from exactly the same place in a French Atlantic coastal port (i) by scraping the biofilm that had formed naturally on the surface of a floating bridge, (ii) by taking marine sediments just under the floating bridge, and (iii) by taking nearby beach sand. Current densities of 2.0 A/m2 were reached using the biofilm sample as inoculum while only 0.4 A/m2 and 0.8 A/m2 were obtained using the underlying sediments and the beach sand, respectively. The structure of bacterial communities forming biofilms was characterized by denaturing gradient gel electrophoresis (DGGE) analysis, and revealed differences between samples with the increase in relative intensities of some bands and the appearance of others. Bacteria close related to Bacteroidetes, Halomonas, and Marinobacterium were retrieved only from the efficient EA-biofilms formed from natural biofilms, whereas, bacteria close related to Mesoflavibacter were predominant on biofilm formed from sediments. The marine biofilm was selected as the inoculum to further optimize the microbial anode. Epifluorescence microscopy and SEM confirmed that maintaining the electrode under constant polarization promoted rapid settlement of the electrode surface by a bacterial monolayer film. The microbial anode was progressively adapted to the consumption of acetate by three serial additions of substrate, thus improving the Coulombic efficiency of acetate consumption from 31 to 89%. The possible oxidation of sulfide played only a very small part in the current production and the biofilm was not able to oxidize hydrogen. Graphite proved to be more efficient than dimensionally stable anode (DSA) or stainless steel butthis result might be due to differences in the surface roughness

Membrane biofouling characterization: effects of sample preparation procedures on biofilm structure and the microbial community

KAUST Repository

Xue, Zheng

2014-07-15

Ensuring the quality and reproducibility of results from biofilm structure and microbial community analysis is essential to membrane biofouling studies. This study evaluated the impacts of three sample preparation factors (ie number of buffer rinses, storage time at 4°C, and DNA extraction method) on the downstream analysis of nitrifying biofilms grown on ultrafiltration membranes. Both rinse and storage affected biofilm structure, as suggested by their strong correlation with total biovolume, biofilm thickness, roughness and the spatial distribution of EPS. Significant variations in DNA yields and microbial community diversity were also observed among samples treated by different rinses, storage and DNA extraction methods. For the tested biofilms, two rinses, no storage and DNA extraction with both mechanical and chemical cell lysis from attached biofilm were the optimal sample preparation procedures for obtaining accurate information about biofilm structure, EPS distribution and the microbial community. © 2014 © 2014 Taylor & Francis.

Antimicrobial action of minocycline microspheres versus 810-nm diode laser on human dental plaque microcosm biofilms.

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Song, Xiaoqing; Yaskell, Tina; Klepac-Ceraj, Vanja; Lynch, Michael C; Soukos, Nikolaos S

2014-02-01

The purpose of this study is to investigate the antimicrobial effects of minocycline hydrochloride microspheres versus infrared light at 810 nm from a diode laser on multispecies oral biofilms in vitro. These biofilms were grown from dental plaque inoculum (oral microcosms) and were obtained from six systemically healthy individuals with generalized chronic periodontitis. Multispecies biofilms were derived using supra- and subgingival plaque samples from mesio-buccal aspects of premolars and molars exhibiting probing depths in the 4- to 5-mm range and 1- to 2-mm attachment loss. Biofilms were developed anaerobically on blood agar surfaces in 96-well plates using a growth medium of prereduced, anaerobically sterilized brain-heart infusion with 2% horse serum. Minocycline HCl 1 mg microspheres were applied on biofilms on days 2 and 5 of their development. Biofilms were also exposed on days 2 and 5 of their growth to 810-nm light for 30 seconds using a power of 0.8 W in a continuous-wave mode. The susceptibility of microorganisms to minocycline or infrared light was evaluated by a colony-forming assay and DNA probe analysis at different time points. At all time points of survival assessment, minocycline was more effective (>2 log10 colony-forming unit reduction) than light treatment (P plaque pathogens to light, and it was not possible after treatment with minocycline due to lack of bacterial growth. The cumulative action of minocycline microspheres on multispecies oral biofilms in vitro led to enhanced killing of microorganisms, whereas a single exposure of light at 810 nm exhibited minimal and non-selective antimicrobial effects.

Association between periodontal condition and subgingival microbiota in women during pregnancy: a longitudinal study

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Priscila Viola BORGO

2014-12-01

Full Text Available Objectivo In this study, the gingival conditions and the quantitative detection for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in pregnant women were determined. Material and Methods Quantitative determinations of periodontal bacteria by using a SyBr green system in women during pregnancy were performed. Women at the 2nd and 3rd trimesters of pregnancy and non-pregnant women were included in this study. A. actinomycetemcomitans was observed in high numbers in women at the 2nd and 3rd trimesters of pregnancy with a significant difference (p<0.05. F. nucleatum and P. intermedia were also observed in high levels. Results and Conclusion Our results show that pregnant women are more susceptible to gingivitis, and the presence of A. actinomycetemcomitans in subgingival biofilm might be taken into account for the treatment of periodontal disease.

Synergy in biofilm formation between Fusobacterium nucleatum and Prevotella species.

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Okuda, Tamaki; Kokubu, Eitoyo; Kawana, Tomoko; Saito, Atsushi; Okuda, Katsuji; Ishihara, Kazuyuki

2012-02-01

The formation of biofilm by anaerobic, Gram-negative bacteria in the subgingival crevice plays an important role in the development of chronic periodontitis. The aim of this study was to characterize the role of coaggregation between Fusobacterium nucleatum and Prevotella species in biofilm formation. Coaggregation between F. nucleatum and Prevotella species was determined by visual assay. Effect of co-culture of the species on biofilm formation was assessed by crystal violet staining. Effect of soluble factor on biofilm formation was also examined using culture supernatant and two-compartment co-culture separated by a porous membrane. Production of autoinducer-2 (AI-2) by the organisms was evaluated using Vibrio harveyi BB170. Cells of all F. nucleatum strains coaggregated with Prevotella intermedia or Prevotella nigrescens with a score of 1-4. Addition of ethylenediamine tetraacetic acid or l-lysine inhibited coaggregation. Coaggregation disappeared after heating of P. intermedia or P. nigrescens cells, or Proteinase K treatment of P. nigrescens cells. Co-culture of F. nucleatum ATCC 25586 with P. intermedia or P. nigrescens strains increased biofilm formation compared with single culture (p culture with culture supernatant of these strains, however, did not enhance biofilm formation by F. nucleatum. Production of AI-2 in Prevotella species was not related to enhancement of biofilm formation by F. nucleatum. These findings indicate that physical contact by coaggregation of F. nucleatum strains with P. intermedia or P. nigrescens plays a key role in the formation of biofilm by these strains. Copyright © 2011 Elsevier Ltd. All rights reserved.

Does pregnancy have an impact on the subgingival microbiota?

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Adriaens, Laurence M; Alessandri, Regina; Spörri, Stefan; Lang, Niklaus P; Persson, G Rutger

2009-01-01

We investigated clinical and subgingival microbiologic changes during pregnancy in 20 consecutive pregnant women > or =18 years not receiving dental care. Bacterial samples from weeks 12, 28, and 36 of pregnancy and at 4 to 6 weeks postpartum were processed for 37 species by checkerboard DNA-DNA hybridization. Clinical periodontal data were collected at week 12 and at 4 to 6 weeks postpartum, and bleeding on probing (BOP) was recorded at sites sampled at the four time points. The mean BOP at week 12 and postpartum was 40.1% +/- 18.2% and 27.4% +/- 12.5%, respectively. The corresponding mean BOP at microbiologic test sites was 15% (week 12) and 21% (postpartum; not statistically significant). Total bacterial counts decreased between week 12 and postpartum (P Streptococcus anginosus, Streptococcus intermedius, Streptococcus mutans, Streptococcus oralis, Streptococcus sanguinis, Selenomonas noxia, and Veillonella parvula. No changes occurred between weeks 12 and 28 of pregnancy. Counts of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, Tannerella forsythia (previously T. forsythensis), and Treponema denticola did not change. Counts of P. gingivalis and T. forsythia at week 12 were associated with gingivitis (P <0.001). Subgingival levels of bacteria associated with periodontitis did not change. P. gingivalis and T. forsythia counts were associated with BOP at week 12. A decrease was found in 17 of 37 species from week 12 to postpartum. Only counts of N. mucosa increased.

Supragingival treatment as an aid to reduce subgingival needs: a 450-day investigation

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Sabrina Carvalho Gomes

2014-01-01

Full Text Available This study investigated the clinical effects of using a supragingival biofilm control regimen (SUPRA as a step prior to scaling and root planing (SRP. A split-mouth clinical trial was performed in which 25 subjects with periodontitis (47.2 ± 6.5 years underwent treatment (days 0-60 and monitoring (days 90-450 phases. At Day 0 (baseline treatments were randomly assigned per quadrant: SUPRA, SRP and S30SRP (SUPRA 30 days before SRP. The full-mouth visible plaque index (VPI, gingival bleeding index (GBI, periodontal probing depth (PPD, bleeding on probing (BOP, and clinical attachment loss (CAL were examined on days 0, 30, 60, 90, 120, 270, and 450. Baseline data were similar among all groups. From days 0 to 60, the groups showed similar significant decreases in VPI and GBI. Reductions in PPD for the SRP (3.39 ± 0.17 to 2.42 ± 0.16 mm and S30SRP (3.31 ± 0.11 to 2.40 ± 0.07 mm groups were greater (p < 0.05 than those for the SUPRA group. This pattern was also observed for BOP. Attachment gain was similar and greater for the SRP (3.34 ± 0.28 to 2.58 ± 0.26 mm and S30SRP (3.25 ± 0.21 to 2.54 ± 0.19 mm groups compared to the SUPRA group. Results were maintained from day 90 forward. Overall, the S30SRP treatment reduced the subgingival treatment needs in 48.16%. Performance of a SUPRA step before SRP decreased subgingival treatment needs and maintained the periodontal stability over time.

Comparison of RNA extraction methods from biofilm samples of Staphylococcus epidermidis

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França Angela

2011-12-01

Full Text Available Abstract Background Microbial biofilms are communities of bacteria adhered to a surface and surrounded by an extracellular polymeric matrix. Biofilms have been associated with increased antibiotic resistance and tolerance to the immune system. Staphylococcus epidermidis is the major bacterial species found in biofilm-related infections on indwelling medical devices. Obtaining high quality mRNA from biofilms is crucial to validate the transcriptional measurements associated with the switching to the biofilm mode of growth. Therefore, we selected three commercially available RNA extraction kits with distinct characteristics, including those using silica membrane or organic extraction methods, and enzymatic or mechanical cell lysis, and evaluated the RNA quality obtained from two distinct S. epidermidis bacterial biofilms. Results RNA extracted using the different kits was evaluated for quantity, purity, integrity, and functionally. All kits were able to extract intact and functional total RNA from the biofilms generated from each S. epidermidis strain. The results demonstrated that the kit based on mechanical lysis and organic extraction (FastRNA® Pro Blue was the only one that was able to isolate pure and large quantities of RNA. Normalized expression of the icaA virulence gene showed that RNA extracted with PureLink™ had a significant lower concentration of icaA mRNA transcripts than the other kits tested. Conclusions When working with complex samples, such as biofilms, that contain a high content extracellular polysaccharide and proteins, special care should be taken when selecting the appropriate RNA extraction system, in order to obtain accurate, reproducible, and biologically significant results. Among the RNA extraction kits tested, FastRNA® Pro Blue was the best option for both S. epidermidis biofilms used.

Subgingival dysbiosis in smoker and non‑smoker patients with chronic periodontitis.

Science.gov (United States)

Coretti, Lorena; Cuomo, Mariella; Florio, Ermanno; Palumbo, Domenico; Keller, Simona; Pero, Raffaela; Chiariotti, Lorenzo; Lembo, Francesca; Cafiero, Carlo

2017-04-01

Periodontitis is one of the most common oral inflammatory diseases, and results in connective tissue degradation and gradual tooth loss. It manifests with formation of periodontal pockets, in which anaerobic and Gram‑negative bacteria proliferate rapidly. Consequently, alteration of the subgingival microbiota is considered the primary etiologic agent of periodontitis. Previous studies have reported that smokers are at increased risk of periodontal disease, in both prevalence and severity, indicating that smoking is a risk factor for the onset and progression of the pathology. In the present study, 16S rRNA sequencing was employed to assess the subgingival microbiota in 6 smoker patients with chronic periodontitis, 6 non‑smoker patients with chronic periodontitis and 8 healthy controls. The results demonstrated significant alterations in the microbial structure of periodontitis patients. High relative abundance of Parvimonans, Desulfubulbus, Paludibacter, Haemophilus, and Sphaerochaeta genera characterized subgingival microbiota of periodontitis patients, both smokers and non‑smokers. Due to the high precision and sensitivity of the 16S rRNA sequencing method, analysis for low‑abundant genera (including Pedobacter, Granulicatella, Paracoccus, Atopobium, Bifidobacterium, Coprococcus, Oridobacteriu, Peptococcus, Oscillospira and Akkermansia) was feasible, and revealed novel phylotypes associated with periodontitis. Of note, a major microbial community alteration was evident in smoker patients, suggesting an association between smoking and severity of subgingival dysbiosis. The present study confirmed that chronic periodontitis is a polymicrobial disease where changes in the equilibrium of subgingival microbiota contribute to severity of pathology.

Subgingival microflora and treatment in prepubertal periodontitis associated with chronic idiopathic neutropenia.

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Kamma, J J; Lygidakis, N A; Nakou, M

1998-09-01

Prepubertal periodontitis affects both primary and permanent dentition. The purpose of this study was to examine the composition of subgingival microflora of the permanent dentition in an 11-year-old Caucasian female, who had premature exfoliation of her deciduous teeth on her 5th year of age, and the response of this condition to the antibiotic therapy and supportive periodontal care. Gingival tissues were highly inflamed and alveolar bone loss was detected radiographically. The girl had experienced frequent upper respiratory tract infections, tonsilitis and recurrent otitis media. Her mother had history of early onset periodontitis associated with chronic idiopathic neutropenia. Blood chemistry tests and immunological examinations were also performed. Subgingival plaque samples were collected from the proximal sites of permanent molars, incisors, canines and maxillary premolars. 27 different microbial species were isolated from the subgingival microflora. Among the predominant species were Porphyromonas gingivalis (17.6%-7.3%), Prevotella intermedia (12.4%-4.7%), Capnocytophaga sputigena (14.4%-10.4%), Capnocytophaga ochracea (13.2%-6.9%) and Actinobacillus actinomycetemcomitans (9.3%-5.5%). Periodontal treatment consisted of scaling, root planing in conjunction with antibiotic administration of Augmentin 312.5 mg and Flagyl 200 mg, each t.i.d. for 10 days. 3 weeks after the antibiotic therapy, bacterial samples were collected from the same sites. All the periodontal pathogens were recovered in lower levels and A.actinomycetemcomitans was almost eliminated in the 3-week period. The evaluation of clinical indices at 3, 6 and 12 months showed that periodontal treatment in conjunction with antibiotics was effective and rapidly followed by marked clinical improvement. The microbiological monitoring at 3, 6 and 12 months after antibiotic treatment and each time prior to supportive periodontal care, revealed that the periodontal pathogens fluctuated in low levels even

Subgingival microbial communities in Leukocyte Adhesion Deficiency and their relationship with local immunopathology.

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Moutsopoulos, Niki M; Chalmers, Natalia I; Barb, Jennifer J; Abusleme, Loreto; Greenwell-Wild, Teresa; Dutzan, Nicolas; Paster, Bruce J; Munson, Peter J; Fine, Daniel H; Uzel, Gulbu; Holland, Steven M

2015-03-01

Leukocyte Adhesion Deficiency I (LAD-I) is a primary immunodeficiency caused by single gene mutations in the CD18 subunit of β2 integrins which result in defective transmigration of neutrophils into the tissues. Affected patients suffer from recurrent life threatening infections and severe oral disease (periodontitis). Microbial communities in the local environment (subgingival plaque) are thought to be the triggers for inflammatory periodontitis, yet little is known regarding the microbial communities associated with LAD-I periodontitis. Here we present the first comprehensive characterization of the subgingival communities in LAD-I, using a 16S rRNA gene-based microarray, and investigate the relationship of this tooth adherent microbiome to the local immunopathology of periodontitis. We show that the LAD subgingival microbiome is distinct from that of health and Localized Aggressive Periodontitits. Select periodontitis-associated species in the LAD microbiome included Parvimonas micra, Porphyromonas endodontalis, Eubacterium brachy and Treponema species. Pseudomonas aeruginosa, a bacterium not typically found in subgingival plaque is detected in LAD-I. We suggest that microbial products from LAD-associated communities may have a role in stimulating the local inflammatory response. We demonstrate that bacterial LPS translocates into the lesions of LAD-periodontitis potentially triggering immunopathology. We also show in in vitro assays with human macrophages and in vivo in animal models that microbial products from LAD-associated subgingival plaque trigger IL-23-related immune responses, which have been shown to dominate in patient lesions. In conclusion, our current study characterizes the subgingival microbial communities in LAD-periodontitis and supports their role as triggers of disease pathogenesis.

Subgingival microbial communities in Leukocyte Adhesion Deficiency and their relationship with local immunopathology.

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Niki M Moutsopoulos

2015-03-01

Full Text Available Leukocyte Adhesion Deficiency I (LAD-I is a primary immunodeficiency caused by single gene mutations in the CD18 subunit of β2 integrins which result in defective transmigration of neutrophils into the tissues. Affected patients suffer from recurrent life threatening infections and severe oral disease (periodontitis. Microbial communities in the local environment (subgingival plaque are thought to be the triggers for inflammatory periodontitis, yet little is known regarding the microbial communities associated with LAD-I periodontitis. Here we present the first comprehensive characterization of the subgingival communities in LAD-I, using a 16S rRNA gene-based microarray, and investigate the relationship of this tooth adherent microbiome to the local immunopathology of periodontitis. We show that the LAD subgingival microbiome is distinct from that of health and Localized Aggressive Periodontitits. Select periodontitis-associated species in the LAD microbiome included Parvimonas micra, Porphyromonas endodontalis, Eubacterium brachy and Treponema species. Pseudomonas aeruginosa, a bacterium not typically found in subgingival plaque is detected in LAD-I. We suggest that microbial products from LAD-associated communities may have a role in stimulating the local inflammatory response. We demonstrate that bacterial LPS translocates into the lesions of LAD-periodontitis potentially triggering immunopathology. We also show in in vitro assays with human macrophages and in vivo in animal models that microbial products from LAD-associated subgingival plaque trigger IL-23-related immune responses, which have been shown to dominate in patient lesions. In conclusion, our current study characterizes the subgingival microbial communities in LAD-periodontitis and supports their role as triggers of disease pathogenesis.

Microbiological Effect of Essential Oils in Combination with Subgingival Ultrasonic Instrumentation and Mouth Rinsing in Chronic Periodontitis Patients

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Toshiya Morozumi

2013-01-01

Full Text Available Thirty chronic periodontitis patients were randomly assigned to 3 groups: control, saline, and essential oil-containing antiseptic (EO. Subgingival plaque was collected from a total of 90 pockets across all subjects. Subsequently, subgingival ultrasonic instrumentation (SUI was performed by using EO or saline as the irrigation agent. After continuous mouth rinsing at home with EO or saline for 7 days, subgingival plaques were sampled again. Periodontopathic bacteria were quantified using the modified Invader PLUS assay. The total bacterial count in shallow pockets (probing pocket depth (PPD = 4-5 mm was significantly reduced in both saline (P<0.05 and EO groups (P<0.01. The total bacterial count (P<0.05 and Porphyromonas gingivalis (P<0.01 and Tannerella forsythia (P<0.05 count in deep pockets (PPD ≥6 mm were significantly reduced only in the EO group. In comparisons of the change ratio relative to baseline value of total bacteria counts across categories, both the saline and EO groups for PPD 4-5 mm and the EO group for PPD 6 mm showed a significantly low ratio (P<0.05. The adjunctive use of EO may be effective in reducing subgingival bacterial counts in both shallow and deep pockets. This trial is registered with UMIN Clinical Trials Registry UMIN000007484.

Multispecies Biofilms and Host Responses: “Discriminating the Trees from the Forest”

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Peyyala, R.; Ebersole, J.L.

2014-01-01

Periodontal diseases reflect a tissue destructive process of the hard and soft tissues of the periodontium that are initiated by the accumulation of multispecies bacterial biofilms in the subgingival sulcus. This accumulation, in both quantity and quality of bacteria, results in a chronic immunoinflammatory response of the host to control this noxious challenge, leading to collateral damage of the tissues. As knowledge of the characteristics of the host-bacterial interactions in the oral cavity has expanded, new knowledge has become available on the complexity of the microbial challenge and the repertoire of host responses to this challenge. Recent results from the Human Microbiome Project continue to extend the array of taxa, genera, and species of bacteria that inhabit the multiple niches in the oral cavity; however, there is rather sparse information regarding variations in how host cells discriminate commensal from pathogenic species, as well as how the host response is affected by the 3-dimensional architecture and interbacterial interactions that occur in the oral biofilms. This review provides some insights into thes- processes by including existing literature on the biology of nonoral bacterial biofilms, and the more recent literature just beginning to document how the oral cavity responds to multispecies biofilms. PMID:23141757

Effect of non-surgical periodontal treatment on the subgingival microbiota of patients with chronic kidney disease

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Hilana Paula Carillo Artese

2012-08-01

Full Text Available This study investigated the effect of non-surgical periodontal therapy on the composition of subgingival microbiota of patients with chronic kidney disease (CKD. Sixteen CKD pre-dialysis individuals (CKD and 14 individuals without clinical evidence of kidney disease (C presenting chronic periodontitis were treated by scaling and root planing. Subgingival samples were collected from each patient and analyzed for their composition by checkerboard at baseline and 3 months post-therapy. Significant differences between groups at baseline were sought by the Mann-Whitney and χ² tests. Changes over time were examined by the Wilcoxon test. At baseline, the CKD group had significantly lower counts of E. faecalis compared to the C group (p < 0.05. After treatment, the levels of a greater number of species were reduced in the C group. Higher levels of A. israelii, C. rectus, F. periodonticum, P. micra, P. nigrescens, T. forsythia, N. mucosa, and S. anginosus (p < 0.05 were found in the CKD group compared to the C group. Also, non-responsive sites in CKD individuals harbored significantly higher levels of pathogenic species (T. forsythia, P. gingivalis, T. denticola, Fusobacterium spp., D. pneumosintes, E. faecalis and S. aureus; p < 0.05 than sites that responded to therapy, as well as non-responsive sites in the C group. The periodontitis-associated subgingival microbiota of CKD and systemically healthy individuals was similar in composition. However, high levels of pathogenic species persisted in the subgingival microbiota of patients with CKD after treatment.

The effectiveness of 0.5–0.7% tetracycline gel to reduced subgingival plaque bacteria

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Ernie Maduratna Setiawati

2008-09-01

Full Text Available Background: The tetracycline was an antimicrobial agent, that a broad spectrum. In addition to the antimicrobial effects, their efficacy was also anticollagenase and removal of the smear layer on the root surface. Purpose: The aim of the study was to evaluate effectiveness tetracycline gel 0.5–0.7% to reduction subgingival plaque bacteria. Method: A laboratory experimental study was conducted to investigate the effectiveness tetracycline gel 0.5–0.7%. Samples were divided into 5 groups with different concentration. The antimicrobial effect was performed using spectrophotometer. The statistical test was used One-Way ANOVA with significant difference 5% and subsequently Tukey-HSD test. Result: The study showed that tetracycline gel 0.5% has the highest antimicrobial has the highest antimicrobial effect. Conclusion: Tetracycline gel with 0.5% concentration is effective in inhibiting the growth of subgingival plaque bacteria.

Periodontal-disease-associated biofilm: A reservoir for pathogens of medical importance.

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Vieira Colombo, Ana Paula; Magalhães, Clarissa Bichara; Hartenbach, Fátima Aparecida Rocha Resende; Martins do Souto, Renata; Maciel da Silva-Boghossian, Carina

2016-05-01

The ecological diversity of the periodontal microenvironment may provide suitable conditions for the colonization of species not usually considered members of the oral microbiota. In this investigation, we aimed to determine the prevalence and levels of pathogenic species of medical relevance in the microbiota of individuals with distinct periodontal clinical status. Subgingival biofilm was obtained from patients with periodontal health (H, n = 81), gingivitis (G, n = 55), generalized aggressive (AgP, n = 36) or chronic periodontitis (CP, n = 98), and analyzed for 39 microbial taxa using a checkerboard DNA-DNA hybridization technique. Microbial differences among groups, as well as associations between clinical and microbiological parameters were sought by non-parametric and univariate correlation tests. Neisseria spp., Peptostreptococus anaerobius, Candida albicans, enterobacteria, Pseudomonas aeruginosa, Eubacterium saphenum, Clostridium difficile and Olsenella uli were detected in high mean prevalence and counts in the subgingival microbiota of the study population. Species that were more related to periodontal inflammation and tissue destruction at the patient and site levels included enterobacteria, C. albicans, Neisseria spp., P. aeruginosa, O. uli, Hafnia alvei, Serratia marcescens and Filifactor alocis (p < 0.05). In contrast, Fusobacterium necrophorum, Lactobacillus acidophilus, Staphylococcus aureus and Streptococcus pneumoniae were associated with periodontal health (p < 0.05). Pathogenic species of medical importance may be detected in high prevalence and levels in the periodontal microbiota. Regardless of their role in periodontal health or disease, the periodontal biofilm may be a source for dissemination and development of systemic infections by these pathogenic microorganisms. Copyright © 2015 Elsevier Ltd. All rights reserved.

Managing sub-gingival fracture by multi-disciplinary approach: Endodontics-forced orthodontic extrusion and prosthetic rehabilitation

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Rakesh Mittal

2013-01-01

Full Text Available Traumatized anterior teeth with sub-gingival crown fractures are a challenge to treat. The management of sub-gingival fractures includes exposing the cervical margin followed by appropriate coronal restoration. The treatment modalities, which involve exposing the cervical margin, are surgical crown lengthening and orthodontic extrusion. This paper reports a case of fractured maxillary anterior tooth at the sub-gingival level that was managed by forced orthodontic extrusion after endodontic treatment followed by esthetic rehabilitation, a much forgotten technique not utilized routinely yet conservative and cost-effective.

Characterization of specimens obtained by different sampling methods for evaluation of periodontal bacteria.

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Okada, Ayako; Sogabe, Kaoru; Takeuchi, Hiroaki; Okamoto, Masaaki; Nomura, Yoshiaki; Hanada, Nobuhiro

2017-12-27

Quantitative analysis of periodontal bacteria is considered useful for clinical diagnosis, evaluation and assessment of the risk of periodontal disease. The purpose of this study was to compare the effectiveness of sampling of saliva, supragingival and subgingival plaque for evaluation of periodontal bacteria. From each of 12 subjects, i) subgingival plaque was collected from the deepest pocket using a sterile paper point, ii) stimulated whole saliva was collected after chewing gum, and iii) supragingival plaque was collected using a tooth brush. These samples were sent to the medical examination laboratory for quantitative analysis of the counts of three periodontal bacterial species: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. The proportions of these bacteria in subgingival plaque were higher than those in saliva or supragingival plaque, but lower in subgingival plaque than in saliva or supragingival plaque. In several cases, periodontal bacteria were below the levels of detection in subgingival plaque. We concluded that samples taken from subgingival plaque may be more useful for evaluating the proportion of periodontal bacteria in deep pockets than is the case for other samples. Therefore, for evaluation of periodontal bacteria, clinicians should consider the characteristics of the specimens obtained using different sampling methods.

Subgingival Microbiome of Gingivitis in Chinese Undergraduates.

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Deng, Ke; Ouyang, Xiang Ying; Chu, Yi; Zhang, Qian

To analyse the microbiome composition of health and gingivitis in Chinese undergraduates with high-throughput sequencing. Sequencing of 16S rRNA gene amplicons was performed with the MiSeq system to compare subgingival bacterial communities from 54 subjects with gingivitis and 12 periodontally healthy controls. A total of 1,967,372 sequences representing 14 phyla, 104 genera, and 96 species were detected. Analysis of similarities (Anosim) test and Principal Component Analysis (PCA) showed significantly different community profiles between the health control and the subjects with gingivitis. Alpha-diversity metrics were significantly higher in the subgingival plaque of the subjects with gingivitis compared with that of the healthy control. Overall, the relative abundance of 35 genera and 46 species were significantly different between the two groups, among them 28 genera and 45 species showed higher relative abundance in the subjects with gingivitis, whereas seven genera and one species showed a higher relative abundance in the healthy control. The genera Porphyromonas, Treponema, and Tannerella showed higher relative abundance in the subjects with gingivitis, while the genera Capnocytophaga showed higher proportions in health controls. Porphyromonas gingivalis, Prevotella intermedia and Porphyromonas endodontalis had higher relative abundance in gingivitis. Among them, Porphyromonas gingivalis was most abundant. Our results revealed significantly different microbial community composition and structures of subgingival plaque between subjects with gingivitis and healthy controls. Subjects with gingivitis showed greater taxonomic diversity compared with periodontally healthy subjects. The proportion of Porphyromonas, especially Porphyromonas gingivalis, may be associated with gingivitis subjects aged between 18 and 21 years old in China. Adults with gingivitis in this age group may have a higher risk of developing periodontitis.

Surface area and volume determination of subgingival calculus using laser fluorescence.

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Shakibaie, Fardad; Walsh, Laurence J

2014-03-01

Visible red (655 nm) laser fluorescence (LF) devices are currently used for identifying deposits of subgingival calculus on the root surfaces of teeth during dental examination and treatment; however, it is not known how the fluorescence readings produced by commercially available LF systems correlate to the nature of the deposits. This laboratory study explored the correlation between LF digital readings and the surface area and volume of subgingival calculus deposits on teeth. A collection of 30 extracted human posterior teeth with various levels of subgingival deposits of calculus across 240 sites were used in a clinical simulation, with silicone impression material used to replicate periodontal soft tissues. The teeth were scored by two examiners by using three commercial LF systems (DIAGNOdent, DIAGNOdent Pen and KEY3). The silicone was removed, and the teeth were removed for photography at × 20 magnification under white or ultraviolet light. The surface area, thickness, and volume were calculated, and both linear least squares regression and nonlinear (Spearman's rank method) correlation coefficients were determined. Visible red LF digital readings showed better correlation to calculus volume than to surface area. Overall, the best performance was found for the KEY3 system (Spearman coefficient 0.59), compared to the Classic DIAGNOdent (0.56) and the DIAGNOdent Pen (0.49). These results indicate that while visible red LF systems vary somewhat in performance, their LF readings provide a useful estimation of the volume of subgingival calculus deposits present on teeth.

The association between subgingival periodontal pathogens and systemic inflammation.

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Winning, Lewis; Patterson, Christopher C; Cullen, Kathy M; Stevenson, Kathryn A; Lundy, Fionnuala T; Kee, Frank; Linden, Gerard J

2015-09-01

To investigate associations between periodontal disease pathogens and levels of systemic inflammation measured by C-reactive protein (CRP). A representative sample of dentate 60-70-year-old men in Northern Ireland had a comprehensive periodontal examination. Men taking statins were excluded. Subgingival plaque samples were analysed by quantitative real time PCR to identify the presence of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia. High-sensitivity CRP (mg/l) was measured from fasting blood samples. Multiple linear regression analysis was performed using log-transformed CRP concentration as the dependent variable, with the presence of each periodontal pathogen as predictor variables, with adjustment for various potential confounders. A total of 518 men (mean age 63.6 SD 3.0 years) were included in the analysis. Multiple regression analysis showed that body mass index (p C-reactive protein. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Tracking of Actinobacillus actinomycetemcomitans in subgingival plaque of aggressive periodontitis patients

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Supriya Kheur

2010-01-01

Full Text Available Background: Actinobacillus actinomycetemcomitans is thought to be one of the etiological agents in aggressive periodontitis as well as indicated in various systemic diseases. Objective: To evaluate the prevalence of A. actinomycetemcomitans in the subgingival plaque of aggressive periodontitis patients. Study Design: Initially, under the selective growth conditions, the isolates were picked from the plaques and their identification was confirmed by polymerase chain reaction using primers specific for A. actinomycetemcomitans subgingival plaque of 15 patients diagnosed clinically and on radiographic criteria as aggressive periodontitis was inoculated on the Tryptic Soy agar with Bacitracin and Vancomycin culture media for 3-5 days under microaerophilic conditions. The positive colonies were selected based on biochemical tests for further analysis using reported primers for A. actinomycetemcomitans. Results: The results showed that 66.67% of aggressive periodontitis patients and 6.67% of control group of normal patients showed evidence of presence of A. actinomycetemcomitans in the subgingival microflora. Conclusion: This is the first study of its kind in an Indian population whereby almost all aggressive periodontitis patients showed evidence of A. actinomycetemcomitans.

Evaluation of biofilm formation by bacterial strains isolated from milking equipment and milk samples from cows with mastitis

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Laura Gonçalves da Silva Chagas

2017-08-01

Full Text Available The presence of biofilm-forming bacteria from the mammary gland of dairy cows adhered to equipment in the milking environment represents one of the major causes of bacterial resistance during mastitis treatment. The aim of this study was to identify strains of Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli in milk samples from cows with mastitis, as well as in the expansion tank and milking set liners. We aimed to quantify the extracellular proteins and polysaccharides in the biofilm produced by each strain. A total of 294 samples were collected from a dairy farm in the municipality of Uberlândia, Minas Gerais. To identify the S. aureus, S. epidermidis and E. coli isolates responsible for biofilm production, we tested the phenotype using the Congo red agar (CRA and microplate adhesion tests. Protein quantification was performed with a Bicinchoninic Acid Protein Assay Kit (BCA kit, and polysaccharides were quantified by the phenol sulfuric acid method. We identified eight strains of S. aureus, one strain of S. epidermidis and 11 strains of E. coli responsible for biofilm production, all of which showed a higher concentration of polysaccharides than proteins in the matrix. Escherichia coli was considered the most prevalent bacterium among the samples, and S. aureus was determined to be the largest biofilm producer. The results of the CRA and microplate adhesion tests were similar in regard to identification of the biofilm-producing strains according to their phenotype and matrix composition. The classification of S. aureus strains as major biofilm producers is of great concern for producers, as such bacteria are considered one of the predominant contagious etiological agents that cause bovine mastitis. In addition, our observation that E. coli and S. epidermidis can produce biofilms highlights the need to reassess prophylactic measures to avoid the adhesion of biofilm-producing bacteria.

Biofilm Risks

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Wirtanen, Gun Linnea; Salo, Satu

2016-01-01

This chapter on biofilm risks deals with biofilm formation of pathogenic microbes, sampling and detection methods, biofilm removal, and prevention of biofilm formation. Several common pathogens produce sticky and/or slimy structures in which the cells are embedded, that is, biofilms, on various...... surfaces in food processing. Biofilms of common foodborne pathogens are reviewed. The issue of persistent and nonpersistent microbial contamination in food processing is also discussed. It has been shown that biofilms can be difficult to remove and can thus cause severe disinfection and cleaning problems...... in food factories. In the prevention of biofilm formation microbial control in process lines should both limit the number of microbes on surfaces and reduce microbial activity in the process. Thus the hygienic design of process equipment and process lines is important in improving the process hygiene...

Matrix-Gla Protein rs4236 [A/G] gene polymorphism and serum and GCF levels of MGP in patients with subgingival dental calculus.

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Doğan, Gülnihal Emrem; Demir, Turgut; Aksoy, Hülya; Sağlam, Ebru; Laloğlu, Esra; Yildirim, Abdulkadir

2016-10-01

Matrix-Gla Protein (MGP) is one of the major Gla-containing protein associated with calcification process. It also has a high affinity for Ca 2+ and hydroxyapatite. In this study we aimed to evaluate the MGP rs4236 [A/G] gene polymorphism in association with subgingival dental calculus. Also a possible relationship between MGP gene polymorphism and serum and GCF levels of MGP were examined. MGP rs4236 [A/G] gene polymorphism was investigated in 110 patients with or without subgingival dental calculus, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques. Additionally, serum and GCF levels of MGP of the patients were compared according to subgingival dental calculus. Comparison of patients with and without subgingival dental calculus showed no statistically significant difference in MGP rs4236 [A/G] gene polymorphism (p=0.368). MGP concentrations in GCF of patients with subgingival dental calculus were statistically higher than those without subgingival dental calculus (p=0.032). However, a significant association was not observed between the genotypes of AA, AG and GG of the MGP rs4236 gene and the serum and GCF concentrations of MGP in subjects. In this study, it was found that MGP rs4236 [A/G] gene polymorphism was not to be associated with subgingival dental calculus. Also, that GCF MGP levels were detected higher in patients with subgingival dental calculus than those without subgingival dental calculus independently of polymorphism, may be the effect of adaptive mechanism to inhibit calculus formation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Detection of odoriferous subgingival and tongue microbiota in diabetic and nondiabetic patients with oral malodor using polymerase chain reaction

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Dinesh R Kamaraj

2011-01-01

Full Text Available Background: Halitosis has been correlated with the concentration of volatile sulfur compounds (VSCs produced in the oral cavity by metabolic activity of bacteria colonizing the periodontal pockets and the dorsum of the tongue. It has been assumed that there is a relationship between periodontal disease and diabetes mellitus. Objectives: The aim of the study was to assess the malodor using the organoleptic method and tanita device; to quantify odoriferous microorganisms of subgingival plaque and tongue coating, such as P. gingivalis (Pg, T. forsythia (Tf, and F. nucleatum (Fn using polymerase chain reaction (PCR in nondiabetic and diabetic chronic periodontitis patients. Patients and Methods: Thirty chronic periodontitis patients (with and without diabetes with 5-7 mm pocket depth, radiographic evidence of bone loss, and presence of oral malodor participated in this study. Subjective assessment of mouth air was done organoleptically and by using a portable sulfide monitor. Tongue coating was also assessed. Results: The scores of plaque index, gingival index, gingival bleeding index, VSC levels, and tongue coating between the nondiabetic and diabetic patients were not significant (P>0.5. In nondiabetic patients, Fn was found to be significantly (P0.5. In diabetic patients, Fn and Tf have shown significant (P<0.5 an increase in subgingival and tongue samples, respectively, whereas Pg has not shown significant difference between subgingival and tongue samples. Interpretation and Conclusion: The results confirm that there is no difference in clinical parameters between nondiabetic and diabetic periodontitis patients, but the odoriferous microbial profiles in tongue samples of diabetic patients were found to be high. However, there is a weak positive correlation between VSC levels, clinical parameters, and odoriferous microbial profiles.

Limitations of Using Propidium Monoazide with qPCR to Discriminate between Live and Dead in Biofilm Samples

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Michael J. Taylor

2014-01-01

Full Text Available Accurately quantifying Legionella for regulatory purposes to protect public health is essential. Real-time PCR (qPCR has been proposed as a better method for detecting and enumerating Legionella in samples than conventional culture method. However, since qPCR amplifies any target DNA in the sample, the technique's inability to discriminate between live and dead cells means that counts are generally significantly overestimated. Propidium monoazide (PMA has been used successfully in qPCR to aid live/dead discrimination. We tested PMA use as a method to count only live Legionella cells in samples collected from a modified chemostat that generates environmentally comparable samples. Counts from PMA-treated samples that were pretreated with either heat or three types of disinfectants (to kill the cells were highly variable, with the only consistent trend being the relationship between biofilm mass and numbers of Legionella cells. Two possibilities explain this result: 1. PMA treatment worked and the subsequent muted response of Legionella to disinfection treatment is a factor of biofilm/microbiological effects; although this does not account for the relationship between the amount of biofilm sampled and the viable Legionella count as determined by PMA-qPCR; or 2. PMA treatment did not work, and any measured decrease or increase in detectable Legionella is because of other factors affecting the method. This is the most likely explanation for our results, suggesting that higher concentrations of PMA might be needed to compensate for the presence of other compounds in an environmental sample or that lower amounts of biofilm need to be sampled. As PMA becomes increasingly toxic at higher concentrations and is very expensive, augmenting the method to include higher PMA concentrations is both counterproductive and cost prohibitive. Conversely, if smaller volumes of biofilm are used, the reproducibility of the method is reduced. Our results suggest that

The microbial changes in subgingival plaques of orthodontic patients: a systematic review and meta-analysis of clinical trials.

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Guo, Runzhi; Lin, Yifan; Zheng, Yunfei; Li, Weiran

2017-06-02

Orthodontic treatment was found to have an impact on the quantity and constitution of subgingival microbiota. However, contradictory findings regarding the effects of fixed appliances on microbial changes were reported. The aim of this systematic review was to investigate the microbial changes in subgingival plaques of orthodontic patients. The PubMed, Cochrane Library, and EMBASE databases were searched up to November 20, 2016. Longitudinal studies observing microbial changes in subgingival plaques at different time points of orthodontic treatment are included. The methodological quality of the included studies was assessed by Methodological index for non-randomized studies (MINORS). The studies that reported the frequency of subgingival periodontopathogens were used for quantitative analysis. Other studies were analysed qualitatively to describe the microbial changes during orthodontic treatment. Thirteen studies were selected, including two controlled clinical trials, three cohort studies and eight self-controlled studies. Four periodontopathogens, including Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi) and Tannerella forsythia (Tf), were analysed. Following orthodontic appliance placement, the frequencies of Pg and Aa showed no significant change (P = 0.97 and P = 0.77), whereas the frequency of Tf significantly increased (P  = 6 months), two studies reported that the levels of subgingival periodontopathogens exhibited a transient increase but decreased to the pretreatment levels afterwards. After removal of the orthodontic appliance, the four periodontopathogens showed no significant difference compared with before removal. The levels of subgingival pathogens presented temporary increases after orthodontic appliance placement, and appeared to return to pretreatment levels several months later. This indicates that orthodontic treatment might not permanently induce periodontal disease by

Relative abundance of total subgingival plaque-specific bacteria in salivary microbiota reflects the overall periodontal condition in patients with periodontitis.

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Kageyama, Shinya; Takeshita, Toru; Asakawa, Mikari; Shibata, Yukie; Takeuchi, Kenji; Yamanaka, Wataru; Yamashita, Yoshihisa

2017-01-01

Increasing attention is being focused on evaluating the salivary microbiota as a promising method for monitoring oral health; however, its bacterial composition greatly differs from that of dental plaque microbiota, which is a dominant etiologic factor of oral diseases. This study evaluated the relative abundance of subgingival plaque-specific bacteria in the salivary microbiota and examined a relationship between the abundance and severity of periodontal condition in patients with periodontitis. Four samples (subgingival and supragingival plaques, saliva, and tongue coating) per each subject were collected from 14 patients with a broad range of severity of periodontitis before periodontal therapy. The bacterial composition was analyzed by 16S rRNA gene amplicon sequencing using Ion PGM. Of the 66 species-level operational taxonomic units (OTUs) representing the mean relative abundance of ≥ 1% in any of the four niches, 12 OTUs corresponding to known periodontal pathogens, including Porphyromonas gingivalis, were characteristically predominant in the subgingival plaque and constituted 37.3 ± 22.9% of the microbiota. The total relative abundance of these OTUs occupied only 1.6 ± 1.2% of the salivary microbiota, but significantly correlated with the percentage of diseased sites (periodontal pocket depth ≥ 4 mm; r = 0.78, P periodontal therapy, the total relative abundance of these 12 OTUs was evaluated as well as before periodontal therapy and reductions of the abundance through periodontal therapy were strongly correlated in saliva and subgingival plaque (r = 0.81, P bacteria representing the present condition of periodontal health.

Investigation of N-acyl homoserine lactone (AHL) molecule production in Gram-negative bacteria isolated from cooling tower water and biofilm samples.

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Haslan, Ezgi; Kimiran-Erdem, Ayten

2013-09-01

In this study, 99 Gram-negative rod bacteria were isolated from cooling tower water, and biofilm samples were examined for cell-to-cell signaling systems, N-acyl homoserine lactone (AHL) signal molecule types, and biofilm formation capacity. Four of 39 (10 %) strains isolated from water samples and 14 of 60 (23 %) strains isolated from biofilm samples were found to be producing a variety of AHL signal molecules. It was determined that the AHL signal molecule production ability and the biofilm formation capacity of sessile bacteria is higher than planktonic bacteria, and there was a statistically significant difference between the AHL signal molecule production of these two groups (p cooling tower water and biofilm samples produced different types of AHL signal molecules and that there were different types of AHL signal molecules in an AHL extract of bacteria. In the present study, it was observed that different isolates of the same strains did not produce the same AHLs or did not produce AHL molecules, and bacteria known as AHL producers did not produce AHL. These findings suggest that detection of signal molecules in bacteria isolated from cooling towers may contribute to prevention of biofilm formation, elimination of communication among bacteria in water systems, and blockage of quorum-sensing controlled virulence of these bacteria.

Evaluation of Two Surface Sampling Methods for Microbiological and Chemical Analyses To Assess the Presence of Biofilms in Food Companies.

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Maes, Sharon; Huu, Son Nguyen; Heyndrickx, Marc; Weyenberg, Stephanie van; Steenackers, Hans; Verplaetse, Alex; Vackier, Thijs; Sampers, Imca; Raes, Katleen; Reu, Koen De

2017-12-01

Biofilms are an important source of contamination in food companies, yet the composition of biofilms in practice is still mostly unknown. The chemical and microbiological characterization of surface samples taken after cleaning and disinfection is very important to distinguish free-living bacteria from the attached bacteria in biofilms. In this study, sampling methods that are potentially useful for both chemical and microbiological analyses of surface samples were evaluated. In the manufacturing facilities of eight Belgian food companies, surfaces were sampled after cleaning and disinfection using two sampling methods: the scraper-flocked swab method and the sponge stick method. Microbiological and chemical analyses were performed on these samples to evaluate the suitability of the sampling methods for the quantification of extracellular polymeric substance components and microorganisms originating from biofilms in these facilities. The scraper-flocked swab method was most suitable for chemical analyses of the samples because the material in these swabs did not interfere with determination of the chemical components. For microbiological enumerations, the sponge stick method was slightly but not significantly more effective than the scraper-flocked swab method. In all but one of the facilities, at least 20% of the sampled surfaces had more than 10 2 CFU/100 cm 2 . Proteins were found in 20% of the chemically analyzed surface samples, and carbohydrates and uronic acids were found in 15 and 8% of the samples, respectively. When chemical and microbiological results were combined, 17% of the sampled surfaces were contaminated with both microorganisms and at least one of the analyzed chemical components; thus, these surfaces were characterized as carrying biofilm. Overall, microbiological contamination in the food industry is highly variable by food sector and even within a facility at various sampling points and sampling times.

Microbiological characteristics of subgingival microbiota in adult periodontitis, localized juvenile periodontitis and rapidly progressive periodontitis subjects.

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Nonnenmacher, C; Mutters, R; de Jacoby, L F

2001-04-01

To describe the prevalence of the cultivable subgingival microbiota in periodontal diseases and to draw attention to the polymicrobial nature of periodontic infections. The study population consisted of 95 patients, 51 females and 44 males, aged 14-62 years. Twenty-nine patients exhibited adult periodontitis (AP), six localized juvenile periodontitis (LJP), and 60 rapidly progressive periodontitis (RPP). Two to four pooled bacterial samples were obtained from each patient. Samples were collected with sterile paper points from the deepest periodontal pockets. The samples were cultured under anaerobic and microaerophilic conditions using selective and non-selective media. Isolates were characterized to species level by conventional biochemical tests and by a commercial rapid test system. Prevotella intermedia and Capnocytophaga spp. were the most frequently detected microorganisms in all diagnostic groups. Porphyromonas gingivalis and Peptostreptococcus micros were found more frequently in AP and RPP patients, while Actinobacillus actinomycetemcomitans and Eikenella corrodens were associated with AP, LJP and RPP patients. The other bacterial species, including Actinomyces spp., Streptococcus spp. and Eubacterium spp., were detected at different levels in the three disease groups. The data show the complexity of the subgingival microbiota associated with different periodontal disease groups, indicating that the detection frequency and levels of recovery of some periodontal pathogens are different in teeth affected by different forms of periodontal disease.

Early supra- and subgingival plaque formation in experimental gingivitis in smokers and never-smokers.

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Branco, Paula; Weidlich, Patricia; Oppermann, Rui Vicente; Rösing, Cassiano Kuchenbecker

2015-01-01

To evaluate supragingival and subgingival plaque formation on the dentogingival area in smokers and never smokers using the experimental gingivitis model and a plaque scoring system that considers the presence of an area free of plaque between plaque and the gingival sulcus called the plaque free zone (PFZ). Male volunteers, 9 current smokers and 10 never-smokers, refrained from oral hygiene procedures in the maxillary incisors and canines (test teeth) for 25 days. Under conditions of clinically healthy gingiva (phase 1) and gingival inflammation (phase 2), the supragingival plaque formation pattern was observed for 4 days in the dentogingival area. Gingival crevicular fluid was also measured. Plaque was dyed with fucsine and its presence was recorded by a calibrated examiner based on a 3-criteria scoring system: 0 - absence of stained plaque; 1 - presence of stained plaque and supragingival PFZ; 2 - presence of stained plaque and absence of PFZ, indicating that subgingival plaque formation has taken place. In both phases, smokers presented a significantly lower relative frequency of sites with subgingival plaque compared to never-smokers (P smokers demonstrated a significantly lower frequency of gingival bleeding than did non-smokers (23.6% vs 66.1%; P Smokers presented significantly lower percentages of sites with subgingival plaque in all experimental periods and presented less gingival inflammation as shown by GBI and gingival crevicular fluid quantification.

Subgingival microbiome in smokers and non-smokers in periodontitis: an exploratory study using traditional targeted techniques and a next-generation sequencing

NARCIS (Netherlands)

Bizzarro, S.; Loos, B.G.; Laine, M.L.; Crielaard, W.; Zaura, E.

2013-01-01

Aim To compare the results of two targeted techniques to an open-ended technique in periodontitis patients, differentiated on the basis of smoking habit. Materials & Methods Thirty periodontitis patients (15 smokers and 15 non-smokers) provided subgingival plaque samples for 16S rRNA gene amplicon

Alterations of the Subgingival Microbiota in Pediatric Crohn's Disease Studied Longitudinally in Discovery and Validation Cohorts.

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Kelsen, Judith; Bittinger, Kyle; Pauly-Hubbard, Helen; Posivak, Leah; Grunberg, Stephanie; Baldassano, Robert; Lewis, James D; Wu, Gary D; Bushman, Frederic D

2015-12-01

Oral manifestations are common in Crohn's disease (CD). Here we characterized the subgingival microbiota in pediatric patients with CD initiating therapy and after 8 weeks to identify microbial community features associated with CD and therapy. Pediatric patients with CD were recruited from The Children's Hospital of Pennsylvania. Healthy control subjects were recruited from primary care or orthopedics clinic. Subgingival plaque samples were collected at initiation of therapy and after 8 weeks. Treatment exposures included 5-ASAs, immunomodulators, steroids, and infliximab. The microbiota was characterized by 16S rRNA gene sequencing. The study was repeated in separate discovery (35 CD, 43 healthy) and validation cohorts (43 CD, 31 healthy). Most subjects in both cohorts demonstrated clinical response after 8 weeks of therapy (discovery cohort 88%, validation cohort 79%). At week 0, both antibiotic exposure and disease state were associated with differences in bacterial community composition. Seventeen genera were identified in the discovery cohort as candidate biomarkers, of which 11 were confirmed in the validation cohort. Capnocytophaga, Rothia, and TM7 were more abundant in CD relative to healthy controls. Other bacteria were reduced in abundance with antibiotic exposure among CD subjects. CD-associated genera were not enriched compared with healthy controls after 8 weeks of therapy. Subgingival microbial community structure differed with CD and antibiotic use. Results in the discovery cohort were replicated in a separate validation cohort. Several potentially pathogenic bacterial lineages were associated with CD but were not diminished in abundance by antibiotic treatment, suggesting targets for additional surveillance.

Subgingival bacteria in Ghanaian adolescents with or without progression of attachment loss

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Gunnar Dahlén

2014-05-01

Full Text Available Objective: This study describes subgingival bacterial profiles associated with clinical periodontal status in Ghanaian adolescents with or without progression of attachment loss. Materials and methods: Among 500 adolescents included in a cohort study, 397 returned 2 years later for a periodontal re-examination, including full-mouth CAL measurements. At follow-up, a subgroup of 98 adolescents was also subjected to bacterial sampling with paper points at four periodontal sites (mesial aspect of 11, 26, 31, and 46 and analyzed with the checkerboard DNA–DNA hybridization technique against DNA-probes from nine periodontitis-associated bacterial species. Results: The 98 Ghanaian adolescents examined in the present study were similar to the entire group examined at the 2-year follow-up with respect to age, gender, and CAL ≥3 mm. A high detection frequency of Fusobacterium nucleatum and Prevotella intermedia (>99% using checkerboard analysis was found, while for Aggregatibacter actinomycetemcomitans the detection frequency was <50%. A strong correlation was found at the individual level between the presence of P. intermedia and the total CAL change, and P. intermedia and Porphyromonas gingivalis were strongly correlated with a change in CAL and probing pocket depth (PPD at the sampled sites. In a linear regression model, a significant discriminating factor for the total CAL change in the dentition during the 2-year follow-up period was obtained for P. intermedia and public school. Conclusion: This study indicates that subgingival bacterial species other than A. actinomycetemcomitans, for example, P. intermedia, have a significant association with periodontal breakdown (change in CAL in Ghanaian adolescents with progression of periodontal attachment loss.

Quantitative real-time PCR combined with propidium monoazide for the selective quantification of viable periodontal pathogens in an in vitro subgingival biofilm model.

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Sánchez, M C; Marín, M J; Figuero, E; Llama-Palacios, A; León, R; Blanc, V; Herrera, D; Sanz, M

2014-02-01

Differentiation of live and dead cells is an important challenge when using molecular diagnosis for microbial identification. This is particularly relevant when bacteria have been exposed to antimicrobial agents. The objective of this study was to test a method using quantitative real-time polymerase chain reaction (qPCR) combined with propidium monoazide (PMA), developed for the selective quantification of viable P. gingivalis, A. actinomycetemcomitans, F. nucleatum and total bacteria in an in vitro biofilm model after antimicrobial treatment. PMA-qPCR method was tested in an in vitro biofilm model, using isopropyl alcohol as the antimicrobial agent. Matured biofilms were exposed for 1, 5, 10 and 30 min to isopropyl alcohol by immersion. Biofilms were disrupted and PMA added (final concentration of 100 μm). After DNA isolation, qPCR was carried out using specific primers and probes for the target bacteria. The differentiation of live and dead cells was tested by analysis of variance. When PMA was used in the presence of viable target bacterial cells, no statistically significant inhibition of qPCR amplification was detected (p > 0.05 in all cases). Conversely, after immersion in isopropyl alcohol of the biofilm, PMA resulted in a significant total reduction of qPCR amplification of about 4 log10 . P. gingivalis showed a vitality reduction in the biofilm of 3 log10 , while A. actinomycetemcomitans and F. nucleatum showed a 2 log10 reduction. These results demonstrate the efficiency of PMA for differentiating viable and dead P. gingivalis, A. actinomycetemcomitans and F. nucleatum cells, as well as total bacteria, in an in vitro biofilm model, after being exposed to an antimicrobial agent. Hence, this PMA-qPCR method may be useful for studying the effect of antimicrobial agents aimed at oral biofilms. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Investigation of in vitro Mineral Forming Bacterial Isolates from Subgingival Calculus

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Turgut Demir

2014-06-01

This is the first report to identify and show that bacteria from subgingival calculus under anaerobic conditions are involved in the formation of dental calculus. [Arch Clin Exp Surg 2014; 3(3.000: 153-160

Oral, intestinal, and skin bacteria in ventral hernia mesh implants

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Odd Langbach

2016-07-01

Full Text Available Background: In ventral hernia surgery, mesh implants are used to reduce recurrence. Infection after mesh implantation can be a problem and rates around 6–10% have been reported. Bacterial colonization of mesh implants in patients without clinical signs of infection has not been thoroughly investigated. Molecular techniques have proven effective in demonstrating bacterial diversity in various environments and are able to identify bacteria on a gene-specific level. Objective: The purpose of this study was to detect bacterial biofilm in mesh implants, analyze its bacterial diversity, and look for possible resemblance with bacterial biofilm from the periodontal pocket. Methods: Thirty patients referred to our hospital for recurrence after former ventral hernia mesh repair, were examined for periodontitis in advance of new surgical hernia repair. Oral examination included periapical radiographs, periodontal probing, and subgingival plaque collection. A piece of mesh (1×1 cm from the abdominal wall was harvested during the new surgical hernia repair and analyzed for bacteria by PCR and 16S rRNA gene sequencing. From patients with positive PCR mesh samples, subgingival plaque samples were analyzed with the same techniques. Results: A great variety of taxa were detected in 20 (66.7% mesh samples, including typical oral commensals and periodontopathogens, enterics, and skin bacteria. Mesh and periodontal bacteria were further analyzed for similarity in 16S rRNA gene sequences. In 17 sequences, the level of resemblance between mesh and subgingival bacterial colonization was 98–100% suggesting, but not proving, a transfer of oral bacteria to the mesh. Conclusion: The results show great bacterial diversity on mesh implants from the anterior abdominal wall including oral commensals and periodontopathogens. Mesh can be reached by bacteria in several ways including hematogenous spread from an oral site. However, other sites such as gut and skin may also

Membrane biofouling characterization: effects of sample preparation procedures on biofilm structure and the microbial community

KAUST Repository

Xue, Zheng; Lu, Huijie; Liu, Wen-Tso

2014-01-01

Ensuring the quality and reproducibility of results from biofilm structure and microbial community analysis is essential to membrane biofouling studies. This study evaluated the impacts of three sample preparation factors (ie number of buffer rinses

In vitro and clinical evaluation of optical coherence tomography for the detection of subgingival calculus and root cementum.

Science.gov (United States)

Tsubokawa, Masaki; Aoki, Akira; Kakizaki, Sho; Taniguchi, Yoichi; Ejiri, Kenichiro; Mizutani, Koji; Koshy, Geena; Akizuki, Tatsuya; Oda, Shigeru; Sumi, Yasunori; Izumi, Yuichi

2018-05-24

This study evaluated the effectiveness of swept-source optical coherence tomography (ss-OCT) for detecting calculus and root cementum during periodontal therapy. Optical coherence tomography (OCT) images were taken before and after removal of subgingival calculus from extracted teeth and compared with non-decalcified histological sections. Porcine gingival sheets of various thicknesses were applied to the root surfaces of extracted teeth with calculus and OCT images were taken. OCT images were also taken before and after scaling and root planing (SRP) in human patients. In vitro, calculus was clearly detected as a white-gray amorphous structure on the root surface, which disappeared after removal. Cementum was identified as a thin, dark-gray layer. The calculus could not be clearly observed when soft tissues were present on the root surface. Clinically, supragingival calculus and cementum could be detected clearly with OCT, and subgingival calculus in the buccal cervical area of the anterior and premolar teeth was identified, which disappeared after SRP. Digital processing of the original OCT images was useful for clarifying the calculus. In conclusion, ss-OCT showed potential as a periodontal diagnostic tool for detecting cementum and subgingival calculus, although the practical applications of subgingival imaging remain limited.

Biofilm-forming activity of bacteria isolated from toilet bowl biofilms and the bactericidal activity of disinfectants against the isolates.

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Mori, Miho; Gomi, Mitsuhiro; Matsumune, Norihiko; Niizeki, Kazuma; Sakagami, Yoshikazu

2013-01-01

To evaluate the sanitary conditions of toilets, the bacterial counts of the toilet bowl biofilms in 5 Kansai area and 11 Kansai and Kanto area homes in Japan were measured in winter and summer seasons, respectively. Isolates (128 strains) were identified by analyzing 16S ribosomal RNA sequences. The number of colonies and bacterial species from biofilms sampled in winter tended to be higher and lower, respectively, than those in summer. Moreover, the composition of bacterial communities in summer and winter samples differed considerably. In summer samples, biofilms in Kansai and Kanto areas were dominated by Blastomonas sp. and Mycobacterium sp., respectively. Methylobacterium sp. was detected in all toilet bowl biofilms except for one sample. Methylobacterium sp. constituted the major presence in biofilms along with Brevundimonas sp., Sphingomonas sp., and/or Pseudomonas sp. The composition ratio of the sum of their genera was 88.0 from 42.9% of the total bacterial flora. The biofilm formation abilities of 128 isolates were investigated, and results suggested that Methylobacterium sp. and Sphingomonas sp. were involved in biofilm formation in toilet bowls. The biofilm formation of a mixed bacteria system that included bacteria with the highest biofilm-forming ability in a winter sample was greater than mixture without such bacteria. This result suggests that isolates possessing a high biofilm-forming activity are involved in the biofilm formation in the actual toilet bowl. A bactericidal test against 25 strains indicated that the bactericidal activities of didecyldimethylammonium chloride (DDAC) tended to be higher than those of polyhexamethylene biguanide (PHMB) and N-benzyl-N,N-dimethyldodecylammonium chloride (ADBAC). In particular, DDAC showed high bactericidal activity against approximately 90% of tested strains under the 5 h treatment.

Effect of sample storage time on detection of hybridization signals in Checkerboard DNA-DNA hybridization.

Science.gov (United States)

do Nascimento, Cássio; Muller, Katia; Sato, Sandra; Albuquerque Junior, Rubens Ferreira

2012-04-01

Long-term sample storage can affect the intensity of the hybridization signals provided by molecular diagnostic methods that use chemiluminescent detection. The aim of this study was to evaluate the effect of different storage times on the hybridization signals of 13 bacterial species detected by the Checkerboard DNA-DNA hybridization method using whole-genomic DNA probes. Ninety-six subgingival biofilm samples were collected from 36 healthy subjects, and the intensity of hybridization signals was evaluated at 4 different time periods: (1) immediately after collecting (n = 24) and (2) after storage at -20 °C for 6 months (n = 24), (3) for 12 months (n = 24), and (4) for 24 months (n = 24). The intensity of hybridization signals obtained from groups 1 and 2 were significantly higher than in the other groups (p  0.05). The Checkerboard DNA-DNA hybridization method was suitable to detect hybridization signals from all groups evaluated, and the intensity of signals decreased significantly after long periods of sample storage.

Efficacy of NVC-422 against Staphylococcus aureus biofilms in a sheep biofilm model of sinusitis.

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Singhal, Deepti; Jekle, Andreas; Debabov, Dmitri; Wang, Lu; Khosrovi, Bez; Anderson, Mark; Foreman, Andrew; Wormald, Peter-John

2012-01-01

Bacterial biofilms are a major obstacle in management of recalcitrant chronic rhinosinusitis. NVC-422 is a potent, fast-acting, broad-spectrum, nonantibiotic, antimicrobial with a new mechanism of action effective against biofilm bacteria in in vitro conditions. The aim of this study was to investigate the safety and efficacy of NVC-422 as local antibiofilm treatment in a sheep model of rhinosinusitis. After accessing and occluding frontal sinus ostia in 24 merino sheep via staged endoscopic procedures, S. aureus clinical isolate was instilled in frontal sinuses. Following biofilm formation, ostial obstruction was removed and sinuses irrigated with 0.1% and 0.5% NVC-422 in 5 mM acetate isotonic saline at pH 4.0. Sheep were monitored for adverse effects and euthanized 24 hours after treatment. Frontal sinuses were assessed for infection and changes in mucosa after the treatment. S. aureus biofilms were identified with Baclight-confocal scanning microscopy protocol and the biofilm biomass assayed by applying the COMSTAT2 program to recorded image stacks. After 2 irrigations with 0.1% NVC-422, S. aureus biofilm biomass was reduced when compared to control sinuses (p = 0.0001), though this effect was variable in samples. NVC-422 0.5% solution irrigations reduced biofilm even more significantly and consistently over all samples (p biofilm biomass (p biofilms, with dose-dependent efficacy in this animal model of biofilm-associated sinusitis. Copyright © 2012 American Rhinologic Society-American Academy of Otolaryngic Allergy, LLC.

New antimicrobial therapies used against fungi present in subgingival sites--a brief review.

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Sardi, Janaina Cássia Orlandi; Almeida, Ana Marisa Fusco; Mendes Giannini, Maria José Soares

2011-10-01

Although the main reservoir of Candida spp. is believed to be the buccal mucosa, these microorganisms can coaggregate with bacteria in subgingival biofilm and adhere to epithelial cells. The treatment of periodontal disease includes scaling and root planning (SRP) associated with proper oral hygiene. However, some patients may have negative responses to different therapeutic procedures, with a continuous loss of insertion, so the use of antimicrobials is needed as an adjuvant to SRP treatment. The use of a broad-spectrum antibiotic, such as tetracycline and metronidazole, as an aid in periodontal treatment has also been a factor for the development of superinfections by resistant bacteria and Candida species, even in patients with HIV. In the dental practice, the most commonly used antifungals are nystatin and fluconazole. However, the introduction of new drugs like the next generation of azoles is essential before the onset of emergent species in periodontal disease. Plants are good options for obtaining a wide variety of drugs. This alternative could benefit a large population that uses plants as a first treatment option. Plants have been used in medicine for a long time and are extensively used in folk medicine, because they represent an economic alternative, are easily accessible and are applicable to various diseases. Herein, we briefly review the literature pertaining the presence of Candida sp. in periodontal pockets, the conventional antifungal resistance and new therapies that include natural antifungal agents are reviewed. Copyright © 2011 Elsevier Ltd. All rights reserved.

Linking community tolerance and structure with low metallic contamination: a field study on 13 biofilms sampled across the Seine river basin.

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Fechner, Lise C; Gourlay-Francé, Catherine; Tusseau-Vuillemin, Marie-Hélène

2014-03-15

It is difficult to assess the biological consequences of diffuse water contamination by micropollutants which are present in rivers at low, even sublethal levels. River biofilms, which respond quickly to changes of environmental parameters, are good candidates to acquire knowledge on the response of aquatic organisms to diffuse chemical contamination in the field. The study was designed as an attempt to link biofilm metal tolerance and metallic contamination in a field survey covering 13 different sampling sites in the Seine river basin (north of France) with low contamination levels. Cd and Zn tolerance of heterotrophic communities was assessed using a short-term toxicity test based on β-glucosidase activity. Metal tolerance levels varied between sites but there was no obvious correlation between tolerance and corresponding water contamination levels for Cd and Zn. Indeed, metallic contamination at the sampling sites remained subtle when compared to water quality standards (only two sampling sites had either Zn or both Cu and Zn concentrations exceeding the Environmental Quality Standards set by the EU Water Framework Directive). Yet, multivariate analysis of the data using Partial Least Squares Regression revealed that both metallic and environmental parameters were important variables explaining the variability of metal tolerance levels. Automated Ribosomal Intergenic Spacer Analysis (ARISA) was also performed on both bacterial and eukaryotic biofilm communities from the 13 sampling sites. Multivariate analysis of ARISA fingerprints revealed that biofilms with similar tolerance levels have similar ARISA profiles. Those results confirm that river biofilms are potential indicators of low, diffuse contamination levels of aquatic systems. Copyright © 2013 Elsevier Ltd. All rights reserved.

Microbiological effects and recolonization patterns after adjunctive subgingival debridement with Er:YAG laser.

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Sanz-Sánchez, Ignacio; Ortiz-Vigón, Alberto; Herrera, David; Sanz, Mariano

2016-07-01

The objective of this study was to assess the microbiological effects and recolonization patterns after non-surgical periodontal treatment protocol based on the adjunctive use of erbium-doped yttrium aluminium garnet (Er:YAG) laser. Patients diagnosed with chronic periodontitis were randomly assigned to two different treatment protocols: test, full-mouth subgingival ultrasonic instrumentation followed by Er-YAG laser application 1 week later to sites with initial probing pocket depth ≥4.5 mm; and control, full-mouth ultrasonic subgingival instrumentation within 1 week. Clinical (at sampled sites) and microbiological (culture-based) parameters were recorded at baseline and 3 and 12 months. Microbiological variables included total counts, frequency of detection, proportions and counts of target species. Results from 19 test and 21 control patients were compared. Minor changes were observed for total colony-forming units, with no differences between groups. For the frequency of detection, a limited and similar impact in both groups was observed for the most prevalent (over 80 %) periodontal pathogens (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum). For proportions, reductions in P. gingivalis occurred at 3 months, both in the test and control groups (from 16.3 to 10 % and 16 to 14.8 %, respectively), although these differences were not statistically significant. At 12 months, the test group showed a statistically significant greater reduction in probing depth for the sampled sites. The adjunctive use of Er:YAG laser when compared with conventional ultrasonic debridement did not provide a microbiological added benefit. Even though some clinical benefits with the adjunctive laser application were identified when comparing both treatment protocols, there were no differences in microbiological outcomes or in the bacterial recolonization patterns.

Inhibitory Effects of Lactoferrin on Growth and Biofilm Formation of Porphyromonas gingivalis and Prevotella intermedia▿

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Wakabayashi, Hiroyuki; Yamauchi, Koji; Kobayashi, Tetsuo; Yaeshima, Tomoko; Iwatsuki, Keiji; Yoshie, Hiromasa

2009-01-01

Lactoferrin (LF) is an iron-binding antimicrobial protein present in saliva and gingival crevicular fluids, and it is possibly associated with host defense against oral pathogens, including periodontopathic bacteria. In the present study, we evaluated the in vitro effects of LF-related agents on the growth and biofilm formation of two periodontopathic bacteria, Porphyromonas gingivalis and Prevotella intermedia, which reside as biofilms in the subgingival plaque. The planktonic growth of P. gingivalis and P. intermedia was suppressed for up to 5 h by incubation with ≥130 μg/ml of human LF (hLF), iron-free and iron-saturated bovine LF (apo-bLF and holo-bLF, respectively), and ≥6 μg/ml of bLF-derived antimicrobial peptide lactoferricin B (LFcin B); but those effects were weak after 8 h. The biofilm formation of P. gingivalis and P. intermedia over 24 h was effectively inhibited by lower concentrations (≥8 μg/ml) of various iron-bound forms (the apo, native, and holo forms) of bLF and hLF but not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia was also reduced by incubation with various iron-bound bLFs, hLF, and LFcin B for 5 h. In an examination of the effectiveness of native bLF when it was used in combination with four antibiotics, it was found that treatment with ciprofloxacin, clarithromycin, and minocycline in combination with native bLF for 24 h reduced the amount of a preformed biofilm of P. gingivalis compared with the level of reduction achieved with each agent alone. These results demonstrate the antibiofilm activity of LF with lower iron dependency against P. gingivalis and P. intermedia and the potential usefulness of LF for the prevention and treatment of periodontal diseases and as adjunct therapy for periodontal diseases. PMID:19451301

[Evaluation of the clinical effect of the teeth with subgingivally involved defect conserved by crown lengthening surgery].

Science.gov (United States)

Wang, Yong; Ding, Xiao-Hong; Yao, Li-Li; Huang, Zhong-Suo; Bian, Hua-Qin

2005-02-01

To evaluate the clinical effect of the teeth with subgingivally involved defect which were conserved by crown lengthening surgery. 62 teeth, with defect subgingivally from 1.5 mm to 4 mm, mobility degree(MD)lengthening surgery by combining flap surgery and osteoectomy, and restored 4 weeks after operation and followed-up for one year. The parameters of MD, sulcus bleeding index (SBI) and maximal defect probing depth (PD) at different times were measured respectively. 46 anterior teeth were divided into two groups based on PD of pre-operation. The groups were as follows: minor defect group (0.05), but a significant increase about MD occurred in the major defect group one year after restoration (Pstage after operation and PD of pre-operation in anterior teeth (r=0.489, 0.526, 0.531, Plengthening surgery may conserve these teeth with subgingivally involved defect, and has a good, long-time clinical effect. But MD showed an increasing trend after operation and significant cor.

Subgingival microbiota in health compared to periodontitis and the influence of smoking.

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Camelo-Castillo, Anny J; Mira, Alex; Pico, Alex; Nibali, Luigi; Henderson, Brian; Donos, Nikolaos; Tomás, Inmaculada

2015-01-01

The etiology of periodontitis has traditionally been associated to a consortium of three bacterial species-the so-called "red-complex" of periodontal disease-which has been the target for most diagnostic and therapeutic strategies. However, other species have also been found to correlate with disease severity. In addition, the influence of smoking on periodontal microbiota is poorly understood. In the current manuscript, the composition of the subgingival microbiota in healthy individuals vs. patients with chronic periodontitis has been investigated using 16S pyrosequencing and the influence of smoking on periodontal composition has been examined. Subgingival bacterial communities were sampled from 82 patients: 22 non-smoking healthy controls, 28 non-smoking periodontal patients, and 32 smoking periodontal patients. Bacterial diversity was higher in periodontal patients than in healthy subjects, which could be interpreted as the consequence of a nutritionally richer environment or a reduced immune competence. Periodontal patients showed a significantly higher prevalence/relative abundance of "established" periopathogens but also other taxa whose role is not well-established and that should be targets for future research. These include Anaeroglobus, Bulleidia, Desulfobulbus, Filifactor, Mogibacterium, Phocaeicola, Schwartzia or TM7. The microbial community of smoking-associated periodontitis is less diverse and distinct from that of non-smokers, indicating that smoking has an influence on periodontal ecology. Interestingly, the high sequencing coverage allowed the detection at low proportions of periodontal pathogens in all healthy individuals, indicating that chronic periodontitis cannot be strictly considered an infectious disease but the outcome of a polymicrobial dysbiosis, where changes in the proportions of microbial consortia trigger the inflammatory and tissue-degradation responses of the host.

Subgingival microbiota in health compared to periodontitis and the influence of smoking

Directory of Open Access Journals (Sweden)

A eCamelo-Castillo

2015-02-01

Full Text Available The aetiology of periodontitis has traditionally been associated to a consortium of three bacterial species -the so-called red-complex of periodontal disease- which has been the target for most diagnostic and therapeutic strategies. However, other species have also been found to correlate with disease severity. In addition, the influence of smoking on periodontal microbiota is poorly understood. In the current manuscript, the composition of the subgingival microbiota in healthy individuals versus patients with chronic periodontitis has been investigated using 16S pyrosequencing and the influence of smoking on periodontal composition has been examined. Subgingival bacterial communities were sampled from 82 patients: 22 non-smoking healthy controls, 28 non-smoking periodontal patients and 32 smoking periodontal patients.Bacterial diversity was higher in periodontal patients than in healthy subjects, which could be interpreted as the consequence of a nutritionally richer environment or a reduced immune competence. Periodontal patients showed a significantly higher prevalence/abundance of established periopathogens but also other taxa whose role is not well-established and that should be targets for future research. These include Anaeroglobus, Bulleidia, Desulfobulbus, Filifactor, Mogibacterium, Phocaeicola, Schwartzia or TM7. The microbial community of smoking-associated periodontitis is less diverse and distinct from that of non-smokers, indicating that smoking has an influence on periodontal ecology. Interestingly, the high sequencing coverage allowed the detection at low proportions of periodontal pathogens in all healthy individuals, indicating that chronic periodontitis cannot be strictly considered an infectious disease but the outcome of a polymicrobial dysbiosis, where changes in the proportions of microbial consortia trigger the inflammatory and tissue-degradation responses of the host.

Altered oral viral ecology in association with periodontal disease.

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Ly, Melissa; Abeles, Shira R; Boehm, Tobias K; Robles-Sikisaka, Refugio; Naidu, Mayuri; Santiago-Rodriguez, Tasha; Pride, David T

2014-05-20

The human oral cavity is home to a large and diverse community of viruses that have yet to be characterized in patients with periodontal disease. We recruited and sampled saliva and oral biofilm from a cohort of humans either periodontally healthy or with mild or significant periodontal disease to discern whether there are differences in viral communities that reflect their oral health status. We found communities of viruses inhabiting saliva and the subgingival and supragingival biofilms of each subject that were composed largely of bacteriophage. While there were homologous viruses common to different subjects and biogeographic sites, for most of the subjects, virome compositions were significantly associated with the oral sites from which they were derived. The largest distinctions between virome compositions were found when comparing the subgingival and supragingival biofilms to those of planktonic saliva. Differences in virome composition were significantly associated with oral health status for both subgingival and supragingival biofilm viruses but not for salivary viruses. Among the differences identified in virome compositions was a significant expansion of myoviruses in subgingival biofilm, suggesting that periodontal disease favors lytic phage. We also characterized the bacterial communities in each subject at each biogeographic site by using the V3 hypervariable segment of the 16S rRNA and did not identify distinctions between oral health and disease similar to those found in viral communities. The significantly altered ecology of viruses of oral biofilm in subjects with periodontal disease compared to that of relatively periodontally healthy ones suggests that viruses may serve as useful indicators of oral health status. Little is known about the role or the constituents of viruses as members of the human microbiome. We investigated the composition of human oral viral communities in a group of relatively periodontally healthy subjects or significant

A novel approach to the use of subgingival controlled-release chlorhexidine delivery in chronic periodontitis: a randomized clinical trial.

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Gonzales, Jose R; Harnack, Lutz; Schmitt-Corsitto, Gabriella; Boedeker, Rolf H; Chakraborty, Trinad; Domann, Eugen; Meyle, Joerg

2011-08-01

We aimed to analyze clinical, microbiologic, and serologic effects of chlorhexidine (CHX) chips used as a subgingival controlled-release delivery device before and immediately after scaling and root planing (SRP). Twenty-four patients presenting with ≥12 teeth with probing depth (PD) ≥5 mm and bleeding on probing were assigned in test or control groups. After prophylaxis, CHX chips (test) or placebo chips (control) were placed in pockets with PD ≥5 mm. Ten days later, SRP was performed in all teeth with PD ≥4 mm in a single appointment. Immediately after SRP, new chips were inserted in all pockets with PD ≥5 mm. Parameters were assessed at baseline; beginning of SRP; and 1, 3, and 6 months after treatment. Subgingival samples were obtained at baseline; beginning of SRP; and at 1 month after treatment. Periodontal pathogens Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Treponema denticola were analyzed. Serum levels of high sensitive C-reactive and lipopolysaccharide-binding proteins were measured. The changes of the parameters between and within the groups were tested by Mann-Whitney U test (P periodontitis.

Differential reflectometry versus tactile sense detection of subgingival calculus in dentistry

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Shakibaie, Fardad; Walsh, Laurence J.

2012-10-01

Detecting dental calculus is clinically challenging in dentistry. This study used typodonts with extracted premolar and molar teeth and simulated gingival tissue to compare the performance of differential reflectometry and periodontal probing. A total of 30 extracted teeth were set in an anatomical configuration in stone to create three typodonts. Clear polyvinyl siloxane impression material was placed to replicate the periodontal soft tissues. Pocket depths ranged from 10 to 15 mm. The three models were placed in a phantom head, and an experienced dentist assessed the presence of subgingival calculus first using the DetecTar (differential reflectometry) and then a periodontal probe. Scores from these two different methods were compared to the gold standard (direct examination of the root surface using 20× magnification) to determine the accuracy and reproducibility. Differential reflectometry was more accurate than tactile assessment (79% versus 60%), and its reproducibility was also higher (Cohen kappa 0.54 versus 0.39). Both methods performed better on single rooted premolar teeth than on multirooted teeth. These laboratory results indicate that differential reflectometry allows more accurate and reproducible detection of subgingival calculus than conventional probing, and supports its use for supplementing traditional periodontal examination methods in dental practice.

AFM Structural Characterization of Drinking Water Biofilm ...

Science.gov (United States)

Due to the complexity of mixed culture drinking water biofilm, direct visual observation under in situ conditions has been challenging. In this study, atomic force microscopy (AFM) revealed the three dimensional morphology and arrangement of drinking water relevant biofilm in air and aqueous solution. Operating parameters were optimized to improve imaging of structural details for a mature biofilm in liquid. By using a soft cantilever (0.03 N/m) and slow scan rate (0.5 Hz), biofilm and individual bacterial cell’s structural topography were resolved and continuously imaged in liquid without loss of spatial resolution or sample damage. The developed methodology will allow future in situ investigations to temporally monitor mixed culture drinking water biofilm structural changes during disinfection treatments. Due to the complexity of mixed culture drinking water biofilm, direct visual observation under in situ conditions has been challenging. In this study, atomic force microscopy (AFM) revealed the three dimensional morphology and arrangement of drinking water relevant biofilm in air and aqueous solution. Operating parameters were optimized to improve imaging of structural details for a mature biofilm in liquid. By using a soft cantilever (0.03 N/m) and slow scan rate (0.5 Hz), biofilm and individual bacterial cell’s structural topography were resolved and continuously imaged in liquid without loss of spatial resolution or sample damage. The developed methodo

The investigation of Helicobacter pylori in the dental biofilm and saliva samples of children with dyspeptic complaints.

Science.gov (United States)

Aksit Bıcak, Damla; Akyuz, Serap; Kıratlı, Binnur; Usta, Merve; Urganci, Nafiye; Alev, Burcin; Yarat, Aysen; Sahin, Fikrettin

2017-03-21

The oral cavity can be an extra-gastric reservoir for Helicobacter pylori (H.pylori). This can play a role in the pathogenesis of halitosis, glossitis, recurrent aphthous stomatitis, and dental caries. The present study was conducted to detect the presence of H.pylori within the dental biofilm and in saliva samples collected from children suffering from dyspepsia and children without any gastrointestinal complaints. Associations with gastric infection, halitosis, and some oral parameters were also evaluated. Seventy children (aged between 5-16) with dyspepsia were selected for the study group and control group composed of 30 healthy children without dyspepsia were also included in the study. After detailed oral and clinical examinations for oral parameters, saliva, and supragingival dental biofilm samples were collected for 16S rRNA and 23S rRNA genes detection by real-time polymerase chain reaction (RT-PCR). The presence of gastric H.pylori was evaluated in endoscopic biopsy specimens histopathologically. Halitosis was evaluated by benzoyl-DL-arginine-naphthylamid (BANA) test. Salivary S.mutans and Lactobacilli sp. counts were also carried out by commercial kits. H.pylori was histopathologically detected amongst 83% of the children with the dyspeptic condition. The detection rate of this bacteria in dental biofilm and saliva samples and halitosis were found relatively higher in the dyspeptic children rather than the control group (p pylori (p > 0.05). In the gastric H.pylori positive group with dyspepsia, DMFT/S and dmft/s numbers and plaque indices were found higher than the control group (p pylori negative group with dyspepsia were found higher than the control group (p pylori positive and negative groups (p > 0.05). Comparing to those with negative for both genes, in children whose dental biofilm and saliva samples were positive for both 16S rRNA and 23S rRNA genes, significantly higher results for halitosis, and DMFS numbers and significantly

Ultraviolet-Absorption Spectroscopic Biofilm Monitor

Science.gov (United States)

Micheels, Ronald H.

2004-01-01

An ultraviolet-absorption spectrometer system has been developed as a prototype instrument to be used in continuous, real-time monitoring to detect the growth of biofilms. Such monitoring is desirable because biofilms are often harmful. For example, biofilms in potable-water and hydroponic systems act as both sources of pathogenic bacteria that resist biocides and as a mechanism for deterioration (including corrosion) of pipes. Biofilms formed from several types of hazardous bacteria can thrive in both plant-growth solutions and low-nutrient media like distilled water. Biofilms can also form in condensate tanks in air-conditioning systems and in industrial heat exchangers. At present, bacteria in potable-water and plant-growth systems aboard the space shuttle (and previously on the Mir space station) are monitored by culture-plate counting, which entails an incubation period of 24 to 48 hours for each sample. At present, there are no commercially available instruments for continuous monitoring of biofilms in terrestrial or spaceborne settings.

Biofilms as bio-indicator for polluted waters? Total reflection X-ray fluorescence analysis of biofilms of the Tisza river (Hungary)

Energy Technology Data Exchange (ETDEWEB)

Mages, Margarete; Ovari, Mihaly; Tuempling, Wolf v. [Department of Inland Water Research Magdeburg, UFZ Centre for Environmental Research Leipzig-Halle, Brueckstrasse 3a, 39114, Magdeburg (Germany); Kroepfl, Krisztina [Department of Chemical Technology and Environmental Chemistry, Eoetvoes University, Pazmany Peter setany 1/A, 1117, Budapest (Hungary)

2004-02-01

The aim of this work was to investigate the heavy metal accumulation by natural biofilms living in the catchment area of the Tisza river in Hungary, as well as in biofilms cultivated in vitro. Laboratory tests have demonstrated that metals can be adsorbed on biofilms, depending on their concentration and on the availability of free sorptive places. Biofilms were cultivated in vitro in natural freshwater from the Saale river, Germany. After reaching the plateau phase, Cu was added to reach a concentration of 100 {mu}g/L. An increase of its mass fraction in the biofilm was observed, which caused the decrease of the concentration in the water phase. Unfortunately, the reactor wall was also found to act as adsorbent for Cu. More detailed results of our in vitro experiments will be published in a forthcoming paper. Naturally grown biofilm samples from exposed as well as background places at the Hungarian rivers Szamos and Tisza were collected in 2000 and 2002 after the cyanide spill, and analysed using total reflection X-ray fluorescence analysis (TXRF). Metal mass fraction differences as high as two orders of magnitude were found between polluted and unpolluted (background) sampling points. Extremely high concentration values, e.g. 5600 {mu}g/g Zn in biofilm, were found at highly polluted sampling points. This means an enrichment factor of ca. 10,000 compared to the water phase. (orig.)

Confocal Raman microscopy for identification of bacterial species in biofilms

Science.gov (United States)

Beier, Brooke D.; Quivey, Robert G.; Berger, Andrew J.

2011-03-01

Implemented through a confocal microscope, Raman spectroscopy has been used to distinguish between biofilm samples of two common oral bacteria species, Streptococcus sanguinis and mutans, which are associated with healthy and cariogenic plaque, respectively. Biofilms of these species are studied as a model of dental plaque. A prediction model has been calibrated and validated using pure biofilms. This model has been used to identify the species of transferred and dehydrated samples (much like a plaque scraping) as well as hydrated biofilms in situ. Preliminary results of confocal Raman mapping of species in an intact two-species biofilm will be shown.

Identification of pink-pigmented bacteria isolated from environmental water samples and their biofilm formation abilities.

Science.gov (United States)

Furuhata, Katsunori; Kato, Yuko; Goto, Keiichi; Saitou, Keiko; Sugiyama, Jun-Ichi; Hara, Motonobu; Fukuyama, Masahumi

2008-06-01

Sixty-seven strains of pink-pigmented bacteria, which were isolated from environmental water samples collected nationwide, were identified by partial 16S rDNA sequence analysis. In addition, the biofilm formation ability of the isolates was experimentally investigated. We could identify only 2 strains at the species level: Pedobacter roseus HS-38 and Runella slithyformis HS-77. The results showed that of the strains tested, 22 strains (32.8%) were Pedobacter spp., which was most frequently identified, followed by 19 strains (28.4%) of Arcicella spp., 16 strains (23.9%) of Deinococcus spp., 5 strains (7.5%) of Roseomonas spp., 4 strains (6.0%) of Flectobacillus spp. and 1 strain (1.5%) of Runella sp. Most isolates showed low similarity values to previously known species, and they were found to be novel species. At a result, it was difficult to identify environmental water-derived pink-pigmented bacteria at the species level. On the other hand, when we measured the absorbance by the crystal violet staining to examine the quantities of biofilm formation of these strains, fifty-five (82.0%) of the 67 isolates formed biofilm. The absorbance of Deinococcus sp. HS-75 was the highest (3.56). When comparing the absorbance values among the genera, Roseomonas spp. showed the highest absorbance (mean:1.62), followed by Deinococcus spp. (mean: 1.03), and Arcicella spp. (mean: 1.01). Strains of Flectobacillus spp. (mean: 0.48) and Pedobacter spp. (mean: 0.42) showed lower absorbance values. As above, it was shown that, at the species level, the pink-pigmented bacteria in the water in the Japanese environment had various levels of ability to form biofilm.

Wound biofilms: lessons learned from oral biofilms

OpenAIRE

Mancl, Kimberly A.; Kirsner, Robert S.; Ajdic, Dragana

2013-01-01

Biofilms play an important role in the development and pathogenesis of many chronic infections. Oral biofilms, more commonly known as dental plaque,are a primary cause of oral diseases including caries, gingivitis and periodontitis. Oral biofilms are commonly studied as model biofilm systems as they are easily accessible, thus biofilm research in oral diseases is advanced with details of biofilm formation and bacterial interactions being well-elucidated. In contrast, wound research has relati...

Selective near-UV ablation of subgingival dental calculus: measurement of removal rates

Science.gov (United States)

Schoenly, J. E.; Seka, W.; Rechmann, P.

2010-02-01

A noncontact profilometer (laser triangulation) was used to measure the removal rates of subgingival dental calculus irradiated with a frequency-doubled Ti:sapphire laser (60-ns pulse duration, 400-nm wavelength, 10-Hz repetition rate, 7-mJ pulse energy). Profilometer traces before and after irradiation were used to create a removal map with 4-μm axial and 15-μm transverse resolution. Twenty-three teeth (15 with calculus and 8 pristine) were irradiated at 90° and 45° under a cooling water spray with a super-Gaussian beam (~300-μm diameter). Subgingival calculus was selectively removed at 5.6 and 4.0 J/cm2 for 90° and 45°, respecetively, within a range of rates, between 2 to 9 μm/pulse. These ablation rates were constant during these exposures. For comparison, pristine cementum irradiated for 10 min at the same peak fluence and pulse repetition rate showed only craters, 15 to 50 μm deep, corresponding to an equivalent removal rate three orders of magnitude smaller than that obtained for calculus. Pristine enamel was not removed under the same irradiation conditions.

Biofilm formation in a hot water system

DEFF Research Database (Denmark)

Bagh, L.K.; Albrechtsen, Hans-Jørgen; Arvin, Erik

2002-01-01

The biofilm formation rate was measured in situ in a hot water system in an apartment building by specially designed sampling equipment, and the net growth of the suspended bacteria was measured by incubation of water samples with the indigeneous bacteria. The biofilm formation rate reached......, in the sludge, or in the water from the distribution system was negligible. This indicated that bacterial growth took place on the inner surfaces in the hot water system and biofilm formation and detachment of bacteria could account for most of the suspended bacteria actually measured in hot water. Therefore...

Neutrophil extracellular trap formation in supragingival biofilms.

Science.gov (United States)

Hirschfeld, Josefine; Dommisch, Henrik; Skora, Philipp; Horvath, Gabor; Latz, Eicke; Hoerauf, Achim; Waller, Tobias; Kawai, Toshihisa; Jepsen, Søren; Deschner, James; Bekeredjian-Ding, Isabelle

2015-01-01

Oral biofilms are the causative agents of the highly prevalent oral diseases periodontitis and caries. Additionally, the host immune response is thought to play a critical role in disease onset. Neutrophils are known to be a key host response factor to bacterial challenge on host surfaces. Release of neutrophil extracellular traps (NETs) as a novel antimicrobial defense strategy has gained increasing attention in the past years. Here, we investigated the influx of neutrophils into the dental plaque and the ability of oral bacteria to trigger intra-biofilm release of NETs and intracellular proteins. Supragingival biofilms and whole saliva were sampled from systemically healthy subjects participating in an experimental gingivitis study. Biofilms were analysed by immunofluorescence followed by confocal and fluorescence microscopy. Moreover, concentrations of cytokines and immune-associated proteins in biofilm suspensions and saliva were assessed by ELISA. Neutrophils obtained from blood were stimulated with twelve bacterial species isolated from cultured biofilms or with lipopolysaccharide to monitor NET formation. Neutrophils, NETs, neutrophil-associated proteins (myeloperoxidase, elastase-2, cathepsin G, cathelicidin LL-37), interleukin-8, interleukin-1β and tumor necrosis factor were detected within plaque samples and saliva. All tested bacterial species as well as the polymicrobial samples isolated from the plaque of each donor induced release of NETs and interleukin-8. The degree of NET formation varied among different subjects and did not correlate with plaque scores or clinical signs of local inflammation. Our findings indicate that neutrophils are attracted towards dental biofilms, in which they become incorporated and where they are stimulated by microbes to release NETs and immunostimulatory proteins. Thus, neutrophils and NETs may be involved in host biofilm control, although their specific role needs to be further elucidated. Moreover, inter

Biofilms.

Science.gov (United States)

López, Daniel; Vlamakis, Hera; Kolter, Roberto

2010-07-01

The ability to form biofilms is a universal attribute of bacteria. Biofilms are multicellular communities held together by a self-produced extracellular matrix. The mechanisms that different bacteria employ to form biofilms vary, frequently depending on environmental conditions and specific strain attributes. In this review, we emphasize four well-studied model systems to give an overview of how several organisms form biofilms: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. Using these bacteria as examples, we discuss the key features of biofilms as well as mechanisms by which extracellular signals trigger biofilm formation.

Diagnosis and understanding of chronic biofilm infections

DEFF Research Database (Denmark)

Thomsen, Trine Rolighed

2016-01-01

Title: Diagnosis and understanding of chronic biofilm infections. Name: Trine Rolighed Thomsen Aalborg University and Danish Technological Institute, Denmark Recent evidence suggests that the microbial community, its spatial distribution and activity play an important role in the prolongation......, anaerobic or unculturable bacteria living in biofilms. Thus, diagnosis of chronic infections is challenged by lack of appropriate sampling strategies and by limitations in microbiological testing methods. The purpose of this study was to improve sampling and diagnosis of chronic infections, especially...... considering the biofilm issue. Systematic and optimized sampling of various specimen types was performed. Extended culture, optimized DNA extraction, quantitative PCR, cloning, next generation sequencing and PNA FISH were applied on different types of specimens for optimized diagnosis. For further...

Occurrence of periodontal pathogens in ethnic groups from a native Brazilian reservation.

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Gaetti-Jardim, Elerson; Pereira, Maurício Fabiano; Vieira, Evanice Menezes Marçal; Schweitzer, Christiane Marie; Okamoto, Ana Cláudia; Ávila-Campos, Mario J

2015-06-01

The present study was designed to evaluate the occurrence of periodontal pathogens in the subgingival biofilm of 100 native Brazilians living at the Umutina Indian Reservation, Mato Grosso State, Brazil. Periodontal clinical examinations were carried out prior to collection of subgingival biofilm, and the presence of 14 periodontal microorganisms was evaluated by polymerase chain reaction (PCR). The prevalence and risk analysis was performed using Cochran and Mantel-Haenszel statistics for dichotomous variables or Pearson's chi-squared test for analysis of proportions when variables had three or more categories. The interrelations between clinical and microbiological parameters were assessed using Fisher's exact test and the Mann-Whitney U test. Individuals with chronic periodontitis were frequently colonized by the association between Porphyromonas gingivalis and Campylobacter rectus, P. gingivalis and Prevotella intermedia, or P. gingivalis and Tannerella forsythia. Patients with chronic periodontitis were also colonized by Porphyromonas gulae and P. intermedia or by the association between P. gulae and T. forsythia. P. gulae was detected only in the subgingival samples from natives on a traditional diet. Gingival bleeding was associated with Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, T. forsythia, P. gingivalis, P. gulae, Porphyromonas endodontalis, P. intermedia, and Prevotella nigrescens. Treponema denticola was uncommon. Peculiar microbiota was demonstrated to be associated with different periodontal disease statuses in native Brazilians, with modest occurrence of certain pathogens, such as T. denticola, and the presence of P. gulae in natives with gingivitis or chronic periodontitis. Copyright © 2015. Published by Elsevier Ltd.

Endoscopic vs. tactile evaluation of subgingival calculus.

Science.gov (United States)

Osborn, Joy B; Lenton, Patricia A; Lunos, Scott A; Blue, Christine M

2014-08-01

Endoscopic technology has been developed to facilitate imagery for use during diagnostic and therapeutic phases of periodontal care. The purpose of this study was to compare the level of subgingival calculus detection using a periodontal endoscope with that of conventional tactile explorer in periodontitis subjects. A convenience sample of 26 subjects with moderate periodontitis in at least 2 quadrants was recruited from the University of Minnesota School of Dentistry to undergo quadrant scaling and root planing. One quadrant from each subject was randomized for tactile calculus detection alone and the other quadrant for tactile detection plus the Perioscope ™ (Perioscopy Inc., Oakland, Cali). A calculus index on a 0 to 3 score was performed at baseline and at 2 post-scaling and root planing visits. Sites where calculus was detected at visit 1 were retreated. T-tests were used to determine within-subject differences between Perioscope™ and tactile measures, and changes in measures between visits. Significantly more calculus was detected using the Perioscope™ vs. tactile explorer for all 3 subject visits (pcalculus detection from baseline to visit 1 were statistically significant for both the Perioscope™ and tactile quadrants (pcalculus detection from visit 1 to visit 2 was only significant for the Perioscope™ quadrant (pcalculus at this visit. It was concluded that the addition of a visual component to calculus detection via the Perioscope™ was most helpful in the re-evaluation phase of periodontal therapy. Copyright © 2014 The American Dental Hygienists’ Association.

Deep sequencing identifies ethnicity-specific bacterial signatures in the oral microbiome.

Directory of Open Access Journals (Sweden)

Matthew R Mason

Full Text Available Oral infections have a strong ethnic predilection; suggesting that ethnicity is a critical determinant of oral microbial colonization. Dental plaque and saliva samples from 192 subjects belonging to four major ethnicities in the United States were analyzed using terminal restriction fragment length polymorphism (t-RFLP and 16S pyrosequencing. Ethnicity-specific clustering of microbial communities was apparent in saliva and subgingival biofilms, and a machine-learning classifier was capable of identifying an individual's ethnicity from subgingival microbial signatures. The classifier identified African Americans with a 100% sensitivity and 74% specificity and Caucasians with a 50% sensitivity and 91% specificity. The data demonstrates a significant association between ethnic affiliation and the composition of the oral microbiome; to the extent that these microbial signatures appear to be capable of discriminating between ethnicities.

Microbial Profile of Supragingival and Subgingival Plaque of Patients With Glycogen Storage Disease

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Chealsea E. Garcia DDS, MS

2016-12-01

Full Text Available Patients with glycogen storage disease (GSD are either orally fed (ORF or gastronomy-tube fed (GTF with cornstarch to maintain normal glucose levels. It is not known whether the use of cornstarch affects the microbiological oral profile of patients with GSD. Thus, the purpose of this study was to compare supragingival and subgingival plaque samples collected from 53 participants with GSD (2-56 years—29 ORF and 24 GTF. The 16S sequence bacterial profiles of plaque DNA were obtained and a total of 768 probes were detected across the plaque groups. Orally fed patients showed higher means of cariogenic species and periodontal health-associated species, whereas GTF patients showed higher means of periopathogenic species ( P < .05. Orally fed patients exhibited high levels of caries pathogens and lower levels of periodontal pathogens possibly due to the acidic environment created by their cornstarch diet, when compared to GTF patients.

16S rDNA-based metagenomic analysis of dental plaque and lung bacteria in patients with severe acute exacerbations of chronic obstructive pulmonary disease.

Science.gov (United States)

Tan, L; Wang, H; Li, C; Pan, Y

2014-12-01

Acute exacerbations of chronic obstructive pulmonary disease (AE-COPD) are leading causes of mortality in hospital intensive care units. We sought to determine whether dental plaque biofilms might harbor pathogenic bacteria that can eventually cause lung infections in patients with severe AE-COPD. Paired samples of subgingival plaque biofilm and tracheal aspirate were collected from 53 patients with severe AE-COPD. Total bacterial DNA was extracted from each sample individually for polymerase chain reaction amplification and/or generation of bacterial 16S rDNA sequences and cDNA libraries. We used a metagenomic approach, based on bacterial 16S rDNA sequences, to compare the distribution of species present in dental plaque and lung. Analysis of 1060 sequences (20 clones per patient) revealed a wide range of aerobic, anaerobic, pathogenic, opportunistic, novel and uncultivable bacterial species. Species indistinguishable between the paired subgingival plaque and tracheal aspirate samples (97-100% similarity in 16S rDNA sequence) were dental plaque pathogens (Aggregatibacter actinomycetemcomitans, Capnocytophaga sputigena, Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola) and lung pathogens (Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa and Streptococcus pneumoniae). Real-time polymerase chain reaction of 16S rDNA indicated lower levels of Pseudomonas aeruginosa and Porphyromonas gingivalis colonizing the dental plaques compared with the paired tracheal aspirate samples. These results support the hypothesis that dental bacteria may contribute to the pathology of severe AE-COPD. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

16S rRNA based microarray analysis of ten periodontal bacteria in patients with different forms of periodontitis.

Science.gov (United States)

Topcuoglu, Nursen; Kulekci, Guven

2015-10-01

DNA microarray analysis is a computer based technology, that a reverse capture, which targets 10 periodontal bacteria (ParoCheck) is available for rapid semi-quantitative determination. The aim of this three-year retrospective study was to display the microarray analysis results for the subgingival biofilm samples taken from patient cases diagnosed with different forms of periodontitis. A total of 84 patients with generalized aggressive periodontitis (GAP,n:29), generalized chronic periodontitis (GCP, n:25), peri-implantitis (PI,n:14), localized aggressive periodontitis (LAP,n:8) and refractory chronic periodontitis (RP,n:8) were consecutively selected from the archives of the Oral Microbiological Diagnostic Laboratory. The subgingival biofilm samples were analyzed by the microarray-based identification of 10 selected species. All the tested species were detected in the samples. The red complex bacteria were the most prevalent with very high levels in all groups. Fusobacterium nucleatum was detected in all samples at high levels. The green and blue complex bacteria were less prevalent compared with red and orange complex, except Aggregatibacter actinomycetemcomitas was detected in all LAP group. Positive correlations were found within all the red complex bacteria and between red and orange complex bacteria especially in GCP and GAP groups. Parocheck enables to monitoring of periodontal pathogens in all forms of periodontal disease and can be alternative to other guiding and reliable microbiologic tests. Copyright © 2015 Elsevier Ltd. All rights reserved.

Untreated periodontal disease in Indonesian adolescents : Subgingival microbiota in relation to experienced progression of periodontitis

NARCIS (Netherlands)

Timmerman, MF; Van der Weijden, GA; Arief, EM; Armand, S; Abbas, F; Winkel, EG; Van Winkelhoff, AJ; Van der Velden, U

Background/aims: In an Indonesian population deprived of regular dental care, the experienced progression of disease between baseline (1987) and follow-up (1994) was investigated in relation to the composition of the subgingival microbiota at follow-up. At baseline the age ranged from 15 to 25

Combating biofilms

DEFF Research Database (Denmark)

Yang, Liang; Liu, Yang; Wu, Hong

2012-01-01

Biofilms are complex microbial communities consisting of microcolonies embedded in a matrix of self-produced polymer substances. Biofilm cells show much greater resistance to environmental challenges including antimicrobial agents than their free-living counterparts. The biofilm mode of life...... is believed to significantly contribute to successful microbial survival in hostile environments. Conventional treatment, disinfection and cleaning strategies do not proficiently deal with biofilm-related problems, such as persistent infections and contamination of food production facilities. In this review......, strategies to control biofilms are discussed, including those of inhibition of microbial attachment, interference of biofilm structure development and differentiation, killing of biofilm cells and induction of biofilm dispersion....

Association of time under immunosuppression and different immunosuppressive medication on periodontal parameters and selected bacteria of patients after solid organ transplantation.

Science.gov (United States)

Schmalz, G; Berisha, L; Wendorff, H; Widmer, F; Marcinkowski, A; Teschler, H; Sommerwerck, U; Haak, R; Kollmar, O; Ziebolz, D

2018-05-01

Aim of this study was to investigate the association of the time under immunosuppression and different immunosuppressive medication on periodontal parameters and selected periodontal pathogenic bacteria of immunosuppressed patients after solid organ transplantation (SOT). 169 Patients after SOT (lung, liver or kidney) were included and divided into subgroups according their time under (0-1, 1-3, 3-6, 6-10 and >10 years) and form of immunosuppression (Tacrolimus, Cyclosporine, Mycophenolate, Glucocorticoids, Sirolimus and monotherapy vs. combination). Periodontal probing depth (PPD) and clinical attachment loss (CAL) were assessed. Periodontal disease severity was classified as healthy/mild, moderate or severe periodontitis. Subgingival biofilm samples were investigated for eleven selected potentially periodontal pathogenic bacteria using polymerasechainreaction. The mean PPD and CAL as well as prevalence of Treponema denticola and Capnocytophaga species was shown to be different but heterogeneous depending on time under immunosuppression (pperiodontal condition compared to patients without Cyclosporine (pperiodontal and microbiological parameters of patients after SOT. Patients under Cyclosporine medication should receive increased attention. Differences in subgingival biofilm, but not in clinical parameters were found for Glucocorticoids, Mycophenolate and combination therapy, making the clinical relevance of this finding unclear.

Supragingival biofilm control and systemic inflammation in patients with type 2 diabetes mellitus

Directory of Open Access Journals (Sweden)

Hilana Paula Carillo ARTESE

2015-01-01

Full Text Available The objective of this study was to evaluate the effect of strict supragingival biofilm control on serum inflammatory markers and on periodontal clinical parameters in type 2 diabetes mellitus (T2DM patients with chronic severe periodontitis. Twenty-four individuals with T2DM and periodontitis were randomly allocated to two treatment groups. The supragingival therapy group (ST, n = 12 received supragingival scaling, whereas the intensive therapy group (IT, n = 12 underwent supra- and subgingival scaling, as well as root planing. Patients from both groups received professional oral hygiene instructions every month. Data regarding visible plaque index (VPI, gingival bleeding index (GBI, bleeding on probing (BOP, probing pocket depth (PPD, clinical attachment level (CAL, serum levels of interleukin (IL-6, IL-17A, IL-8, tumor necrosis factor α (TNF-α, monocyte chemoattractant protein (MCP-1 enzyme-linked immunosorbent assay (ELISA, and glycated hemoglobin (HbA1c levels were obtained at baseline and at 6 months post-therapy. Both therapies resulted in the improvement of almost all clinical periodontal parameters (p 0.05, between the two periods. However, MCP-1 levels were significantly reduced in both the ST (p = 0.034 and the IT (p = 0.016 groups, whereas the serum IL-6 levels were significantly reduced only in the IT group (p = 0.001. Strict control of supragingival biofilm has a limited effect on systemic inflammatory markers, and a moderate effect on periodontal clinical parameters.

The presence of biofilm forming microorganisms on hydrotherapy equipment and facilities.

Science.gov (United States)

Jarząb, Natalia; Walczak, Maciej

2017-10-01

Hydrotherapy equipment provides a perfect environment for the formation and growth of microbial biofilms. Biofilms may reduce the microbiological cleanliness of hydrotherapy equipment and harbour opportunistic pathogens and pathogenic bacteria. The aims of this study were to investigate the ability of microorganisms that colonize hydrotherapy equipment to form biofilms, and to assess the influence of temperature and nutrients on the rate of biofilm formation. Surface swab samples were collected from the whirlpool baths, inhalation equipment and submerged surfaces of a brine pool at the spa center in Ciechocinek, Poland. We isolated and identified microorganisms from the swab samples and measured their ability to form biofilms. Biofilm formation was observed at a range of temperatures, in both nutrient-deficient and nutrient-rich environments. We isolated and identified microorganisms which are known to form biofilms on medical devices (e.g. Stenotrophomonas maltophilia). All isolates were classified as opportunistic pathogens, which can cause infections in humans with weakened immunity systems. All isolates showed the ability to form biofilms in the laboratory conditions. The potential for biofilm formation was higher in the presence of added nutrients. In addition, the hydrolytic activity of the biofilm was connected with the presence of nutrients.

Subgingival microbiota in health compared to periodontitis and the influence of smoking

OpenAIRE

Camelo-Castillo, Anny J.; Mira, Alex; Pico, Alex; Nibali, Luigi; Henderson, Brian; Donos, Nikolaos; Tomás, Inmaculada

2015-01-01

The etiology of periodontitis has traditionally been associated to a consortium of three bacterial species-the so-called "red-complex" of periodontal disease-which has been the target for most diagnostic and therapeutic strategies. However, other species have also been found to correlate with disease severity. In addition, the influence of smoking on periodontal microbiota is poorly understood. In the current manuscript, the composition of the subgingival microbiota in healthy individuals vs....

PCR detection of four periodontopathogens from subgingival clinical samples Detecção por PCR de quatro periodontopatógenos de pacientes com doença periodontal e de indivíduos sadios

Directory of Open Access Journals (Sweden)

Mario Julio Avila-Campos

2003-04-01

Full Text Available In this study, A. actinomycetemcomitans, B. forsythus, P. gingivalis and F. nucleatum were identified from subgingival plaque from 50 periodontal patients and 50 healthy subjects. Subgingival clinical samples were collected with sterilized paper points and transported in VMGA III. From all the diluted clinical samples (1:10, DNA was obtained by boiling, and after centrifugation the supernatant was used as template. Specific primers for each bacterial species were used in PCR. PCR amplification was sensitive to identify these organisms. PCR products from each species showed a single band and can be used to identify periodontal organisms from clinical specimens. PCR detection odds ratio values for A. actinomycetemcomitans and B. forsythus were significantly associated with disease showing a higher OR values for B. forsythus (2.97, 95% CI 1.88 - 4.70. These results suggest a strong association among the studied species and the periodontal lesion.Em nosso estudo quatro periodontopatógenos foram isolados e identificados de placas subgengivais de 50 pacientes com doença periodontal e de 50 indivíduos sadios. As placas subgengivais foram coletadas com pontas de papel e transportadas em VMGA III. Foram realizadas diluições seriadas das amostras clínicas (1:10, e os DNA foram obtidos por fervura. Iniciadores específicos para cada bactéria foram usados no PCR. As amplificações mostraram-se sensíveis na identificação de A. actinomycetemcomitans, B. forsythus, P. gingivalis e F. nucleatum. As reações de PCR produziram bandas específicas para cada espécie e podem ser usadas na identificação desses organismos periodontais diretamente das amostras clínicas. Os valores de odds ratio para a detecção de A. actinomycetemcomitans e B. forsythus foram significativamente associados com a doença periodontal mostrando altos valores de OR para B. forsythus (2,97, 95% CI 1,88 - 4,70. Esses resultados sugerem uma forte associação entre os

Antibiotic resistance and biofilm formation among coagulase-negative staphylococci isolated from clinical samples at a tertiary care hospital of eastern Nepal.

Science.gov (United States)

Shrestha, Lok Bahadur; Bhattarai, Narayan Raj; Khanal, Basudha

2017-01-01

Coagulase negative staphylococci were long regarded non-pathogenic as they are the commensals of human skin and mucosa but the recent changes in the medical practice and changes in underlying host populations, they are being considered significant pathogens associated with number of nosocomial infections. The objective of the study was to determine the species, antimicrobial susceptibility pattern, biofilm forming ability of the clinically significant CoNS isolates and to compare the different methods for the detection of biofilm formation. A total of 52 clinically significant CoNS isolates obtained from different units during a year period were studied. Characterization was done using standard microbiological guidelines and antimicrobial susceptibility was done following CLSI guidelines. Biofilm formation was detected by using three methods i.e. tissue culture plate method, congo red agar method and tube adherence method. Among 52 isolates , S. epidermidis (52%) was the most common species which was followed by S. saprophyticus (18%) and S. haemolyticus (14%). Antimicrobial susceptibility pattern of CoNS documented resistance of 80% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 58% of the isolates. Biofilm formation was observed in 65.38% of the isolates. The accuracy of Congo red agar and tube adherence method for the detection of biofilm formation was 82% and 76% respectively. CoNS isolates obtained from clinical samples should be processed routinely and antimicrobial susceptibility testing should be performed. Multidrug-resistant CoNS are prevalent. All the three methods i.e. tissue culture plate, Congo red agar and tube adherence method can be used in detecting biofilm formation.

Biofilms

OpenAIRE

López, Daniel; Vlamakis, Hera; Kolter, Roberto

2010-01-01

The ability to form biofilms is a universal attribute of bacteria. Biofilms are multicellular communities held together by a self-produced extracellular matrix. The mechanisms that different bacteria employ to form biofilms vary, frequently depending on environmental conditions and specific strain attributes. In this review, we emphasize four well-studied model systems to give an overview of how several organisms form biofilms: Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and ...

Biofilms and their modifications by laser irradiation

International Nuclear Information System (INIS)

Richter, Asta; Gonpot, Preethee; Smith, Roger

2001-01-01

Biofilms are grown on different materials with various surface morphology and are investigated by light and scanning force microscopy. The growth patterns, coverage and adherence of the biofilm are shown to depend on the type of the substrate and its roughness as well as on the type of micro-organisms. Here we present investigations of Eschericia coli bacterial biofilms grown on the polymer material polyetheretherketone and also on titanium films on glass substrates. A Monte Carlo simulation of the growth process is developed which takes into account the aspect ratio of the micro-organisms and the diffusion of nutrient over the surface to feed them. A pulsed nitrogen laser has been applied to the samples and the interaction of the laser beam with the biofilm and the underlying substrate has been studied. Because of the inhomogeneity of the biofilms the ablated areas are different. With increasing number of laser pulses more biofilm material is removed but there appears also damage of the substrate. The threshold energy fluence for the biofilm ablation is estimated and depends on the sticking power of the bacteria. Ablation rates for the removal of the biofilms are also obtained

QUALITATIVE EVALUATION OF SUBGINGIVAL MICROFLORA AFTER THE CHEWING OF BETEL LEAF

OpenAIRE

K Vani; Jose Maji; Rao Srinivasa

2011-01-01

Betel a branching vine, scientifically called as Piper betel, is used in a number of traditional remedies and known to have immune boosting as well as antibacterial properties. This study was conducted to assess the qualitative changes in the sub-gingival micro flora, after the chewing of betel leaves, in order to evaluate the antimicrobial effect of this medicinal plant on oral pathogens.Forty volunteers were made to chew betel leaves daily for 5-10 minutes for a period of two weeks and the ...

Oral cavity anaerobic pathogens in biofilm formation on voice prostheses

NARCIS (Netherlands)

Bertl, Kristina; Zijnge, Vincent; Zatorska, Beata; Leonhard, Matthias; Schneider-Stickler, Berit; Harmsen, Hermie J. M.

BACKGROUND: A polymerase chain reaction (PCR)-based method has been used to identify oral anaerobic pathogens in biofilms on voice prostheses. The purpose of the present study was to determine the location of those pathogens inside the biofilms. METHODS: Biofilms of 15 voice prostheses were sampled

Nanoindentation of Pseudomonas aeruginosa bacterial biofilm using atomic force microscopy

International Nuclear Information System (INIS)

Baniasadi, Mahmoud; Xu, Zhe; Du, Yingjie; Lu, Hongbing; Minary-Jolandan, Majid; Gandee, Leah; Zimmern, Philippe

2014-01-01

Bacterial biofilms are a source of many chronic infections. Biofilms and their inherent resistance to antibiotics are attributable to a range of health issues including affecting prosthetic implants, hospital-acquired infections, and wound infection. Mechanical properties of biofilm, in particular, at micro- and nano-scales, are governed by microstructures and porosity of the biofilm, which in turn may contribute to their inherent antibiotic resistance. We utilize atomic force microscopy (AFM)-based nanoindentation and finite element simulation to investigate the nanoscale mechanical properties of Pseudomonas aeruginosa bacterial biofilm. This biofilm was derived from human samples and represents a medically relevant model. (paper)

Mucosal biofilm detection in chronic otitis media

DEFF Research Database (Denmark)

Wessman, Marcus; Bjarnsholt, Thomas; Eickhardt-Sørensen, Steffen Robert

2015-01-01

The objectives of this study were to examine middle ear biopsies from Greenlandic patients with chronic otitis media (COM) for the presence of mucosal biofilms and the bacteria within the biofilms. Thirty-five middle ear biopsies were obtained from 32 Greenlandic COM patients admitted to ear...... of the patients served as controls. PNA-FISH showed morphological signs of biofilms in 15 out of 35 (43 %) middle ear biopsies. In the control skin biopsies, there were signs of biofilms in eight out of 23 biopsies (30 %), probably representing skin flora. PCR and 16s sequencing detected bacteria in seven out...... of 20 (35 %) usable middle ear biopsies, and in two out of ten (20 %) usable control samples. There was no association between biofilm findings and PCR and 16s sequencing. Staphylococci were the most common bacteria in bacterial culture. We found evidence of bacterial biofilms in 43 % of middle ear...

Quantification of diatoms in biofilms: Standardisation of methods

Digital Repository Service at National Institute of Oceanography (India)

Patil, J.S.; Anil, A.C.

of the difficulty in sampling and enumeration. Scraping or brushing are the traditional methods used for removal of diatoms from biofilms developed on solid substrata. The method of removal is the most critical step in enumerating the biofilm diatom community...

Role of bacterial biofilm in development of middle ear effusion.

Science.gov (United States)

Tawfik, Sedeek Abd El-Salam; Ibrahim, Ahmed Aly; Talaat, Iman Mamdoh; El-Alkamy, Soliman Samy Abd El-Raouf; Youssef, Ahmed

2016-11-01

Biofilms have been implicated in the development of several chronic upper respiratory tract infections. Role of bacterial biofilms has been well studied in the pathogenesis of chronic rhinosinusitis. However, its impact on development of middle ear effusion is still a matter of debate. To study the extent of surface adenoid biofilm and evaluate its role in the pathogenesis of chronic otitis media with effusion in children. The study was carried out on 40 children in Alexandria Main University Hospital between 1 and 16 years of age without sex predilection, who were divided into two groups. The first group (20 children) had otitis media with effusion associated with adenoid hypertrophy, whereas the second group (20 children) had adenoid hypertrophy without middle ear effusion. Adenoidectomy with ventilation tube insertion was done for group 1 cases, whereas, only Adenoidectomy was done for group 2 cases. The samples were processed for the detection of biofilms by scanning electron microscopy. The biofilm formation was graded according to extension. Biofilm formation was detected on all samples for group 1. Adenoids removed from patients with otitis media with effusion had higher-grade biofilm formation than the other group (P 0.0001). No correlation was found between adenoid size and biofilm formation. In pediatric population, adenoid surface biofilm formation may be involved in the pathogenesis otitis media with effusion.

Biofilm architecture in a novel pressurized biofilm reactor.

Science.gov (United States)

Jiang, Wei; Xia, Siqing; Duan, Liang; Hermanowicz, Slawomir W

2015-01-01

A novel pure-oxygen pressurized biofilm reactor was operated at different organic loading, mechanical shear and hydrodynamic conditions to understand the relationships between biofilm architecture and its operation. The ultimate goal was to improve the performance of the biofilm reactor. The biofilm was labeled with seven stains and observed with confocal laser scanning microscopy. Unusual biofilm architecture of a ribbon embedded between two surfaces with very few points of attachment was observed. As organic loading increased, the biofilm morphology changed from a moderately rough layer into a locally smoother biomass with significant bulging protuberances, although the chemical oxygen demand (COD) removal efficiency remained unchanged at about 75%. At higher organic loadings, biofilms contained a larger fraction of active cells distributed uniformly within a proteinaceous matrix with decreasing polysaccharide content. Higher hydrodynamic shear in combination with high organic loading resulted in the collapse of biofilm structure and a substantial decrease in reactor performance (a COD removal of 16%). Moreover, the important role of proteins for the spatial distribution of active cells was demonstrated quantitatively.

Antibiotic resistance and biofilm formation of some bacteria isolated from sediment, water and fish farms in Malaysia

Science.gov (United States)

Faja, Orooba Meteab; Usup, Gires; Ahmad, Asmat

2018-04-01

A total of 90 isolates of bacteria were isolated, from sediment (10) samples, water (10) samples and fish (12) samples (Sea bass, Snapper, Grouper and Tilapia). These include 22 isolates of bacteria from sediment, 28 isolates from water and 40 isolates from fish. All the isolates were tested for sensitivity to 13 antibiotics using disc diffusion method. The isolates showed high resistance to some antibiotics based on samples source. Isolates from sediment showed highest resistance toward novobiocin, kanamycin, ampicillin and streptomycin while isolates from water showed highest resistance against vancomycin, penicillin, streptomycin and tetracycline, in contrast, in fish sample showed highest resistance toward vancomycin, ampicillin, streptomycin and tetracycline. Most of the isolates showed biofilm formation ability with different degrees. Out of 22 bacteria isolates from water, two isolates were weak biofilm formers, six isolates moderate biofilm formers and fourteen isolates strong biofilm formers. While, out of 28 bacteria isolates from water one isolate was weak biofilm former, five isolates moderate biofilm formers and 22 strong biofilm formers Fish isolate showed three isolates (8%) moderate biofilm formers and 27 isolates strong biofilm formers. Biofilm formation was one of the factors that lead to antibiotic resistance of the bacterial isolates from these samples.

Activity of disinfectants and biofilm production of Corynebacterium pseudotuberculosis

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Maria da C.A. Sá

2013-11-01

Full Text Available To verify the occurrence of caseous lymphadenitis in sheep and goats on farms of Pernambuco, Brazil, and in animals slaughtered in two Brazilian cities (Petrolina/PE and Juazeiro/BA, and to characterize the susceptibility profile of Corynebacterium pseudotuberculosis to disinfectants and antimicrobials, and its relationship with biofilm production were the objectives of this study. 398 samples were tested for sensitivity to antimicrobial drugs, disinfectants, and biofilm production. Among the 108 samples collected on the properties, 75% were positive for C. pseudotuberculosis. Slaughterhouse samples indicated an occurrence of caseous lymphadenitis in 15.66% and 6.31% for animals slaughtered in Petrolina and Juazeiro respectively. With respect to antimicrobials, the sensitivity obtained was 100% for florfenicol and tetracycline; 99.25% for enrofloxacin, ciprofloxacin and lincomycin; 98.99% for cephalothin; 98.74% for norfloxacin and sulfazotrim; 97.74% for gentamicin; 94.22% for ampicillin; 91.71% for amoxicillin; 91.21% for penicillin G; 89.19% for neomycin and 0% for novobiocin. In analyzes with disinfectants, the efficiency for chlorhexidine was 100%, 97.20% for quaternary ammonium, 87.40% for chlorine and 84.40% for iodine. 75% of the isolates were weak or non-biofilm producers. For the consolidated biofilm, found that iodine decreased biofilm formation in 13 isolates and quaternary ammonia in 11 isolates. The reduction of the biofilm formation was observed for iodine and quaternary ammonium in consolidated biofilm formation in 33% and 28% of the isolates, respectively. The results of this study highlight the importance of establishing measures to prevent and control the disease.

Antibiotic resistance and biofilm formation among coagulase-negative staphylococci isolated from clinical samples at a tertiary care hospital of eastern Nepal

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Lok Bahadur Shrestha

2017-08-01

Full Text Available Abstract Background Coagulase negative staphylococci were long regarded non-pathogenic as they are the commensals of human skin and mucosa but the recent changes in the medical practice and changes in underlying host populations, they are being considered significant pathogens associated with number of nosocomial infections. The objective of the study was to determine the species, antimicrobial susceptibility pattern, biofilm forming ability of the clinically significant CoNS isolates and to compare the different methods for the detection of biofilm formation. Methods A total of 52 clinically significant CoNS isolates obtained from different units during a year period were studied. Characterization was done using standard microbiological guidelines and antimicrobial susceptibility was done following CLSI guidelines. Biofilm formation was detected by using three methods i.e. tissue culture plate method, congo red agar method and tube adherence method. Results Among 52 isolates, S. epidermidis (52% was the most common species which was followed by S. saprophyticus (18% and S. haemolyticus (14%. Antimicrobial susceptibility pattern of CoNS documented resistance of 80% to ampicillin. Resistance to cefoxitin and ceftriaxone was observed in 58% of the isolates. Biofilm formation was observed in 65.38% of the isolates. The accuracy of Congo red agar and tube adherence method for the detection of biofilm formation was 82% and 76% respectively. Conclusion CoNS isolates obtained from clinical samples should be processed routinely and antimicrobial susceptibility testing should be performed. Multidrug-resistant CoNS are prevalent. All the three methods i.e. tissue culture plate, Congo red agar and tube adherence method can be used in detecting biofilm formation.

Staphylococcus aureus biofilm removal by targeting biofilm-associated extracellular proteins

Directory of Open Access Journals (Sweden)

Sudhir K Shukla

2017-01-01

Methods: Biofilm assay was done in 96-well microtitre plate to evaluate the effect of proteinase K on biofilms of bovine mastitis S. Aureus isolates. Extracellular polymeric substances were extracted and evaluated for their composition (protein, polysaccharides and extracellular DNA, before and after the proteinase K treatment. Results: Biofilm assay showed that 2 μg/ml proteinase K significantly inhibited biofilm development in bap-positive S. aureus V329 as well as other S. aureus isolates (SA7, SA10, SA33, SA352, but not in bap-mutant M556 and SA392 (a weak biofilm-producing strain. Proteinase K treatment on S. aureus planktonic cells showed that there was no inhibition of planktonic growth up to 32 μg/ml of proteinase K. Proteinase K treatment on 24 h old preformed biofilms showed an enhanced dispersion of bap-positive V329 and SA7, SA10, SA33 and SA352 biofilms; however, proteinase K did not affect the bap-mutant S. aureus M556 and SA392 biofilms. Biofilm compositions study before and after proteinase K treatment indicated that Bap might also be involved in eDNA retention in the biofilm matrix that aids in biofilm stability. When proteinase K was used in combination with antibiotics, a synergistic effect in antibiotic efficacy was observed against all biofilm-forming S. aureus isolates. Interpretation & conclusions: Proteinase K inhibited biofilms growth in S. aureus bovine mastitis isolates but did not affect their planktonic growth. An enhanced dispersion of preformed S. aureus biofilms was observed on proteinase K treatment. Proteinase K treatment with antibiotics showed a synergistic effect against S. aureus biofilms. The study suggests that dispersing S. aureus by protease can be of use while devising strategies againstS. aureus biofilms.

Membrane biofilm communities in full-scale membrane bioreactors are not randomly assembled and consist of a core microbiome

KAUST Repository

Matar, Gerald Kamil

2017-06-21

Finding efficient biofouling control strategies requires a better understanding of the microbial ecology of membrane biofilm communities in membrane bioreactors (MBRs). Studies that characterized the membrane biofilm communities in lab-and pilot-scale MBRs are numerous, yet similar studies in full-scale MBRs are limited. Also, most of these studies have characterized the mature biofilm communities with very few studies addressing early biofilm communities. In this study, five full-scale MBRs located in Seattle (Washington, U.S.A.) were selected to address two questions concerning membrane biofilm communities (early and mature): (i) Is the assembly of biofilm communities (early and mature) the result of random immigration of species from the source community (i.e. activated sludge)? and (ii) Is there a core membrane biofilm community in full-scale MBRs? Membrane biofilm (early and mature) and activated sludge (AS) samples were collected from the five MBRs, and 16S rRNA gene sequencing was applied to investigate the bacterial communities of AS and membrane biofilms (early and mature). Alpha and beta diversity measures revealed clear differences in the bacterial community structure between the AS and biofilm (early and mature) samples in the five full-scale MBRs. These differences were mainly due to the presence of large number of unique but rare operational taxonomic units (∼13% of total reads in each MBR) in each sample. In contrast, a high percentage (∼87% of total reads in each MBR) of sequence reads was shared between AS and biofilm samples in each MBR, and these shared sequence reads mainly belong to the dominant taxa in these samples. Despite the large fraction of shared sequence reads between AS and biofilm samples, simulated biofilm communities from random sampling of the respective AS community revealed that biofilm communities differed significantly from the random assemblages (P < 0.001 for each MBR), indicating that the biofilm communities (early

Bacteriophage use to control Salmonella biofilm on surfaces present in chicken slaughterhouses.

Science.gov (United States)

Garcia, Keila Carolina de Ornellas Dutka; Corrêa, Isadora Mainieri de Oliveira; Pereira, Larissa Quinto; Silva, Tarcísio Macedo; Mioni, Mateus de Souza Ribeiro; Izidoro, Ana Carolina de Moraes; Bastos, Igor Henrique Vellano; Gonçalves, Guilherme Augusto Marietto; Okamoto, Adriano Sakai; Andreatti Filho, Raphael Lucio

2017-09-01

Foodborne diseases represent a major risk to public health worldwide. Pathogenic bacteria can live in the form of biofilm within the food industry, providing a permanent source of contamination. The aim of this study was to evaluate the influence of the types of adhesion surfaces on Salmonella biofilm formation at eight different times, and analyze the action time of a bacteriophage pool on established biofilms. Most of the samples used were classified as weak biofilm producers, with serovars Enteritidis and Heidelberg showing the highest frequency of biofilm formation. Glass and stainless steel surfaces significantly favored biofilm formation at 60 and 36 h of incubation respectively, but the polyvinyl chloride surface did not favor biofilm production, suggesting that the type of material may interfere with production. The bacteriophage pool action period focused on 3 h, but treatment of 9 h on glass surface biofilms was superior to other treatments because it affected the largest number of samples. These results suggests that some surface types and Salmonella serotypes may promote biofilm formation and indicate bacteriophages as an alternative to control biofilms. But further studies are required to prove the effectiveness and safety of bacteriophage therapy as an alternative in the antimicrobial control in the processing plants. © 2017 Poultry Science Association Inc.

Antibiotic Susceptibility of Periodontal Streptococcus Constellatus and Streptococcus Intermedius Clinical Isolates

NARCIS (Netherlands)

Rams, Thomas E; Feik, Diane; Mortensen, Joel E; Degener, John E; van Winkelhoff, Arie J

2014-01-01

Background: Streptococcus constellatus and Streptococcus intermedius in subgingival dental plaque biofilms may contribute to forms of periodontitis that resist treatment with conventional mechanical root debridement/surgical procedures and may additionally participate in some extraoral infections.

Study of biofilm in bacteria from water pipelines.

Science.gov (United States)

Mahapatra, Ashoka; Padhi, Nupur; Mahapatra, Dharitri; Bhatt, Mamta; Sahoo, Debasish; Jena, Swetlina; Dash, Debabrata; Chayani, Nirupama

2015-03-01

A biofilm is a layer of microorganisms contained in a matrix (slime layer), which forms on surfaces in contact with water. Their presence in drinking water pipe networks can be responsible for a wide range of water quality and operational problems. To identify the bacterial isolates, obtained from water pipelines of kitchens, to evaluate the water quality & to study the biofilm producing capacity of the bacterial isolates from various sources. A prospective study using water samples from aqua guard & pipelines to kitchens of S.C.B Medical College hostels. Standard biochemical procedures for bacterial identification, multiple tube culture & MPN count to evaluate water quality & tissue culture plate (TCP) method for biofilm detection was followed. STATA software version 9.2 from STATA Corporation, College station road, 90 Houston, Texas was used for statistical analysis. One hundred eighty seven isolates were obtained from 45 water samples cultured. The isolates were Acinetobacter spp. (44), Pseudomonas spp.(41), Klebsiella spp.(36) & others . Biofilm was detected in (37) 19.78 % of the isolates (95% CI 30.08% -43.92%) including Acinetobacter spp.-10, Klebsiella spp. - 9, Pseudomonas spp. - 9, & others, majority (34) of which were from kitchen pipelines. Water from pipeline sources was unsatisfactory for consumption as the MPN counts were > 10. Most of the biofilm producers were gram negative bacilli & Pseudomonas & Acinetobacter spp. were strong (4+) biofilm producers.

Absolute quantification of Aggregatibacter actinomycetemcomitans in patients carrying haplotypes associated with susceptibility to chronic periodontitis: multifaceted evaluation with periodontitis covariants.

Science.gov (United States)

Cirelli, Thamiris; Finoti, Livia S; Corbi, Sâmia C T; Anovazzi, Giovana; Nepomuceno, Rafael; Orrico, Silvana R P; Cirelli, Joni A; Mayer, Márcia P A; Scarel-Caminaga, Raquel M

2017-09-29

This study aimed to evaluate the association between haplotypes in the interleukin 8 (IL8) and IL4 genes previously associated to chronic periodontitis (CP) and the levels of Aggregatibacter actinomycetemcomitans (A.a.) in subgingival sites of patients with and without CP. Moreover, multifaceted evaluations were made to search associations among patients' genetic background with the A.a. levels and previous clinical/immunological/microbiological findings. Subgingival sites (n = 596) of 104 patients were divided into susceptible to CP by the IL8 haplotype ATC/TTC (IL8+); non-susceptible to CP by the IL8 AGT/TTC (IL8-); susceptible to CP by the IL4 TCI/CCI (IL4+); protection against CP by the IL4 TTD/CTI (IL4-). Subgingival biofilm samples from diseased and healthy sites of CP patients and from control sites of health patients were obtained for absolute quantification of A.a. by quantitative real-time polymerase chain reaction. For diseased sites, samples were collected before and 45 days after periodontal treatment. The IL4 but not the IL8 haplotypes were associated with levels of A.a. (in both periods). After periodontal treatment, higher levels of A.a. were found in subgingival sites of (IL4-) patients, and higher levels of IL-4 were associated with deeper probing pockets in these same patients. Significant correlations were found among genetic (patients carrying IL8 or IL4 haplotypes), microbiological and immunological data showing the interrelationship of different factors in the CP. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Bacterial biofilm in chronic lesions of Hidradenitis Suppurativa

DEFF Research Database (Denmark)

Ring, H C; Bay, L; Nilsson, M

2017-01-01

BACKGROUND: Chronic non-healing or recurrent inflammatory lesions, reminiscent of infection but recalcitrant to antibiotic therapy generally characterize biofilm driven-diseases. Chronic lesions of Hidradenitis Suppurativa (HS) exhibit several aspects, which are compatible with well-known biofilm...... infections. OBJECTIVE: To determine and quantify the potential presence of bacterial aggregates in chronic HS lesions. METHODS: In 42 consecutive HS patients suffering from chronic lesions, biopsies were obtained from lesional as well as from perilesional skin. Samples were investigated using Peptide Nucleic...... Acid (PNA) - Fluorescence in situ Hybridization (FISH) in combination with Confocal Laser Scanning Microscopy (CLSM). In addition, corresponding histopathological analysis in hematoxylin and eosin slides were performed. RESULTS: Biofilms were seen in 67% of the samples of chronic lesions and in 75...

Biofilm roughness determines Cryptosporidium parvum retention in environmental biofilms.

Science.gov (United States)

DiCesare, E A Wolyniak; Hargreaves, B R; Jellison, K L

2012-06-01

The genus Cryptosporidium is a group of waterborne protozoan parasites that have been implicated in significant outbreaks of gastrointestinal infections throughout the world. Biofilms trap these pathogens and can contaminate water supplies through subsequent release. Biofilm microbial assemblages were collected seasonally from three streams in eastern Pennsylvania and used to grow biofilms in laboratory microcosms. Daily oocyst counts in the influx and efflux flow allowed the calculation of daily oocyst retention in the biofilm. Following the removal of oocysts from the influx water, oocyst attachment to the biofilm declined to an equilibrium state within 5 days that was sustained for at least 25 days. Varying the oocyst loading rate for the system showed that biofilm retention could be saturated, suggesting that discrete binding sites determined the maximum number of oocysts retained. Oocyst retention varied seasonally but was consistent across all three sites; however, seasonal oocyst retention was not consistent across years at the same site. No correlation between oocyst attachment and any measured water quality parameter was found. However, oocyst retention was strongly correlated with biofilm surface roughness and roughness varied among seasons and across years. We hypothesize that biofilm roughness and oocyst retention are dependent on environmentally driven changes in the biofilm community rather than directly on water quality conditions. It is important to understand oocyst transport dynamics to reduce risks of human infection. Better understanding of factors controlling biofilm retention of oocysts should improve our understanding of oocyst transport at different scales.

Acinetobacter baumannii in Southern Croatia: clonal lineages, biofilm formation, and resistance patterns.

Science.gov (United States)

Kaliterna, Vanja; Kaliterna, Mariano; Hrenović, Jasna; Barišić, Zvonimir; Tonkić, Marija; Goic-Barisic, Ivana

2015-01-01

Acinetobacter baumannii is one of the most prevalent causes of severe hospital-acquired infections and is responsible for the dramatic increase in carbapenem resistance in Croatia in the last 5 years. Such data have encouraged multicenter research focused on the organism's ability to form biofilm, susceptibility to antibiotics, and particular genotype lineage. Biofilm formation in 109 unrelated clinical isolates of A. baumannii recovered in six cities of Southern Croatia was investigated. Genotyping was performed by pulsed-field gel electrophoresis and antibiotic profile was tested by applying the disc diffusion method and confirmed by determining the minimum inhibitory concentrations. The ability to form biofilm in vitro was determined from overnight cultures of the collected isolates on microtiter plates, after staining with crystal violet, and quantified at 570 nm after solubilization with ethanol. The statistical relevance was calculated in an appropriate program with level of statistical confidence. There was no significant difference in biofilm formation due to the genotype lineage. Isolates collected from intensive care units (ICUs) and isolated from respiratory samples were more likely to create a biofilm compared with isolates from other departments and other samples. There was a significant difference in the ability to produce biofilm in relation to antibiotic resistance pattern. A large proportion of A. baumannii isolates that were resistant to ampicillin/sulbactam, carbapenems, and amikacin were found to be biofilm-negative. In contrast, isolates susceptible and intermediately susceptible to ampicillin/sulbactam, carbapenems, and amikacin were biofilm producers. Clinical isolates of A. baumannii from respiratory samples in ICUs with a particular susceptibility pattern are more prone to form biofilm.

The investigation of Helicobacter pylori in the dental biofilm and saliva samples of children with dyspeptic complaints

OpenAIRE

Aksit B?cak, Damla; Akyuz, Serap; K?ratl?, Binnur; Usta, Merve; Urganci, Nafiye; Alev, Burcin; Yarat, Aysen; Sahin, Fikrettin

2017-01-01

Background The oral cavity can be an extra-gastric reservoir for Helicobacter pylori (H.pylori). This can play a role in the pathogenesis of halitosis, glossitis, recurrent aphthous stomatitis, and dental caries. The present study was conducted to detect the presence of H.pylori within the dental biofilm and in saliva samples collected from children suffering from dyspepsia and children without any gastrointestinal complaints. Associations with gastric infection, halitosis, and some oral para...

Clinical implications of microbial biofilms in chronic rhinosinusitis and orbital cellulitis.

Science.gov (United States)

Nayak, Niranjan; Satpathy, Gita; Prasad, Sujata; Thakar, Alok; Chandra, Mahesh; Nag, T C

2016-09-21

Discovery of sessile mode of microbial existence (Biofilm state) focussed much interest, during the recent years, on the study of biofilms in many recurring and chronic infections. However, the exact role of microbial biofilms in chronic rhinosinusitis and orbital cellulitis were not elucidated earlier. The purpose of the present study was to look for the adherent property and biofilm producing ability of the clinical isolates in chronic rhinosinusitis and orbital cellulitis, and to look for the effects of antimicrobial agents on these biofilms by colorimetric assay and ultrastructural analysis. Organisms were isolated and identified from various clinical samples in patients with chronic sinusitis and orbital cellulitis. Antimicrobial sensitivity testing was carried out by the standard protocol. Biofilms were developed; quantified and antimicrobial drug perfusion through the biofilm model was evaluated by the earlier devised procedure. Electronmicroscopic study of the biofilm was performed by the recommended technique. Of the total of 70 clinical samples processed, 48 i.e. 68.5 % grew bacteria and 13 i.e.(18.6 %) fungi. Staphylococcus aureus (20), S epidermidis (16) and Pseudomonas aeruginosa (6) accounted for the majority of the bacterial isolates. Aspergillus flavus (8), however was the commonest amongst the fungi. A total of 40 bacteria and 8 fungi could be tested for biofilm production. Eighteen (45 %) of the 40 bacterial isolates and 4(50 %) out of the 8 A flavus isolates were found to be biofilm producers. In vitro adherence testing revealed that majority i.e. 16 (88.8 %) of the 18 biofilm positive bacteria were adherent to artificial surfaces. Antimicrobial drug perfusion through the biofilm model was poor. Antimicrobial treatment was totally ineffective against strong biofilm producers, whose electron microscopic picture was quite similar to that observed for biofilm producers without any antimicrobial pre-treatment. Filamentous fungi, like bacteria

Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran

Directory of Open Access Journals (Sweden)

Elahe Tajbakhsh

2016-04-01

Full Text Available Abstract Background Uropathogenic Escherichia coli O- Serogroups with their virulence factors are the most prevalent causes of UTIs. The present research performed to track common uropathogenic E.coli serogroups, antibiotic resistance pattern of strains and prevalence of virulence genes in isolations having the ability to constitute biofilm. Methods In this research 130 E.coli isolation from patients having UTI symptoms were collected and antimicrobial resistance pattern was performed by Kirby-Bauer method. Polymerase chain reaction was done using primer pairs to identify common serogroups of uropathogenic E.coli and studying virulence genes in isolations creating biofilm. Results Among 130 E.coli isolates, 80 (61.53 % were able to make biofilm that 15 isolates (18.75 % indicated strong reaction, 20 (25 % of medium and 45 (56.25 % of weak biofilm reaction. Among isolations creating biofilm, the highest resistance reported to Ampicillin (87.5 % and the lowest to Nitrofurantoin (3.75 %. The frequency of fimH, pap, sfa and afa genes in isolations having the ability to create strong biofilm reported 93.33 %, 86.66 %, 86.66 % and 66.66 %, respectively. Conclusions The findings indicated the importance of virulence genes in serogroups producing uropathogenic E.coli biofilm. It is recommended that strains producing biofilm before antibiotic use should be studied.

Temporal Variations in the Abundance and Composition of Biofilm Communities Colonizing Drinking Water Distribution Pipes

Science.gov (United States)

Kelly, John J.; Minalt, Nicole; Culotti, Alessandro; Pryor, Marsha; Packman, Aaron

2014-01-01

Pipes that transport drinking water through municipal drinking water distribution systems (DWDS) are challenging habitats for microorganisms. Distribution networks are dark, oligotrophic and contain disinfectants; yet microbes frequently form biofilms attached to interior surfaces of DWDS pipes. Relatively little is known about the species composition and ecology of these biofilms due to challenges associated with sample acquisition from actual DWDS. We report the analysis of biofilms from five pipe samples collected from the same region of a DWDS in Florida, USA, over an 18 month period between February 2011 and August 2012. The bacterial abundance and composition of biofilm communities within the pipes were analyzed by heterotrophic plate counts and tag pyrosequencing of 16S rRNA genes, respectively. Bacterial numbers varied significantly based on sampling date and were positively correlated with water temperature and the concentration of nitrate. However, there was no significant relationship between the concentration of disinfectant in the drinking water (monochloramine) and the abundance of bacteria within the biofilms. Pyrosequencing analysis identified a total of 677 operational taxonomic units (OTUs) (3% distance) within the biofilms but indicated that community diversity was low and varied between sampling dates. Biofilms were dominated by a few taxa, specifically Methylomonas, Acinetobacter, Mycobacterium, and Xanthomonadaceae, and the dominant taxa within the biofilms varied dramatically between sampling times. The drinking water characteristics most strongly correlated with bacterial community composition were concentrations of nitrate, ammonium, total chlorine and monochloramine, as well as alkalinity and hardness. Biofilms from the sampling date with the highest nitrate concentration were the most abundant and diverse and were dominated by Acinetobacter. PMID:24858562

Biofilm Development

DEFF Research Database (Denmark)

Tolker-Nielsen, Tim

2015-01-01

During the past decade we have gained much knowledge about the molecular mechanisms that are involved in initiation and termination of biofilm formation. In many bacteria, these processes appear to occur in response to specific environmental cues and result in, respectively, induction or terminat......During the past decade we have gained much knowledge about the molecular mechanisms that are involved in initiation and termination of biofilm formation. In many bacteria, these processes appear to occur in response to specific environmental cues and result in, respectively, induction...... or termination of biofilm matrix production via the second messenger molecule c-di-GMP. In between initiation and termination of biofilm formation we have defined specific biofilm stages, but the currently available evidence suggests that these transitions are mainly governed by adaptive responses......, and not by specific genetic programs. It appears that biofilm formation can occur through multiple pathways and that the spatial structure of the biofilms is species dependent as well as dependent on environmental conditions. Bacterial subpopulations, e.g., motile and nonmotile subpopulations, can develop...

Biofilm Infections

DEFF Research Database (Denmark)

Bjarnsholt, Thomas; Jensen, Peter Østrup; Moser, Claus Ernst

A still increasing interest and emphasis on the sessile bacterial lifestyle biofilms has been seen since it was realized that the vast majority of the total microbial biomass exists as biofilms. Aggregation of bacteria was first described by Leeuwenhoek in 1677, but only recently recognized...... as being important in chronic infection. In 1993 the American Society for Microbiology (ASM) recognized that the biofilm mode of growth was relevant to microbiology. This book covers both the evidence for biofilms in many chronic bacterial infections as well as the problems facing these infections...... such as diagnostics, pathogenesis, treatment regimes and in vitro and in vivo models for studying biofilms. This is the first scientific book on biofilm infections, chapters written by the world leading scientist and clinicians. The intended audience of this book is scientists, teachers at university level as well...

Biofilm Formation on Different Materials Used in Oral Rehabilitation

OpenAIRE

Souza, Júlio C. M.; Mota, Raquel R. C.; Sordi, Mariane B.; Passoni, Bernardo B.; Benfatti, Cesar A. M.; Magini, Ricardo S.

2016-01-01

Abstract The aim of this study was to evaluate the density and the morphological aspects of biofilms adhered to different materials applied in oral rehabilitation supported by dental implants. Sixty samples were divided into four groups: feldspar-based porcelain, CoCr alloy, commercially pure titanium grade IV and yttria-stabilized zirconia. Human saliva was diluted into BHI supplemented with sucrose to grow biofilms for 24 or 48 h. After this period, biofilm was removed by 1% protease treatm...

Antibiotic resistance and ndvB gene expression among biofilm ...

African Journals Online (AJOL)

A novel antibiotic resistant mechanism among biofilms is glucan-mediated sequestration in which ndvB gene encodes a glucosyltransferase involved in the formation of this glucans. We studied the biofilm formation and antibiotic susceptibility pattern of P. aeruginosa isolated from clinical samples, and measured the ...

Biophysics of biofilm infection.

Science.gov (United States)

Stewart, Philip S

2014-04-01

This article examines a likely basis of the tenacity of biofilm infections that has received relatively little attention: the resistance of biofilms to mechanical clearance. One way that a biofilm infection persists is by withstanding the flow of fluid or other mechanical forces that work to wash or sweep microorganisms out of the body. The fundamental criterion for mechanical persistence is that the biofilm failure strength exceeds the external applied stress. Mechanical failure of the biofilm and release of planktonic microbial cells is also important in vivo because it can result in dissemination of infection. The fundamental criterion for detachment and dissemination is that the applied stress exceeds the biofilm failure strength. The apparent contradiction for a biofilm to both persist and disseminate is resolved by recognizing that biofilm material properties are inherently heterogeneous. There are also mechanical aspects to the ways that infectious biofilms evade leukocyte phagocytosis. The possibility of alternative therapies for treating biofilm infections that work by reducing biofilm cohesion could (1) allow prevailing hydrodynamic shear to remove biofilm, (2) increase the efficacy of designed interventions for removing biofilms, (3) enable phagocytic engulfment of softened biofilm aggregates, and (4) improve phagocyte mobility and access to biofilm. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Dental Investigations: Efficiency of Nonsurgical Periodontal Therapy in Moderate Chronic Periodontitis

OpenAIRE

Mlachkova Antoaneta M.; Popova Christina L.

2014-01-01

INTRODUCTION: Chronic periodontitis is defined as an inflammatory disease of the supporting tissues of teeth caused by microorganisms in the dental biofilm, resulting in progressive destruction of the periodontal ligament and alveolar bone with pocket formation and gingival recession. Treatment of chronic periodontitis aims at arresting the inflammation and stopping the loss of attachment by removal and control of the supra- and subgingival biofilm and establishing a local environment and mic...

THE INFLUENCE OF SURFACE-FREE ENERGY ON SUPRAGINGIVAL AND SUBGINGIVAL PLAQUE MICROBIOLOGY - AN IN-VIVO STUDY ON IMPLANTS

NARCIS (Netherlands)

QUIRYNEN, M; VANDERMEI, HC; BOLLEN, CML; VANDENBOSSCHE, LH; DOORNBUSCH, GI; VANSTEENBERGHE, D; BUSSCHER, HJ

THE INFLUENCE OF SURFACE FREE ENERGY on supra- and subgingival plaque microbiology was examined in 9 patients with functional fixed prostheses supported by endosseous titanium implants. Two abutments (trans-mucosal part of the 2 stage implant) were replaced by either a new titanium abutment or a

Treatment of Oral Multispecies Biofilms by an Anti-Biofilm Peptide.

Science.gov (United States)

Wang, Zhejun; de la Fuente-Núñez, Cesar; Shen, Ya; Haapasalo, Markus; Hancock, Robert E W

2015-01-01

Human oral biofilms are multispecies microbial communities that exhibit high resistance to antimicrobial agents. Dental plaque gives rise to highly prevalent and costly biofilm-related oral infections, which lead to caries or other types of oral infections. We investigated the ability of the recently identified anti-biofilm peptide 1018 to induce killing of bacterial cells present within oral multispecies biofilms. At 10 μg/ml (6.5 μM), peptide 1018 was able to significantly (pbiofilm formation over 3 days. The activity of the peptide on preformed biofilms was found to be concentration-dependent since more than 60% of the total plaque biofilm cell population was killed by 10 μg/ml of peptide 1018 in 3 days, while at 5 μg/ml 50% of cells were dead and at 1 μg/ml the peptide triggered cell death in around 30% of the total bacterial population, as revealed by confocal microscopy. The presence of saliva did not affect peptide activity, since no statistically significant difference was found in the ability of peptide 1018 to kill oral biofilms using either saliva coated and non-saliva coated hydroxyapatite surfaces. Scanning electron microscopy experiments indicated that peptide 1018 induced cell lysis in plaque biofilms. Furthermore, combined treatment using peptide 1018 and chlorhexidine (CHX) increased the anti-biofilm activity of each compound compared to when these were used alone, resulting in >50% of the biofilm being killed and >35% being dispersed in only 3 minutes. Peptide 1018 may potentially be used by itself or in combination with CHX as a non-toxic and effective anti-biofilm agent for plaque disinfection in clinical dentistry.

Detection of Intracellular Adhesion (ica and Biofilm Formation Genes in Staphylococcus aureus Isolates from Clinical Samples

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Khadije Rezaie Keikhaie

2017-02-01

Full Text Available Introduction: Nosocomial infections that result in the formation of biofilms on the surfaces of biomedical implants are a leading cause of sepsis and are often associated with colonization of the implants by Staphylococcus epidermidis. Biofilm formation is thought to require two sequential steps: adhesion of cells to a solid substrate followed by cell-cell adhesion, creating multiple layers of cells. Intercellular adhesion requires the polysaccharide intercellular adhesion (PIA, which is composed of linear β-1, 6-linked glucosaminylglycans and can be synthesized in vitro from UDP-N-acetylglucosamine by products of the intercellular adhesion (ica locus. We have investigated a variety of Staphylococcus aureus strains and find that all strains tested contain the ica locus and that several can form biofilms in vitro. Material and Method: A total of 31 clinical S. aureus isolates were collected from Zabol, Iran. In vitro biofilm formation ability was determined by microliter tissue culture plates. All clinical isolates were examined for determination the ica locus by using PCR method. Result: The results of this study showed that 40 strains of Staphylococcus aureus, 12 strains carrying the gene Cocos icaA (30% and 8 strains carrying the gene icaD (20% and the number of five strains (12.5% containing both genes ica A and has been ica D. Conclusions:  S. aureus clinical isolates have different ability to form biofilm. This may be caused by the differences in the expression of biofilm related genes, genetic make-up and physiological conditions.

Unravelling the core microbiome of biofilms in cooling tower systems.

Science.gov (United States)

Di Gregorio, L; Tandoi, V; Congestri, R; Rossetti, S; Di Pippo, F

2017-11-01

In this study, next generation sequencing and catalyzed reporter deposition fluorescence in situ hybridization, combined with confocal microscopy, were used to provide insights into the biodiversity and structure of biofilms collected from four full-scale European cooling systems. Water samples were also analyzed to evaluate the impact of suspended microbes on biofilm formation. A common core microbiome, containing members of the families Sphingomonadaceae, Comamonadaceae and Hyphomicrobiaceae, was found in all four biofilms, despite the water of each coming from different sources (river and groundwater). This suggests that selection of the pioneer community was influenced by abiotic factors (temperature, pH) and tolerances to biocides. Members of the Sphingomonadaceae were assumed to play a key role in initial biofilm formation. Subsequent biofilm development was driven primarily by light availability, since biofilms were dominated by phototrophs in the two studied 'open' systems. Their interactions with other microbial populations then shaped the structure of the mature biofilm communities analyzed.

Staphylococcus aureus biofilms: recent developments in biofilm dispersal.

Science.gov (United States)

Lister, Jessica L; Horswill, Alexander R

2014-01-01

Staphylococcus aureus is a major cause of nosocomial and community-acquired infections and represents a significant burden on the healthcare system. S. aureus attachment to medical implants and host tissue, and the establishment of a mature biofilm, play an important role in the persistence of chronic infections. The formation of a biofilm, and encasement of cells in a polymer-based matrix, decreases the susceptibility to antimicrobials and immune defenses, making these infections difficult to eradicate. During infection, dispersal of cells from the biofilm can result in spread to secondary sites and worsening of the infection. In this review, we discuss the current understanding of the pathways behind biofilm dispersal in S. aureus, with a focus on enzymatic and newly described broad-spectrum dispersal mechanisms. Additionally, we explore potential applications of dispersal in the treatment of biofilm-mediated infections.

Treatment of Oral Multispecies Biofilms by an Anti-Biofilm Peptide.

Directory of Open Access Journals (Sweden)

Zhejun Wang

Full Text Available Human oral biofilms are multispecies microbial communities that exhibit high resistance to antimicrobial agents. Dental plaque gives rise to highly prevalent and costly biofilm-related oral infections, which lead to caries or other types of oral infections. We investigated the ability of the recently identified anti-biofilm peptide 1018 to induce killing of bacterial cells present within oral multispecies biofilms. At 10 μg/ml (6.5 μM, peptide 1018 was able to significantly (p50% of the biofilm being killed and >35% being dispersed in only 3 minutes. Peptide 1018 may potentially be used by itself or in combination with CHX as a non-toxic and effective anti-biofilm agent for plaque disinfection in clinical dentistry.

The in vivo biofilm

DEFF Research Database (Denmark)

Bjarnsholt, Thomas; Alhede, Maria; Alhede, Morten

2013-01-01

Bacteria can grow and proliferate either as single, independent cells or organized in aggregates commonly referred to as biofilms. When bacteria succeed in forming a biofilm within the human host, the infection often becomes very resistant to treatment and can develop into a chronic state. Biofilms...... have been studied for decades using various in vitro models, but it remains debatable whether such in vitro biofilms actually resemble in vivo biofilms in chronic infections. In vivo biofilms share several structural characteristics that differ from most in vitro biofilms. Additionally, the in vivo...... experimental time span and presence of host defenses differ from chronic infections and the chemical microenvironment of both in vivo and in vitro biofilms is seldom taken into account. In this review, we discuss why the current in vitro models of biofilms might be limited for describing infectious biofilms...

Dose requirements for UVC disinfection of catheter biofilms

DEFF Research Database (Denmark)

Bak, Jimmy; Ladefoged, Søren D.; Tvede, Michael

2009-01-01

Bacterial biofilms on permanent catheters are the major sources of infection. Exposure to ultraviolet-C (UVC) light has been proposed as a method for disinfecting the inner surface of catheters. Specification of a UVC-based device for in vivo disinfection is based on the knowledge of the required...... doses to kill catheter biofilm. Given these doses and the power of available UVC light sources, calculation of the necessary treatment times is then possible. To determine the required doses, contaminated urinary catheters were used as test samples and UVC treated in vitro. Patient catheters (n = 67......) were collected and cut into segments of equal size and treated with various UVC doses. After treatment, the biofilm was removed by scraping and quantified by counting colony forming units. Percentage killing rates were determined by calculating ratios between UVC-treated samples and controls (no UVC...

Depth-Resolved Nanospray Desorption Electrospray Ionization Mass Spectrometry in Biofilms

Science.gov (United States)

2015-11-16

metabolites and possibly proteins from biofilms at different depth. A microcapillary system was developed and tested for this goal. The...demonstrated that a dual microcapillary system can be used to collect nanoliter sample from the biofilms at 1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND...Spectrometry in Biofilms The views, opinions and/or findings contained in this report are those of the author(s) and should not contrued as an official

Salmonella biofilms

NARCIS (Netherlands)

Castelijn, G.A.A.

2013-01-01

Biofilm formation by Salmonellaspp. is a problem in the food industry, since biofilms may act as a persistent source of product contamination. Therefore the aim of this study was to obtain more insight in the processes involved and the factors contributing to Salmonellabiofilm

Visualizing Antimicrobials in Bacterial Biofilms: Three-Dimensional Biochemical Imaging Using TOF-SIMS.

Science.gov (United States)

Davies, Sarah K; Fearn, Sarah; Allsopp, Luke P; Harrison, Freya; Ware, Ecaterina; Diggle, Stephen P; Filloux, Alain; McPhail, David S; Bundy, Jacob G

2017-01-01

Bacterial biofilms are groups of bacteria that exist within a self-produced extracellular matrix, adhering to each other and usually to a surface. They grow on medical equipment and inserts such as catheters and are responsible for many persistent infections throughout the body, as they can have high resistance to many antimicrobials. Pseudomonas aeruginosa is an opportunistic pathogen that can cause both acute and chronic infections and is used as a model for research into biofilms. Direct biochemical methods of imaging of molecules in bacterial biofilms are of high value in gaining a better understanding of the fundamental biology of biofilms and biochemical gradients within them. Time of flight-secondary-ion mass spectrometry (TOF-SIMS) is one approach, which combines relatively high spatial resolution and sensitivity and can perform depth profiling analysis. It has been used to analyze bacterial biofilms but has not yet been used to study the distribution of antimicrobials (including antibiotics and the antimicrobial metal gallium) within biofilms. Here we compared two methods of imaging of the interior structure of P. aeruginosa in biological samples using TOF-SIMS, looking at both antimicrobials and endogenous biochemicals: cryosectioning of tissue samples and depth profiling to give pseudo-three-dimensional (pseudo-3D) images. The sample types included both simple biofilms grown on glass slides and bacteria growing in tissues in an ex vivo pig lung model. The two techniques for the 3D imaging of biofilms are potentially valuable complementary tools for analyzing bacterial infection. IMPORTANCE Modern analytical techniques are becoming increasingly important in the life sciences; imaging mass spectrometry offers the opportunity to gain unprecedented amounts of information on the distribution of chemicals in samples-both xenobiotics and endogenous compounds. In particular, simultaneous imaging of antibiotics (and other antimicrobial compounds) and bacterium

Physics of biofilms: the initial stages of biofilm formation and dynamics

International Nuclear Information System (INIS)

Lambert, Guillaume; Bergman, Andrew; Zhang, Qiucen; Bortz, David; Austin, Robert

2014-01-01

One of the physiological responses of bacteria to external stress is to assemble into a biofilm. The formation of a biofilm greatly increases a bacterial population's resistance to a hostile environment by shielding cells, for example, from antibiotics. In this paper, we describe the conditions necessary for the emergence of biofilms in natural environments and relate them to the emergence of biofilm formation inside microfluidic devices. We show that competing species of Escherichia coli bacteria form biofilms to spatially segregate themselves in response to starvation stress, and use in situ methods to characterize the physical properties of the biofilms. Finally, we develop a microfluidic platform to study the inter-species interactions and show how biofilm-mediated genetic interactions can improve a species’ resistance to external stress. (paper)

Role of Sediment Size and Biostratinomy on the Development of Biofilms in Recent Avian Vertebrate Remains

Directory of Open Access Journals (Sweden)

Joseph E. Peterson

2017-04-01

Full Text Available Microscopic soft tissues have been identified in fossil vertebrate remains collected from various lithologies. However, the diagenetic mechanisms to preserve such tissues have remained elusive. While previous studies have described infiltration of biofilms in Haversian and Volkmann's canals, biostratinomic alteration (e.g., trampling, and iron derived from hemoglobin as playing roles in the preservation processes, the influence of sediment texture has not previously been investigated. This study uses a Kolmogorov Smirnov Goodness-of-Fit test to explore the influence of biostratinomic variability and burial media against the infiltration of biofilms in bone samples. Controlled columns of sediment with bone samples were used to simulate burial and subsequent groundwater flow. Sediments used in this study include clay-, silt-, and sand-sized particles modeled after various fluvial facies commonly associated with fossil vertebrates. Extant limb bone samples obtained from Gallus gallus domesticus (Domestic Chicken buried in clay-rich sediment exhibit heavy biofilm infiltration, while bones buried in sands and silts exhibit moderate levels. Crushed bones exhibit significantly lower biofilm infiltration than whole bone samples. Strong interactions between biostratinomic alteration and sediment size are also identified with respect to biofilm development. Sediments modeling crevasse splay deposits exhibit considerable variability; whole-bone crevasse splay samples exhibit higher frequencies of high-level biofilm infiltration, and crushed-bone samples in modeled crevasse splay deposits display relatively high frequencies of low-level biofilm infiltration. These results suggest that sediment size, depositional setting, and biostratinomic condition play key roles in biofilm infiltration in vertebrate remains, and may influence soft tissue preservation in fossil vertebrates.

Role of sediment size and biostratinomy on the development of biofilms in recent avian vertebrate remains

Science.gov (United States)

Peterson, Joseph E.; Lenczewski, Melissa E.; Clawson, Steven R.; Warnock, Jonathan P.

2017-04-01

Microscopic soft tissues have been identified in fossil vertebrate remains collected from various lithologies. However, the diagenetic mechanisms to preserve such tissues have remained elusive. While previous studies have described infiltration of biofilms in Haversian and Volkmann’s canals, biostratinomic alteration (e.g., trampling), and iron derived from hemoglobin as playing roles in the preservation processes, the influence of sediment texture has not previously been investigated. This study uses a Kolmogorov Smirnov Goodness-of-Fit test to explore the influence of biostratinomic variability and burial media against the infiltration of biofilms in bone samples. Controlled columns of sediment with bone samples were used to simulate burial and subsequent groundwater flow. Sediments used in this study include clay-, silt-, and sand-sized particles modeled after various fluvial facies commonly associated with fossil vertebrates. Extant limb bone samples obtained from Gallus gallus domesticus (Domestic Chicken) buried in clay-rich sediment exhibit heavy biofilm infiltration, while bones buried in sands and silts exhibit moderate levels. Crushed bones exhibit significantly lower biofilm infiltration than whole bone samples. Strong interactions between biostratinomic alteration and sediment size are also identified with respect to biofilm development. Sediments modeling crevasse splay deposits exhibit considerable variability; whole-bone crevasse splay samples exhibit higher frequencies of high-level biofilm infiltration, and crushed-bone samples in modeled crevasse splay deposits display relatively high frequencies of low-level biofilm infiltration. These results suggest that sediment size, depositional setting, and biostratinomic condition play key roles in biofilm infiltration in vertebrate remains, and may influence soft tissue preservation in fossil vertebrates.

FISHing for gutta-percha-adhered biofilms in purulent post-treatment apical periodontitis.

Science.gov (United States)

Zehnder, M; Rechenberg, D-K; Thurnheer, T; Lüthi-Schaller, H; Belibasakis, G N

2017-06-01

This study investigated the possibility of depicting individual taxa in clinically relevant biofilms using fluorescent in situ hybridization (FISH). Gutta-percha samples were collected from the apical aspect of root canals associated with a chronic apical abscess (test samples, n = 8). Corresponding control samples were obtained from previously filled root canals with apparently normal periapical tissues (n = 3). The transport medium was investigated for detached biofilm fragments using FISH staining and conventional epifluorescence microscopy. Gutta-percha samples were stained by multiplex FISH, and inspected using confocal laser scanning microscopy. FISH of the transport medium confirmed the presence of the main species formerly identified by conventional methods in post-treatment purulent endodontic infections, most prominently Fusobacterium spp., Bacteroidetes and Prevotellaceae. Treponemes were identified in five of eight cases associated with purulent infections, but Enterococcus faecalis and Staphylococcus spp. were not identified. The biofilms on gutta-percha from root canals associated with apical periodontitis showed dense aggregates of variable composition. Control samples contained few, if any, bacteria in the transport medium, and featured no biofilms on the respective gutta-percha specimens. The current study revealed some direct, visual in situ information on the nature of biofilms associated with purulent periapical infections in man. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Fungal Biofilms: In Vivo Models for Discovery of Anti-Biofilm Drugs.

Science.gov (United States)

Nett, Jeniel E; Andes, David R

2015-06-01

During infection, fungi frequently transition to a biofilm lifestyle, proliferating as communities of surface-adherent aggregates of cells. Phenotypically, cells in a biofilm are distinct from free-floating cells. Their high tolerance of antifungals and ability to withstand host defenses are two characteristics that foster resilience. Biofilm infections are particularly difficult to eradicate, and most available antifungals have minimal activity. Therefore, the discovery of novel compounds and innovative strategies to treat fungal biofilms is of great interest. Although many fungi have been observed to form biofilms, the most well-studied is Candida albicans. Animal models have been developed to simulate common Candida device-associated infections, including those involving vascular catheters, dentures, urinary catheters, and subcutaneous implants. Models have also reproduced the most common mucosal biofilm infections: oropharyngeal and vaginal candidiasis. These models incorporate the anatomical site, immune components, and fluid dynamics of clinical niches and have been instrumental in the study of drug resistance and investigation of novel therapies. This chapter describes the significance of fungal biofilm infections, the animal models developed for biofilm study, and how these models have contributed to the development of new strategies for the eradication of fungal biofilm infections.

Effectiveness of adjunctive subgingival administration of amino acids and sodium hyaluronate gel on clinical and immunological parameters in the treatment of chronic periodontitis

Science.gov (United States)

Bevilacqua, Lorenzo; Eriani, Jessica; Serroni, Ilde; Liani, Giuliana; Borelli, Violetta; Castronovo, Gaetano; Di Lenarda, Roberto

2012-01-01

Summary Aims The aim of this clinical trial was to compare clinical and biochemical healing outcomes following ultrasonic mechanical instrumentation versus ultrasonic mechanical instrumentation associated with topical subgingival application of amino acids and sodium hyaluronate gel. Methods Eleven systemically healthy subjects with moderate-severe chronic periodontitis, who had four sites with pocket probing depth and clinical attachment level greater than or equal to 5 mm were randomly assigned to two different types of treatment: two pockets were treated with ultrasonic debridement (Control Group) and two pockets with ultrasonic mechanical instrumentation associated with 0,5 ml of amino acids and sodium hyaluronate gel (Test Group). Probing depth, clinical attachment level, plaque index and bleeding on probing were recorded at baseline, 45 and 90 days. Levels of calprotectin and myeloperoxidase activity in gingival crevicular fluid were assessed at baseline and on day 7 and 45. Results Statistical significance was found between baseline and day 45 in relation to probing depth reduction and bleeding on probing between groups for both of the tested treatments. Significant reductions in μg/sample of calprotectin and myeloperoxidase were found after 1-week and an increase at 45 days in both groups. There were no statistically significant differences between other variables evaluated in this study. Conclusions These data suggest that subgingival application of hyaluronic acid following ultrasonic mechanical instrumentation is beneficial for improving periodontal parameters. PMID:23087790

Biofilm Formation on Different Materials Used in Oral Rehabilitation.

Science.gov (United States)

Souza, Júlio C M; Mota, Raquel R C; Sordi, Mariane B; Passoni, Bernardo B; Benfatti, Cesar A M; Magini, Ricardo S

2016-01-01

The aim of this study was to evaluate the density and the morphological aspects of biofilms adhered to different materials applied in oral rehabilitation supported by dental implants. Sixty samples were divided into four groups: feldspar-based porcelain, CoCr alloy, commercially pure titanium grade IV and yttria-stabilized zirconia. Human saliva was diluted into BHI supplemented with sucrose to grow biofilms for 24 or 48 h. After this period, biofilm was removed by 1% protease treatment and then analyzed by spectrophotometry (absorbance), colony forming unit method (CFU.cm-2) and field-emission guns scanning electron microscopy (FEG-SEM). The highest values of absorbance and CFU.cm-2 were recorded on biofilms grown on CoCr alloys when compared to the other test materials for 24 or 48 h. Also, FEG-SEM images showed a high biofilm density on CoCr. There were no significant differences in absorbance and CFU.cm-2 between biofilms grown on zirconia, porcelain and titanium (p<0.05). Microbiological assays associated with microscopic analyses detected a higher accumulation of oral biofilms on CoCr-based materials than that on titanium or zirconia that are used for prosthetic structures.

Treatment of Oral Biofilms by a D-Enantiomeric Peptide.

Science.gov (United States)

Zhang, Tian; Wang, Zhejun; Hancock, Robert E W; de la Fuente-Núñez, César; Haapasalo, Markus

2016-01-01

Almost all dental diseases are caused by biofilms that consist of multispecies communities. DJK-5, which is a short D-enantiomeric, protease-resistant peptide with broad-spectrum anti-biofilm activity, was tested for its effect on oral multispecies biofilms. Peptide DJK-5 at 10 μg/mL effectively prevented the growth of these microbes in culture media in a time-dependent manner. In addition to the prevention of growth, peptide DJK-5 completely killed both Streptococcus mutans and Enterococcus faecalis suspended from biofilms after 30 minutes of incubation in liquid culture media. DJK-5 also led to the effective killing of microbes in plaque biofilm. The proportion of bacterial cells killed by 10 μg/mL of DJK-5 was similar after 1 and 3 days, both exceeding 85%. DJK-5 was able to significantly prevent biofilm formation over 3 days (P = 0.000). After 72 hours of exposure, DJK-5 significantly reduced and almost completely prevented plaque biofilm production by more than 90% of biovolume compared to untreated controls (P = 0.000). The proportion of dead biofilm bacteria at the 10 μg/mL DJK-5 concentration was similar for 1- and 3-day-old biofilms, whereby >86% of the bacteria were killed. DJK-5 was also able to kill >79% and >85% of bacteria, respectively, after one-time and three brief treatments of 3-day-old biofilms. The combination of DJK-5 and chlorhexidine showed the best bacterial killing among all treatments, with ~83% and >88% of bacterial cells killed after 1 and 3 minutes, respectively. No significant difference was found in the percentage of biofilm killing amongst three donor plaque samples after DJK-5 treatment. In particular, DJK-5 showed strong performance in inhibiting biofilm development and eradicating pre-formed oral biofilms compared to L-enantiomeric peptide 1018. DJK-5 was very effective against oral biofilms when used alone or combined with chlorhexidine, and may be a promising agent for use in oral anti-biofilm strategies in the future.

Pseudomonas aeruginosa Biofilm Infections

DEFF Research Database (Denmark)

Rybtke, Morten; Hultqvist, Louise Dahl; Givskov, Michael

2015-01-01

Studies of biopsies from infectious sites, explanted tissue and medical devises have provided evidence that biofilms are the underlying cause of a variety of tissue-associated and implant-associated recalcitrant human infections. With a need for novel anti-biofilm treatment strategies, research...... in biofilm infection microbiology, biofilm formation mechanisms and biofilm-associated antimicrobial tolerance has become an important area in microbiology. Substantial knowledge about biofilm formation mechanisms, biofilm-associated antimicrobial tolerance and immune evasion mechanisms has been obtained...... through work with biofilms grown in in vitro experimental setups, and the relevance of this information in the context of chronic infections is being investigated by the use of animal models of infection. Because our current in vitro experimental setups and animal models have limitations, new advanced...

Evaluation of microbial biofilm communities from an Alberta oil sands tailings pond.

Science.gov (United States)

Golby, Susanne; Ceri, Howard; Gieg, Lisa M; Chatterjee, Indranil; Marques, Lyriam L R; Turner, Raymond J

2012-01-01

Bitumen extraction from the oil sands of Alberta has resulted in millions of cubic meters of waste stored on-site in tailings ponds. Unique microbial ecology is expected in these ponds, which may be key to their bioremediation potential. We considered that direct culturing of microbes from a tailings sample as biofilms could lead to the recovery of microbial communities that provide good representation of the ecology of the tailings. Culturing of mixed species biofilms in vitro using the Calgary Biofilm Device (CBD) under aerobic, microaerobic, and anaerobic growth conditions was successful both with and without the addition of various growth nutrients. Denaturant gradient gel electrophoresis and 16S rRNA gene pyrotag sequencing revealed that unique mixed biofilm communities were recovered under each incubation condition, with the dominant species belonging to Pseudomonas, Thauera, Hydrogenophaga, Rhodoferax, and Acidovorax. This work used an approach that allowed organisms to grow as a biofilm directly from a sample collected of their environment, and the biofilms cultivated in vitro were representative of the endogenous environmental community. For the first time, representative environmental mixed species biofilms have been isolated and grown under laboratory conditions from an oil sands tailings pond environment and a description of their composition is provided.

Effect of engineered environment on microbial community structure in biofilter and biofilm on reverse osmosis membrane.

Science.gov (United States)

Jeong, Sanghyun; Cho, Kyungjin; Jeong, Dawoon; Lee, Seockheon; Leiknes, TorOve; Vigneswaran, Saravanamuthu; Bae, Hyokwan

2017-11-01

Four dual media filters (DMFs) were operated in a biofiltration mode with different engineered environments (DMF I and II: coagulation with/without acidification and DMF III and IV: without/with chlorination). Designed biofilm enrichment reactors (BERs) containing the removable reverse osmosis (RO) coupons, were connected at the end of the DMFs in parallel to analyze the biofilm on the RO membrane by DMF effluents. Filtration performances were evaluated in terms of dissolved organic carbon (DOC) and assimilable organic carbon (AOC). Organic foulants on the RO membrane were also quantified and fractionized. The bacterial community structures in liquid (seawater and effluent) and biofilm (DMF and RO) samples were analyzed using 454-pyrosequencing. The DMF IV fed with the chlorinated seawater demonstrated the highest reductions of DOC including LMW-N as well as AOC among the other DMFs. The DMF IV was also effective in reducing organic foulants on the RO membrane surface. The bacterial community structure was grouped according to the sample phase (i.e., liquid and biofilm samples), sampling location (i.e., DMF and RO samples), and chlorination (chlorinated and non-chlorinated samples). In particular, the biofilm community in the DMF IV differed from the other DMF treatments, suggesting that chlorination exerted as stronger selective pressure than pH adjustment or coagulation on the biofilm community. In the DMF IV, several chemoorganotrophic chlorine-resistant biofilm-forming bacteria such as Hyphomonas, Erythrobacter, and Sphingomonas were predominant, and they may enhance organic carbon degradation efficiency. Diverse halophilic or halotolerant organic degraders were also found in other DMFs (i.e., DMF I, II, and III). Various kinds of dominant biofilm-forming bacteria were also investigated in RO membrane samples; the results provided possible candidates that cause biofouling when DMF process is applied as the pretreatment option for the RO process. Copyright �

Effect of engineered environment on microbial community structure in biofilter and biofilm on reverse osmosis membrane

KAUST Repository

Jeong, Sanghyun

2017-07-25

Four dual media filters (DMFs) were operated in a biofiltration mode with different engineered environments (DMF I and II: coagulation with/without acidification and DMF III and IV: without/with chlorination). Designed biofilm enrichment reactors (BERs) containing the removable reverse osmosis (RO) coupons, were connected at the end of the DMFs in parallel to analyze the biofilm on the RO membrane by DMF effluents. Filtration performances were evaluated in terms of dissolved organic carbon (DOC) and assimilable organic carbon (AOC). Organic foulants on the RO membrane were also quantified and fractionized. The bacterial community structures in liquid (seawater and effluent) and biofilm (DMF and RO) samples were analyzed using 454-pyrosequencing. The DMF IV fed with the chlorinated seawater demonstrated the highest reductions of DOC including LMW-N as well as AOC among the other DMFs. The DMF IV was also effective in reducing organic foulants on the RO membrane surface. The bacterial community structure was grouped according to the sample phase (i.e., liquid and biofilm samples), sampling location (i.e., DMF and RO samples), and chlorination (chlorinated and non-chlorinated samples). In particular, the biofilm community in the DMF IV differed from the other DMF treatments, suggesting that chlorination exerted as stronger selective pressure than pH adjustment or coagulation on the biofilm community. In the DMF IV, several chemoorganotrophic chlorine-resistant biofilm-forming bacteria such as Hyphomonas, Erythrobacter, and Sphingomonas were predominant, and they may enhance organic carbon degradation efficiency. Diverse halophilic or halotolerant organic degraders were also found in other DMFs (i.e., DMF I, II, and III). Various kinds of dominant biofilm-forming bacteria were also investigated in RO membrane samples; the results provided possible candidates that cause biofouling when DMF process is applied as the pretreatment option for the RO process.

The synergistic effect of ultrasonic activation and irrigation on Enterococcus faecalis biofilm

Directory of Open Access Journals (Sweden)

Abeer A Al-Mahdi

2016-01-01

Full Text Available Aim: The aim of this investigation was to compare the efficacy of passive ultrasonic irrigation (PUI with either 2.5% sodium hypochlorite (NaOCl or saline, with that of conventional syringe irrigation on intraradicular Enterococcus faecalis biofilm. Materials and Methods: Biofilms of E. faecalis were established over 21 days in 80 single roots that had undergone biomechanical preparation followed by gamma radiation. Biofilms were treated for 1 min with 2.5% NaOCl/PUI (Group 1, 2.5% NaOCl (Group 2, sterile saline/PUI (Group 3, and sterile saline (Group 4. The positive control (n = 4 was used to confirm the presence of biofilm before various treatments. Additional four samples that served as a negative control were used to confirm the sterility of the samples. Biofilm eradication was evaluated by Colony Forming Unit (CFU quantification and scanning electron microscopy (SEM. Results: The median of CFUs of S1 was significantly higher than that of S2 in all experimental groups. SEM examination showed a significant difference between the positive control and the experimental groups (P < 0.001, with the highest score of biofilm in the positive control group followed by Group 4 and both groups were not statistically significant from each other (P = 0.067. Following various treatments, the highest scores of biofilm were observed in the coronal third and the least were in the apical third. Conclusions: PUI did not increase the effectiveness of NaOCl irrigation on biofilm removal, however, PUI enhanced biofilm disturbance when used with saline. The least mean score of remaining biofilm was in the apical third of all treatment groups compared to other thirds.

Visualizing the dental biofilm matrix by means of fluorescence lectin-binding analysis

DEFF Research Database (Denmark)

Tawakoli, Pune Nina; Neu, Thomas R; Busck, Mette Marie

2017-01-01

lectins to visualize and quantify extracellular glycoconjugates in dental biofilms. Lectin binding was screened on pooled supragingival biofilm samples collected from 76 subjects using confocal microscopy. FLBA was then performed with 10 selected lectins on biofilms grown in situ for 48 h in the absence......The extracellular matrix is a poorly studied, yet important component of dental biofilms. Fluorescence lectin-binding analysis (FLBA) is a powerful tool to characterize glycoconjugates in the biofilm matrix. This study aimed to systematically investigate the ability of 75 fluorescently labeled......-biofilms: Aleuria aurantia (AAL), Calystega sepiem (Calsepa), Lycopersicon esculentum (LEA), Morniga-G (MNA-G) and Helix pomatia (HPA). No significant correlation between the binding of specific lectins and bacterial composition was found. Fluorescently labeled lectins enable the visualization of glycoconjugates...

Comparison of the Candida albicans and biofilm formation amount on natural tooth,

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Serdar Polat

2012-03-01

Full Text Available Objective: This study compared the retention of the Candidaalbicans and biofilm formation on natural teeth, porcelainand acrylic resin.Materials and methods: Samples are taken with thesterile ecuvion sticks from the buccal embrasures of thenatural tooth, porcelain and acrylic. The biofilm productionof candida reproducing strains was determined withmicroplate method. Samples are settled in 0.5 ml sterilephosphate buffered saline. Reproduction detected coloniesdefined to species in accordance with their macroscopicand microscopic features and germ tube test inmicrobiology laboratory.Results: There was significant differences for retention ofCandida albicans and biofilm formation on the surface oftooth, porcelain and acrylic (p<0.05.Conclusion: Adherence of Candida albicans and biofilmformation on the porcelain significantly less than naturaltooth and acrylic, and retention and biofilm formation onthe tooth less than acrylic.

Biofilm-forming ability profiling of Staphylococcus aureus and Staphylococcus epidermidis mastitis isolates

DEFF Research Database (Denmark)

Oliveira, M; Bexiga, R; Nunes, S F

2006-01-01

of FISH to artificially contaminated milk samples allowed the direct observation of biofilm production by 37.5% isolates, showing total agreement with the CRA results. This method better mimics the in vivo conditions, especially in terms of the presence of calcium and iron, which in high concentrations...... hybridisation (FISH) protocol that would allow the direct observation of biofilm formation in milk samples. The analysis of phenotypic expression in Congo Red Agar (CRA) and by FISH, showed that 37.5% of the S. aureus isolates produced biofilm, while by optical density measurement only 18.75% isolates revealed...

Non-Tuberculous Mycobacteria multispecies biofilms in cystic fibrosis: development of an in vitro Mycobacterium abscessus and Pseudomonas aeruginosa dual species biofilm model.

Science.gov (United States)

Rodríguez-Sevilla, Graciela; García-Coca, Marta; Romera-García, David; Aguilera-Correa, John Jairo; Mahíllo-Fernández, Ignacio; Esteban, Jaime; Pérez-Jorge, Concepción

2018-04-01

Lung disease in cystic fibrosis (CF) is characterized by the progressive colonization of the respiratory tract by different bacteria, which develop polymicrobial biofilms. In the past decades, there has been an increase in the number of CF patients infected with Non-Tuberculous Mycobacteria (NTM). Although Mycobacterium abscessus is the main NTM isolated globally, little is known about M. abscessus multispecies biofilm formation. In the present study we developed an in vitro model to study the phenotypic characteristics of biofilms formed by M. abscessus and Pseudomonas aeruginosa, a major pathogen in CF. For that purpose, dual species biofilms were grown on polycarbonate membranes with a fixed concentration of P. aeruginosa and different inoculums of M. abscessus. The biofilms were sampled at 24, 48, and 72 h and bacteria were quantified in specific media. The results revealed that the increasing initial concentration of M. abscessus in dual species biofilms had an effect on its population only at 24 and 48 h, whereas P. aeruginosa was not affected by the different concentrations used of M. abscessus. Time elapsed increased biofilm formation of both species, specially between 24 and 48 h. According to the results, the conditions to produce a mature dual species biofilm in which the relative species distribution remained stable were 72 h growth of the mixed microbial culture at a 1:1 ratio. A significant decrease in mycobacterial population in dual compared to single species biofilms was found, suggesting that P. aeruginosa has a negative influence on M. abscessus. Finally, in a proof of concept experiment, young and mature dual species biofilms were exposed to clarithromycin. Copyright © 2018 Elsevier GmbH. All rights reserved.

Inferring energy sources in constructed wetlands through stable isotope analysis of microbial biofilms

Energy Technology Data Exchange (ETDEWEB)

Jurkowski, K.; Ciborowski, J. [Windsor Univ., ON (Canada); Daly, C. [Suncor Energy, Fort McMurray, AB (Canada)

2010-07-01

This study presented a novel method of sequestering the microbial biofilm in constructed wetland ecosystems. Artificial substrates were fixed within 8 wetlands differing in age and construction materials over a 2 year period at oil sands lease sites in northeastern Alberta. Autotrophic and heterotrophic biofilm samples were collected from both the subsurface and epibenthic zones of the pipe surfaces of each submerged substrate assembly. A mixing model of d13C, d15N and d34S isotopic signatures was used to assess the contribution of 4 potential nutrient sources of the biofilm. Samples included dominant living and senescent emergent as well as submergent macrophytes, particulate organic matter, dissolved organic carbon, and invertebrates. The samples were collected to compare the biofilm signatures of each wetland in relation to the heterotrophic processes caused by the assimilation of oil sands-derived hydrocarbons and autochthonous detrital pools.

Inferring energy sources in constructed wetlands through stable isotope analysis of microbial biofilms

International Nuclear Information System (INIS)

Jurkowski, K.; Ciborowski, J.; Daly, C.

2010-01-01

This study presented a novel method of sequestering the microbial biofilm in constructed wetland ecosystems. Artificial substrates were fixed within 8 wetlands differing in age and construction materials over a 2 year period at oil sands lease sites in northeastern Alberta. Autotrophic and heterotrophic biofilm samples were collected from both the subsurface and epibenthic zones of the pipe surfaces of each submerged substrate assembly. A mixing model of d13C, d15N and d34S isotopic signatures was used to assess the contribution of 4 potential nutrient sources of the biofilm. Samples included dominant living and senescent emergent as well as submergent macrophytes, particulate organic matter, dissolved organic carbon, and invertebrates. The samples were collected to compare the biofilm signatures of each wetland in relation to the heterotrophic processes caused by the assimilation of oil sands-derived hydrocarbons and autochthonous detrital pools.

Association between pathogens from tracheal aspirate and oral biofilm of patients on mechanical ventilation

Directory of Open Access Journals (Sweden)

Luana Carneiro Diniz SOUZA

2017-06-01

Full Text Available Abstract The aim of this study was to detect possible associations between respiratory pathogens from tracheal aspirate and oral biofilm samples in intubated patients in an intensive care unit (ICU, and to identify the most common respiratory pathogens in oral biofilm, particularly in patients that developed ventilator-associated pneumonia (VAP. Two oral biofilm samples were collected from the tongue of intubated patients (at admission and after 48 hours and analyzed by culture with the Antibiotic Sensitivity Test. The results from the tongue biofilm samples were compared with the tracheal secretions samples. A total of 59.37% of patients exhibited the same species of pathogens in their tracheal aspirate and oral biofilm, of which 8 (42.1% developed VAP, 10 (52.63% did not develop pneumonia and one (5.26% had aspiration pneumonia. There was a statistically significant association between presence of microorganisms in the tracheal and mouth samples for the following pathogens: Klebsiella pneumoniae, Candida albicans, Pseudomonas aeruginosa, Enterobacter gergoviae, Streptococcus spp and Serratia marcescens (p < 0.05. Pathogens that are present in tracheal aspirates of intubated patients can be detected in their oral cavity, especially in those who developed VAP or aspiration pneumonia. Thus, the results indicate that an improved oral care in these patients could decrease ICU pneumonia rates.

Association between pathogens from tracheal aspirate and oral biofilm of patients on mechanical ventilation.

Science.gov (United States)

Souza, Luana Carneiro Diniz; Mota, Vanise Barros Rodrigues da; Carvalho, Alícia Valéria Dos Santos Zaranza de; Corrêa, Rita da Graça Carvalhal Frazão; Libério, Silvana Amado; Lopes, Fernanda Ferreira

2017-06-05

The aim of this study was to detect possible associations between respiratory pathogens from tracheal aspirate and oral biofilm samples in intubated patients in an intensive care unit (ICU), and to identify the most common respiratory pathogens in oral biofilm, particularly in patients that developed ventilator-associated pneumonia (VAP). Two oral biofilm samples were collected from the tongue of intubated patients (at admission and after 48 hours) and analyzed by culture with the Antibiotic Sensitivity Test. The results from the tongue biofilm samples were compared with the tracheal secretions samples. A total of 59.37% of patients exhibited the same species of pathogens in their tracheal aspirate and oral biofilm, of which 8 (42.1%) developed VAP, 10 (52.63%) did not develop pneumonia and one (5.26%) had aspiration pneumonia. There was a statistically significant association between presence of microorganisms in the tracheal and mouth samples for the following pathogens: Klebsiella pneumoniae, Candida albicans, Pseudomonas aeruginosa, Enterobacter gergoviae, Streptococcus spp and Serratia marcescens (p aspirates of intubated patients can be detected in their oral cavity, especially in those who developed VAP or aspiration pneumonia. Thus, the results indicate that an improved oral care in these patients could decrease ICU pneumonia rates.

Biofilm antifungal susceptibility of Candida urine isolated from ambulatory patients

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Débora da Luz Becker

2016-07-01

Full Text Available Background and Objectives: the association between the biofilm formations an antifungal resistance has been suggested to be an important factor in the pathogenesis of several Candida species. Besides, studies have included invasive candidiasis from hospitalized patients; however there are few studies that evaluated the species distribution, antifungal susceptibility and biofilm formation of Candida species isolated from ambulatory patients. Thus, the aim of this study was to evaluate whether biofilm producing contributes to antifungal resistance in Candida isolates from urine sample obtained from ambulatory patients. Methods: During one year, 25 urine samples positive for yeast were collected, stored and plated on agar supplemented with chloramphenicol and Sabouread left at room temperature for 5 days for subsequent: 52% (13/25 were C. albicans, 36% (9/25 C. tropicalis, 8% (2/25 C. krusei and 4% (1/25 C. parapsilosis. Results: The ability to form biofilm was detected in 23 (92% of the yeast studied and 15.4% (2/13 of C. albicans were fluconazole (FLU and ketoconazole (KET resistant, while 11.1% (1/9 of C. tropicalis were ketoconazole resistant and were anidulafungin (ANI non-susceptible. Conclusion: our results showed the high capacity for biofilm formation among Candida isolates from ambulatory patients.

Performance of fast-setting impression materials in the reproduction of subgingival tooth surfaces without soft tissue retraction.

Science.gov (United States)

Rudolph, Heike; Röhl, Andreas; Walter, Michael H; Luthardt, Ralph G; Quaas, Sebastian

2014-01-01

Fast-setting impression materials may be prone to inaccuracies due to accidental divergence from the recommended mixing protocol. This prospective randomized clinical trial aimed to assess three-dimensional (3D) deviations in the reproduction of subgingival tooth surfaces and to determine the effect of either following or purposely diverging from the recommended mixing procedure for a fast-setting addition-curing silicone (AS) and fast-setting polyether (PE). After three impressions each were taken from 96 participants, sawcut gypsum casts were fabricated with a standardized procedure and then optically digitized. Data were assessed with a computer-aided 3D analysis. For AS impressions, multivariate analysis of variance revealed a significant influence of the individual tooth and the degree to which the recommended mixing protocol was violated. For PE impressions, the ambient air temperature and individual tooth showed significant effects, while divergence from the recommended mixing protocol was not of significance. The fast-setting PE material was not affected by changes in the recommended mixing protocol. For the two fast-setting materials examined, no divergences from the recommended mixing protocol of less than 2 minutes led to failures in the reproduction of the subgingival tooth surfaces.

Artificial biofilms establish the role of matrix interactions in staphylococcal biofilm assembly and disassembly

Science.gov (United States)

Stewart, Elizabeth J.; Ganesan, Mahesh; Younger, John G.; Solomon, Michael J.

2015-01-01

We demonstrate that the microstructural and mechanical properties of bacterial biofilms can be created through colloidal self-assembly of cells and polymers, and thereby link the complex material properties of biofilms to well understood colloidal and polymeric behaviors. This finding is applied to soften and disassemble staphylococcal biofilms through pH changes. Bacterial biofilms are viscoelastic, structured communities of cells encapsulated in an extracellular polymeric substance (EPS) comprised of polysaccharides, proteins, and DNA. Although the identity and abundance of EPS macromolecules are known, how these matrix materials interact with themselves and bacterial cells to generate biofilm morphology and mechanics is not understood. Here, we find that the colloidal self-assembly of Staphylococcus epidermidis RP62A cells and polysaccharides into viscoelastic biofilms is driven by thermodynamic phase instability of EPS. pH conditions that induce phase instability of chitosan produce artificial S. epidermidis biofilms whose mechanics match natural S. epidermidis biofilms. Furthermore, pH-induced solubilization of the matrix triggers disassembly in both artificial and natural S. epidermidis biofilms. This pH-induced disassembly occurs in biofilms formed by five additional staphylococcal strains, including three clinical isolates. Our findings suggest that colloidal self-assembly of cells and matrix polymers produces biofilm viscoelasticity and that biofilm control strategies can exploit this mechanism. PMID:26272750

Morphological evidence of biofilm formation in Greenlanders with chronic suppurative otitis media

DEFF Research Database (Denmark)

Homøe, Preben; Bjarnsholt, Thomas; Wessman, Marcus

2009-01-01

Biofilm may explain the recurrences and recalcitrant episodes of otorrhea in chronic suppurative otitis media (CSOM). This study investigates bacterial biofilm in Greenlanders with CSOM and chronic otitis media with effusion (COME). The study is partly blinded, prospective and retrospective. Six...... were analyzed with microscopy and peptide nucleic acid fluorescence in situ hybridization (PNA-FISH). Biofilm was confirmed in 83% of CSOM smears but in none of the COME smears. Mucosal biofilm was confirmed in 80% of the biopsies from adults with CSOM. This study provides direct morphological evidence...... of biofilm in samples from human CSOM. This may help to explain the microbiological mechanisms of the disease and alter the treatment strategy in the future....

Biofilms on Hospital Shower Hoses: Characterization and ...

Science.gov (United States)

Although the source of drinking water used in hospitals is commonly, biofilms on water pipelines are refuge to bacteria that survive different disinfection strategies. Drinking water (DW) biofilms are well known to harbor opportunistic pathogens, however, these biofilm communities remain poorly characterized by culture-independent approaches that circumvent the limitations of conventional monitoring efforts. Hence, the frequency of pathogens in DW biofilms and how biofilm members withstand high doses of disinfectants and/or chlorine residuals in the water supply remain speculative, but directly impact public health. The aim of this study was to characterize the composition of microbial communities growing on five hospital shower hoses using both culture-dependent and culture-independent techniques. Two different sequence-based methods were used to characterize the bacterial fractions: 16S rRNA gene sequencing of bacterial cultures and next generation sequencing of metagenomes. Based on the metagenomic data, we found that Mycobacterium-like species was the abundant bacterial taxa that overlapped among the five samples. We also recovered the draft genome of a novel Mycobacterium species, closely related to opportunistic pathogenic nontuberculous mycobacteria, M. rhodesiae and M. tusciae, in addition to other, less abundant species. In contrast, the cultured fraction was mostly affiliated to Proteobacteria, such as members of the Sphingomonas, Blastomonas and Porph

Hydraulic resistance of biofilms

KAUST Repository

Dreszer, C.

2013-02-01

Biofilms may interfere with membrane performance in at least three ways: (i) increase of the transmembrane pressure drop, (ii) increase of feed channel (feed-concentrate) pressure drop, and (iii) increase of transmembrane passage. Given the relevance of biofouling, it is surprising how few data exist about the hydraulic resistance of biofilms that may affect the transmembrane pressure drop and membrane passage. In this study, biofilms were generated in a lab scale cross flow microfiltration system at two fluxes (20 and 100Lm-2h-1) and constant cross flow (0.1ms-1). As a nutrient source, acetate was added (1.0mgL-1 acetate C) besides a control without nutrient supply. A microfiltration (MF) membrane was chosen because the MF membrane resistance is very low compared to the expected biofilm resistance and, thus, biofilm resistance can be determined accurately. Transmembrane pressure drop was monitored. As biofilm parameters, thickness, total cell number, TOC, and extracellular polymeric substances (EPS) were determined, it was demonstrated that no internal membrane fouling occurred and that the fouling layer actually consisted of a grown biofilm and was not a filter cake of accumulated bacterial cells. At 20Lm-2h-1 flux with a nutrient dosage of 1mgL-1 acetate C, the resistance after 4 days reached a value of 6×1012m-1. At 100Lm-2h-1 flux under the same conditions, the resistance was 5×1013m-1. No correlation of biofilm resistance to biofilm thickness was found; Biofilms with similar thickness could have different resistance depending on the applied flux. The cell number in biofilms was between 4×107 and 5×108 cellscm-2. At this number, bacterial cells make up less than a half percent of the overall biofilm volume and therefore did not hamper the water flow through the biofilm significantly. A flux of 100Lm-2h-1 with nutrient supply caused higher cell numbers, more biomass, and higher biofilm resistance than a flux of 20Lm-2h-1. However, the biofilm thickness

Effect of biofilm formation, and biocorrosion on denture base fractures.

Science.gov (United States)

Sahin, Cem; Ergin, Alper; Ayyildiz, Simel; Cosgun, Erdal; Uzun, Gulay

2013-05-01

The aim of this study was to investigate the destructive effects of biofilm formation and/or biocorrosive activity of 6 different oral microorganisms. Three different heat polymerized acrylic resins (Ivocap Plus, Lucitone 550, QC 20) were used to prepare three different types of samples. Type "A" samples with "V" type notch was used to measure the fracture strength, "B" type to evaluate the surfaces with scanning electron microscopy and "C" type for quantitative biofilm assay. Development and calculation of biofilm covered surfaces on denture base materials were accomplished by SEM and quantitative biofilm assay. According to normality assumptions ANOVA or Kruskal-Wallis was selected for statistical analysis (α=0.05). Significant differences were obtained among the adhesion potential of 6 different microorganisms and there were significant differences among their adhesion onto 3 different denture base materials. Compared to the control groups after contamination with the microorganisms, the three point bending test values of denture base materials decreased significantly (P.05). All the tested microorganisms had destructive effect over the structure and composition of the denture base materials.

beta-lactamase producing bacteria in the subgingival microflora of adult patients with periodontitis. A comparison between Spain and The Netherlands

NARCIS (Netherlands)

Herrera, D; van Winkelhoff, AJ; Dellemijn-Kippuw, N; Winkel, EG; Sanz, M

Background/aims: Countries with a high per capita antibiotic use frequently demonstrate a high level of drug resistance. The aim of this study was to compare the prevalence and levels of beta-lactamase producing bacteria in the subgingival microflora in adult patients with periodontitis in Spain and

Antibacterial effect of taurolidine (2%) on established dental plaque biofilm.

Science.gov (United States)

Arweiler, Nicole Birgit; Auschill, Thorsten Mathias; Sculean, Anton

2012-04-01

Preliminary data have suggested that taurolidine may bear promising disinfectant properties for the therapy of bacterial infections. However, at present, the potential antibacterial effect of taurolidine on the supragingival plaque biofilm is unknown. To evaluate the antibacterial effect of taurolidine on the supragingival plaque biofilm using the vital fluorescence technique and to compare it with the effect of NaCl and chlorhexidine (CHX), 18 subjects had to refrain from all mechanical and chemical hygiene measures for 24 h. A voluminous supragingival plaque sample was taken from the buccal surfaces of the lower molars and wiped on an objective slide. The sample was then divided into three equal parts and mounted with one of the three test or control preparations (a) NaCl, (b) taurolidine 2% and (c) CHX 0.2%. After a reaction time of 2 min, the test solutions were sucked of. Subsequently, the plaque biofilm was stained with fluorescence dye and vitality of the plaque flora was evaluated under the fluorescence microscope (VF%). Plaque samples treated with NaCl showed a mean VF of 82.42 ± 6.04%. Taurolidine affected mean VF with 47.57 ± 16.60% significantly (p plaque biofilm which was, however, not as pronounced as that of CHX.

Biofilm Fixed Film Systems

Directory of Open Access Journals (Sweden)

Dipesh Das

2011-09-01

Full Text Available The work reviewed here was published between 2008 and 2010 and describes research that involved aerobic and anoxic biofilm treatment of water pollutants. Biofilm denitrification systems are covered when appropriate. References catalogued here are divided on the basis of fundamental research area or reactor types. Fundamental research into biofilms is presented in two sections, Biofilm Measurement and Characterization and Growth and Modeling. The reactor types covered are: trickling filters, rotating biological contactors, fluidized bed bioreactors, submerged bed biofilm reactors, biological granular activated carbon, membrane bioreactors, and immobilized cell reactors. Innovative reactors, not easily classified, are then presented, followed by a section on biofilms on sand, soil and sediment.

Antibiotic resistance in Pseudomonas aeruginosa biofilms: towards the development of novel anti-biofilm therapies.

Science.gov (United States)

Taylor, Patrick K; Yeung, Amy T Y; Hancock, Robert E W

2014-12-10

The growth of bacteria as structured aggregates termed biofilms leads to their protection from harsh environmental conditions such as physical and chemical stresses, shearing forces, and limited nutrient availability. Because of this highly adapted ability to survive adverse environmental conditions, bacterial biofilms are recalcitrant to antibiotic therapies and immune clearance. This is particularly problematic in hospital settings where biofilms are a frequent cause of chronic and device-related infections and constitute a significant burden on the health-care system. The major therapeutic strategy against infections is the use of antibiotics, which, due to adaptive resistance, are often insufficient to clear biofilm infections. Thus, novel biofilm-specific therapies are required. Specific features of biofilm development, such as surface adherence, extracellular matrix formation, quorum sensing, and highly regulated biofilm maturation and dispersal are currently being studied as targets to be exploited in the development of novel biofilm-specific treatments. Using Pseudomonas aeruginosa for illustrative purposes, this review highlights the antibiotic resistance mechanisms of biofilms, and discusses current research into novel biofilm-specific therapies. Copyright © 2014 Elsevier B.V. All rights reserved.

The Biofilm Challenge

DEFF Research Database (Denmark)

Alhede, Maria; Alhede, Morten

2014-01-01

The concept of biofilms has emerged in the clinical setting during the last decade. Infections involving biofilms have been documented in all parts of the human body, and it is currently believed that the presence of biofilm-forming bacteria is equivalent to chronic infection. A quick Pubmed search...

Compaction and relaxation of biofilms

KAUST Repository

Valladares Linares, R.

2015-06-18

Operation of membrane systems for water treatment can be seriously hampered by biofouling. A better characterization of biofilms in membrane systems and their impact on membrane performance may help to develop effective biofouling control strategies. The objective of this study was to determine the occurrence, extent and timescale of biofilm compaction and relaxation (decompaction), caused by permeate flux variations. The impact of permeate flux changes on biofilm thickness, structure and stiffness was investigated in situ and non-destructively with optical coherence tomography using membrane fouling monitors operated at a constant crossflow velocity of 0.1 m s−1 with permeate production. The permeate flux was varied sequentially from 20 to 60 and back to 20 L m−2 h−1. The study showed that the average biofilm thickness on the membrane decreased after elevating the permeate flux from 20 to 60 L m−2 h−1 while the biofilm thickness increased again after restoring the original flux of 20 L m−2 h−1, indicating the occurrence of biofilm compaction and relaxation. Within a few seconds after the flux change, the biofilm thickness was changed and stabilized, biofilm compaction occurred faster than the relaxation after restoring the original permeate flux. The initial biofilm parameters were not fully reinstated: the biofilm thickness was reduced by 21%, biofilm stiffness had increased and the hydraulic biofilm resistance was elevated by 16%. Biofilm thickness was related to the hydraulic biofilm resistance. Membrane performance losses are related to the biofilm thickness, density and morphology, which are influenced by (variations in) hydraulic conditions. A (temporarily) permeate flux increase caused biofilm compaction, together with membrane performance losses. The impact of biofilms on membrane performance can be influenced (increased and reduced) by operational parameters. The article shows that a (temporary) pressure increase leads to more

Proteus mirabilis biofilm - qualitative and quantitative colorimetric methods-based evaluation.

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Kwiecinska-Piróg, Joanna; Bogiel, Tomasz; Skowron, Krzysztof; Wieckowska, Ewa; Gospodarek, Eugenia

2014-01-01

Proteus mirabilis strains ability to form biofilm is a current topic of a number of research worldwide. In this study the biofilm formation of P. mirabilis strains derived from urine of the catheterized and non-catheterized patients has been investigated. A total number of 39 P. mirabilis strains isolated from the urine samples of the patients of dr Antoni Jurasz University Hospital No. 1 in Bydgoszcz clinics between 2011 and 2012 was used. Biofilm formation was evaluated using two independent quantitative and qualitative methods with TTC (2,3,5-triphenyl-tetrazolium chloride) and CV (crystal violet) application. The obtained results confirmed biofilm formation by all the examined strains, except quantitative method with TTC, in which 7.7% of the strains did not have this ability. It was shown that P. mirabilis rods have the ability to form biofilm on the surfaces of both biomaterials applied, polystyrene and polyvinyl chloride (Nelaton catheters). The differences in ability to form biofilm observed between P. mirabilis strains derived from the urine of the catheterized and non-catheterized patients were not statistically significant.

Proteus mirabilis biofilm - Qualitative and quantitative colorimetric methods-based evaluation

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Joanna Kwiecinska-Piróg

2014-12-01

Full Text Available Proteus mirabilis strains ability to form biofilm is a current topic of a number of research worldwide. In this study the biofilm formation of P. mirabilis strains derived from urine of the catheterized and non-catheterized patients has been investigated. A total number of 39 P. mirabilis strains isolated from the urine samples of the patients of dr Antoni Jurasz University Hospital No. 1 in Bydgoszcz clinics between 2011 and 2012 was used. Biofilm formation was evaluated using two independent quantitative and qualitative methods with TTC (2,3,5-triphenyl-tetrazolium chloride and CV (crystal violet application. The obtained results confirmed biofilm formation by all the examined strains, except quantitative method with TTC, in which 7.7% of the strains did not have this ability. It was shown that P. mirabilis rods have the ability to form biofilm on the surfaces of both biomaterials applied, polystyrene and polyvinyl chloride (Nelaton catheters. The differences in ability to form biofilm observed between P. mirabilis strains derived from the urine of the catheterized and non-catheterized patients were not statistically significant.

Manipulatiaon of Biofilm Microbial Ecology

Energy Technology Data Exchange (ETDEWEB)

Burkhalter, R.; Macnaughton, S.J.; Palmer, R.J.; Smith, C.A.; Whitaker, K.W.; White, D.C.; Zinn, M.; kirkegaard, R.

1998-08-09

The Biofilm mode of growth provides such significant advantages to the members of the consortium that most organisms in important habitats are found in biofilms. The study of factors that allow manipulation of biofilm microbes in the biofilm growth state requires that reproducible biofilms by generated. The most effective monitoring of biofilm formation, succession and desquamation is with on-line monitoring of microbial biofilms with flowcell for direct observation. The biofilm growth state incorporates a second important factor, the heterogeneity in the distribution in time and space of the component members of the biofilm consortium. This heterogeneity is reflected not only in the cellular distribution but in the metabolic activity within a population of cells. Activity and cellular distribution can be mapped in four dimensions with confocal microscopy, and function can be ascertained by genetically manipulated reporter functions for specific genes or by vital stains. The methodology for understanding the microbial ecology of biofilms is now much more readily available and the capacity to manipulate biofilms is becoming an important feature of biotechnology.

Manipulation of Biofilm Microbial Ecology

Energy Technology Data Exchange (ETDEWEB)

White, D.C.; Palmer, R.J., Jr.; Zinn, M.; Smith, C.A.; Burkhalter, R.; Macnaughton, S.J.; Whitaker, K.W.; Kirkegaard, R.D.

1998-08-15

The biofilm mode of growth provides such significant advantages to the members of the consortium that most organisms in important habitats are found in biofilms. The study of factors that allow manipulation of biofilm microbes in the biofilm growth state requires that reproducible biofilms be generated. The most effective monitoring of biofilm formation, succession and desaturation is with on-line monitoring of microbial biofilms with flowcell for direct observation. The biofilm growth state incorporates a second important factor, the heterogeneity in distribution in time and space of the component members of the biofilm consortium. This heterogeneity is reflected not only in the cellular distribution but in the metabolic activity within a population of cells. Activity and cellular distribution can be mapped in four dimensions with confocal microscopy, and function can be ascertained by genetically manipulated reporter functions for specific genes or by vital stains. The methodology for understanding the microbial ecology of biofilms is now much more readily available and the capacity to manipulate biofilms is becoming an important feature of biotechnology.

Biofilms in chronic infections - a matter of opportunity - monospecies biofilms in multispecies infections

DEFF Research Database (Denmark)

Burmølle, Mette; Thomsen, Trine Rolighed; Fazli, Mustafa

2010-01-01

It has become evident that aggregation or biofilm formation is an important survival mechanism for bacteria in almost any environment. In this review, we summarize recent visualizations of bacterial aggregates in several chronic infections (chronic otitis media, cystic fibrosis, infection due...... to permanent tissue fillers and chronic wounds) both as to distribution (such as where in the wound bed) and organization (monospecies or multispecies microcolonies). We correlate these biofilm observations to observations of commensal biofilms (dental and intestine) and biofilms in natural ecosystems (soil......). The observations of the chronic biofilm infections point toward a trend of low bacterial diversity and sovereign monospecies biofilm aggregates even though the infection in which they reside are multispecies. In contrast to this, commensal and natural biofilm aggregates contain multiple species that are believed...

Molecular-level evaluation of selected periodontal pathogens from subgingival regions in canines and humans with periodontal disease.

Science.gov (United States)

Gołyńska, Magdalena; Polkowska, Izabela; Bartoszcze-Tomaszewska, Małgorzata; Sobczyńska-Rak, Aleksandra; Matuszewski, Łukasz

2017-03-30

Dogs commonly serve as a model for various human conditions, including periodontal diseases. The aim of this study was to identify the anaerobic bacteria that colonize the subgingival areas in dogs and humans by using rapid real-time polymerase chain reaction (RT-PCR)-based tests and to compare the results obtained in each species. Bacterial microflora evaluations, both quantitative and qualitative, were performed by applying ready-made tests on twelve dogs and twelve humans. Five samples were collected from each subject's deepest gingival pockets and joined to form a collective sample. The results of the study revealed interspecies similarities in the prevalences of Porphyromonas ( P .) gingivalis, Treponema denticola, Tannerella forsythia , and Fusobacterium nucleatum . Red complex bacteria comprised the largest portion of the studied bacterial complexes in all study groups, with P. gingivalis being the most commonly isolated bacterium. The results show similarities in the prevalence of bacterial microflora in dogs and humans. Microbiological analysis of gingival pockets by using rapid real-time PCR-based tests in clinical practice, both veterinary and human, can facilitate the choice of appropriate pharmacological treatment and can provide a basis for subsequent verification of the treatment's effectiveness.

Mycobacterium biofilms: factors involved in development, dispersal, and therapeutic strategies against biofilm-relevant pathogens.

Science.gov (United States)

Xiang, Xiaohong; Deng, Wanyan; Liu, Minqiang; Xie, Jianping

2014-01-01

Many bacteria can develop biofilm (BF), a multicellular structure largely combining bacteria and their extracellular polymeric substances (EPS). The formation of biofilm results in an alternative existence in which microbes ensure their survival in adverse environments. Biofilm-relevant infections are more persistent, resistant to most antibiotics, and more recalcitrant to host immunity. Mycobacterium tuberculosis, the causative agent of tuberculosis, can develop biofilm, though whether M. tuberculosis can form biofilm within tuberculosis patients has yet to be determined. Here, we summarize the factors involved in the development and dispersal of mycobacterial biofilms, as well as underlying regulatory factors and inhibitors against biofilm to deepen our understanding of their development and to elucidate potential novel modes of action for future antibiotics. Key factors in biofilm formation identified as drug targets represent a novel and promising avenue for developing better antibiotics.

Characterization and fluorescence of yellow biofilms in karst caves, southwest Slovenia

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Janez Mulec

2015-05-01

Full Text Available Biofilms of different colours that colonize surfaces within karst caves represent a source of nutrients. They occur commonly and abundantly at sites with sediments, and close to seepages or underground rivers. Golden-yellow subaerial biofilms are particularly well observed because of their contrast with their surroundings, the characteristics of the pigment and recently, even more, due to the characteristics of light-emitting diode (LED illumination. Yellow microbial biofilms were sampled from three caves in southwestern Slovenia, Dimnice, Križna jama and Sveta jama. The highest concentration of cultivable microbes (2.33×108 CFU/g and the biggest number of identified bacteria (66.0% were retrieved from a sample from Sveta jama. Using MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight for bacterial identification showed that different species of Pseudomonas prevailed in all samples. Yellow biofilms showed an absorption peak around 400 nm, and two emission peaks, a major in the blue (~460 nm and a minor in the orange (~600 nm parts of the spectrum when excited at 405 nm. Microbial mats that colonize surfaces are probably frequently overlooked in caves because they are difficult to observe when they have no pigmentation and the contrast with their surroundings is low. Additional studies are needed to aid the understanding of the role of pigmented biofilms and their interactions with underlying substrata in respect of the evolution of substrate micromorphology.

Meningococcal biofilm formation

DEFF Research Database (Denmark)

Lappann, M.; Haagensen, Janus Anders Juul; Claus, H.

2006-01-01

We show that in a standardized in vitro flow system unencapsulated variants of genetically diverse lineages of Neisseria meningitidis formed biofilms, that could be maintained for more than 96 h. Biofilm cells were resistant to penicillin, but not to rifampin or ciprofloxacin. For some strains......, microcolony formation within biofilms was observed. Microcolony formation in strain MC58 depended on a functional copy of the pilE gene encoding the pilus subunit pilin, and was associated with twitching of cells. Nevertheless, unpiliated pilE mutants formed biofilms showing that attachment and accumulation......X alleles was identified among genetically diverse meningococcal strains. PilX alleles differed in their propensity to support autoaggregation of cells in suspension, but not in their ability to support microcolony formation within biofilms in the continuous flow system....

Biofilms in wounds

DEFF Research Database (Denmark)

Cooper, R A; Bjarnsholt, Thomas; Alhede, M

2014-01-01

Following confirmation of the presence of biofilms in chronic wounds, the term biofilm became a buzzword within the wound healing community. For more than a century pathogens have been successfully isolated and identified from wound specimens using techniques that were devised in the nineteenth...... extracellular polymeric substances (EPS). Cells within such aggregations (or biofilms) display varying physiological and metabolic properties that are distinct from those of planktonic cells, and which contribute to their persistence. There are many factors that influence healing in wounds and the discovery...... of biofilms in chronic wounds has provided new insight into the reasons why. Increased tolerance of biofilms to antimicrobial agents explains the limited efficacy of antimicrobial agents in chronic wounds and illustrates the need to develop new management strategies. This review aims to explain the nature...

Biofilm extracellular DNA enhances mixed species biofilms of Staphylococcus epidermidis and Candida albicans.

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Pammi, Mohan; Liang, Rong; Hicks, John; Mistretta, Toni-Ann; Versalovic, James

2013-11-14

Polymicrobial infections are responsible for significant mortality and morbidity in adults and children. Staphylococcus epidermidis and Candida albicans are the most frequent combination of organisms isolated from polymicrobial infections. Vascular indwelling catheters are sites for mixed species biofilm formation and pose a significant risk for polymicrobial infections. We hypothesized that enhancement of biofilms in a mixed species environment increases patient mortality and morbidity. Mixed species biofilms of S. epidermidis and C. albicans were evaluated in vitro and in a subcutaneous catheter infection model in vivo. Mixed species biofilms were enhanced compared to single species biofilms of either S. epidermidis or C. albicans. A mixed species environment increased catheter infection and increased dissemination of S. epidermidis in mice. Microarrays were used to explore differential gene expression of S. epidermidis in the mixed species biofilms. In mixed species biofilms, compared to single species S. epidermidis biofilms, 2.7% of S. epidermidis genes were upregulated and 6% were down regulated. Staphylococcal autolysis repressors lrgA and lrgB were down regulated 36-fold and 27-fold respectively. The role of biofilm extracellular DNA was investigated by quantitation and by evaluating the effects of DNAse in a concentration and time dependent manner. S. epidermidis specific eDNA was increased in mixed species biofilms and further confirmed by degradation with DNAse. Mixed-species biofilms are enhanced and associated with increased S. epidermidis-specific eDNA in vitro and greater systemic dissemination of S. epidermidis in vivo. Down regulation of the lrg operon, a repressor of autolysis, associated with increased eDNA suggests a possible role for bacterial autolysis in mixed species biofilms. Enhancement and systemic dissemination of S. epidermidis may explain adverse outcomes after clinical polymicrobial infections of S. epidermidis and C. albicans.

Biofilm extracellular DNA enhances mixed species biofilms of Staphylococcus epidermidis and Candida albicans

Science.gov (United States)

2013-01-01

Background Polymicrobial infections are responsible for significant mortality and morbidity in adults and children. Staphylococcus epidermidis and Candida albicans are the most frequent combination of organisms isolated from polymicrobial infections. Vascular indwelling catheters are sites for mixed species biofilm formation and pose a significant risk for polymicrobial infections. We hypothesized that enhancement of biofilms in a mixed species environment increases patient mortality and morbidity. Results Mixed species biofilms of S. epidermidis and C. albicans were evaluated in vitro and in a subcutaneous catheter infection model in vivo. Mixed species biofilms were enhanced compared to single species biofilms of either S. epidermidis or C. albicans. A mixed species environment increased catheter infection and increased dissemination of S. epidermidis in mice. Microarrays were used to explore differential gene expression of S. epidermidis in the mixed species biofilms. In mixed species biofilms, compared to single species S. epidermidis biofilms, 2.7% of S. epidermidis genes were upregulated and 6% were down regulated. Staphylococcal autolysis repressors lrgA and lrgB were down regulated 36-fold and 27-fold respectively. The role of biofilm extracellular DNA was investigated by quantitation and by evaluating the effects of DNAse in a concentration and time dependent manner. S. epidermidis specific eDNA was increased in mixed species biofilms and further confirmed by degradation with DNAse. Conclusions Mixed-species biofilms are enhanced and associated with increased S. epidermidis-specific eDNA in vitro and greater systemic dissemination of S. epidermidis in vivo. Down regulation of the lrg operon, a repressor of autolysis, associated with increased eDNA suggests a possible role for bacterial autolysis in mixed species biofilms. Enhancement and systemic dissemination of S. epidermidis may explain adverse outcomes after clinical polymicrobial infections of S

Biofilm formation by clinical isolates and the implications in chronic infections

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Sanchez Carlos J

2013-01-01

Full Text Available Abstract Background Biofilm formation is a major virulence factor contributing to the chronicity of infections. To date few studies have evaluated biofilm formation in infecting isolates of patients including both Gram-positive and Gram-negative multidrug-resistant (MDR species in the context of numerous types of infectious syndromes. Herein, we investigated the biofilm forming capacity in a large collection of single patient infecting isolates and compared the relationship between biofilm formation to various strain characteristics. Methods The biofilm-forming capacity of 205 randomly sampled clinical isolates from patients, collected from various anatomical sites, admitted for treatment at Brooke Army Medical Center (BAMC from 2004–2011, including methicillin-resistant/methicillin susceptible Staphylococcus aureus (MRSA/MSSA (n=23, Acinetobacter baumannii (n=53, Pseudomonas aeruginosa (n=36, Klebsiella pneumoniae (n=54, and Escherichia coli (n=39, were evaluated for biofilm formation using the high-throughput microtiter plate assay and scanning electron microscopy (SEM. Relationships between biofilm formation to clonal type, site of isolate collection, and MDR phenotype were evaluated. Furthermore, in patients with relapsing infections, serial strains were assessed for their ability to form biofilms in vitro. Results Of the 205 clinical isolates tested, 126 strains (61.4% were observed to form biofilms in vitro at levels greater than or equal to the Staphylococcus epidermidis, positive biofilm producing strain, with P. aeruginosa and S. aureus having the greatest number of biofilm producing strains. Biofilm formation was significantly associated with specific clonal types, the site of isolate collection, and strains positive for biofilm formation were more frequently observed to be MDR. In patients with relapsing infections, the majority of serial isolates recovered from these individuals were observed to be strong biofilm producers in vitro

Pseudomonas aeruginosa biofilm infections

DEFF Research Database (Denmark)

Tolker-Nielsen, Tim

2014-01-01

Bacteria in natural, industrial and clinical settings predominantly live in biofilms, i.e., sessile structured microbial communities encased in self-produced extracellular matrix material. One of the most important characteristics of microbial biofilms is that the resident bacteria display...... a remarkable increased tolerance toward antimicrobial attack. Biofilms formed by opportunistic pathogenic bacteria are involved in devastating persistent medical device-associated infections, and chronic infections in individuals who are immune-compromised or otherwise impaired in the host defense. Because...... the use of conventional antimicrobial compounds in many cases cannot eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections. The present review is focussed on the important opportunistic pathogen and biofilm model organism Pseudomonas aeruginosa. Initially...

Molecular Techniques Revealed Highly Diverse Microbial Communities in Natural Marine Biofilms on Polystyrene Dishes for Invertebrate Larval Settlement

KAUST Repository

Lee, On On

2014-01-09

Biofilm microbial communities play an important role in the larval settlement response of marine invertebrates. However, the underlying mechanism has yet to be resolved, mainly because of the uncertainties in characterizing members in the communities using traditional 16S rRNA gene-based molecular methods and in identifying the chemical signals involved. In this study, pyrosequencing was used to characterize the bacterial communities in intertidal and subtidal marine biofilms developed during two seasons. We revealed highly diverse biofilm bacterial communities that varied with season and tidal level. Over 3,000 operational taxonomic units with estimates of up to 8,000 species were recovered in a biofilm sample, which is by far the highest number recorded in subtropical marine biofilms. Nineteen phyla were found, of which Cyanobacteria and Proteobacteria were the most dominant one in the intertidal and subtidal biofilms, respectively. Apart from these, Actinobacteria, Bacteroidetes, and Planctomycetes were the major groups recovered in both intertidal and subtidal biofilms, although their relative abundance varied among samples. Full-length 16S rRNA gene clone libraries were constructed for the four biofilm samples and showed similar bacterial compositions at the phylum level to those revealed by pyrosequencing. Laboratory assays confirmed that cyrids of the barnacle Balanus amphitrite preferred to settle on the intertidal rather than subtidal biofilms. This preference was independent of the biofilm bacterial density or biomass but was probably related to the biofilm community structure, particularly, the Proteobacterial and Cyanobacterial groups. © 2014 Springer Science+Business Media New York.

Biofilm-Associated Gene Expression in Staphylococcus pseudintermedius on a Variety of Implant Materials.

Science.gov (United States)

Crawford, Evan C; Singh, Ameet; Gibson, Thomas W G; Scott Weese, J

2016-05-01

To evaluate the expression of biofilm-associated genes in Staphylococcus pseudintermedius on multiple clinically relevant surfaces. In vitro experimental study. Two strains of methicillin-resistant S. pseudintermedius isolated from clinical infections representing the most common international isolates. A quantitative polymerase chain reaction (qPCR) assay for expression of genes related to biofilm initial adhesion, formation/maturation, antimicrobial resistance, and intracellular communication was developed and validated. S. pseudintermedius biofilms were grown on 8 clinically relevant surfaces (polymethylmethacrylate, stainless steel, titanium, latex, silicone, polydioxanone, polystyrene, and glass) and samples of logarithmic and stationary growth phases were collected. Gene expression in samples was measured by qPCR. Significant differences in gene expression were identified between surfaces and between bacterial strains for most gene/strain/surface combinations studied. Expression of genes responsible for production of extracellular matrix were increased in biofilms. Expression of genes responsible for initial adhesion and intracellular communication was markedly variable. Antimicrobial resistance gene expression was increased on multiple surfaces, including stainless steel and titanium. A method for evaluation of expression of multiple biofilm-associated genes in S. pseudintermedius was successfully developed and applied to the study of biofilms on multiple surfaces. Variations in expression of these genes have a bearing on understanding the development and treatment of implant-associated biofilm infections and will inform future clinical research. © Copyright 2016 by The American College of Veterinary Surgeons.

Correlative Imaging of Structural and Elemental Composition of Bacterial Biofilms

International Nuclear Information System (INIS)

Yang, Y; Heine, R; Xu, F; Helfen, L; Baumbach, T; Suhonen, H; Rosenhahn, A; Gorniak, T; Kirchen, S; Schwartz, T

2013-01-01

Synchrotron-based phase contrast tomography (holotomography) and scanning hard X-ray fluorescence microscopy (SXFM) are combined to characterize the three-dimensional (3D) structural and corresponding elemental distribution of bacterial biofilms of Pseudomonas aeruginosa. Samples were fixed without contrast agents or microtomal sectioning. Within an intact microbial community single bacteria are clearly resolved, and their morphology can be directly visualized together with the elemental content. Such 3D set of complementary information at cellular level is essential for gaining a deeper understanding of biofilm evolution aiming to develop potential strategies on biofilm growth control and prevention

Association of biofilm production with colonization among clinical isolates of Acinetobacter baumannii.

Science.gov (United States)

Ryu, Seong Yeol; Baek, Won-Ki; Kim, Hyun Ah

2017-03-01

The pathogen Acinetobacter baumannii is increasingly causing healthcare-associated infections worldwide, particularly in intensive care units. Biofilm formation, a factor contributing to the virulence of A. baumannii , is associated with long-term persistence in hospital environments. The present study investigates the clinical impact of biofilm production on colonization and acquisition after patient admission. Forty-nine A. baumannii isolates were obtained between August and November 2013 from Keimyung University Dongsan Medical Center, Daegu, Korea. All isolates were obtained from sputum samples of new patients infected or colonized by A. baumannii . The microtiter plate assay was used to determine biofilm formation. Twenty-four A. baumannii isolates (48%) demonstrated enhanced biofilm formation capacity than that of the standard A. baumannii strain (ATCC 19606). All isolates were resistant to carbapenem, 38 isolates (77%) were collected from patients in an intensive care unit, and 47 isolates (95%) were from patients who had been exposed to antibiotics in the previous month. The median duration of colonization was longer for biofilm-producing isolates than that of the biofilm non-biofilm producing isolates (18 days vs. 12 days, p < 0.05). Simultaneous colonization with other bacteria was more common for biofilm-producing isolates than that for the non-biofilm producing isolates. The most prevalent co-colonizing bacteria was Staphylococcus aureus . Biofilm-producing isolates seem to colonize the respiratory tract for longer durations than the non-biofilm producing isolates. During colonization, biofilm producers promote co-colonization by other bacteria, particularly S. aureus . Additional research is required to determine possible links between biofilm formation and nosocomial infection.

Investigate Nasal Colonize Staphylococcus Species Biofilm Produced

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Cemil Demir

2014-03-01

Full Text Available Aim: 127 S.aureus and 65 CoNS strains were isolated from patients noses%u2019. To produce a biofilm ability was investigated using three different methods. Slime-positive and negative staphylococcies%u2019 resistance were evaluated against different antibiotics. Material and Method: Swap samples puted 7% blood agar. Staphylococcus aureus and coagulase-negative staphylococci (CoNS isolates biofilm produced ability were investigated using Congo Red Agar (CRA, microplates (MP and Standard Tube (ST methods. In addition to that, presence of antibiotic resistance of the staphylococcal isolates are determined agar disc diffusion method. Results: The rate of biofilm producing Staphylococcus spp strains was found to be 72.4%, 67.7%, and 62.9%, respectively with CRA, MP, and ST tests. There was no significant relationship among the tests (p>0.05. In addition, antibiotic resistance of Staphylococcus spp. against various antibiotics was also determined by the agar disk diffusion method. Resistance rates of biofilm positive (BP Staphylococcus spp for penicilin G, ampicilin, amocycilin/clavulanic acid, tetracyclin, eritromycin, gentamycin, and enrofloxacin 71.7%, 69.7%, 6.2%, 20.7%, 21.4%, 1.4%, and 0.7%, respectively. Resistance rates of biofilm negative (BN spp for 42.6%, 23.4%, 4.3%, 14.9%, 19.1%, 0.0%, 0.0% respectively. All Staphylococcus isolates were found to be susceptible to vancomycin and teicaplonin. Although BP strains antibiotic resistance rates were observed higher than BN strains. But resistance rates were not found statistically significant (p>0.05. Discussion: CRA is the reliablity and specifity method to determine Staphylococcus spp. biofilm produce ability.

Molecular Analysis of Bacterial Communities in Biofilms of a Drinking Water Clearwell

Science.gov (United States)

Zhang, Minglu; Liu, Wenjun; Nie, Xuebiao; Li, Cuiping; Gu, Junnong; Zhang, Can

2012-01-01

Microbial community structures in biofilms of a clearwell in a drinking water supply system in Beijing, China were examined by clone library, terminal restriction fragment length polymorphism (T-RFLP) and 454 pyrosequencing of the amplified 16S rRNA gene. Six biofilm samples (designated R1–R6) collected from six locations (upper and lower sites of the inlet, middle and outlet) of the clearwell revealed similar bacterial patterns by T-RFLP analysis. With respect to the dominant groups, the phylotypes detected by clone library and T-RFLP generally matched each other. A total of 9,543 reads were obtained from samples located at the lower inlet and the lower outlet sites by pyrosequencing. The bacterial diversity of the two samples was compared at phylum and genus levels. Alphaproteobacteria dominated the communities in both samples and the genus of Sphingomonas constituted 75.1%–99.6% of this phylum. A high level of Sphingomonas sp. was first observed in the drinking water biofilms with 0.6–1.0 mg L−1 of chlorine residual. Disinfectant-resistant microorganisms deserve special attention in drinking water management. This study provides novel insights into the microbial populations in drinking water systems and highlights the important role of Sphingomonas species in biofilm formation. PMID:23059725

Cyanobacteria, algae and microfungi present in biofilm from Božana Cave (Serbia

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Slađana Popović

2015-05-01

Full Text Available Phototrophic microorganisms (cyanobacteria and algae and microfungi, were identified from biofilm on the walls of the entrance of BožanaCavein west Serbia. Temperature, relative humidity and light intensity were measured, and chlorophyll a content determined. Light intensity differed from the entrance inwards. However, Chl a content was not proportional to light intensity, instead it was positively correlated to biofilm weight. Biofilm samples from two sites were also observed using a scanning electron microscope. Coccoid forms of cyanobacteria were abundant at the sampling site with the lowest light intensity, while members of the order Nostocales were predominant at the sampling site with the highest light intensity measured. Cyanobacteria were the dominant group of phototrophs colonizing cave walls (29 taxa, with the order Chroococcales prevailing (21 taxa. The most frequently documented cyanobacteria were species from genera Gloeocapsa, Scytonema, Aphanocapsa and Chroococcus. Desmococcus olivaceus and Trentepohlia aurea were the only green algae documented on cave walls. Ascomycetes were common (e.g. Alternaria, Aspergillus, Cladosporium, Epicoccum, Penicillum and Trichoderma, while zygomycetes and oomycetes were less frequent. The different color of each biofilm sample was ascribed to the presence of various different species of cyanobacteria and algae.

Periodontal microbiota of Sardinian children: comparing 200-year-old samples to present-day ones

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Germano Orrù

2017-02-01

Full Text Available Introduction: The microrganisms of the human oral cavity include more than 700 species or phenotypes of bacteria. Some “diseases of civilization” are strictly correlated to changes in the microbiome following the food revolution that occurred after WWII. For that reason, a precise recognition of the microbiome profile before and after this period should be useful to determine the health-compatible model of microbiome. The aim of this study was to compare the microbiome profiles (number of total cells, and pathogen types of dental samples obtained from two distinct groups of children, a 200-year-old retrieved one and a present one.Methods: Two different groups of samples have been studied. The first group was a set of 50 recent subgingival plaque samples obtained from children of age 2-8 years, 14 males and 36 females. They were enrolled by the Department of Dental Disease Prevention (University of Cagliari, in Sardinia, Italy during standard dental care procedures. None reported periodontal disease and none had been under antibiotic therapy during the previous 6 months. The second group was an old retrieved group that included 24 teeth from 6 different 6- to 8-year-old crania fragments; they were obtained from a 200-year-old charnel-house located in Villaputzu, a city close to Cagliari. Representative periodontal bacteria have been identified by a previously published real-time PCR procedure (Sokransky et al., 1998 in which P. gingivalis and T. forsythia (red complex, A. actinomycetemcomitans (green complex and F. nucleatum (orange complex were detected. In addition, the title of each pathogen was expressed as a percentage of the total bacteria (biofilm in the sample.Results and discussion: The profile of periodontal microbiomes, between recent/ancient samples showed a significant difference relative to Sokransky’s red complex bacteria (p < 0.05. In all analyzed periodontal strains, the pathogenic bacteria P. gingivalis and T. forsythia

Hydraulic resistance of biofilms

KAUST Repository

Dreszer, C.; Vrouwenvelder, Johannes S.; Paulitsch-Fuchs, Astrid H.; Zwijnenburg, Arie; Kruithof, Joop C.; Flemming, Hans Curt

2013-01-01

resistance is very low compared to the expected biofilm resistance and, thus, biofilm resistance can be determined accurately. Transmembrane pressure drop was monitored. As biofilm parameters, thickness, total cell number, TOC, and extracellular polymeric

Efficacy of a marine bacterial nuclease against biofilm forming microorganisms isolated from chronic rhinosinusitis.

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Robert C Shields

Full Text Available BACKGROUND: The persistent colonization of paranasal sinus mucosa by microbial biofilms is a major factor in the pathogenesis of chronic rhinosinusitis (CRS. Control of microorganisms within biofilms is hampered by the presence of viscous extracellular polymers of host or microbial origin, including nucleic acids. The aim of this study was to investigate the role of extracellular DNA in biofilm formation by bacteria associated with CRS. METHODS/PRINCIPAL FINDINGS: Obstructive mucin was collected from patients during functional endoscopic sinus surgery. Examination of the mucous by transmission electron microscopy revealed an acellular matrix punctuated occasionally with host cells in varying states of degradation. Bacteria were observed in biofilms on mucosal biopsies, and between two and six different species were isolated from each of 20 different patient samples. In total, 16 different bacterial genera were isolated, of which the most commonly identified organisms were coagulase-negative staphylococci, Staphylococcus aureus and α-haemolytic streptococci. Twenty-four fresh clinical isolates were selected for investigation of biofilm formation in vitro using a microplate model system. Biofilms formed by 14 strains, including all 9 extracellular nuclease-producing bacteria, were significantly disrupted by treatment with a novel bacterial deoxyribonuclease, NucB, isolated from a marine strain of Bacillus licheniformis. Extracellular biofilm matrix was observed in untreated samples but not in those treated with NucB and extracellular DNA was purified from in vitro biofilms. CONCLUSION/SIGNIFICANCE: Our data demonstrate that bacteria associated with CRS form robust biofilms which can be reduced by treatment with matrix-degrading enzymes such as NucB. The dispersal of bacterial biofilms with NucB may offer an additional therapeutic target for CRS sufferers.

Biofilm in endodontics: A review

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Jhajharia, Kapil; Parolia, Abhishek; Shetty, K Vikram; Mehta, Lata Kiran

2015-01-01

Endodontic disease is a biofilm-mediated infection, and primary aim in the management of endodontic disease is the elimination of bacterial biofilm from the root canal system. The most common endodontic infection is caused by the surface-associated growth of microorganisms. It is important to apply the biofilm concept to endodontic microbiology to understand the pathogenic potential of the root canal microbiota as well as to form the basis for new approaches for disinfection. It is foremost to understand how the biofilm formed by root canal bacteria resists endodontic treatment measures. Bacterial etiology has been confirmed for common oral diseases such as caries and periodontal and endodontic infections. Bacteria causing these diseases are organized in biofilm structures, which are complex microbial communities composed of a great variety of bacteria with different ecological requirements and pathogenic potential. The biofilm community not only gives bacteria effective protection against the host's defense system but also makes them more resistant to a variety of disinfecting agents used as oral hygiene products or in the treatment of infections. Successful treatment of these diseases depends on biofilm removal as well as effective killing of biofilm bacteria. So, the fundamental to maintain oral health and prevent dental caries, gingivitis, and periodontitis is to control the oral biofilms. From these aspects, the formation of biofilms carries particular clinical significance because not only host defense mechanisms but also therapeutic efforts including chemical and mechanical antimicrobial treatment measures have the most difficult task of dealing with organisms that are gathered in a biofilm. The aim of this article was to review the mechanisms of biofilms’ formation, their roles in pulpal and periapical pathosis, the different types of biofilms, the factors influencing biofilm formation, the mechanisms of their antimicrobial resistance, techniques to

Bacteriophages and Biofilms

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David R. Harper

2014-06-01

Full Text Available Biofilms are an extremely common adaptation, allowing bacteria to colonize hostile environments. They present unique problems for antibiotics and biocides, both due to the nature of the extracellular matrix and to the presence within the biofilm of metabolically inactive persister cells. Such chemicals can be highly effective against planktonic bacterial cells, while being essentially ineffective against biofilms. By contrast, bacteriophages seem to have a greater ability to target this common form of bacterial growth. The high numbers of bacteria present within biofilms actually facilitate the action of bacteriophages by allowing rapid and efficient infection of the host and consequent amplification of the bacteriophage. Bacteriophages also have a number of properties that make biofilms susceptible to their action. They are known to produce (or to be able to induce enzymes that degrade the extracellular matrix. They are also able to infect persister cells, remaining dormant within them, but re-activating when they become metabolically active. Some cultured biofilms also seem better able to support the replication of bacteriophages than comparable planktonic systems. It is perhaps unsurprising that bacteriophages, as the natural predators of bacteria, have the ability to target this common form of bacterial life.

Reliability of Haemophilus influenzae biofilm measurement via static method, and determinants of in vitro biofilm production.

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Obaid, Najla A; Tristram, Stephen; Narkowicz, Christian K; Jacobson, Glenn A

2016-12-01

Information is lacking regarding the precision of microtitre plate (MTP) assays used to measure biofilm. This study investigated the precision of an MTP assay to measure biofilm production by nontypeable Haemophilus influenzae (NTHi) and the effects of frozen storage and inoculation technique on biofilm production. The density of bacterial final growth was determined by absorbance after 18-20 h incubation, and biofilm production was then measured by absorbance after crystal violet staining. Biofilm formation was categorised as high and low for each strain. For the high biofilm producing strains of NTHi, interday reproducibility of NTHi biofilm formation measured using the MTP assay was excellent and met the acceptance criteria, but higher variability was observed in low biofilm producers. Method of inoculum preparation was a determinant of biofilm formation with inoculum prepared directly from solid media showing increased biofilm production for at least one of the high producing strains. In general, storage of NTHi cultures at -80 °C for up to 48 weeks did not have any major effect on their ability to produce biofilm.

Effect of Splinting on Dimensional Accuracy of Impressions Made of Implants with Different Subgingival Alignments.

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Beyabanaki, Elaheh; Shamshiri, Ahmed Reza; Alikhasi, Marzieh; Monzavi, Abbas

2017-01-01

Placement of implants at deeper levels of gingiva is sometimes inevitable because of issues like esthetics or bone availability. The accuracy of impressions may be affected in these situations. The aim of this in vitro study was to evaluate the effects of splinting and length of impression copings on the accuracy of impressions made of deeply placed implants. A metal model with two parallel implants (Implantium; Dentium) was fabricated. One hundred and twenty impressions were made using the direct impression technique with and without splinting the impression copings (using short and long impression copings). Impressions were made of implants at three subgingival levels (1, 3, and 6 mm) using regular viscosity poly(vinyl siloxane). The impressions were poured with type IV dental stone. Displacements in the x, y, and z axes, as well as rotational discrepancies and interimplant distances were measured with a coordinate measuring machine. Data were analyzed with Kruskal-Wallis, Mann-Whitney, and nonparametric adjusted rank transform tests. There was less rotational displacement using longer impression copings at different subgingival positions of the implants, either with splinted or nonsplinted direct technique (p impressions at different apico-coronal levels of implants than the splinted technique using short impression copings (p impression copings yielded better results than shorter ones in both splinted and nonsplinted techniques. Also, nonsplinted short impression copings produced more accurate impressions than splinted short impression copings. © 2015 by the American College of Prosthodontists.

Bacterial biofilm and associated infections

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Muhsin Jamal

2018-01-01

Full Text Available Microscopic entities, microorganisms that drastically affect human health need to be thoroughly investigated. A biofilm is an architectural colony of microorganisms, within a matrix of extracellular polymeric substance that they produce. Biofilm contains microbial cells adherent to one-another and to a static surface (living or non-living. Bacterial biofilms are usually pathogenic in nature and can cause nosocomial infections. The National Institutes of Health (NIH revealed that among all microbial and chronic infections, 65% and 80%, respectively, are associated with biofilm formation. The process of biofilm formation consists of many steps, starting with attachment to a living or non-living surface that will lead to formation of micro-colony, giving rise to three-dimensional structures and ending up, after maturation, with detachment. During formation of biofilm several species of bacteria communicate with one another, employing quorum sensing. In general, bacterial biofilms show resistance against human immune system, as well as against antibiotics. Health related concerns speak loud due to the biofilm potential to cause diseases, utilizing both device-related and non-device-related infections. In summary, the understanding of bacterial biofilm is important to manage and/or to eradicate biofilm-related diseases. The current review is, therefore, an effort to encompass the current concepts in biofilm formation and its implications in human health and disease.

Orthodontic extrusion of subgingivally fractured tooth using a removable appliance: an alternative treatment to reestablish biological width.

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Verma, Kanika Gupta; Juneja, Suruchi; Kumar, Sandeep; Goyal, Tanya

2014-01-01

Restoration of a traumatically injured tooth presents a clinical challenge for a predictable aesthetic outcome. This case report describes a multidisciplinary approach of a subgingivally fractured permanent maxillary central incisor. A removable orthodontic appliance was used for orthodontic extrusion of root, and surgical gingival recontouring was done with electrocautery to reestablish the biological width. Form and function were restored establishing biological width and esthetics was repaired with porcelain fused to metal crown.

Inhibitory effect of zinc oxide nanoparticles on pseudomonas aeruginosa biofilm formation

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Mohammad Hassani Sangani

2015-04-01

Full Text Available Objective(s: Bacterial biofilm formation causes many persistent and chronic infections. The matrix protects biofilm bacteria from exposure to innate immune defenses and antibiotic treatments. The purpose of this study was to evaluate the biofilm formation of clinical isolates of Pseudomonas aeruginosa and the activity of zinc oxide nanoparticles (ZnO NPs on biofilm. Materials and Methods: After collecting bacteria from clinical samples of hospitalized patients, the ability of organisms were evaluated to create biofilm by tissue culture plate (TCP assay. ZnO NPs were synthesized by sol gel method and the efficacy of different concentrations (50- 350 µg/ml of ZnO NPs was assessed on biofilm formation and also elimination of pre-formed biofilm by using TCP method. Results:The average diameter of synthesized ZnO NPs was 20 nm. The minimum inhibitory concentration of nanoparticles was 150- 158 μg/ml and the minimum bactericidal concentration was higher (325 µg/ml. All 15 clinical isolates of P. aeruginosa were able to produce biofilm. Treating the organisms with nanoparticles at concentrations of 350 μg/ml resulted in more than 94% inhibition in OD reduction%. Molecular analysis showed that the presence of mRNA of pslA gene after treating bacteria with ZnO NPs for 30 minutes. Conclusion: The results showed that ZnO NPs can inhibit the establishment of P. aeruginosa biofilms and have less effective in removing pre-formed biofilm. However the tested nanoparticles exhibited anti-biofilm effect, but mRNA of pslA gene could be still detected in the medium by RT-PCR technique after 30 minutes treatment with ZnO.

Experimental study of cadmium interaction with periphytic biofilms

International Nuclear Information System (INIS)

Pokrovsky, O.S.; Feurtet-Mazel, A.; Martinez, R.E.; Morin, S.; Baudrimont, M.; Duong, T.; Coste, M.

2010-01-01

This study addresses the interaction of Cd with natural biofilms of periphytic diatoms grown during different seasons in metal-contaminated and metal-non-contaminated streams, along a tributary of the Lot River, France. Specifically, it aims to test whether the biofilms from contaminated sites have developed a protective mechanism due to high Cd exposure. Towards this goal, reversible adsorption experiments on untreated biofilms were performed in 0.01 M NaNO 3 with a pH ranging from 2 to 8, Cd concentration from 0.5 to 10,000 μg/L and exposure time from 1 to 24 h. Two types of experiments, pH-dependent adsorption edge and constant-pH 'Langmuirian'-type isotherms were conducted. Results were adequately modeled using a Linear Programming Model. It was found that the adsorption capacities of natural biofilm consortia with respect to Cd do not depend on season and are not directly linked to the growth environment. The biofilms grown in non-contaminated (4.6 ppb Cd in solid) and contaminated (570 ppb Cd in solid) settings exhibit similar adsorption capacities in the Cd concentration range in solution of 100-10,000 μg/L but quite different capacities at low Cd concentration (0.5-100 μg/L); unexpectedly, the non-contaminated biofilm adsorbs approximately 10 times more Cd than the contaminated one. It is therefore possible that the strong low-abundant ligands (for example, phosphoryl or sulfhydryls) are already metal-saturated on surfaces of biofilm grown in the contaminated site whereas these sites are still available for metal adsorption in samples grown in non-contaminated sites.

Experimental study of cadmium interaction with periphytic biofilms

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Pokrovsky, O.S., E-mail: oleg@lmtg.obs-mip.fr [Geochimie et Biogeochimie Experimentale, UMR 5563, CNRS-OMP-Universite Toulouse, 14 Avenue Edouard Belin, 31400 Toulouse (France); Feurtet-Mazel, A. [Universite de Bordeaux 1, CNRS, UMR 5805 EPOC, Place du Dr Peyneau, 33120 Arcachon (France); Martinez, R.E. [Center for Applied Geosciences, Universitat Tuebingen, Sigwartstrasse 10, Tuebingen 72076 (Germany); Morin, S. [Unite de Recherche Reseaux, Epuration et Qualite des Eaux REQE, Cemagref, 50 Avenue de Verdun, F-33612 Cestas Cedex (France); Baudrimont, M. [Universite de Bordeaux 1, CNRS, UMR 5805 EPOC, Place du Dr Peyneau, 33120 Arcachon (France); Duong, T. [Universite de Bordeaux 1, CNRS, UMR 5805 EPOC, Place du Dr Peyneau, 33120 Arcachon (France)] [Institute of Environmental Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Cau Giay, Hanoi (Viet Nam); Coste, M. [Unite de Recherche Reseaux, Epuration et Qualite des Eaux REQE, Cemagref, 50 Avenue de Verdun, F-33612 Cestas Cedex (France)

2010-03-15

This study addresses the interaction of Cd with natural biofilms of periphytic diatoms grown during different seasons in metal-contaminated and metal-non-contaminated streams, along a tributary of the Lot River, France. Specifically, it aims to test whether the biofilms from contaminated sites have developed a protective mechanism due to high Cd exposure. Towards this goal, reversible adsorption experiments on untreated biofilms were performed in 0.01 M NaNO{sub 3} with a pH ranging from 2 to 8, Cd concentration from 0.5 to 10,000 {mu}g/L and exposure time from 1 to 24 h. Two types of experiments, pH-dependent adsorption edge and constant-pH 'Langmuirian'-type isotherms were conducted. Results were adequately modeled using a Linear Programming Model. It was found that the adsorption capacities of natural biofilm consortia with respect to Cd do not depend on season and are not directly linked to the growth environment. The biofilms grown in non-contaminated (4.6 ppb Cd in solid) and contaminated (570 ppb Cd in solid) settings exhibit similar adsorption capacities in the Cd concentration range in solution of 100-10,000 {mu}g/L but quite different capacities at low Cd concentration (0.5-100 {mu}g/L); unexpectedly, the non-contaminated biofilm adsorbs approximately 10 times more Cd than the contaminated one. It is therefore possible that the strong low-abundant ligands (for example, phosphoryl or sulfhydryls) are already metal-saturated on surfaces of biofilm grown in the contaminated site whereas these sites are still available for metal adsorption in samples grown in non-contaminated sites.

DNase I and proteinase K impair Listeria monocytogenes biofilm formation and induce dispersal of pre-existing biofilms.

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Nguyen, Uyen T; Burrows, Lori L

2014-09-18

Current sanitation methods in the food industry are not always sufficient for prevention or dispersal of Listeria monocytogenes biofilms. Here, we determined if prevention of adherence or dispersal of existing biofilms could occur if biofilm matrix components were disrupted enzymatically. Addition of DNase during biofilm formation reduced attachment (biofilms with 100μg/ml of DNase for 24h induced incomplete biofilm dispersal, with biofilm remaining compared to control. In contrast, addition of proteinase K completely inhibited biofilm formation, and 72h biofilms-including those grown under stimulatory conditions-were completely dispersed with 100μg/ml proteinase K. Generally-regarded-as-safe proteases bromelain and papain were less effective dispersants than proteinase K. In a time course assay, complete dispersal of L. monocytogenes biofilms from both polystyrene and type 304H food-grade stainless steel occurred within 5min at proteinase K concentrations above 25μg/ml. These data confirm that both DNA and proteins are required for L. monocytogenes biofilm development and maintenance, and that these components of the biofilm matrix can be targeted for effective prevention and removal of biofilms. Copyright © 2014 Elsevier B.V. All rights reserved.

Candida/Candida biofilms. First description of dual-species Candida albicans/C. rugosa biofilm.

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Martins, Carlos Henrique Gomes; Pires, Regina Helena; Cunha, Aline Oliveira; Pereira, Cristiane Aparecida Martins; Singulani, Junya de Lacorte; Abrão, Fariza; Moraes, Thais de; Mendes-Giannini, Maria José Soares

2016-04-01

Denture liners have physical properties that favour plaque accumulation and colonization by Candida species, irritating oral tissues and causing denture stomatitis. To isolate and determine the incidence of oral Candida species in dental prostheses, oral swabs were collected from the dental prostheses of 66 patients. All the strains were screened for their ability to form biofilms; both monospecies and dual-species combinations were tested. Candida albicans (63 %) was the most frequently isolated microorganism; Candida tropicalis (14 %), Candida glabrata (13 %), Candida rugosa (5 %), Candida parapsilosis (3 %), and Candida krusei (2 %) were also detected. The XTT assay showed that C. albicans SC5314 possessed a biofilm-forming ability significantly higher (p biofilm was less than the total CFU of a monospecies C. albicans biofilm. In contrast to the profuse hyphae verified in monospecies C. albicans biofilms, micrographies showed that the C. albicans/non-albicans Candida biofilms consisted of sparse yeast forms and profuse budding yeast cells that generated a network. These results suggested that C. albicans and the tested Candida species could co-exist in biofilms displaying apparent antagonism. The study provide the first description of C. albicans/C. rugosa mixed biofilm. Copyright © 2016 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Surface proteins and the formation of biofilms by Staphylococcus aureus.

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Kim, Sung Joon; Chang, James; Rimal, Binayak; Yang, Hao; Schaefer, Jacob

2018-03-01

Staphylococcus aureus biofilms pose a serious clinical threat as reservoirs for persistent infections. Despite this clinical significance, the composition and mechanism of formation of S. aureus biofilms are unknown. To address these problems, we used solid-state NMR to examine S. aureus (SA113), a strong biofilm-forming strain. We labeled whole cells and cell walls of planktonic cells, young biofilms formed for 12-24h after stationary phase, and more mature biofilms formed for up to 60h after stationary phase. All samples were labeled either by (i) [ 15 N]glycine and l-[1- 13 C]threonine, or in separate experiments, by (ii) l-[2- 13 C, 15 N]leucine. We then measured 13 C- 15 N direct bonds by C{N} rotational-echo double resonance (REDOR). The increase in peptidoglycan stems that have bridges connected to a surface protein was determined directly by a cell-wall double difference (biofilm REDOR difference minus planktonic REDOR difference). This procedure eliminates errors arising from differences in 15 N isotopic enrichments and from the routing of 13 C label from threonine degradation to glycine. For both planktonic cells and the mature biofilm, 20% of pentaglycyl bridges are not cross-linked and are potential surface-protein attachment sites. None of these sites has a surface protein attached in the planktonic cells, but one-fourth have a surface protein attached in the mature biofilm. Moreover, the leucine-label shows that the concentration of β-strands in leucine-rich regions doubles in the mature biofilm. Thus, a primary event in establishing a S. aureus biofilm is extensive decoration of the cell surface with surface proteins that are linked covalently to the cell wall and promote cell-cell adhesion. Copyright © 2017 Elsevier B.V. All rights reserved.

Biofilm formation and determination of minimum biofilm eradication concentration of antibiotics in Mycoplasma hyopneumoniae.

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Tassew, Dereje Damte; Mechesso, Abraham Fikru; Park, Na-Hye; Song, Ju-Beom; Shur, Joo-Woon; Park, Seung-Chun

2017-10-20

The study was aimed to investigate biofilm forming ability of Mycoplasma hyopneumoniae and to determine the minimum biofilm eradication concentrations of antibiotics. Biofilm forming ability of six strains of M. hyopneumoniae was examined using crystal violet staining on coverslips. The results demonstrated an apparent line of biofilm growth in 3 of the strains isolated from swine with confirmed cases of enzootic pneumonia. BacLight bacterial viability assay revealed that the majority of the cells were viable after 336 hr of incubation. Moreover, M. hyopneumoniae persists in the biofilm after being exposed to 10 fold higher concentration of antibiotics than the minimum inhibitory concentrations in planktonic cells. To the best of our knowledge, this is the first report of biofilm formation in M. hyopneumoniae. However, comprehensive studies on the mechanisms of biofilm formation are needed to combat swine enzootic pneumonia caused by resistant M. hyopneumoniae.

CLINICAL AND MICROBIOLOGICAL DATA IN PATIENTS WITH CHRONIC PERIODONTITIS.

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Christina Popova

2013-07-01

Full Text Available Periodontitis is an infectious disease concerning supporting tissues of the teeth. The primary etiological agent for disease development and progression is the subgingival biofilm, but recently it is known that host factors may modify the pathological process or may affect the severity and /or extent. The increasing levels of some specific pathogenic subgingival bacteria such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and others can result in periodontal destruction and possibly correlate with disease severity. Data from controlled studies show high prevalence of P. gingivalis, T. forsythia and Tr. denticola which represent the red complex (coexistence of these three species in patients with moderate and severe chronic periodontitis. Parallel investigation of probing depth (PD and clinical attachment level (CAL with the microbiological testing may give a confirmation of relation between subgingival pathogenic bacteria and severity of periodontitis.

Molecular analysis of long-term biofilm formation on PVC and cast iron surfaces in drinking water distribution system.

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Liu, Ruyin; Zhu, Junge; Yu, Zhisheng; Joshi, DevRaj; Zhang, Hongxun; Lin, Wenfang; Yang, Min

2014-04-01

To understand the impacts of different plumbing materials on long-term biofilm formation in water supply system, we analyzed microbial community compositions in the bulk water and biofilms on faucets with two different materials-polyvinyl chloride (PVC) and cast iron, which have been frequently used for more than10 years. Pyrosequencing was employed to describe both bacterial and eukaryotic microbial compositions. Bacterial communities in the bulk water and biofilm samples were significantly different from each other. Specific bacterial populations colonized on the surface of different materials. Hyphomicrobia and corrosion associated bacteria, such as Acidithiobacillus spp., Aquabacterium spp., Limnobacter thiooxidans, and Thiocapsa spp., were the most dominant bacteria identified in the PVC and cast iron biofilms, respectively, suggesting that bacterial colonization on the material surfaces was selective. Mycobacteria and Legionella spp. were common potential pathogenic bacteria occurred in the biofilm samples, but their abundance was different in the two biofilm bacterial communities. In contrast, the biofilm samples showed more similar eukaryotic communities than the bulk water. Notably, potential pathogenic fungi, i.e., Aspergillus spp. and Candida parapsilosis, occurred in similar abundance in both biofilms. These results indicated that microbial community, especially bacterial composition was remarkably affected by the different pipe materials (PVC and cast iron). Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters.

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Holling, Nina; Dedi, Cinzia; Jones, Caroline E; Hawthorne, Joseph A; Hanlon, Geoffrey W; Salvage, Jonathan P; Patel, Bhavik A; Barnes, Lara M; Jones, Brian V

2014-06-01

Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation. © 2014 The Authors. FEMS Microbiology Letters published by John Wiley & Sons Ltd on behalf of Federation of European Microbiological Societies.

Patterned biofilm formation reveals a mechanism for structural heterogeneity in bacterial biofilms.

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Gu, Huan; Hou, Shuyu; Yongyat, Chanokpon; De Tore, Suzanne; Ren, Dacheng

2013-09-03

Bacterial biofilms are ubiquitous and are the major cause of chronic infections in humans and persistent biofouling in industry. Despite the significance of bacterial biofilms, the mechanism of biofilm formation and associated drug tolerance is still not fully understood. A major challenge in biofilm research is the intrinsic heterogeneity in the biofilm structure, which leads to temporal and spatial variation in cell density and gene expression. To understand and control such structural heterogeneity, surfaces with patterned functional alkanthiols were used in this study to obtain Escherichia coli cell clusters with systematically varied cluster size and distance between clusters. The results from quantitative imaging analysis revealed an interesting phenomenon in which multicellular connections can be formed between cell clusters depending on the size of interacting clusters and the distance between them. In addition, significant differences in patterned biofilm formation were observed between wild-type E. coli RP437 and some of its isogenic mutants, indicating that certain cellular and genetic factors are involved in interactions among cell clusters. In particular, autoinducer-2-mediated quorum sensing was found to be important. Collectively, these results provide missing information that links cell-to-cell signaling and interaction among cell clusters to the structural organization of bacterial biofilms.

Factors affecting dental biofilm in patients wearing fixed orthodontic appliances.

Science.gov (United States)

Mei, Li; Chieng, Joyce; Wong, Connie; Benic, Gareth; Farella, Mauro

2017-12-01

The aim of this study is to investigate the amount and the distribution of biofilm in patients wearing fixed appliances and its relation with age, gender, frequency of tooth brushing, and patient motivation. The sample comprised 52 patients (15.5 ± 3.6 years old, 30 females and 22 males) wearing fixed orthodontic appliances. Dental biofilm was assessed using a modified plaque index (PI). A questionnaire was used to collect patient's information, including gender, age, treatment motivation, and frequency of tooth brushing. Gingival (PI score = 0.9 ± 0.7), mesial (0.8 ± 0.6), and distal (0.8 ± 0.5) areas accumulated more biofilm than occlusal areas (0.3 ± 0.3) (P appliances have the highest biofilm accumulation on the maxillary lateral incisors and maxillary canines, particularly in the gingival area and areas behind arch wires. Less biofilm was observed in female and adult patients and in those who were self-motivated and brushed their teeth more often.

Focusing on Environmental Biofilms With Variable-Pressure Scanning Electron Microscopy

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Joubert, L.; Wolfaardt, G. M.; Du Plessis, K.

2006-12-01

Since the term biofilm has been coined almost 30 years ago, visualization has formed an integral part of investigations on microbial attachment. Electron microscopic (EM) biofilm studies, however, have been limited by the hydrated extracellular matrix which loses structural integrity with conventional preparative techniques, and under required high-vacuum conditions, resulting in a loss of information on spatial relationships and distribution of biofilm microbes. Recent advances in EM technology enable the application of Variable Pressure Scanning Electron Microscopy (VP SEM) to biofilms, allowing low vacuum and hydrated chamber atmosphere during visualization. Environmental biofilm samples can be viewed in situ, unfixed and fully hydrated, with application of gold-sputter-coating only, to increase image resolution. As the impact of microbial biofilms can be both hazardous and beneficial to man and his environment, recognition of biofilms as a natural form of microbial existence is needed to fully assess the potential role of microbial communities on technology. The integration of multiple techniques to elucidate biofilm processes has become imperative for unraveling complex phenotypic adaptations of this microbial lifestyle. We applied VP SEM as integrative technique with traditional and novel analytical techniques to (1)localize lignocellulosic microbial consortia applied for producing alternative bio-energy sources in the mining wastewater industry, (2) characterize and visualize wetland microbial communities in the treatment of winery wastewater, and (3)determine the impact of recombinant technology on yeast biofilm behavior. Visualization of microbial attachment to a lignocellulose substrate, and degradation of exposed plant tissue, gave insight into fiber degradation and volatile fatty acid production for biological sulphate removal from mining wastewater. Also, the 3D-architecture of complex biofilms developing in constructed wetlands was correlated with

A personal history of research on microbial biofilms and biofilm infections.

Science.gov (United States)

Høiby, Niels

2014-04-01

The observation of aggregated microorganisms surrounded by a self-produced matrix adhering to surfaces or located in tissues or secretions is as old as microbiology, with both Leeuwenhoek and Pasteur describing the phenomenon. In environmental and technical microbiology, biofilms were already shown 80-90 years ago to be important for biofouling on submerged surfaces, e.g. ships. The concept of biofilm infections and their importance in medicine is, however, biofilm was introduced into medicine in 1985 by Costerton. In the following decades, it became obvious that biofilm infections are widespread in medicine, and their importance is now generally accepted. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Design of a dynamic biofilm imaging cell for white-light interferometric microscopy

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Larimer, Curtis; Brann, Michelle; Suter, Jonathan D.; Addleman, R. Shane

2017-11-01

In microbiology research, there is a strong need for next-generation imaging and sensing instrumentation that will enable minimally invasive and label-free investigation of soft, hydrated structures, such as in bacterial biofilms. White-light interferometry (WLI) can provide high-resolution images of surface topology without the use of fluorescent labels but is not typically used to image biofilms because there is insufficient refractive index contrast to induce reflection from the biofilm's interface. The soft structure and water-like bulk properties of hydrated biofilms make them difficult to characterize in situ, especially in a nondestructive manner. We build on our prior description of static biofilm imaging and describe the design of a dynamic growth flow cell that enables monitoring of the thickness and topology of live biofilms over time using a WLI microscope. The microfluidic system is designed to grow biofilms in dynamic conditions and to create a reflective interface on the surface while minimizing disruption of fragile structures. The imaging cell was also designed to accommodate limitations imposed by the depth of focus of the microscope's objective lens. Example images of live biofilm samples are shown to illustrate the ability of the flow cell and WLI instrument to (1) support bacterial growth and biofilm development, (2) image biofilm structure that reflects growth in flow conditions, and (3) monitor biofilm development over time nondestructively. In future work, the apparatus described here will enable surface metrology measurements (roughness, surface area, etc.) of biofilms and may be used to observe changes in biofilm structure in response to changes in environmental conditions (e.g., flow velocity, availability of nutrients, and presence of biocides). This development will open opportunities for the use of WLI in bioimaging.

Stratified growth in Pseudomonas aeruginosa biofilms

DEFF Research Database (Denmark)

Werner, E.; Roe, F.; Bugnicourt, A.

2004-01-01

In this study, stratified patterns of protein synthesis and growth were demonstrated in Pseudomonas aeruginosa biofilms. Spatial patterns of protein synthetic activity inside biofilms were characterized by the use of two green fluorescent protein (GFP) reporter gene constructs. One construct...... synthesis was restricted to a narrow band in the part of the biofilm adjacent to the source of oxygen. The zone of active GFP expression was approximately 60 Am wide in colony biofilms and 30 Am wide in flow cell biofilms. The region of the biofilm in which cells were capable of elongation was mapped...... by treating colony biofilms with carbenicillin, which blocks cell division, and then measuring individual cell lengths by transmission electron microscopy. Cell elongation was localized at the air interface of the biofilm. The heterogeneous anabolic patterns measured inside these biofilms were likely a result...

Physiological differentiation within a single-species biofilm fueled by serpentinization.

Science.gov (United States)

Brazelton, William J; Mehta, Mausmi P; Kelley, Deborah S; Baross, John A

2011-01-01

Carbonate chimneys at the Lost City hydrothermal field are coated in biofilms dominated by a single phylotype of archaea known as Lost City Methanosarcinales. In this study, we have detected surprising physiological complexity in single-species biofilms, which is typically indicative of multispecies biofilm communities. Multiple cell morphologies were visible within the biofilms by transmission electron microscopy, and some cells contained intracellular membranes that may facilitate methane oxidation. Both methane production and oxidation were detected at 70 to 80°C and pH 9 to 10 in samples containing the single-species biofilms. Both processes were stimulated by the presence of hydrogen (H(2)), indicating that methane production and oxidation are part of a syntrophic interaction. Metagenomic data included a sequence encoding AMP-forming acetyl coenzyme A synthetase, indicating that acetate may play a role in the methane-cycling syntrophy. A wide range of nitrogen fixation genes were also identified, many of which were likely acquired via lateral gene transfer (LGT). Our results indicate that cells within these single-species biofilms may have differentiated into multiple physiological roles to form multicellular communities linked by metabolic interactions and LGT. Communities similar to these Lost City biofilms are likely to have existed early in the evolution of life, and we discuss how the multicellular characteristics of ancient hydrogen-fueled biofilm communities could have stimulated ecological diversification, as well as unity of biochemistry, during the earliest stages of cellular evolution. Our previous work at the Lost City hydrothermal field has shown that its carbonate chimneys host microbial biofilms dominated by a single uncultivated "species" of archaea. In this paper, we integrate evidence from these previous studies with new data on the metabolic activity and cellular morphology of these archaeal biofilms. We conclude that the archaeal biofilm

Detection of fusobacterium nucleatum and fadA adhesin gene in patients with orthodontic gingivitis and non-orthodontic periodontal inflammation.

Science.gov (United States)

Liu, Ping; Liu, Yi; Wang, Jianning; Guo, Yang; Zhang, Yujie; Xiao, Shuiqing

2014-01-01

Fusobacterium nucleatum is one of the most abundant gram-negative bacilli colonizing the subgingival plaque and closely associated with periodontal disease. However it is unclear whether F. nucleatum is involved in gingival inflammation under orthodontic appliance. A novel adhesin, FadA, which is unique to oral Fusobacteria, is required for F. nucleatum binding and invasion to epithelial cells and thus may play an important role in colonization of Fusobacterium in the host. In this study, we evaluated the prevalence of F. nucleatum and its virulence factor FadA adhesion gene (fadA) in 169 subgingival biofilm samples from 55 cases of gingivitis patients with orthodontic appliances, 49 cases of gingivitis patients without orthodontic treatment, 35 cases of periodontitis patients and 30 cases of periodontally healthy people via PCR. The correlations between the F. nucleatum/fadA and gingivitis index(GI)was also analyzed. The detection rate of F. nucleatum/fadA in periodontitis group and non-orthodontic gingivitis group was higher than the other two groups (pgingivitis group than in health people (pgingivitis and periodontal disease compared with orthodontic gingivitis.

Experimental evolution in biofilm populations

Science.gov (United States)

Steenackers, Hans P.; Parijs, Ilse; Foster, Kevin R.; Vanderleyden, Jozef

2016-01-01

Biofilms are a major form of microbial life in which cells form dense surface associated communities that can persist for many generations. The long-life of biofilm communities means that they can be strongly shaped by evolutionary processes. Here, we review the experimental study of evolution in biofilm communities. We first provide an overview of the different experimental models used to study biofilm evolution and their associated advantages and disadvantages. We then illustrate the vast amount of diversification observed during biofilm evolution, and we discuss (i) potential ecological and evolutionary processes behind the observed diversification, (ii) recent insights into the genetics of adaptive diversification, (iii) the striking degree of parallelism between evolution experiments and real-life biofilms and (iv) potential consequences of diversification. In the second part, we discuss the insights provided by evolution experiments in how biofilm growth and structure can promote cooperative phenotypes. Overall, our analysis points to an important role of biofilm diversification and cooperation in bacterial survival and productivity. Deeper understanding of both processes is of key importance to design improved antimicrobial strategies and diagnostic techniques. PMID:26895713

Biofilm development in fixed bed biofilm reactors: experiments and simple models for engineering design purposes.

Science.gov (United States)

Szilágyi, N; Kovács, R; Kenyeres, I; Csikor, Zs

2013-01-01

Biofilm development in a fixed bed biofilm reactor system performing municipal wastewater treatment was monitored aiming at accumulating colonization and maximum biofilm mass data usable in engineering practice for process design purposes. Initially a 6 month experimental period was selected for investigations where the biofilm formation and the performance of the reactors were monitored. The results were analyzed by two methods: for simple, steady-state process design purposes the maximum biofilm mass on carriers versus influent load and a time constant of the biofilm growth were determined, whereas for design approaches using dynamic models a simple biofilm mass prediction model including attachment and detachment mechanisms was selected and fitted to the experimental data. According to a detailed statistical analysis, the collected data have not allowed us to determine both the time constant of biofilm growth and the maximum biofilm mass on carriers at the same time. The observed maximum biofilm mass could be determined with a reasonable error and ranged between 438 gTS/m(2) carrier surface and 843 gTS/m(2), depending on influent load, and hydrodynamic conditions. The parallel analysis of the attachment-detachment model showed that the experimental data set allowed us to determine the attachment rate coefficient which was in the range of 0.05-0.4 m d(-1) depending on influent load and hydrodynamic conditions.

Experimental model of biofilm implant-related osteomyelitis to test combination biomaterials using biofilms as initial inocula.

Science.gov (United States)

Williams, Dustin L; Haymond, Bryan S; Woodbury, Kassie L; Beck, J Peter; Moore, David E; Epperson, R Tyler; Bloebaum, Roy D

2012-07-01

Currently, the majority of animal models that are used to study biofilm-related infections use planktonic bacterial cells as initial inocula to produce positive signals of infection in biomaterials studies. However, the use of planktonic cells has potentially led to inconsistent results in infection outcomes. In this study, well-established biofilms of methicillin-resistant Staphylococcus aureus were grown and used as initial inocula in an animal model of a Type IIIB open fracture. The goal of the work was to establish, for the first time, a repeatable model of biofilm implant-related osteomyelitis, wherein biofilms were used as initial inocula to test combination biomaterials. Results showed that 100% of animals that were treated with biofilms developed osteomyelitis, whereas 0% of animals not treated with biofilm developed infection. The development of this experimental model may lead to an important shift in biofilm and biomaterials research by showing that when biofilms are used as initial inocula, they may provide additional insights into how biofilm-related infections in the clinic develop and how they can be treated with combination biomaterials to eradicate and/or prevent biofilm formation. Copyright © 2012 Wiley Periodicals, Inc.

Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation

DEFF Research Database (Denmark)

Stapper, A.P.; Narasimhan, G.; Oman, D.E.

2004-01-01

of their biofilm formation using confocal laser scanning microscopy. Biofilm Image Processing (BIP) and Community Statistics (COMSTAT) software programs were used to provide quantitative measurements of the two-dimensional biofilm images. All three strains formed distinguishable biofilm architectures, indicating...

Current understanding of multi-species biofilms

DEFF Research Database (Denmark)

Yang, Liang; Liu, Yang; Wu, Hong

2011-01-01

every year worldwide to deal with damage to equipment, contaminations of products, energy losses, and infections in human beings resulted from microbial biofilms. Microorganisms compete, cooperate, and communicate with each other in multi-species biofilms. Understanding the mechanisms of multi......Direct observation of a wide range of natural microorganisms has revealed the fact that the majority of microbes persist as surface-attached communities surrounded by matrix materials, called biofilms. Biofilms can be formed by a single bacterial strain. However, most natural biofilms are actually......-species biofilm formation will facilitate the development of methods for combating bacterial biofilms in clinical, environmental, industrial, and agricultural areas. The most recent advances in the understanding of multi-species biofilms are summarized and discussed in the review....

Prevention of biofilm formation and removal of existing biofilms by extracellular DNases of Campylobacter jejuni.

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Brown, Helen L; Reuter, Mark; Hanman, Kate; Betts, Roy P; van Vliet, Arnoud H M

2015-01-01

The fastidious nature of the foodborne bacterial pathogen Campylobacter jejuni contrasts with its ability to survive in the food chain. The formation of biofilms, or the integration into existing biofilms by C. jejuni, is thought to contribute to food chain survival. As extracellular DNA (eDNA) has previously been proposed to play a role in C. jejuni biofilms, we have investigated the role of extracellular DNases (eDNases) produced by C. jejuni in biofilm formation. A search of 2791 C. jejuni genomes highlighted that almost half of C. jejuni genomes contains at least one eDNase gene, but only a minority of isolates contains two or three of these eDNase genes, such as C. jejuni strain RM1221 which contains the cje0256, cje0566 and cje1441 eDNase genes. Strain RM1221 did not form biofilms, whereas the eDNase-negative strains NCTC 11168 and 81116 did. Incubation of pre-formed biofilms of NCTC 11168 with live C. jejuni RM1221 or with spent medium from a RM1221 culture resulted in removal of the biofilm. Inactivation of the cje1441 eDNase gene in strain RM1221 restored biofilm formation, and made the mutant unable to degrade biofilms of strain NCTC 11168. Finally, C. jejuni strain RM1221 was able to degrade genomic DNA from C. jejuni NCTC 11168, 81116 and RM1221, whereas strain NCTC 11168 and the RM1221 cje1441 mutant were unable to do so. This was mirrored by an absence of eDNA in overnight cultures of C. jejuni RM1221. This suggests that the activity of eDNases in C. jejuni affects biofilm formation and is not conducive to a biofilm lifestyle. These eDNases do however have a potential role in controlling biofilm formation by C. jejuni strains in food chain relevant environments.

Biofilm bacterial communities in urban drinking water distribution systems transporting waters with different purification strategies.

Science.gov (United States)

Wu, Huiting; Zhang, Jingxu; Mi, Zilong; Xie, Shuguang; Chen, Chao; Zhang, Xiaojian

2015-02-01

Biofilm formation in drinking water distribution systems (DWDS) has many adverse consequences. Knowledge of microbial community structure of DWDS biofilm can aid in the design of an effective control strategy. However, biofilm bacterial community in real DWDS and the impact of drinking water purification strategy remain unclear. The present study investigated the composition and diversity of biofilm bacterial community in real DWDSs transporting waters with different purification strategies (conventional treatment and integrated treatment). High-throughput Illumina MiSeq sequencing analysis illustrated a large shift in the diversity and structure of biofilm bacterial community in real DWDS. Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Nitrospirae, and Cyanobacteria were the major components of biofilm bacterial community. Proteobacteria (mainly Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria) predominated in each DWDS biofilm, but the compositions of the dominant proteobacterial classes and genera and their proportions varied among biofilm samples. Drinking water purification strategy could shape DWDS biofilm bacterial community. Moreover, Pearson's correlation analysis indicated that Actinobacteria was positively correlated with the levels of total alkalinity and dissolved organic carbon in tap water, while Firmicutes had a significant positive correlation with nitrite nitrogen.

In vivo biofilm formation on different dental ceramics.

Science.gov (United States)

Bremer, Felicia; Grade, Sebastian; Kohorst, Philipp; Stiesch, Meike

2011-01-01

To investigate the formation of oral biofilm on various dental ceramics in vivo. Five different ceramic materials were included: a veneering glass- ceramic, a lithium disilicate glass-ceramic, a yttrium-stabilized zirconia (Y-TZP), a hot isostatically pressed (HIP) Y-TZP ceramic, and an HIP Y-TZP ceramic with 25% alumina. Test specimens were attached to individually designed acrylic appliances; five volunteers wore these appliances for 24 hours in the maxillary arch. After intraoral exposure, the samples were removed from the appliances and the adhering biofilms vitally stained. Then, the two-dimensional surface coating and thickness of the adhering biofilm were determined by confocal laser scanning microscopy. Statistical analysis was performed using one-way ANOVA with the level of significance set at .05. Significant differences (P ceramic materials. The lowest surface coating (19.0%) and biofilm thickness (1.9 Μm) were determined on the HIP Y-TZP ceramic; the highest mean values were identified with the lithium disilicate glass-ceramic (46.8%, 12.6 Μm). Biofilm formation on various types of dental ceramics differed significantly; in particular, zirconia exhibited low plaque accumulation. In addition to its high strength, low plaque accumulation makes zirconia a promising material for various indications (including implant abutments and telescopic crowns) that previously were met only with metal-based materials.

Disinfection of Pseudomonas aeruginosa biofilm contaminated tube lumens with ultraviolet C light emitting diodes

DEFF Research Database (Denmark)

Bak, Jimmy; Ladefoged, Søren D; Tvede, Michael

2010-01-01

with biofilm was investigated. It was shown that UVC light propagation was possible through both Teflon and catheter tubes (silicone). The disinfection efficiency of the diodes was demonstrated on tubes contaminated artificially with a Pseudomonas aeruginosa biofilm. The tubes were connected to a flow system...... and biofilms were produced during a 3 day period. Tubes in lengths of 10 (Teflon, silicone) and 20 cm (Teflon) were contaminated. Tubes for control and for UVC treatment were contaminated in parallel. Biofilms were sampled from the total inner surface of the tubes. Colony counts on the control samples were...... in the range of 5 x 10(5)-1.3 x 10(9) CFU ml(-1), with disinfection rates in the range 96-100%. The applied UVC doses corresponded to treatment times between 15 and 300 min. Disinfection (100%) was obtained in 10 cm Teflon tubes exposed for 30 min (detection limit...

Disinfection of Pseudomonas aeruginosa biofilm contaminated tube lumens with ultraviolet C light emitting diodes

DEFF Research Database (Denmark)

Bak, Jimmy; Ladefoged, S.D.; Tvede, M.

2010-01-01

with biofilm was investigated. It was shown that UVC light propagation was possible through both Teflon and catheter tubes ( silicone). The disinfection efficiency of the diodes was demonstrated on tubes contaminated artificially with a Pseudomonas aeruginosa biofilm. The tubes were connected to a flow system...... and biofilms were produced during a 3 day period. Tubes in lengths of 10 ( Teflon, silicone) and 20 cm ( Teflon) were contaminated. Tubes for control and for UVC treatment were contaminated in parallel. Biofilms were sampled from the total inner surface of the tubes. Colony counts on the control samples were...... in the range of 5 x 10(5)-1.3 x 10(9) CFU ml(-1), with disinfection rates in the range 96-100%. The applied UVC doses corresponded to treatment times between 15 and 300 min. Disinfection (100%) was obtained in 10 cm Teflon tubes exposed for 30 min (detection limit...

Evaluation of intraspecies interactions in biofilm formation by Methylobacterium species isolated from pink-pigmented household biofilms.

Science.gov (United States)

Xu, Fang-Fang; Morohoshi, Tomohiro; Wang, Wen-Zhao; Yamaguchi, Yuka; Liang, Yan; Ikeda, Tsukasa

2014-01-01

Concern regarding household biofilms has grown due to their widespread existence and potential to threaten human health by serving as pathogen reservoirs. Previous studies identified Methylobacterium as one of the dominant genera found in household biofilms. In the present study, we examined the mechanisms underlying biofilm formation by using the bacterial consortium found in household pink slime. A clone library analysis revealed that Methylobacterium was the predominant genus in household pink slime. In addition, 16 out of 21 pink-pigmented bacterial isolates were assigned to the genus Methylobacterium. Although all of the Methylobacterium isolates formed low-level biofilms, the amount of the biofilms formed by Methylobacterium sp. P-1M and P-18S was significantly increased by co-culturing with other Methylobacterium strains that belonged to a specific phylogenetic group. The single-species biofilm was easily washed from the glass surface, whereas the dual-species biofilm strongly adhered after washing. A confocal laser scanning microscopy analysis showed that the dual-species biofilms were significantly thicker and tighter than the single-species biofilms.

Comparisons of Subgingival Microbial Profiles of Refractory Periodontitis, Severe Periodontitis and Periodontal Health using the Human Oral Microbe Identification Microarray (HOMIM)

Science.gov (United States)

Colombo, Ana Paula V.; Boches, Susan K.; Cotton, Sean L.; Goodson, J. Max; Kent, Ralph; Haffajee, Anne D.; Socransky, Sigmund S.; Hasturk, Hatice; Van Dyke, Thomas E.; Paster, Bruce J.

2013-01-01

Aim This study compared the subgingival microbiota of subjects with refractory periodontitis (RP) to those in subjects with treatable periodontitis (GR) or periodontal health (PH) using the Human Oral Microbe Identification Microarray (HOMIM). Methods At baseline, subgingival plaque samples were taken from 47 periodontitis and 20 PH individuals, and analyzed for the presence of 300 species by HOMIM. The periodontitis subjects were classified as RP (n=17) based on mean attachment loss (AL) and/or >3 sites with AL ≥2.5 mm after SRP, surgery and systemically administered amoxicillin and metronidazole or as GR (n=30) based on mean attachment gain and no sites with AL ≥2.5 mm after treatment. Significant differences in taxa among groups were sought using the Kruskal Wallis and Chi-square tests. Results More species were detected in diseased patients (GR or RP) than those without disease (PH). RP subjects were distinguished from GR and PH by a significantly high frequency of putative periodontal pathogens such as, Parvimonas micra, Campylobacter gracilis, Eubacterium nodatum, Selenomonas noxia, Tannerella forsythia, Porphyromonas gingivalis, Prevotella spp., Treponema spp., Eikenella corrodens, as well as “unusual” species (Pseudoramibacter alactolyticus, TM7 spp. oral taxon (OT) 346/356, Bacteroidetes spp. OT 272/274, Solobacterium moorei, Desulfobulbus sp. OT 041, Brevundimonas diminuta, Sphaerocytophaga sp. OT 337, Shuttleworthia satelles, Filifactor alocis, Dialister invisus/pneumosintes, Granulicatella adiacens, Mogibacterium tidmidum, Veillonella atypica, Mycoplasma salivarium, Synergistes sp. cluster II, Acidaminococcaceae [G-1] sp. OT 132/150/155/148/135) [pspp. cluster I, Capnocytophaga sputigena, Cardiobacterium hominis, Haemophilus parainfluenzae, Lautropia mirabilis, Propionibacterium propionicum, Rothia dentocariosa/mucilagenosa, Streptococcus sanguinis (p<0.05). Conclusion RP patients present a distinct microbial profile compared to patients in the

Anti-biofilm Properties of the Fecal Probiotic Lactobacilli Against Vibrio spp.

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Sumanpreet Kaur

2018-04-01

Full Text Available Diarrheal disease caused by Vibrio cholerae is endemic in developing countries including India and is associated with high rate of mortality especially in children. V. cholerae is known to form biofilms on the gut epithelium, and the biofilms once formed are resistant to the action of antibiotics. Therefore agents that prevent the biofilm formation and disperse the preformed biofilms are associated with therapeutic benefits. The use of antibiotics for the treatment of cholera is associated with side effects such as gut dysbiosis due to depletion of gut microflora, and the increasing problem of antibiotic resistance. Thus search for safe alternative therapeutic agents is warranted. Herein, we screened the lactobacilli spp. isolated from the fecal samples of healthy children for their abilities to prevent biofilm formation and to disperse the preformed biofilms of V. cholerae and V. parahaemolyticus by using an in vitro assay. The results showed that the culture supernatant (CS of all the seven isolates of Lactobacillus spp. used in the study inhibited the biofilm formation of V. cholerae by more than 90%. Neutralization of pH of CS completely abrogated their antimicrobial activities against V. cholera, but had negligible effects on their biofilm inhibitory potential. Further, CS of all the lactobacilli isolates caused the dispersion of preformed V. cholerae biofilms in the range 62–85%; however, pH neutralization of CS reduced the biofilm dispersal potential of the 4 out of 7 isolates by 19–57%. Furthermore, the studies showed that CS of none of the lactobacilii isolates had antimicrobial activity against V. parahaemolyticus, but 5 out of 7 isolates inhibited the formation of its biofilm in the range 62–82%. However, none of the CS dispersed the preformed biofilms of V. parahaemolyticus. The ability of CS to inhibit the adherence of Vibrio spp. to the epithelial cell line was also determined. Thus, we conclude that the biofilm dispersive

Antimicrobial Photoinactivation Using Visible Light Plus Water-Filtered Infrared-A (VIS + wIRA Alters In Situ Oral Biofilms.

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A Al-Ahmad

Full Text Available Recently, growing attention has been paid to antimicrobial photodynamic therapy (aPDT in dentistry. Changing the microbial composition of initial and mature oral biofilm by aPDT using visible light plus water-filtered infrared-A wavelengths (VIS + wIRA has not yet been investigated. Moreover, most aPDT studies have been conducted on planktonic bacterial cultures. Therefore, in the present clinical study we cultivated initial and mature oral biofilms in six healthy volunteers for 2 hours or 3 days, respectively. The biofilms were treated with aPDT using VIS+wIRA (200 mW cm(-2, toluidine blue (TB and chlorine e6 (Ce6 for 5 minutes. Chlorhexidine treated biofilm samples served as positive controls, while untreated biofilms served as negative controls. After aPDT treatment the colony forming units (CFU of the biofilm samples were quantified, and the surviving bacteria were isolated in pure cultures and identified using MALDI-TOF, biochemical tests and 16S rDNA-sequencing. aPDT killed more than 99.9% of the initial viable bacterial count and 95% of the mature oral biofilm in situ, independent of the photosensitizer. The number of surviving bacterial species was highly reduced to 6 (TB and 4 (Ce6 in the treated initial oral biofilm compared to the 20 different species of the untreated biofilm. The proportions of surviving bacterial species were also changed after TB- and Ce6-mediated aPDT of the mature oral biofilm, resulting in a shift in the microbial composition of the treated biofilm compared to that of the control biofilm. In conclusion, aPDT using VIS + wIRA showed a remarkable potential to eradicate both initial and mature oral biofilms, and also to markedly alter the remaining biofilm. This encourages the clinical use of aPDT with VIS + wIRA for the treatment of periimplantitis and periodontitis.

Extracellular matrix assembly in extreme acidic eukaryotic biofilms and their possible implications in heavy metal adsorption

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Aguilera, Angeles [Centro de Astrobiologia (INTA-CSIC), Carretera de Ajalvir Km 4, Torrejon de Ardoz, 28850 Madrid (Spain)], E-mail: aguileraba@inta.es; Souza-Egipsy, Virginia [Centro de Astrobiologia (INTA-CSIC), Carretera de Ajalvir Km 4, Torrejon de Ardoz, 28850 Madrid (Spain); San Martin-Uriz, Patxi [Centro de Biologia Molecular (UAM-CSIC), Universidad Autonoma de Madrid, Cantoblanco, 28049 Madrid (Spain); Amils, Ricardo [Centro de Astrobiologia (INTA-CSIC), Carretera de Ajalvir Km 4, Torrejon de Ardoz, 28850 Madrid (Spain); Centro de Biologia Molecular (UAM-CSIC), Universidad Autonoma de Madrid, Cantoblanco, 28049 Madrid (Spain)

2008-07-30

To evaluate the importance of the extracellular matrix in relation to heavy metal binding capacity in extreme acidic environments, the extracellular polymeric substances (EPS) composition of 12 biofilms isolated from Rio Tinto (SW, Spain) was analyzed. Each biofilm was composed mainly by one or two species of eukaryotes, although other microorganisms were present. EPS ranged from 130 to 439 mg g{sup -1} biofilm dry weight, representing between 15% and the 40% of the total biofilm dry weight (DW). Statistically significant differences (p < 0.05) were found in the amount of total EPS extracted from biofilms dominated by the same organism at different sampling points. The amount of EPS varied among different biofilms collected from the same sampling location. Colloidal EPS ranged from 42 to 313 mg g{sup -1} dry weight; 10% to 30% of the total biofilm dry weight. Capsular EPS ranged from 50 to 318 mg g{sup -1} dry weight; 5% to 30% of the total biofilm dry weight. Seven of the 12 biofilms showed higher amounts of capsular than colloidal EPS (p < 0.05). Total amount of EPS decreased when total cell numbers and pH increased. There was a positive correlation between EPS concentration and heavy metal concentration in the water. Observations by low temperature scanning electron microscopy (LTSEM) revealed the mineral adsorption in the matrix of EPS and onto the cell walls. EPS in all biofilms were primarily composed of carbohydrates, heavy metals and humic acid, plus small quantities of proteins and DNA. After carbohydrates, heavy metals were the second main constituents of the extracellular matrix. Their total concentrations ranged from 3 to 32 mg g{sup -1} biofilm dry weight, reaching up to 16% of the total composition. In general, the heavy metal composition of the EPS extracted from the biofilms closely resembled the metal composition of the water from which the biofilms were collected.

Bacterial Biofilm Infection Detected in Breast Implant-Associated Anaplastic Large-Cell Lymphoma.

Science.gov (United States)

Hu, Honghua; Johani, Khalid; Almatroudi, Ahmad; Vickery, Karen; Van Natta, Bruce; Kadin, Marshall E; Brody, Garry; Clemens, Mark; Cheah, Chan Yoon; Lade, Stephen; Joshi, Preeti Avinash; Prince, H Miles; Deva, Anand K

2016-06-01

A recent association between breast implants and the development of anaplastic large-cell lymphoma (ALCL) has been observed. The purpose of this study was to identify whether bacterial biofilm is present in breast implant-associated ALCL and, if so, to compare the bacterial microbiome to nontumor capsule samples from breast implants with contracture. Twenty-six breast implant-associated ALCL samples were analyzed for the presence of biofilm by real-time quantitative polymerase chain reaction, next-generation sequencing, fluorescent in situ hybridization, and scanning electron microscopy, and compared to 62 nontumor capsule specimens. Both the breast implant-associated ALCL and nontumor capsule samples yielded high mean numbers of bacteria (breast implant-associated ALCL, 4.7 × 10 cells/mg of tissue; capsule, 4.9 × 10 cells/mg of tissue). Analysis of the microbiome in breast implant-associated ALCL specimens showed significant differences with species identified in nontumor capsule specimens. There was a significantly greater proportion of Ralstonia spp. present in ALCL specimens compared with nontumor capsule specimens (p capsule specimens compared with breast implant-associated ALCL specimens (p < 0.001). Bacterial biofilm was visualized both on scanning electron microscopy and fluorescent in situ hybridization. This novel finding of bacterial biofilm and a distinct microbiome in breast implant-associated ALCL samples points to a possible infectious contributing cause. Breast implants are widely used in both reconstructive and aesthetic surgery, and strategies to reduce their contamination should be more widely studied and practiced. Risk, V.

Establishing a laboratory model of dental unit waterlines bacterial biofilms using a CDC biofilm reactor.

Science.gov (United States)

Yoon, Hye Young; Lee, Si Young

2017-11-01

In this study, a laboratory model to reproduce dental unit waterline (DUWL) biofilms was developed using a CDC biofilm reactor (CBR). Bacteria obtained from DUWLs were filtered and cultured in Reasoner's 2A (R2A) for 10 days, and were subsequently stored at -70°C. This stock was cultivated on R2A in batch mode. After culturing for five days, the bacteria were inoculated into the CBR. Biofilms were grown on polyurethane tubing for four days. Biofilm accumulation and thickness was 1.3 × 10 5  CFU cm -2 and 10-14 μm respectively, after four days. Bacteria in the biofilms included cocci and rods of short and medium lengths. In addition, 38 bacterial genera were detected in biofilms. In this study, the suitability and reproducibility of the CBR model for DUWL biofilm formation were demonstrated. The model provides a foundation for the development of bacterial control methods for DUWLs.

Effects of pesticides and pharmaceuticals on biofilms in a highly impacted river

International Nuclear Information System (INIS)

Proia, L.; Osorio, V.; Soley, S.; Köck-Schulmeyer, M.; Pérez, S.; Barceló, D.; Romaní, A.M.; Sabater, S.

2013-01-01

We investigated the effects of pharmaceuticals and pesticides detected in a Mediterranean river, on fluvial biofilms by translocation experiments performed under controlled conditions. Water was sampled from three sites along a pollution gradient. Biofilms grown in mesocosms containing relatively clean water were translocated to heavily polluted water. Several biofilm descriptors were measured before and after translocations. Fifty-seven pharmaceuticals and sixteen pesticides compounds were detected in river waters. The translocation from less to more polluted site was the most effective. Autotrophic biomass and peptidase increased while phosphatase and photosynthetic efficiency decreased. Multivariate analysis revealed that analgesics and anti-inflammatories significantly affected biofilm responses. Ibuprofen and paracetamol were associated with negative effects on photosynthesis, and with the decrease of the green algae/cyanobacteria ratio, while diclofenac was associated with phosphatase activity. The effects of these emerging compounds on biofilms structure and function may cause important alterations in river ecosystem functioning. -- Highlights: •We investigated the pharmaceuticals and pesticides occurrence in the Llobregat River. •We studied the effects on biofilm communities by translocation experiments. •Both authotrophs and heterotrophs responded when translocated to more polluted sites. •Analgesics and anti-inflammatories significantly affected biofilm responses. •Ibuprofen, Paracetamol and Diclofenac are the most effective compounds. -- Analgesics and anti-inflammatories detected in river water significantly affected responses of biofilm communities when translocated along a pollution gradient

New Technologies for Studying Biofilms

Science.gov (United States)

FRANKLIN, MICHAEL J.; CHANG, CONNIE; AKIYAMA, TATSUYA; BOTHNER, BRIAN

2016-01-01

Bacteria have traditionally been studied as single-cell organisms. In laboratory settings, aerobic bacteria are usually cultured in aerated flasks, where the cells are considered essentially homogenous. However, in many natural environments, bacteria and other microorganisms grow in mixed communities, often associated with surfaces. Biofilms are comprised of surface-associated microorganisms, their extracellular matrix material, and environmental chemicals that have adsorbed to the bacteria or their matrix material. While this definition of a biofilm is fairly simple, biofilms are complex and dynamic. Our understanding of the activities of individual biofilm cells and whole biofilm systems has developed rapidly, due in part to advances in molecular, analytical, and imaging tools and the miniaturization of tools designed to characterize biofilms at the enzyme level, cellular level, and systems level. PMID:26350329

BACTERIAL PROFILES FOR CHRONIC AND AGGRESSIVE PERIODONTITIS IN A SAMPLE POPULATION GROUP. A CROSS-SECTIONAL STUDY

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Alexandra-Cornelia TEODORESCU

2017-06-01

Full Text Available Aim. The study aims at determining some possible significant differences in the subgingival microbial profiles of patients with generalized chronic periodontitis (GCP and generalized aggressive periodontitis (GAP, as a tool in helping with differential diagnostic. Materials and methods. 20 subgingival fluid samples (10 from GAP and 10 from GCP patients were subjected to a Real-Time Polymerase Chain Reaction technique in order to determine the prevalence and the counts of 9 periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tanerella forsythia, Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Eubacterium nodatum and Capnocytophaga gingivalis. Results and discussion. Fusobacterium nucleatum was singnificantly correlated with the aggressive periodontitis group, but no significant differences were found for the other 8 periodontal bacteria. Conclusions. The prevalence or count of some periodontal pathogens could help clinicians make an easier differential diagnostic between GCP and GAP, however further studies, conducted on larger population samples, are still needed.

Dynamic interactions of neutrophils and biofilms

Directory of Open Access Journals (Sweden)

Josefine Hirschfeld

2014-12-01

Full Text Available Background: The majority of microbial infections in humans are biofilm-associated and difficult to treat, as biofilms are highly resistant to antimicrobial agents and protect themselves from external threats in various ways. Biofilms are tenaciously attached to surfaces and impede the ability of host defense molecules and cells to penetrate them. On the other hand, some biofilms are beneficial for the host and contain protective microorganisms. Microbes in biofilms express pathogen-associated molecular patterns and epitopes that can be recognized by innate immune cells and opsonins, leading to activation of neutrophils and other leukocytes. Neutrophils are part of the first line of defense and have multiple antimicrobial strategies allowing them to attack pathogenic biofilms. Objective/design: In this paper, interaction modes of neutrophils with biofilms are reviewed. Antimicrobial strategies of neutrophils and the counteractions of the biofilm communities, with special attention to oral biofilms, are presented. Moreover, possible adverse effects of neutrophil activity and their biofilm-promoting side effects are discussed. Results/conclusion: Biofilms are partially, but not entirely, protected against neutrophil assault, which include the processes of phagocytosis, degranulation, and formation of neutrophil extracellular traps. However, virulence factors of microorganisms, microbial composition, and properties of the extracellular matrix determine whether a biofilm and subsequent microbial spread can be controlled by neutrophils and other host defense factors. Besides, neutrophils may inadvertently contribute to the physical and ecological stability of biofilms by promoting selection of more resistant strains. Moreover, neutrophil enzymes can degrade collagen and other proteins and, as a result, cause harm to the host tissues. These parameters could be crucial factors in the onset of periodontal inflammation and the subsequent tissue breakdown.

Determining productivity of transferred benthic biofilms within wetlands differing in anthropogenic stressors

International Nuclear Information System (INIS)

Frederick, K.; Foote, L.; Ciborowski, J.

2010-01-01

Algal biofilms are a fundamental contributor to wetland productivity. The films maintain high turnover rates, nutrient uptake and storage capacities are prevalent in shallow water as well as over large littoral zones. This study investigated biofilm transfer techniques as a means of accelerating carbon capture, plant production, and colonization in reclaimed oil sand wetlands affected by process water. The study examined the productivity of transferred biofilms and their ability to accelerate succession; methods of transferring biofilms; and the community composition of algae in relation to other substrates. Microcosms with 4 types of substrates were submersed in experimental trenches containing either process water or natural water. Dissolved oxygen, chlorophyll, and biomass standing crop sampling was conducted at intervals throughout a 1-year period. Analysis of variance (ANOVA) was conducted to compare the substrate types. Results of the study will be used to assess the impacts of oil sands process affected materials (OSPM) on benthic biofilm productivity and the initial carbon accumulation process.

Determining productivity of transferred benthic biofilms within wetlands differing in anthropogenic stressors

Energy Technology Data Exchange (ETDEWEB)

Frederick, K.; Foote, L. [Alberta Univ., Edmonton, AB (Canada); Ciborowski, J. [Windsor Univ., ON (Canada)

2010-07-01

Algal biofilms are a fundamental contributor to wetland productivity. The films maintain high turnover rates, nutrient uptake and storage capacities are prevalent in shallow water as well as over large littoral zones. This study investigated biofilm transfer techniques as a means of accelerating carbon capture, plant production, and colonization in reclaimed oil sand wetlands affected by process water. The study examined the productivity of transferred biofilms and their ability to accelerate succession; methods of transferring biofilms; and the community composition of algae in relation to other substrates. Microcosms with 4 types of substrates were submersed in experimental trenches containing either process water or natural water. Dissolved oxygen, chlorophyll, and biomass standing crop sampling was conducted at intervals throughout a 1-year period. Analysis of variance (ANOVA) was conducted to compare the substrate types. Results of the study will be used to assess the impacts of oil sands process affected materials (OSPM) on benthic biofilm productivity and the initial carbon accumulation process.

Effects of ceftazidime and ciprofloxacin on biofilm formation in Proteus mirabilis rods.

Science.gov (United States)

Kwiecińska-Piróg, Joanna; Bogiel, Tomasz; Gospodarek, Eugenia

2013-10-01

Proteus mirabilis rods are one of the most commonly isolated species of the Proteus genus from human infections, mainly those from the urinary tract and wounds. They are often related to biofilm structure formation. The bacterial cells of the biofilm are less susceptible to routinely used antimicrobials, making the treatment more difficult. The aim of this study was to evaluate quantitatively the influence of ceftazidime and ciprofloxacin on biofilm formation on the polyvinyl chloride surface by 42 P. mirabilis strains isolated from urine, purulence, wound swab and bedsore samples. It has been shown that ceftazidime and ciprofloxacin at concentrations equal to 1/4, 1/2 and 1 times their MIC values for particular Proteus spp. strains decrease their ability to form biofilms. Moreover, ciprofloxacin at concentrations equal to 1/4, 1/2 and 1 times their MIC values for particular P. mirabilis strains reduces biofilm formation more efficiently than ceftazidime at the corresponding concentration values.

Maggot excretions inhibit biofilm formation on biomaterials.

Science.gov (United States)

Cazander, Gwendolyn; van de Veerdonk, Mariëlle C; Vandenbroucke-Grauls, Christina M J E; Schreurs, Marco W J; Jukema, Gerrolt N

2010-10-01

Biofilm-associated infections in trauma surgery are difficult to treat with conventional therapies. Therefore, it is important to develop new treatment modalities. Maggots in captured bags, which are permeable for larval excretions/secretions, aid in healing severe, infected wounds, suspect for biofilm formation. Therefore we presumed maggot excretions/secretions would reduce biofilm formation. We studied biofilm formation of Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella oxytoca, Enterococcus faecalis, and Enterobacter cloacae on polyethylene, titanium, and stainless steel. We compared the quantities of biofilm formation between the bacterial species on the various biomaterials and the quantity of biofilm formation after various incubation times. Maggot excretions/secretions were added to existing biofilms to examine their effect. Comb-like models of the biomaterials, made to fit in a 96-well microtiter plate, were incubated with bacterial suspension. The formed biofilms were stained in crystal violet, which was eluted in ethanol. The optical density (at 595 nm) of the eluate was determined to quantify biofilm formation. Maggot excretions/secretions were pipetted in different concentrations to (nonstained) 7-day-old biofilms, incubated 24 hours, and finally measured. The strongest biofilms were formed by S. aureus and S. epidermidis on polyethylene and the weakest on titanium. The highest quantity of biofilm formation was reached within 7 days for both bacteria. The presence of excretions/secretions reduced biofilm formation on all biomaterials. A maximum of 92% of biofilm reduction was measured. Our observations suggest maggot excretions/secretions decrease biofilm formation and could provide a new treatment for biofilm formation on infected biomaterials.

A Nonbactericidal Zinc-Complexing Ligand as a Biofilm Inhibitor: Structure-Guided Contrasting Effects on Staphylococcus aureus Biofilm.

Science.gov (United States)

Kapoor, Vidushi; Rai, Rajanikant; Thiyagarajan, Durairaj; Mukherjee, Sandipan; Das, Gopal; Ramesh, Aiyagari

2017-08-04

Zinc-complexing ligands are prospective anti-biofilm agents because of the pivotal role of zinc in the formation of Staphylococcus aureus biofilm. Accordingly, the potential of a thiosemicarbazone (compound C1) and a benzothiazole-based ligand (compound C4) in the prevention of S. aureus biofilm formation was assessed. Compound C1 displayed a bimodal activity, hindering biofilm formation only at low concentrations and promoting biofilm growth at higher concentrations. In the case of C4, a dose-dependent inhibition of S. aureus biofilm growth was observed. Atomic force microscopy analysis suggested that at higher concentrations C1 formed globular aggregates, which perhaps formed a substratum that favored adhesion of cells and biofilm formation. In the case of C4, zinc supplementation experiments validated zinc complexation as a plausible mechanism of inhibition of S. aureus biofilm. Interestingly, C4 was nontoxic to cultured HeLa cells and thus has promise as a therapeutic anti-biofilm agent. The essential understanding of the structure-driven implications of zinc-complexing ligands acquired in this study might assist future screening regimes for identification of potent anti-biofilm agents. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Adenoid Reservoir for Pathogenic Biofilm Bacteria▿

Science.gov (United States)

Nistico, L.; Kreft, R.; Gieseke, A.; Coticchia, J. M.; Burrows, A.; Khampang, P.; Liu, Y.; Kerschner, J. E.; Post, J. C.; Lonergan, S.; Sampath, R.; Hu, F. Z.; Ehrlich, G. D.; Stoodley, P.; Hall-Stoodley, L.

2011-01-01

Biofilms of pathogenic bacteria are present on the middle ear mucosa of children with chronic otitis media (COM) and may contribute to the persistence of pathogens and the recalcitrance of COM to antibiotic treatment. Controlled studies indicate that adenoidectomy is effective in the treatment of COM, suggesting that the adenoids may act as a reservoir for COM pathogens. To investigate the bacterial community in the adenoid, samples were obtained from 35 children undergoing adenoidectomy for chronic OM or obstructive sleep apnea. We used a novel, culture-independent molecular diagnostic methodology, followed by confocal microscopy, to investigate the in situ distribution and organization of pathogens in the adenoids to determine whether pathogenic bacteria exhibited criteria characteristic of biofilms. The Ibis T5000 Universal Biosensor System was used to interrogate the extent of the microbial diversity within adenoid biopsy specimens. Using a suite of 16 broad-range bacterial primers, we demonstrated that adenoids from both diagnostic groups were colonized with polymicrobial biofilms. Haemophilus influenzae was present in more adenoids from the COM group (P = 0.005), but there was no significant difference between the two patient groups for Streptococcus pneumoniae or Staphylococcus aureus. Fluorescence in situ hybridization, lectin binding, and the use of antibodies specific for host epithelial cells demonstrated that pathogens were aggregated, surrounded by a carbohydrate matrix, and localized on and within the epithelial cell surface, which is consistent with criteria for bacterial biofilms. PMID:21307211

Portable hyperspectral fluorescence imaging system for detection of biofilms on stainless steel surfaces

Science.gov (United States)

Jun, Won; Lee, Kangjin; Millner, Patricia; Sharma, Manan; Chao, Kuanglin; Kim, Moon S.

2008-04-01

A rapid nondestructive technology is needed to detect bacterial contamination on the surfaces of food processing equipment to reduce public health risks. A portable hyperspectral fluorescence imaging system was used to evaluate potential detection of microbial biofilm on stainless steel typically used in the manufacture of food processing equipment. Stainless steel coupons were immersed in bacterium cultures, such as E. coli, Pseudomonas pertucinogena, Erwinia chrysanthemi, and Listeria innocula. Following a 1-week exposure, biofilm formations were assessed using fluorescence imaging. In addition, the effects on biofilm formation from both tryptic soy broth (TSB) and M9 medium with casamino acids (M9C) were examined. TSB grown cells enhance biofilm production compared with M9C-grown cells. Hyperspectral fluorescence images of the biofilm samples, in response to ultraviolet-A (320 to 400 nm) excitation, were acquired from approximately 416 to 700 nm. Visual evaluation of individual images at emission peak wavelengths in the blue revealed the most contrast between biofilms and stainless steel coupons. Two-band ratios compared with the single-band images increased the contrast between the biofilm forming area and stainless steel coupon surfaces. The 444/588 nm ratio images exhibited the greatest contrast between the biofilm formations and stainless coupon surfaces.

Mechanisms of Candida biofilm drug resistance

Science.gov (United States)

Taff, Heather T; Mitchell, Kaitlin F; Edward, Jessica A; Andes, David R

2013-01-01

Candida commonly adheres to implanted medical devices, growing as a resilient biofilm capable of withstanding extraordinarily high antifungal concentrations. As currently available antifungals have minimal activity against biofilms, new drugs to treat these recalcitrant infections are urgently needed. Recent investigations have begun to shed light on the mechanisms behind the profound resistance associated with the biofilm mode of growth. This resistance appears to be multifactorial, involving both mechanisms similar to conventional, planktonic antifungal resistance, such as increased efflux pump activity, as well as mechanisms specific to the biofilm lifestyle. A unique biofilm property is the production of an extracellular matrix. Two components of this material, β-glucan and extracellular DNA, promote biofilm resistance to multiple antifungals. Biofilm formation also engages several stress response pathways that impair the activity of azole drugs. Resistance within a biofilm is often heterogeneous, with the development of a subpopulation of resistant persister cells. In this article we review the molecular mechanisms underlying Candida biofilm antifungal resistance and their relative contributions during various growth phases. PMID:24059922

Oral Biofilm Architecture on Natural Teeth

NARCIS (Netherlands)

Zijnge, Vincent; van Leeuwen, M. Barbara M.; Degener, John E.; Abbas, Frank; Thurnheer, Thomas; Gmuer, Rudolf; Harmsen, Hermie J. M.

2010-01-01

Periodontitis and caries are infectious diseases of the oral cavity in which oral biofilms play a causative role. Moreover, oral biofilms are widely studied as model systems for bacterial adhesion, biofilm development, and biofilm resistance to antibiotics, due to their widespread presence and

Candida Biofilms: Development, Architecture, and Resistance

Science.gov (United States)

CHANDRA, JYOTSNA; MUKHERJEE, PRANAB K.

2015-01-01

Intravascular device–related infections are often associated with biofilms (microbial communities encased within a polysaccharide-rich extracellular matrix) formed by pathogens on the surfaces of these devices. Candida species are the most common fungi isolated from catheter-, denture-, and voice prosthesis–associated infections and also are commonly isolated from contact lens–related infections (e.g., fungal keratitis). These biofilms exhibit decreased susceptibility to most antimicrobial agents, which contributes to the persistence of infection. Recent technological advances have facilitated the development of novel approaches to investigate the formation of biofilms and identify specific markers for biofilms. These studies have provided extensive knowledge of the effect of different variables, including growth time, nutrients, and physiological conditions, on biofilm formation, morphology, and architecture. In this article, we will focus on fungal biofilms (mainly Candida biofilms) and provide an update on the development, architecture, and resistance mechanisms of biofilms. PMID:26350306

Anti-Biofilm and Immunomodulatory Activities of Peptides That Inhibit Biofilms Formed by Pathogens Isolated from Cystic Fibrosis Patients

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César de la Fuente-Núñez

2014-10-01

Full Text Available Cystic fibrosis (CF patients often acquire chronic respiratory tract infections due to Pseudomonas aeruginosa and Burkholderia cepacia complex (Bcc species. In the CF lung, these bacteria grow as multicellular aggregates termed biofilms. Biofilms demonstrate increased (adaptive resistance to conventional antibiotics, and there are currently no available biofilm-specific therapies. Using plastic adherent, hydroxyapatite and flow cell biofilm models coupled with confocal and scanning electron microscopy, it was demonstrated that an anti-biofilm peptide 1018 prevented biofilm formation, eradicated mature biofilms and killed biofilms formed by a wide range of P. aeruginosa and B. cenocepacia clinical isolates. New peptide derivatives were designed that, compared to their parent peptide 1018, showed similar or decreased anti-biofilm activity against P. aeruginosa biofilms, but increased activity against biofilms formed by the Gram-positive bacterium methicillin resistant Staphylococcus aureus. In addition, some of these new peptide derivatives retained the immunomodulatory activity of 1018 since they induced the production of the chemokine monocyte chemotactic protein-1 (MCP-1 and suppressed lipopolysaccharide-mediated tumor necrosis factor-α (TNF-α production by human peripheral blood mononuclear cells (PBMC and were non-toxic towards these cells. Peptide 1018 and its derivatives provide promising leads for the treatment of chronic biofilm infections and hyperinflammatory lung disease in CF patients.

Taxonomic and chemical assessment of exceptionally abundant rock mine biofilm

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Karolina Tomczyk-Żak

2017-08-01

Full Text Available Background An exceptionally thick biofilm covers walls of ancient gold and arsenic Złoty Stok mine (Poland in the apparent absence of organic sources of energy. Methods and Results We have characterized this microbial community using culture-dependent and independent methods. We sequenced amplicons of the 16S rRNA gene obtained using generic primers and additional primers targeted at Archaea and Actinobacteria separately. Also, we have cultured numerous isolates from the biofilm on different media under aerobic and anaerobic conditions. We discovered very high biodiversity, and no single taxonomic group was dominant. The majority of almost 4,000 OTUs were classified above genus level indicating presence of novel species. Elemental analysis, performed using SEM-EDS and X-ray, of biofilm samples showed that carbon, sulphur and oxygen were not evenly distributed in the biofilm and that their presence is highly correlated. However, the distribution of arsenic and iron was more flat, and numerous intrusions of elemental silver and platinum were noted, indicating that microorganisms play a key role in releasing these elements from the rock. Conclusions Altogether, the picture obtained throughout this study shows a very rich, complex and interdependent system of rock biofilm. The chemical heterogeneity of biofilm is a likely explanation as to why this oligotrophic environment is capable of supporting such high microbial diversity.

Biofilm Induced Tolerance Towards Antimicrobial Peptides

DEFF Research Database (Denmark)

Folkesson, Anders; Haagensen, Janus Anders Juul; Zampaloni, Claudia

2008-01-01

Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We address the question of how biofilm organization affects antibiotic susceptibility. We established Escherichia coli biofilms with differential structural organization due...... to the presence of IncF plasmids expressing altered forms of the transfer pili in two different biofilm model systems. The mature biofilms were subsequently treated with two antibiotics with different molecular targets, the peptide antibiotic colistin and the fluoroquinolone ciprofloxacin. The dynamics...... of microbial killing were monitored by viable count determination, and confocal laser microscopy. Strains forming structurally organized biofilms show an increased bacterial survival when challenged with colistin, compared to strains forming unstructured biofilms. The increased survival is due to genetically...

Conductive properties of methanogenic biofilms.

Science.gov (United States)

Li, Cheng; Lesnik, Keaton Larson; Liu, Hong

2018-02-01

Extracellular electron transfer between syntrophic partners needs to be efficiently maintained in methanogenic environments. Direct extracellular electron transfer via electrical current is an alternative to indirect hydrogen transfer but requires construction of conductive extracellular structures. Conductive mechanisms and relationship between conductivity and the community composition in mixed-species methanogenic biofilms are not well understood. The present study investigated conductive behaviors of methanogenic biofilms and examined the correlation between biofilm conductivity and community composition between different anaerobic biofilms enriched from the same inoculum. Highest conductivity observed in methanogenic biofilms was 71.8±4.0μS/cm. Peak-manner response of conductivity upon changes over a range of electrochemical potentials suggests that electron transfer in methanogenic biofilms occurs through redox driven super-exchange. The strong correlation observed between biofilm conductivity and Geobacter spp. in the metabolically diverse anaerobic communities suggests that the efficiency of DEET may provide pressure for microbial communities to select for species that can produce electrical conduits. Copyright © 2017 Elsevier B.V. All rights reserved.

Silver against Pseudomonas aeruginosa biofilms

DEFF Research Database (Denmark)

Bjarnsholt, Thomas; Kirketerp-Møller, K.; Kristiansen, S.

2007-01-01

bacteria in both the planktonic and biofilm modes of growth. The action of silver on mature in vitro biofilms of Pseudomonas aeruginosa, a primary pathogen of chronic infected wounds, was investigated. The results show that silver is very effective against mature biofilms of P. aeruginosa......, but that the silver concentration is important. A concentration of 5-10 ig/mL silver sulfadiazine eradicated the biofilm whereas a lower concentration (1 ig/mL) had no effect. The bactericidal concentration of silver required to eradicate the bacterial biofilm was 10-100 times higher than that used to eradicate...... planktonic bacteria. These observations strongly indicate that the concentration of silver in currently available wound dressings is much too low for treatment of chronic biofilm wounds. It is suggested that clinicians and manufacturers of the said wound dressings consider whether they are treating wounds...

Dynamics of Biofilm Regrowth in Drinking Water Distribution Systems.

Science.gov (United States)

Douterelo, I; Husband, S; Loza, V; Boxall, J

2016-07-15

The majority of biomass within water distribution systems is in the form of attached biofilm. This is known to be central to drinking water quality degradation following treatment, yet little understanding of the dynamics of these highly heterogeneous communities exists. This paper presents original information on such dynamics, with findings demonstrating patterns of material accumulation, seasonality, and influential factors. Rigorous flushing operations repeated over a 1-year period on an operational chlorinated system in the United Kingdom are presented here. Intensive monitoring and sampling were undertaken, including time-series turbidity and detailed microbial analysis using 16S rRNA Illumina MiSeq sequencing. The results show that bacterial dynamics were influenced by differences in the supplied water and by the material remaining attached to the pipe wall following flushing. Turbidity, metals, and phosphate were the main factors correlated with the distribution of bacteria in the samples. Coupled with the lack of inhibition of biofilm development due to residual chlorine, this suggests that limiting inorganic nutrients, rather than organic carbon, might be a viable component in treatment strategies to manage biofilms. The research also showed that repeat flushing exerted beneficial selective pressure, giving another reason for flushing being a viable advantageous biofilm management option. This work advances our understanding of microbiological processes in drinking water distribution systems and helps inform strategies to optimize asset performance. This research provides novel information regarding the dynamics of biofilm formation in real drinking water distribution systems made of different materials. This new knowledge on microbiological process in water supply systems can be used to optimize the performance of the distribution network and to guarantee safe and good-quality drinking water to consumers. Copyright © 2016 Douterelo et al.

Anti-Biofilm and Antivirulence Activities of Metabolites from Plectosphaerella cucumerina against Pseudomonas aeruginosa

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Jinwei Zhou

2017-05-01

Full Text Available This study reported the efficacy of the metabolites of Plectosphaerella cucumerina, one phyllosphere fungus from Orychophragmus violaceus, against Pseudomonas aeruginosa quorum sensing (QS and QS-regulated biofilms. The minimum inhibitory concentration (MIC of the ethyl acetate (EtOAc extract from P. cucumerina against P. aeruginosa PAO1 was 1.25 mg mL−1. At sub-MIC concentrations, P. cucumerina extract (0.25–1 mg mL−1 not only inhibited biofilm formation but also disrupted preformed biofilms of P. aeruginosa PAO1 without affecting its growth. Fluorescence and scanning electron microscope (SEM showed architectural disruption of the biofilms when treated with P. cucumerina metabolites. Further investigation demonstrated that metabolites in P. cucumerina attenuated the QS-dependent virulence factors. LC-MS/MS spectra coupled with experimentally standard samples suggested that patulin and emodin might act as the principal components possessing anti-biofilm and antivirulence activities. This is the first report of (1 the isolation of P. cucumerina from the phyllosphere of O. violaceus and (2 anti-biofilm, antivirulence, and biofilm disruption activities of this fungus. Thus, this study provides fascinating new pathways for screening antipathogenic agents.

Nitritation performance and biofilm development of co- and counter-diffusion biofilm reactors: Modeling and experimental comparison

DEFF Research Database (Denmark)

Wang, Rongchang; Terada, Akihiko; Lackner, Susanne

2009-01-01

A comparative study was conducted on the start-up performance and biofilm development in two different biofilm reactors with aim of obtaining partial nitritation. The reactors were both operated under oxygen limited conditions, but differed in geometry. While substrates (O-2, NH3) co......-diffused in one geometry, they counter-diffused in the other. Mathematical simulations of these two geometries were implemented in two 1-D multispecies biofilm models using the AQUASIM software. Sensitivity analysis results showed that the oxygen mass transfer coefficient (K-i) and maximum specific growth rate...... results showed that the counter-diffusion biofilms developed faster and attained a larger maximum biofilm thickness than the co-diffusion biofilms. Under oxygen limited condition (DO

Strategies for combating bacterial biofilms: A focus on anti-biofilm agents and their mechanisms of action

Science.gov (United States)

Roy, Ranita; Tiwari, Monalisa; Donelli, Gianfranco; Tiwari, Vishvanath

2018-01-01

ABSTRACT Biofilm refers to the complex, sessile communities of microbes found either attached to a surface or buried firmly in an extracellular matrix as aggregates. The biofilm matrix surrounding bacteria makes them tolerant to harsh conditions and resistant to antibacterial treatments. Moreover, the biofilms are responsible for causing a broad range of chronic diseases and due to the emergence of antibiotic resistance in bacteria it has really become difficult to treat them with efficacy. Furthermore, the antibiotics available till date are ineffective for treating these biofilm related infections due to their higher values of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), which may result in in-vivo toxicity. Hence, it is critically important to design or screen anti-biofilm molecules that can effectively minimize and eradicate biofilm related infections. In the present article, we have highlighted the mechanism of biofilm formation with reference to different models and various methods used for biofilm detection. A major focus has been put on various anti-biofilm molecules discovered or tested till date which may include herbal active compounds, chelating agents, peptide antibiotics, lantibiotics and synthetic chemical compounds along with their structures, mechanism of action and their respective MICs, MBCs, minimum biofilm inhibitory concentrations (MBICs) as well as the half maximal inhibitory concentration (IC50) values available in the literature so far. Different mode of action of anti biofilm molecules addressed here are inhibition via interference in the quorum sensing pathways, adhesion mechanism, disruption of extracellular DNA, protein, lipopolysaccharides, exopolysaccharides and secondary messengers involved in various signaling pathways. From this study, we conclude that the molecules considered here might be used to treat biofilm-associated infections after significant structural modifications, thereby

Transcriptome Analysis of Porphyromonas gingivalis and Acinetobacter baumannii in Polymicrobial Communities.

Science.gov (United States)

Miller, Daniel P; Wang, Qian; Weinberg, Aaron; Lamont, Richard J

2018-06-25

Acinetobacter baumannii is a nosocomial, opportunistic pathogen that causes several serious conditions such as meningitis, septicemia, endocarditis and pneumonia. It can be found in the oral biofilm, which may be a reservoir for pneumonia and chronic obstructive pulmonary disease. Subgingival colonization by A. baumannii is associated with chronic and aggressive periodontitis as well as refractory periodontal disease. Porphyromonas gingivalis, a keystone periodontal pathogen localized to subgingival plaque, is also implicated in several chronic conditions including aspiration pneumonia. While both bacteria are found together in subgingival plaque and can cause multiple polymicrobial infections, nothing is known about the interactions between these two important human pathogens. In this study, we used RNA sequencing to understand the transcriptional response of both species as they adapt to heterotypic communities. Among the differentially regulated genes were those encoding a number of important virulence factors for both species including adhesion, biofilm formation, and protein secretion. Additionally, the presence of A. baumannii increased the abundance of P. gingivalis in model dual species communities Collectively these results suggest that both P. gingivalis and A. baumannii adapt to each other and have synergistic potential for increased pathogenicity. In identifying the mechanisms that promote pathogenicity and refractory disease, novel approaches to mitigate polymicrobial synergistic interactions may be developed to treat or prevent associated diseases. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Extracellular DNA Contributes to Dental Biofilm Stability

DEFF Research Database (Denmark)

Schlafer, Sebastian; Meyer, Rikke Louise; Dige, Irene

2017-01-01

dental biofilms. This study aimed to determine whether eDNA was part of the matrix in biofilms grown in situ in the absence of sucrose and whether treatment with DNase dispersed biofilms grown for 2.5, 5, 7.5, 16.5, or 24 h. Three hundred biofilms from 10 study participants were collected and treated...... the amount of biofilm in very early stages of growth (up to 7.5 h), but the treatment effect decreased with increasing biofilm age. This study proves the involvement of eDNA in dental biofilm formation and its importance for biofilm stability in the earliest stages. Further research is required to uncover...

Biofilm Formation by Mycobacterium bovis: Influence of Surface Kind and Temperatures of Sanitizer Treatments on Biofilm Control

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Victoria O. Adetunji

2014-01-01

Full Text Available Mycobacterium bovis causes classic bovine tuberculosis, a zoonosis which is still a concern in Africa. Biofilm forming ability of two Mycobacterium bovis strains was assessed on coupons of cement, ceramic, or stainless steel in three different microbiological media at 37°C with agitation for 2, 3, or 4 weeks to determine the medium that promotes biofilm. Biofilm mass accumulated on coupons was treated with 2 sanitizers (sanitizer A (5.5 mg L−1 active iodine and sanitizer B (170.6 g1 alkyl dimethylbenzyl ammonium chloride, 78 g−1 didecyldimethyl ammonium chloride, 107.25 g L−1 glutaraldehyde, 146.25 g L−1 isopropanol, and 20 g L−1 pine oil at 28 and 45°C and in hot water at 85°C for 5 min. Residual biofilms on treated coupons were quantified using crystal violet binding assay. The two strains had a similar ability to form biofilms on the three surfaces. More biofilms were developed in media containing 5% liver extract. Biofilm mass increased as incubation time increased till the 3rd week. More biofilms were formed on cement than on ceramic and stainless steel surfaces. Treatment with hot water at 85°C reduced biofilm mass, however, sanitizing treatments at 45°C removed more biofilms than at 28°C. However, neither treatment completely eliminated the biofilms. The choice of processing surface and temperatures used for sanitizing treatments had an impact on biofilm formation and its removal from solid surfaces.

Biofilm characteristics and evaluation of the sanitation procedures of thermophilic Aeribacillus pallidus E334 biofilms.

Science.gov (United States)

Kilic, Tugba; Karaca, Basar; Ozel, Beste Piril; Ozcan, Birgul; Cokmus, Cumhur; Coleri Cihan, Arzu

2017-04-01

The ability of Aeribacillus pallidus E334 to produce pellicle and form a biofilm was studied. Optimal biofilm formation occurred at 60 °C, pH 7.5 and 1.5% NaCl. Extra polymeric substances (EPS) were composed of proteins and eDNA (21.4 kb). E334 formed biofilm on many surfaces, but mostly preferred polypropylene and glass. Using CLSM analysis, the network-like structure of the EPS was observed. The A. pallidus biofilm had a novel eDNA content. DNaseI susceptibility (86.8% removal) of eDNA revealed its importance in mature biofilms, but the purified eDNA was resistant to DNaseI, probably due to its extended folding outside the matrix. Among 15 cleaning agents, biofilms could be removed with alkaline protease and sodium dodecyl sulphate (SDS). The removal of cells from polypropylene and biomass on glass was achieved with combined SDS/alkaline protease treatment. Strong A. pallidus biofilms could cause risks for industrial processes and abiotic surfaces must be taken into consideration in terms of sanitation procedures.

Levels of Candidate Periodontal Pathogens in Subgingival Biofilm

OpenAIRE

Oliveira, R.R.D.S.; Fermiano, D.; Feres, M.; Figueiredo, L.C.; Teles, F.R.F.; Soares, G.M.S.; Faveri, M.

2016-01-01

In recent years, several new periodontal taxa have been associated with the etiology of periodontitis. A recent systematic review provides further support for the pathogenic role of 17 species/phylotypes. Thus, the aim of this study was to assess the prevalence and levels of these species in subjects with generalized chronic periodontitis (GChP; n = 30), generalized aggressive periodontitis (GAgP; n = 30), and periodontal health (PH; n = 30). All subjects underwent clinical and microbiologica...

Metal concentrations in stream biofilm and sediments and their potential to explain biofilm microbial community structure

International Nuclear Information System (INIS)

Ancion, Pierre-Yves; Lear, Gavin; Dopheide, Andrew; Lewis, Gillian D.

2013-01-01

Concentrations of metals associated with sediments have traditionally been analysed to assess the extent of heavy metal contamination in freshwater environments. Stream biofilms present an alternative medium for this assessment which may be more relevant to the risk incurred by stream ecosystems as they are intensively grazed by aquatic organisms at a higher trophic level. Therefore, we investigated zinc, copper and lead concentrations in biofilms and sediments of 23 stream sites variously impacted by urbanisation. Simultaneously, biofilm bacterial and ciliate protozoan community structure was analysed by Automated Ribosomal Intergenic Spacer Analysis and Terminal Restriction Fragment Length Polymorphism, respectively. Statistical analysis revealed that biofilm associated metals explained a greater proportion of the variations observed in bacterial and ciliate communities than did sediment associated-metals. This study suggests that the analysis of metal concentrations in biofilms provide a good assessment of detrimental effects of metal contaminants on aquatic biota. - Highlights: ► Zn, Cu and Pb concentrations in biofilm and sediments from 23 streams were assessed. ► Bacteria and ciliate protozoa were simultaneously used as biological indicators. ► Zn and Cu were generally enriched in biofilm compared to sediments. ► Metals in biofilm provide a useful assessment of freshwater ecosystem contamination. ► Results highlight the likely ecological importance of biofilm associated metals. - Metal concentrations in stream biofilms provide a good assessment of the effects of trace metal contaminants on freshwater ecosystems.

The clinical impact of bacterial biofilms

DEFF Research Database (Denmark)

Høiby, Niels; Ciofu, Oana; Johansen, Helle Krogh

2011-01-01

Bacteria survive in nature by forming biofilms on surfaces and probably most, if not all, bacteria (and fungi) are capable of forming biofilms. A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and extracellular DNA....... Bacterial biofilms are resistant to antibiotics, disinfectant chemicals and to phagocytosis and other components of the innate and adaptive inflammatory defense system of the body. It is known, for example, that persistence of staphylococcal infections related to foreign bodies is due to biofilm formation....... Likewise, chronic Pseudomonas aeruginosa lung infections in cystic fibrosis patients are caused by biofilm growing mucoid strains. Gradients of nutrients and oxygen exist from the top to the bottom of biofilms and the bacterial cells located in nutrient poor areas have decreased metabolic activity...

Calcium-Phosphate-Osteopontin Particles Reduce Biofilm Formation and pH Drops in in situ Grown Dental Biofilms.

Science.gov (United States)

Schlafer, Sebastian; Ibsen, Casper J S; Birkedal, Henrik; Nyvad, Bente

2017-01-01

This 2-period crossover study investigated the effect of calcium-phosphate-osteopontin particles on biofilm formation and pH in 48-h biofilms grown in situ. Bovine milk osteopontin is a highly phosphorylated glycoprotein that has been shown to interfere with bacterial adhesion to salivary-coated surfaces. Calcium-phosphate-osteopontin particles have been shown to reduce biofilm formation and pH drops in a 5-species laboratory model of dental biofilm without affecting bacterial viability. Here, smooth surface biofilms from 10 individuals were treated ex vivo 6 times/day for 30 min with either calcium-phosphate-osteopontin particles or sterile saline. After growth, the amount of biofilm formed was determined by confocal microscopy, and pH drops upon exposure to glucose were monitored using confocal-microscopy-based pH ratiometry. A total of 160 biofilms were analysed. No adverse effects of repeated ex vivo treatment with calcium-phosphate-osteopontin particles were observed. Particle treatment resulted in a 32% lower amount of biofilm formed (p Biofilm pH was significantly higher upon particle treatment, both shortly after the addition of glucose and after 30 min of incubation with glucose (p biofilms as well as the remineralizing potential of the particles. © 2016 S. Karger AG, Basel.

pH, redox potential and local biofilm potential microenvironments within Geobacter sulfurreducens biofilms and their roles in electron transfer.

Science.gov (United States)

Babauta, Jerome T; Nguyen, Hung Duc; Harrington, Timothy D; Renslow, Ryan; Beyenal, Haluk

2012-10-01

The limitation of pH inside electrode-respiring biofilms is a well-known concept. However, little is known about how pH and redox potential are affected by increasing current inside biofilms respiring on electrodes. Quantifying the variations in pH and redox potential with increasing current is needed to determine how electron transfer is tied to proton transfer within the biofilm. In this research, we quantified pH and redox potential variations in electrode-respiring Geobacter sulfurreducens biofilms as a function of respiration rates, measured as current. We also characterized pH and redox potential at the counter electrode. We concluded that (1) pH continued to decrease in the biofilm through different growth phases, showing that the pH is not always a limiting factor in a biofilm and (2) decreasing pH and increasing redox potential at the biofilm electrode were associated only with the biofilm, demonstrating that G. sulfurreducens biofilms respire in a unique internal environment. Redox potential inside the biofilm was also compared to the local biofilm potential measured by a graphite microelectrode, where the tip of the microelectrode was allowed to acclimatize inside the biofilm. Copyright © 2012 Wiley Periodicals, Inc.

Exploring the secrets of the three-dimensional architecture of phototrophic biofilms in caves

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Roldàn Monica

2009-01-01

Full Text Available Caves with dim natural light, and lighted hypogean environments, have been found to host phototrophic microorganisms from varioustaxonomic groups. These microorganisms group themselves into assemblies known as communities or biofilms, which are associated withrock surfaces. In this work, the phototrophic biofilms that colonise speleothems, walls and floors in three tourist caves (Spain were studied.Confocal laser scanning microscopy (CLSM and scanning electron microscopy (SEM were used to study these organisms and acquirethree-dimensional data on their biofilm structure. CLSM was used in a multi-channel mode whereby the different channels map individualbiofilm components. Cyanobacteria, green microalgae, diatoms, mosses and lichens were found to be grouped as biofilms that differedaccording to the sampling sites. The biofilms were classified into six types regarding their environmental conditions. These types weredefined by their constituent organisms, the thickness of their photosynthetic layers and their structure. Light-related stress is associated with lower biofilm thickness and species diversity, as is low humidity, and, in the case of artificially illuminated areas, the duration of lightexposure.

Cleaning and Disinfection of Bacillus cereus Biofilm.

Science.gov (United States)

Deal, Amanda; Klein, Dan; Lopolito, Paul; Schwarz, John Spencer

2016-01-01

Methodology has been evolving for the testing of disinfectants against bacterial single-species biofilms, as the difficulty of biofilm remediation continues to gain much-needed attention. Bacterial single-species biofilm contamination presents a real risk to good manufacturing practice-regulated industries. However, mixed-species biofilms and biofilms containing bacterial spores remain an even greater challenge for cleaning and disinfection. Among spore-forming microorganisms frequently encountered in pharmaceutical manufacturing areas, the spores of Bacillus cereus are often determined to be the hardest to disinfect and eradicate. One of the reasons for the low degree of susceptibility to disinfection is the ability of these spores to be encapsulated within an exopolysachharide biofilm matrix. In this series of experiments, we evaluated the disinfectant susceptibility of B. cereus biofilms relative to disassociated B. cereus spores and biofilm from a non-spore-forming species. Further, we assessed the impact that pre-cleaning has on increasing that susceptibility. Methodology has been evolving for the testing of disinfectants against bacterial single-species biofilms, as the difficulty of biofilm remediation continues to gain much-needed attention. Bacterial single-species biofilm contamination presents a real risk to good manufacturing practice-regulated industries. However, mixed-species biofilms and biofilms containing bacterial spores remain an even greater challenge for cleaning and disinfection. Among spore-forming microorganisms frequently encountered in pharmaceutical manufacturing areas, the spores of Bacillus cereus are often determined to be the hardest to disinfect and eradicate. One of the reasons for the low degree of susceptibility to disinfection is the ability of these spores to be encapsulated within an exopolysachharide biofilm matrix. In this series of experiments, we evaluated the disinfectant susceptibility of B. cereus biofilms relative to

A Commensal Strain of Staphylococcus epidermidis Overexpresses Membrane Proteins Associated with Pathogenesis When Grown in Biofilms.

Science.gov (United States)

Águila-Arcos, S; Ding, S; Aloria, K; Arizmendi, J M; Fearnley, I M; Walker, J E; Goñi, F M; Alkorta, I

2015-06-01

Staphylococcus epidermidis has emerged as one of the major nosocomial pathogens associated with infections of implanted medical devices. The most important factor in the pathogenesis of these infections is the formation of bacterial biofilms. Bacteria grown in biofilms are more resistant to antibiotics and to the immune defence system than planktonic bacteria. In these infections, the antimicrobial therapy usually fails and the removal of the biofilm-coated implanted device is the only effective solution. In this study, three proteomic approaches were performed to investigate membrane proteins associated to biofilm formation: (i) sample fractionation by gel electrophoresis, followed by isotopic labelling and LC-MS/MS analysis, (ii) in-solution sample preparation, followed by isotopic labelling and LC-MS/MS analysis and (iii) in-solution sample preparation and label-free LC-MS/MS analysis. We found that the commensal strain S. epidermidis CECT 231 grown in biofilms expressed higher levels of five membrane and membrane-associated proteins involved in pathogenesis: accumulation-associated protein, staphylococcal secretory antigen, signal transduction protein TRAP, ribonuclease Y and phenol soluble modulin beta 1 when compared with bacteria grown under planktonic conditions. These results indicate that a commensal strain can acquire a pathogenic phenotype depending on the mode of growth.

Antibiotic tolerance and microbial biofilms

DEFF Research Database (Denmark)

Folkesson, Anders

Increased tolerance to antimicrobial agents is thought to be an important feature of microbes growing in biofilms. We study the dynamics of antibiotic action within hydrodynamic flow chamber biofilms of Escherichia coli and Pseudomonas aeruginosa using isogenic mutants and fluorescent gene...... expression reporters and we address the question of how biofilm organization affects antibiotic susceptibility. The dynamics of microbial killing is monitored by viable count determination, and confocal laser microscopy. Our work shows that the apparent increased antibiotic tolerance is due to the formation...... of antibiotic tolerant subpopulations within the biofilm. The formation of these subpopulations is highly variable and dependent on the antibiotic used, the biofilm structural organization and the induction of specific tolerance mechanisms....

Biofilm formation on abiotic surfaces

DEFF Research Database (Denmark)

Tang, Lone

2011-01-01

Bacteria can attach to any surface in contact with water and proliferate into complex communities enclosed in an adhesive matrix, these communities are called biofilms. The matrix makes the biofilm difficult to remove by physical means, and bacteria in biofilm can survive treatment with many...

Role of biofilm roughness and hydrodynamic conditions in Legionella pneumophila adhesion to and detachment from simulated drinking water biofilms.

Science.gov (United States)

Shen, Yun; Monroy, Guillermo L; Derlon, Nicolas; Janjaroen, Dao; Huang, Conghui; Morgenroth, Eberhard; Boppart, Stephen A; Ashbolt, Nicholas J; Liu, Wen-Tso; Nguyen, Thanh H

2015-04-07

Biofilms in drinking water distribution systems (DWDS) could exacerbate the persistence and associated risks of pathogenic Legionella pneumophila (L. pneumophila), thus raising human health concerns. However, mechanisms controlling adhesion and subsequent detachment of L. pneumophila associated with biofilms remain unclear. We determined the connection between L. pneumophila adhesion and subsequent detachment with biofilm physical structure characterization using optical coherence tomography (OCT) imaging technique. Analysis of the OCT images of multispecies biofilms grown under low nutrient condition up to 34 weeks revealed the lack of biofilm deformation even when these biofilms were exposed to flow velocity of 0.7 m/s, typical flow for DWDS. L. pneumophila adhesion on these biofilm under low flow velocity (0.007 m/s) positively correlated with biofilm roughness due to enlarged biofilm surface area and local flow conditions created by roughness asperities. The preadhered L. pneumophila on selected rough and smooth biofilms were found to detach when these biofilms were subjected to higher flow velocity. At the flow velocity of 0.1 and 0.3 m/s, the ratio of detached cell from the smooth biofilm surface was from 1.3 to 1.4 times higher than that from the rough biofilm surface, presumably because of the low shear stress zones near roughness asperities. This study determined that physical structure and local hydrodynamics control L. pneumophila adhesion to and detachment from simulated drinking water biofilm, thus it is the first step toward reducing the risk of L. pneumophila exposure and subsequent infections.

Microbial analysis of in situ biofilm formation in drinking water distribution systems: implications for monitoring and control of drinking water quality.

Science.gov (United States)

Douterelo, Isabel; Jackson, M; Solomon, C; Boxall, J

2016-04-01

Biofilm formation in drinking water distribution systems (DWDS) is influenced by the source water, the supply infrastructure and the operation of the system. A holistic approach was used to advance knowledge on the development of mixed species biofilms in situ, by using biofilm sampling devices installed in chlorinated networks. Key physico-chemical parameters and conventional microbial indicators for drinking water quality were analysed. Biofilm coverage on pipes was evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The microbial community structure, bacteria and fungi, of water and biofilms was assessed using pyrosequencing. Conventional wisdom leads to an expectation for less microbial diversity in groundwater supplied systems. However, the analysis of bulk water showed higher microbial diversity in groundwater site samples compared with the surface water site. Conversely, higher diversity and richness were detected in biofilms from the surface water site. The average biofilm coverage was similar among sites. Disinfection residual and other key variables were similar between the two sites, other than nitrates, alkalinity and the hydraulic conditions which were extremely low at the groundwater site. Thus, the unexpected result of an exceptionally low diversity with few dominant genera (Pseudomonas and Basidiobolus) in groundwater biofilm samples, despite the more diverse community in the bulk water, is attributed to the low-flow hydraulic conditions. This finding evidences that the local environmental conditions are shaping biofilm formation, composition and amount, and hence managing these is critical for the best operation of DWDS to safeguard water quality.

Biofilm formation in surface and drinking water distribution systems in Mafikeng, South Africa

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Suma George Mulamattathil

2014-11-01

Full Text Available Poor quality source water and poorly treated reused wastewater may result in poor quality drinking water that has a higher potential to form biofilms. A biofilm is a group of microorganisms which adhere to a surface. We investigated biofilm growth in the drinking water distribution systems in the Mafikeng area, in the North- West Province of South Africa. Analysis was conducted to determine the presence of faecal coliforms, total coliforms, Pseudomonas spp. and Aeromonas spp. in the biofilms. Biofilms were grown on a device that contained copper and galvanised steel coupons. A mini tap filter – a point-of-use treatment device which can be used at a single faucet – was also used to collect samples. Scanning electron microscopy demonstrated that multi-species biofilms developed on all the coupons as well as on the point-of-use filters. Galvanised steel and carbon filters had the highest density of biofilm. Total coliforms, faecal coliforms and Pseudomonas spp. were isolated from raw water biofilm coupons only. Aeromonas spp. and Pseudomonas spp. were isolated from filters. The susceptibility of selected isolates was tested against 11 antibiotics of clinical interest. The most prevalent antibiotic resistance phenotype observed was KF-AP-C-E-OT-K-TM-A. The presence of virulence genes was determined using the polymerase chain reaction. These results indicate that bacteria present in the water have the ability to colonise as biofilms and drinking water biofilms may be a reservoir for opportunistic bacteria including Pseudomonas and Aeromonas species.

Exopolymeric substances from drinking water biofilms: Dynamics of production and relation with disinfection by products.

Science.gov (United States)

Lemus Pérez, M F; Rodríguez Susa, M

2017-06-01

Exopolymeric substances (EPS) as an external matrix of biofilm could react with disinfectants in drinking water networks forming disinfection by-products (DBP). Based on an experimental setup using two chlorine conditions-biofilm 1 (2.6 ± 0.8 mgCl/L) and biofilm 2 (0.7 ± 0.2 mg Cl/L)-samples of biofilms were recovered during 9 campaigns and EPS were extracted. Analyses of SUVA, fluorescence and amino acid (AA) content were carried out on the EPS to observe variation over time and correlations with DBP formation potential (DBP fp ) after chlorination. SUVA values were under 2 L/mgC*m showing that both EPS were hydrophilic. Slightly higher SUVA in biofilm 2 with low variation over time was observed. Fluorescence showed that aromatic proteins and fulvic like substances were the principal components and increased in biofilm 1 over time. AA decreased with time, and higher values of alanine, threonine, proline and isoleucine were observed in biofilm 2. Based on general associations, the SUVA of biofilm 2 correlated well with chloroform (CF) (r = 0.80). Generally, in both biofilms, tryptophan-like substances were negatively correlated with DBP while humic acid-like substances correlated positively, but with low indexes (r = 0.3-0.6). Correlations of data from individual sampling increased the indices (r over 0.8), suggesting a temporal influence of other factors on DBP fp such as inorganics, filtered water and the structural composition of EPS. In biofilm 1, Br-haloacetic acids (Br-HAA), dibromoacetonitrile and bromochloro acetonitrile were inversely associated with arginine and valine, as were di and trichloropropanone to arginine. On the contrary, in biofilm 2, the following amino acids correlated positively with DBP: alanine with Br-HAA, alanine with CF, alanine with N-DBP (chloropicrin, di and tri-chloro acetonitrile), and valine with CF. As this is the first report about the relation between temporal variation of EPS and DBP fp of biofilms in two

Optimized candidal biofilm microtiter assay

NARCIS (Netherlands)

Krom, Bastiaan P.; Cohen, Jesse B.; Feser, Gail E. McElhaney; Cihlar, Ronald L.

Microtiter based candidal biofilm formation is commonly being used. Here we describe the analysis of factors influencing the development of candidal biofilms such as the coating with serum, growth medium and pH. The data reported here show that optimal candidal biofilm formation is obtained when

Identification of anti-biofilm components in Withania somnifera and their effect on virulence of Streptococcus mutans biofilms.

Science.gov (United States)

Pandit, S; Cai, J N; Song, K Y; Jeon, J G

2015-08-01

The aim of this study was to identify components of the Withania somnifera that could show anti-virulence activity against Streptococcus mutans biofilms. The anti-acidogenic activity of fractions separated from W. somnifera was compared, and then the most active anti-acidogenic fraction was chemically characterized using gas chromatography-mass spectroscopy. The effect of the identified components on the acidogenicity, aciduricity and extracellular polymeric substances (EPS) formation of S. mutans UA159 biofilms was evaluated. The change in accumulation and acidogenicity of S. mutans UA159 biofilms by periodic treatments (10 min per treatment) with the identified components was also investigated. Of the fractions, n-hexane fraction showed the strongest anti-acidogenic activity and was mainly composed of palmitic, linoleic and oleic acids. Of the identified components, linoleic and oleic acids strongly affected the acid production rate, F-ATPase activity and EPS formation of the biofilms. Periodic treatment with linoleic and oleic acids during biofilm formation also inhibited the biofilm accumulation and acid production rate of the biofilms without killing the biofilm bacteria. These results suggest that linoleic and oleic acids may be effective agents for restraining virulence of S. mutans biofilms. Linoleic and oleic acids may be promising agents for controlling virulence of cariogenic biofilms and subsequent dental caries formation. © 2015 The Society for Applied Microbiology.

Biofilm Surface Density Determines Biocide Effectiveness

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Sara Bas

2017-12-01

Full Text Available High resistance of biofilms for chemical challenges is a serious industrial and medical problem. In this work a gradient of surface covered with biofilm has been produced and correlated to the effectiveness of different commercially available oxidative biocides. The results for thin Escherichia coli biofilms grown in rich media supplemented with glucose or lactose on glass or poly methyl methacrylate surfaces indicate that the effectiveness of hydrogen peroxide or chlorine dioxide and quaternary ammonium compounds is inversely proportional to the fraction of the surface covered with the biofilm. In areas where biofilm covered more than 90% of the available surface the biocide treatment was inefficient after 60 min of incubation. The combined effect of oxidant and surfactant increased the effectiveness of the biocide. On the other hand, the increased biofilm viscoelasticity reduced biocide effectiveness. The results emphasize differential biocide effectiveness depending on the fraction of the attached bacterial cells. The results suggest that biofilm biocide resistance is an acquired property that increases with biofilm maturation. The more dense sessile structures present lower log reductions compared to less dense ones.

Modelling of toluene biodegradation and biofilm growth in a fixed biofilm reactor

DEFF Research Database (Denmark)

Arcangeli, Jean-Pierre; Arvin, Erik

1992-01-01

The modelling of aerobic biodegradation of toluene and the associated biofilm growth in a fixed biofilm system is presented. The model includes four biomass fractions, three dissolved components, and seven processes. It is assumed that part of the active biomass is composed of filamentous bacteria...... which grow relatively fast and detach easily, leading to a biomass growth delayed with respect to substrate degradation. The non-filamentous bacteria inside the biofilm also degrade toluene but with a slower rate compared to the filamentous bacteria. Because the nonfilamentous bacteria do not detach......, they are primarily responsible for the biofilm growth. The active biomass decays into biodegradable and ``inert'' dead biomass which is hydrolyzed into soluble products at two different rates. These products are partly degradable by the biomass and constitute the endogenous respiration. The dynamic growth phase...

Modelling of toluene biodegradation and biofilm growth in a fixed biofilm reactor

DEFF Research Database (Denmark)

Arcangeli, Jean-Pierre; Arvin, Erik

1992-01-01

The modelling of aerobic biodegradation of toluene and the associated biofilm growth in a fixed biofilm system is presented. The model includes four biomass fractions, three dissolved components, and seven processes. It is assumed that part of the active biomass is composed of filamentous bacteria......, they are primarily responsible for the biofilm growth. The active biomass decays into biodegradable and ``inert'' dead biomass which is hydrolyzed into soluble products at two different rates. These products are partly degradable by the biomass and constitute the endogenous respiration. The dynamic growth phase...... which grow relatively fast and detach easily, leading to a biomass growth delayed with respect to substrate degradation. The non-filamentous bacteria inside the biofilm also degrade toluene but with a slower rate compared to the filamentous bacteria. Because the nonfilamentous bacteria do not detach...

Biofilms and planktonic cells of Deinococcus geothermalis in extreme environments

Science.gov (United States)

Panitz, Corinna; Reitz, Guenther; Rabbow, Elke; Rettberg, Petra; Flemming, Hans-Curt; Wingender, Jost; Froesler, Jan

In addition to the several extreme environments on Earth, Space can be considered as just another exceptional environment with a unique mixture of stress factors comprising UV radiation, vacuum, desiccation, temperature, ionizing radiation and microgravity. Life that processes in these environments can depend on the life forms and their state of living. The question is whether there are different strategies for individual microorganisms compared to communities of the same organisms to cope with the different factors of their surroundings. Comparative studies of the survi-val of these communities called biofilms and planktonic cell samples of Deinococcus geothermalis stand at the focal point of the presented investigations. A biofilm is a structured community of microorganisms that live encapsulated in a matrix of extracellular polymeric substances on a surface. Microorganisms living in a biofilm usually have significantly different properties to cooperate than individually living microorganisms of the same species. An advantage of the biofilm is increased resistance to various chemical and physical effects, while the dense extracellular matrix and the outer layer of the cells protect the interior of the microbial consortium. The space experiment BOSS (Biofilm organisms surfing Space) as part the ESA experimental unit EXPOSE R-2 with a planned launch date in July 2014 will be subsequently mounted on the Russian Svesda module outside the ISS. An international team of scientists coordinated by Dr. P. Rettberg will investigate the hypothesis whether microorganisms organized as biofilm outmatch the same microorganisms exposed individually in the long-term survival of the harsh environmental conditions as they occur in space and on Mars. Another protective function in the samples could be dust par-ticles for instance Mars regolith simulant contained inside the biofilms or mixed with the planktonic cells, as additional shelter especially against the extraterrestrial UV

Paired methods to measure biofilm killing and removal: a case study with Penicillin G treatment of Staphylococcus aureus biofilm.

Science.gov (United States)

Ausbacher, D; Lorenz, L; Pitts, B; Stewart, P S; Goeres, D M

2018-03-01

Biofilms are microbial aggregates that show high tolerance to antibiotic treatments in vitro and in vivo. Killing and removal are both important in biofilm control, therefore methods that measure these two mechanisms were evaluated in a parallel experimental design. Kill was measured using the single tube method (ASTM method E2871) and removal was determined by video microscopy and image analysis using a new treatment flow cell. The advantage of the parallel test design is that both methods used biofilm covered coupons harvested from a CDC biofilm reactor, a well-established and standardized biofilm growth method. The control Staphylococcus aureus biofilms treated with growth medium increased by 0·6 logs during a 3-h contact time. Efficacy testing showed biofilms exposed to 400 μmol l -1 penicillin G decreased by only 0·3 logs. Interestingly, time-lapse confocal scanning laser microscopy revealed that penicillin G treatment dispersed the biofilm despite being an ineffective killing agent. In addition, no biofilm removal was detected when assays were performed in 96-well plates. These results illustrate that biofilm behaviour and impact of treatments can vary substantially when assayed by different methods. Measuring both killing and removal with well-characterized methods will be crucial for the discovery of new anti-biofilm strategies. Biofilms are tolerant to antimicrobial treatments and can lead to persistent infections. Finding new anti-biofilm strategies and understanding their mode-of-action is therefore of high importance. Historically, antimicrobial testing has focused on measuring the decrease in viability. While kill data are undeniably important, measuring biofilm disruption provides equally useful information. Starting with biofilm grown in the same reactor, we paired assessment of biofilm removal using a new treatment-flow-cell and real-time microscopy with kill data collected using the single tube method (ASTM E2871). Pairing these two methods

Novel method for quantitative estimation of biofilms

DEFF Research Database (Denmark)

Syal, Kirtimaan

2017-01-01

Biofilm protects bacteria from stress and hostile environment. Crystal violet (CV) assay is the most popular method for biofilm determination adopted by different laboratories so far. However, biofilm layer formed at the liquid-air interphase known as pellicle is extremely sensitive to its washing...... and staining steps. Early phase biofilms are also prone to damage by the latter steps. In bacteria like mycobacteria, biofilm formation occurs largely at the liquid-air interphase which is susceptible to loss. In the proposed protocol, loss of such biofilm layer was prevented. In place of inverting...... and discarding the media which can lead to the loss of the aerobic biofilm layer in CV assay, media was removed from the formed biofilm with the help of a syringe and biofilm layer was allowed to dry. The staining and washing steps were avoided, and an organic solvent-tetrahydrofuran (THF) was deployed...

Calcium-Phosphate-Osteopontin Particles Reduce Biofilm Formation and pH Drops in in situ-Grown Dental Biofilms

DEFF Research Database (Denmark)

Schlafer, Sebastian; Ibsen, Casper Jon Steenberg; Birkedal, Henrik

2017-01-01

This 2-period crossover study investigated the effect of calcium-phosphate-osteopontin particles on biofilm formation and pH in 48-h biofilms grown in situ. Bovine milk osteopontin is a highly phosphorylated glycoprotein that has been shown to interfere with bacterial adhesion to salivary......-coated surfaces. Calcium-phosphate-osteopontin particles have been shown to reduce biofilm formation and pH drops in a 5-species laboratory model of dental biofilm without affecting bacterial viability. Here, smooth surface biofilms from 10 individuals were treated ex vivo 6 times/day for 30 min with either...... calcium-phosphate-osteopontin particles or sterile saline. After growth, the amount of biofilm formed was determined by confocal microscopy, and pH drops upon exposure to glucose were monitored using confocal-microscopy-based pH ratiometry. A total of 160 biofilms were analysed. No adverse effects...

Aspartate inhibits Staphylococcus aureus biofilm formation.

Science.gov (United States)

Yang, Hang; Wang, Mengyue; Yu, Junping; Wei, Hongping

2015-04-01

Biofilm formation renders Staphylococcus aureus highly resistant to conventional antibiotics and host defenses. Four D-amino acids (D-Leu, D-Met, D-Trp and D-Tyr) have been reported to be able to inhibit biofilm formation and disassemble established S. aureus biofilms. We report here for the first time that both D- and L-isoforms of aspartate (Asp) inhibited S. aureus biofilm formation on tissue culture plates. Similar biofilm inhibition effects were also observed against other staphylococcal strains, including S. saprophyticus, S. equorum, S. chromogenes and S. haemolyticus. It was found that Asp at high concentrations (>10 mM) inhibited the growth of planktonic N315 cells, but at subinhibitory concentrations decreased the cellular metabolic activity without influencing cell growth. The decreased cellular metabolic activity might be the reason for the production of less protein and DNA in the matrix of the biofilms formed in the presence of Asp. However, varied inhibition efficacies of Asp were observed for biofilms formed by clinical staphylococcal isolates. There might be mechanisms other than decreasing the metabolic activity, e.g. the biofilm phenotypes, affecting biofilm formation in the presence of Asp. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Biofilm models of polymicrobial infection.

Science.gov (United States)

Gabrilska, Rebecca A; Rumbaugh, Kendra P

2015-01-01

Interactions between microbes are complex and play an important role in the pathogenesis of infections. These interactions can range from fierce competition for nutrients and niches to highly evolved cooperative mechanisms between different species that support their mutual growth. An increasing appreciation for these interactions, and desire to uncover the mechanisms that govern them, has resulted in a shift from monomicrobial to polymicrobial biofilm studies in different disease models. Here we provide an overview of biofilm models used to study select polymicrobial infections and highlight the impact that the interactions between microbes within these biofilms have on disease progression. Notable recent advances in the development of polymicrobial biofilm-associated infection models and challenges facing the study of polymicrobial biofilms are addressed.

Arsenate Retention by Epipsammic Biofilms Developed on Streambed Sediments: Influence of Phosphate

Directory of Open Access Journals (Sweden)

D. M. Prieto

2013-01-01

Full Text Available Natural geological conditions together with the impact of human activities could produce environmental problems due to high As concentrations. The aim of this study was to assess the role of epipsammic biofilm-sediment systems onto As (V sorption and to evaluate the effect of the presence of equimolar P concentrations on As retention. A natural biofilm was grown on sediment samples in the laboratory, using river water as nutrient supplier. Sorption experiments with initial As concentrations 0, 5, 25, 50, 100, 250, and 500 μg L−1 were performed. The average percentage of As sorbed was 78.9±3.5 and 96.9±6.6% for the sediment and biofilm-sediment systems, respectively. Phosphate decreased by 25% the As sorption capactity in the sediment devoid of biofilm, whereas no significant effect was observed in the systems with biofilm. Freundlich, Sips, and Toth models were the best to describe experimental data. The maximum As sorption capacity of the sediment and biofilm-sediment systems was, respectively, 6.6 and 6.8 μg g−1 and 4.5 and 7.8 μg g−1 in the presence of P. In conclusion, epipsammic biofilms play an important role in the environmental quality of river systems, increasing As retention by the system, especially in environments where both As and P occur simultaneously.

Study on E. coli and Salmonella biofilms from fresh fruits and vegetables.

Science.gov (United States)

Amrutha, Balagopal; Sundar, Kothandapani; Shetty, Prathapkumar Halady

2017-04-01

Foodborne outbreaks associated with fresh fruits and vegetables are on the rise worldwide. Biofilm formation is one of the important traits of pathogens making them strongly attached to substrates as well as express virulence phenotypes. Present study investigates the biofilm forming ability of E. coli and Salmonella sp. isolated from fresh fruits and vegetables. A total of 53 strains, including 35 E. coli and 18 Salmonella sp. isolated from different fruit and vegetable samples were taken into account for the study. Initial screening for biofilm formation was done using Congo Red agar plate test. Results revealed that 22.8% E. coli and 22.2% Salmonella sp. were potential biofilm formers. However, the MTP (Micro-Titre Plate) assay suggested more isolates of both E. coli and Salmonella sp. were moderate to strong biofilm producers. Agar plate diffusion assay with Agrobacterium tumefaciens NTL-4 showed the production of quorum signaling molecules (AHLs) by three isolates of E. coli and one Salmonella sp. Two E. coli isolates showed a significant amount of EPS production indicating higher biofilm forming potential. The Presence of LUX R homologue gene ( sdi A) in two of the Salmonella isolates were confirmed by PCR which demonstrated their potential pathogenicity. Results of the work underline the biofilm forming and potentially virulent capacities of isolates from the surface of fruits and vegetables.

Mycobacterium fortuitum and Mycobacterium chelonae biofilm formation under high and low nutrient conditions.

Science.gov (United States)

Hall-Stoodley, L; Keevil, C W; Lappin-Scott, H M

1998-12-01

The rapidly growing mycobacteria (RGM) are broadly disbursed in the environment. They have been recovered from freshwater, seawater, wastewater and even potable water samples and are increasingly associated with non-tuberculous mycobacterial disease. There is scant evidence that non-tuberculous mycobacteria (NTM) and RGM form biofilms. Therefore, an experimental system was designed to assess the ability of RGM to form biofilms under controlled laboratory conditions. A flat plate reactor flow cell was attached to either a high or low nutrient reservoir and monitored by image analysis over time. Two surfaces were chosen for assessment of biofilm growth: silastic which is commonly used in medical settings and high density polyethylene (HDPE) which is prevalent in water distribution systems. The results show that Mycobacterium fortuitum and M. chelonae formed biofilms under both high and low nutrient conditions on both surfaces studied. These results suggest that RGM may form biofilms under a variety of conditions in industrial and medical environments. 1998 Society of Applied Microbiology.

Standardization and classification of In vitro biofilm formation by clinical isolates of Staphylococcus aureus

Directory of Open Access Journals (Sweden)

Ashish Kumar Singh

2017-01-01

Full Text Available Background: Staphylococcus aureus is Gram-positive bacterium commonly associated with nosocomial infections. The development of biofilm exhibiting drug resistance especially in foreign body associated infections has enabled the bacterium to draw considerable attention. However, till date, consensus guidelines for in vitro biofilm quantitation and categorization criterion for the bacterial isolates based on biofilm-forming capacity are lacking. Therefore, it was intended to standardize in vitro biofilm formation by clinical isolates of S. aureus and then to classify them on the basis of their biofilm-forming capacity. Materials and Methods: A study was conducted for biofilm quantitation by tissue culture plate (TCP assay employing 61 strains of S. aureus isolated from clinical samples during May 2015– December 2015 wherein several factors influencing the biofilm formation were optimized. Therefore, it was intended to propose a biofilm classification criteria based on the standard deviation multiples of the control differentiating them into non, low, medium, and high biofilm formers. Results: Brain-heart infusion broth was found to be more effective in biofilm formation compared to trypticase soy broth. Heat fixation was more effective than chemical fixation. Although, individually, glucose, sucrose, and sodium chloride (NaCl had no significant effect on biofilm formation, a statistically significant increase in absorbance was observed after using the supplement mix consisting of 222.2 mM glucose, 116.9 mM sucrose, and 1000 mM NaCl (P = 0.037. Conclusions: The present study puts forth a standardized in vitro TCP assay for biofilm biomass quantitation and categorization criteria for clinical isolates of S. aureus based on their biofilm-forming capacity. The proposed in vitro technique may be further evaluated for its usefulness in the management of persistent infections caused by the bacterium.

Exposure to Crude Oil and Chemical Dispersant May Impact Marine Microbial Biofilm Composition and Steel Corrosion

Directory of Open Access Journals (Sweden)

Jennifer L. Salerno

2018-06-01

Full Text Available The release of hydrocarbons and chemical dispersant in marine environments may disrupt benthic ecosystems, including artificial reefs, formed by historic steel shipwrecks, and their associated organisms. Experiments were performed to determine the impacts of crude oil, dispersed crude oil, and dispersant on the community structure and function of microorganisms in seawater (SW and biofilms formed on carbon steel, a common ship hull construction material. Steel corrosion was also monitored to illustrate how oil spills may impact preservation of steel shipwrecks. Microcosms were filled with seawater (SW and incubated at 4°C. Carbon steel disks (CSDs were placed in each tank, and tanks were amended with crude oil and/or dispersant or no treatment. SW and CSD biofilms were sampled biweekly for genetic analysis using Illumina sequencing of 16S ribosomal RNA gene amplicons. Predicted and sequenced bacterial metagenomes were analyzed to examine impacts of oil and dispersant on metabolic function. Gammaproteobacteria, Alphaproteobacteria, and Flavobacteriia dominated SW and biofilms. Bacterial community structure differed significantly between treatments for SW and biofilms. OTUs affiliated with known (Pseudomonas and potential (Marinomonas hydrocarbon-degraders were roughly twice as abundant in biofilms treated with oil and dispersed oil, and steel corrosion of CSDs in these treatments was higher compared to control and dispersant treatments. OTUs affiliated with the Rhodobacteraceae family (biofilm formers and potential oil degraders were less abundant in the dispersant treatment compared to other treatments in biofilm and SW samples, but OTUs affiliated with the Pseudoalteromonas genus (biofilm formers and proposed hydrocarbon degraders were more abundant in dispersant-treated biofilms. Overall, functional gene analyses revealed a decrease in genes (predicted using PICRUSt and observed in sequenced metagenomes associated with hydrocarbon degradation

Fremmedlegemeinfektioner--nyt om biofilm og quorum sensing

DEFF Research Database (Denmark)

Høiby, Niels; Johansen, Helle Krogh; Ciofu, Oana

2007-01-01

Biofilms are structured consortia of bacteria embedded in self-produced polymer matrix. Biofilms are resistant to antibiotics, disinfectives and phagocytosis. The persistence of foreign body infections is due to biofilms. Chronic P. aeruginosa lung infection in cystic fibrosis patients is a biofilm....... Bacteria in biofilms communicate by means of quorum sensing which activates genes for virulence factors. Biofilms can be prevented by antibiotic prophylaxis or early therapy or by quorum sensing inhibitors which make them susceptible to antibiotics and phagocytosis....

Understanding the fundamental mechanisms of biofilms development and dispersal: BIAM (Biofilm Intensity and Architecture Measurement), a new tool for studying biofilms as a function of their architecture and fluorescence intensity.

Science.gov (United States)

Baudin, Marine; Cinquin, Bertrand; Sclavi, Bianca; Pareau, Dominique; Lopes, Filipa

2017-09-01

Confocal laser scanning microscopy (CLSM) is one of the most relevant technologies for studying biofilms in situ. Several tools have been developed to investigate and quantify the architecture of biofilms. However, an approach to quantify correctly the evolution of intensity of a fluorescent signal as a function of the structural parameters of a biofilm is still lacking. Here we present a tool developed in the ImageJ open source software that can be used to extract both structural and fluorescence intensity from CLSM data: BIAM (Biofilm Intensity and Architecture Measurement). This is of utmost significance when studying the fundamental mechanisms of biofilm growth, differentiation and development or when aiming to understand the effect of external molecules on biofilm phenotypes. In order to provide an example of the potential of such a tool in this study we focused on biofilm dispersion. cis-2-Decenoic acid (CDA) is a molecule known to induce biofilm dispersion of multiple bacterial species. The mechanisms by which CDA induces dispersion are still poorly understood. To investigate the effects of CDA on biofilms, we used a reporter strain of Escherichia coli (E. coli) that expresses the GFPmut2 protein under control of the rrnBP1 promoter. Experiments were done in flow cells and image acquisition was made with CLSM. Analysis carried out using the new tool, BIAM, indicates that CDA affects the fluorescence intensity of the biofilm structures as well as biofilm architectures. Indeed, our results demonstrate that CDA removes more than 35% of biofilm biovolume and suggest that it results in an increase of the biofilm's mean fluorescence intensity (MFI) by more than 26% compared to the control biofilm in the absence of CDA. Copyright © 2017. Published by Elsevier B.V.

Physicochemical characteristics and microbial community evolution of biofilms during the start-up period in a moving bed biofilm reactor.

Science.gov (United States)

Zhu, Yan; Zhang, Yan; Ren, Hong-Qiang; Geng, Jin-Ju; Xu, Ke; Huang, Hui; Ding, Li-Li

2015-03-01

This study aimed to investigate biofilm properties evolution coupled with different ages during the start-up period in a moving bed biofilm reactor system. Physicochemical characteristics including adhesion force, extracellular polymeric substances (EPS), morphology as well as volatile solid and microbial community were studied. Results showed that the formation and development of biofilms exhibited four stages, including (I) initial attachment and young biofilm formation, (II) biofilms accumulation, (III) biofilm sloughing and updating, and (IV) biofilm maturation. During the whole start-up period, adhesion force was positively and significantly correlated with the contents of EPS, especially the content of polysaccharide. In addition, increased adhesion force and EPS were beneficial for biofilm retention. Gram-negative bacteria mainly including Sphaerotilus, Zoogloea and Haliscomenobacter were predominant in the initial stage. Actinobacteria was beneficial to resist sloughing. Furthermore, filamentous bacteria were dominant in maturation biofilm. Copyright © 2015 Elsevier Ltd. All rights reserved.

Comparisons of subgingival microbial profiles of refractory periodontitis, severe periodontitis, and periodontal health using the human oral microbe identification microarray.

Science.gov (United States)

Colombo, Ana Paula V; Boches, Susan K; Cotton, Sean L; Goodson, J Max; Kent, Ralph; Haffajee, Anne D; Socransky, Sigmund S; Hasturk, Hatice; Van Dyke, Thomas E; Dewhirst, Floyd; Paster, Bruce J

2009-09-01

This study compared the subgingival microbiota of subjects with refractory periodontitis (RP) to those in subjects with treatable periodontitis (GRs = good responders) or periodontal health (PH) using the Human Oral Microbe Identification Microarray (HOMIM). At baseline, subgingival plaque samples were taken from 47 subjects with periodontitis and 20 individuals with PH and analyzed for the presence of 300 species by HOMIM. The subjects with periodontitis were classified as having RP (n = 17) based on mean attachment loss (AL) and/or more than three sites with AL >or=2.5 mm after scaling and root planing, surgery, and systemically administered amoxicillin and metronidazole or as GRs (n = 30) based on mean attachment gain and no sites with AL >or=2.5 mm after treatment. Significant differences in taxa among the groups were sought using the Kruskal-Wallis and chi(2) tests. More species were detected in patients with disease (GR or RP) than in those without disease (PH). Subjects with RP were distinguished from GRs or those with PH by a significantly higher frequency of putative periodontal pathogens, such as Parvimonas micra (previously Peptostreptococcus micros or Micromonas micros), Campylobacter gracilis, Eubacterium nodatum, Selenomonas noxia, Tannerella forsythia (previously T. forsythensis), Porphyromonas gingivalis, Prevotella spp., Treponema spp., and Eikenella corrodens, as well as unusual species (Pseudoramibacter alactolyticus, TM7 spp. oral taxon [OT] 346/356, Bacteroidetes sp. OT 272/274, Solobacterium moorei, Desulfobulbus sp. OT 041, Brevundimonas diminuta, Sphaerocytophaga sp. OT 337, Shuttleworthia satelles, Filifactor alocis, Dialister invisus/pneumosintes, Granulicatella adiacens, Mogibacterium timidum, Veillonella atypica, Mycoplasma salivarium, Synergistes sp. cluster II, and Acidaminococcaceae [G-1] sp. OT 132/150/155/148/135) (P spp. cluster I, Capnocytophaga sputigena, Cardiobacterium hominis, Haemophilus parainfluenzae, Lautropia mirabilis

Studying bacterial multispecies biofilms

DEFF Research Database (Denmark)

Røder, Henriette Lyng; Sørensen, Søren Johannes; Burmølle, Mette

2016-01-01

The high prevalence and significance of multispecies biofilms have now been demonstrated in various bacterial habitats with medical, industrial, and ecological relevance. It is highly evident that several species of bacteria coexist and interact in biofilms, which highlights the need for evaluating...... the approaches used to study these complex communities. This review focuses on the establishment of multispecies biofilms in vitro, interspecies interactions in microhabitats, and how to select communities for evaluation. Studies have used different experimental approaches; here we evaluate the benefits...... and drawbacks of varying the degree of complexity. This review aims to facilitate multispecies biofilm research in order to expand the current limited knowledge on interspecies interactions. Recent technological advances have enabled total diversity analysis of highly complex and diverse microbial communities...

Calcium transcriptionally regulates the biofilm machinery of Xylella fastidiosa to promote continued biofilm development in batch cultures.

Science.gov (United States)

Parker, Jennifer K; Chen, Hongyu; McCarty, Sara E; Liu, Lawrence Y; De La Fuente, Leonardo

2016-05-01

The functions of calcium (Ca) in bacteria are less characterized than in eukaryotes, where its role has been studied extensively. The plant-pathogenic bacterium Xylella fastidiosa has several virulence features that are enhanced by increased Ca concentrations, including biofilm formation. However, the specific mechanisms driving modulation of this feature are unclear. Characterization of biofilm formation over time showed that 4 mM Ca supplementation produced denser biofilms that were still developing at 96 h, while biofilm in non-supplemented media had reached the dispersal stage by 72 h. To identify changes in global gene expression in X. fastidiosa grown in supplemental Ca, RNA-Seq of batch culture biofilm cells was conducted at three 24-h time intervals. Results indicate that a variety of genes are differentially expressed in response to Ca, including genes related to attachment, motility, exopolysaccharide synthesis, biofilm formation, peptidoglycan synthesis, regulatory functions, iron homeostasis, and phages. Collectively, results demonstrate that Ca supplementation induces a transcriptional response that promotes continued biofilm development, while biofilm cells in nonsupplemented media are driven towards dispersion of cells from the biofilm structure. These results have important implications for disease progression in planta, where xylem sap is the source of Ca and other nutrients for X. fastidiosa. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

Candida albicans biofilm development in vitro for photodynamic therapy study

International Nuclear Information System (INIS)

Suzuki, Luis Claudio

2009-01-01

Photodynamic therapy (PDT) is a phototherapy based on the use of a photo sensitizer (PS) in the presence of low intensity light with resonant wavelength of absorption of the PS and biological systems that can raise awareness, generating reactive oxygen species. Studies show that PDT has a lethal effect on Candida albicans. The biofilm formed by C. albicans is the cause of infections associated with medical devices such as catheters, with a proven resistance to antifungal agents, and the removal of the catheter colonized almost always is necessary. However, few studies in literature report the behavior and response of biofilm organized by C. albicans against PDT. The aims of this study were to develop a methodology for in vitro biofilm formation of C. albicans, evaluate the sensitivity of the biofilm of C. albicans to antimicrobial photodynamic therapy using PS as the methylene blue (MB) and hypocrellin B: La +3 (HBL a+3 ) and analyze the biofilm by Optical Coherence Tomography (OCT). For biofilm formation, discs were made from elastomeric silicone catheters. The PS were dissolved in solution of PBS, and the MB had two different concentrations tested in the biofilm: 100μM and 1mM; HBLa +3 only one of 10μM. The irradiation of both dyes with the microorganism was done by two different LEDs, one with red emission at λ = 630nm ± 20nm and the other one blue emission at λ = 460nm ± 30nm. We performed a curve of survival fraction versus time of irradiation of each sample with biofilm and suspension of the microorganism in the yeast form to verify the susceptibility of the front PDT. The yeast showed 100% reduction using both PS, but at different times of irradiation (30s to HBLa +3 and 6 min for the MB at 100μM). When the therapy was applied in biofilm, the MB 100μM did not show any significant reduction, while at concentration of 1mM was reduced by 100% after 6 min of irradiation. The HBLa +3 biofilm group showed a lower reduction in the concentration of 10μM in

A novel technique using potassium permanganate and reflectance confocal microscopy to image biofilm extracellular polymeric matrix reveals non-eDNA networks in Pseudomonas aeruginosa biofilms.

Science.gov (United States)

Swearingen, Matthew C; Mehta, Ajeet; Mehta, Amar; Nistico, Laura; Hill, Preston J; Falzarano, Anthony R; Wozniak, Daniel J; Hall-Stoodley, Luanne; Stoodley, Paul

2016-02-01

Biofilms are etiologically important in the development of chronic medical and dental infections. The biofilm extracellular polymeric substance (EPS) determines biofilm structure and allows bacteria in biofilms to adapt to changes in mechanical loads such as fluid shear. However, EPS components are difficult to visualize microscopically because of their low density and molecular complexity. Here, we tested potassium permanganate, KMnO4, for use as a non-specific EPS contrast-enhancing stain using confocal laser scanning microscopy in reflectance mode. We demonstrate that KMnO4 reacted with EPS components of various strains of Pseudomonas, Staphylococcus and Streptococcus, yielding brown MnO2 precipitate deposition on the EPS, which was quantifiable using data from the laser reflection detector. Furthermore, the MnO2 signal could be quantified in combination with fluorescent nucleic acid staining. COMSTAT image analysis indicated that KMnO4 staining increased the estimated biovolume over that determined by nucleic acid staining alone for all strains tested, and revealed non-eDNA EPS networks in Pseudomonas aeruginosa biofilm. In vitro and in vivo testing indicated that KMnO4 reacted with poly-N-acetylglucosamine and Pseudomonas Pel polysaccharide, but did not react strongly with DNA or alginate. KMnO4 staining may have application as a research tool and for diagnostic potential for biofilms in clinical samples. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Bacterial biofilms: prokaryotic adventures in multicellularity

DEFF Research Database (Denmark)

Webb, J.S.; Givskov, Michael Christian; Kjelleberg, S.

2003-01-01

The development of bacterial biofilms includes both the initial social behavior of undifferentiated cells, as well as cell death and differentiation in the mature biofilm, and displays several striking similarities with higher organisms. Recent advances in the field provide new insight...... into differentiation and cell death events in bacterial biofilm development and propose that biofilms have an unexpected level of multicellularity....

Comparison of the efficacy of subgingival irrigation with 2% povidone-iodine and tetracycline HCl in subjects with chronic moderate periodontitis: A clinico microbiological study

Directory of Open Access Journals (Sweden)

Jayachandran Perayil

2016-01-01

Full Text Available Background: This study was performed to evaluate and compare the clinical and antimicrobial efficacy of subgingival irrigation with tetracycline and povidone-iodine as an adjunct to nonsurgical periodontal therapy. Materials and Methods: Twenty subjects with chronic moderate periodontitis were recruited in this split-mouth study with probing pocket depth of >3 and ≤5 mm and clinical attachment loss of 3-4 mm in relation to 16, 36, and 46. In each subject, three selected periodontal pockets were assigned to receive one out of three irrigants (1 sterile water (control in 16; (2 tetracycline at 10 mg/ml in 36; (3 2% povidone-iodine in 46, and these sites were designated as Group A, Group B, and Group C, respectively. Plaque score, gingival score, pocket probing depth, and clinical attachment level were evaluated before treatment and at 1 and 3 months posttreatment. Multiplex polymerase chain reaction was used to detect Porphyromonas gingivalis and Tannerella forsythensis which have been implicated as the major risk factors for periodontal disease. Subgingival plaque collected before treatment and at 1 and 3 months posttreatment. Data were analysed using ANOVA and repeated measure ANOVA. Results were considered significant if P < 0.05. Results: Clinical and microbiological parameters were reduced posttreatment, the reduction being significantly higher in Group B compared to Group C. Conclusion: It can be concluded that chemical and mechanical therapies were of slight benefit in the treatment of chronic moderate periodontitis, and there was an adjunctive effect of significance when scaling and root planing was combined with a single subgingival irrigation with tetracycline or povidone-iodine in lower concentration.

Fremmedlegemeinfektioner--nyt om biofilm og quorum sensing

DEFF Research Database (Denmark)

Høiby, Niels; Johansen, Helle Krogh; Ciofu, Oana

2007-01-01

Biofilms are structured consortia of bacteria embedded in self-produced polymer matrix. Biofilms are resistant to antibiotics, disinfectives and phagocytosis. The persistence of foreign body infections is due to biofilms. Chronic P. aeruginosa lung infection in cystic fibrosis patients is a biofilm....... Bacteria in biofilms communicate by means of quorum sensing which activates genes for virulence factors. Biofilms can be prevented by antibiotic prophylaxis or early therapy or by quorum sensing inhibitors which make them susceptible to antibiotics and phagocytosis. Udgivelsesdato: 2007-Nov-26...

Determination of carbon in natural freshwater biofilms with total reflection X-ray fluorescence spectrometry

International Nuclear Information System (INIS)

Ovari, M.; Streli, C.; Wobrauschek, P.; Zaray, Gy.

2009-01-01

There is a growing interest in determination of low Z elements, i.e., carbon to phosphorus, in biological samples. Total reflection X-ray fluorescence spectrometry (TXRF) has been already established as suitable trace element analytical method with low sample demand and quite good quantification limits. Recently, the determinable element range was extended towards Z = 6 (carbon). Biofilms can be used for biomonioring purposes in the aquatic environment. Besides the trace metals, especially the determination of the carbon content is important for the better understanding of the early stage of biofilm formation. For this, an ATI low Z spectrometer equipped with Cr-anode X-ray tube, multilayer monochromator, vacuum chamber, and a Si(Li) detector with ultra thin window was used. Biofilms were grown on two different artificial supports (granite and plexiglass), freeze dried, suspended in high purity water and analyzed. As an internal standard the natural titanium content of the biofilms was used. The accuracy of the method was checked by total carbon measurement using a combusting carbon analyzer.

Determination of carbon in natural freshwater biofilms with total reflection X-ray fluorescence spectrometry

Energy Technology Data Exchange (ETDEWEB)

Ovari, M. [Department of Analytical Chemistry, Eoetvoes University, Budapest, H-1117, Budapest, Pazmany Peter stny. 1/a. (Hungary)], E-mail: ovari@chem.elte.hu; Streli, C.; Wobrauschek, P. [Atominstitut of the Austrian Universities, TU-Wien, Stadionallee 2, A-1020, Wien (Austria); Zaray, Gy. [Department of Analytical Chemistry, Eoetvoes University, Budapest, H-1117, Budapest, Pazmany Peter stny. 1/a. (Hungary); Cooperative Research Centre of Environmental Chemistry, Eoetvoes University, Budapest, H-1117, Budapest, Pazmany Peter stny. 1/a. (Hungary)

2009-08-15

There is a growing interest in determination of low Z elements, i.e., carbon to phosphorus, in biological samples. Total reflection X-ray fluorescence spectrometry (TXRF) has been already established as suitable trace element analytical method with low sample demand and quite good quantification limits. Recently, the determinable element range was extended towards Z = 6 (carbon). Biofilms can be used for biomonioring purposes in the aquatic environment. Besides the trace metals, especially the determination of the carbon content is important for the better understanding of the early stage of biofilm formation. For this, an ATI low Z spectrometer equipped with Cr-anode X-ray tube, multilayer monochromator, vacuum chamber, and a Si(Li) detector with ultra thin window was used. Biofilms were grown on two different artificial supports (granite and plexiglass), freeze dried, suspended in high purity water and analyzed. As an internal standard the natural titanium content of the biofilms was used. The accuracy of the method was checked by total carbon measurement using a combusting carbon analyzer.

The innovation of cryo-SEM freeze-fracturing methodology demonstrated on high pressure frozen biofilm.

Science.gov (United States)

Hrubanova, Kamila; Nebesarova, Jana; Ruzicka, Filip; Krzyzanek, Vladislav

2018-04-22

In this study we present an innovative method for the preparation of fully hydrated samples of microbial biofilms of cultures Staphylococcus epidermidis, Candida parapsilosis and Candida albicans. Cryo-scanning electron microscopy (cryo-SEM) and high-pressure freezing (HPF) rank among cutting edge techniques in the electron microscopy of hydrated samples such as biofilms. However, the combination of these techniques is not always easily applicable. Therefore, we present a method of combining high-pressure freezing using EM PACT2 (Leica Microsystems), which fixes hydrated samples on small sapphire discs, with a high resolution SEM equipped with the widely used cryo-preparation system ALTO 2500 (Gatan). Using a holder developed in house, a freeze-fracturing technique was applied to image and investigate microbial cultures cultivated on the sapphire discs. In our experiments, we focused on the ultrastructure of the extracellular matrix produced during cultivation and the relationships among microbial cells in the biofilm. The main goal of our investigations was the detailed visualization of areas of the biofilm where the microbial cells adhere to the substrate/surface. We show the feasibility of this technique, which is clearly demonstrated in experiments with various freeze-etching times. Copyright © 2018 Elsevier Ltd. All rights reserved.

Antibiotic resistance of bacterial biofilms

DEFF Research Database (Denmark)

Hoiby, N.; Bjarnsholt, T.; Givskov, M.

2010-01-01

A biofilm is a structured consortium of bacteria embedded in a self-produced polymer matrix consisting of polysaccharide, protein and DNA. Bacterial biofilms cause chronic infections because they show increased tolerance to antibiotics and disinfectant chemicals as well as resisting phagocytosis...... and other components of the body's defence system. The persistence of, for example, staphylococcal infections related to foreign bodies is due to biofilm formation. Likewise, chronic Pseudomonas aeruginosa lung infection in cystic fibrosis patients is caused by biofilm-growing mucoid strains....... Characteristically, gradients of nutrients and oxygen exist from the top to the bottom of biofilms and these gradients are associated with decreased bacterial metabolic activity and increased doubling times of the bacterial cells; it is these more or less dormant cells that are responsible for some of the tolerance...

Inactivation of Efflux Pumps Abolishes Bacterial Biofilm Formation

DEFF Research Database (Denmark)

Kvist, Malin; Hancock, Viktoria; Klemm, Per

2008-01-01

Bacterial biofilms cause numerous problems in health care and industry; notably, biofilms are associated with a large number of infections. Biofilm-dwelling bacteria are particularly resistant to antibiotics, making it hard to eradicate biofilm-associated infections. Bacteria rely on efflux pumps...... to get rid of toxic substances. We discovered that efflux pumps are highly active in bacterial biofilms, thus making efflux pumps attractive targets for antibiofilm measures. A number of efflux pump inhibitors (EPIs) are known. EPIs were shown to reduce biofilm formation, and in combination they could...... abolish biofilm formation completely. Also, EPIs were able to block the antibiotic tolerance of biofilms. The results of this feasibility study might pave the way for new treatments for biofilm-related infections and may be exploited for prevention of biofilms in general....

Differential effects of antifungal agents on expression of genes related to formation of Candida albicans biofilms.

Science.gov (United States)

Chatzimoschou, Athanasios; Simitsopoulou, Maria; Antachopoulos, Charalampos; Walsh, Thomas J; Roilides, Emmanuel

2016-01-01

The purpose of this study was to analyse specific molecular mechanisms involved in the intrinsic resistance of C. albicans biofilms to antifungals. We investigated the transcriptional profile of three genes (BGL2, SUN41, ECE1) involved in Candida cell wall formation in response to voriconazole or anidulafungin after the production of intermediate and mature biofilms. C. albicans M61, a well-documented biofilm producer strain, was used for the development of intermediate (12 h and 18 h) and completely mature biofilms (48 h). After exposure of cells from each biofilm growth mode to voriconazole (128 and 512 mg l(-1)) or anidulafungin (0.25 and 1 mg l(-1)) for 12-24 h, total RNA samples extracted from biofilm cells were analysed by RT-PCR. The voriconazole and anidulafungin biofilm MIC was 512 and 0.5 mg l(-1) respectively. Anidulafungin caused significant up-regulation of SUN41 (3.7-9.3-fold) and BGL2 (2.2-2.8 fold) in intermediately mature biofilms; whereas, voriconazole increased gene expression in completely mature biofilms (SUN41 2.3-fold, BGL2 2.1-fold). Gene expression was primarily down-regulated by voriconazole in intermediately, but not completely mature biofilms. Both antifungals caused down-regulation of ECE1 in intermediately mature biofilms. © 2015 Blackwell Verlag GmbH.

Casein Phosphopeptide-Amorphous Calcium Phosphate Reduces Streptococcus mutans Biofilm Development on Glass Ionomer Cement and Disrupts Established Biofilms.

Science.gov (United States)

Dashper, Stuart G; Catmull, Deanne V; Liu, Sze-Wei; Myroforidis, Helen; Zalizniak, Ilya; Palamara, Joseph E A; Huq, N Laila; Reynolds, Eric C

2016-01-01

Glass ionomer cements (GIC) are dental restorative materials that are suitable for modification to help prevent dental plaque (biofilm) formation. The aim of this study was to determine the effects of incorporating casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) into a GIC on the colonisation and establishment of Streptococcus mutans biofilms and the effects of aqueous CPP-ACP on established S mutans biofilms. S. mutans biofilms were either established in flow cells before a single ten min exposure to 1% w/v CPP-ACP treatment or cultured in static wells or flow cells with either GIC or GIC containing 3% w/w CPP-ACP as the substratum. The biofilms were then visualised using confocal laser scanning microscopy after BacLight LIVE/DEAD staining. A significant decrease in biovolume and average thickness of S. mutans biofilms was observed in both static and flow cell assays when 3% CPP-ACP was incorporated into the GIC substratum. A single ten min treatment with aqueous 1% CPP-ACP resulted in a 58% decrease in biofilm biomass and thickness of established S. mutans biofilms grown in a flow cell. The treatment also significantly altered the structure of these biofilms compared with controls. The incorporation of 3% CPP-ACP into GIC significantly reduced S. mutans biofilm development indicating another potential anticariogenic mechanism of this material. Additionally aqueous CPP-ACP disrupted established S. mutans biofilms. The use of CPP-ACP containing GIC combined with regular CPP-ACP treatment may lower S. mutans challenge.

Multiscale Investigation on Biofilm Distribution and Its Impact on Macroscopic Biogeochemical Reaction Rates: BIOFILM DISTRIBUTION AND RATE SCALING

Energy Technology Data Exchange (ETDEWEB)

Yan, Zhifeng [Institute of Surface-Earth System Science, Tianjin University, Tianjin China; Pacific Northwest National Laboratory, Richland WA USA; Liu, Chongxuan [Pacific Northwest National Laboratory, Richland WA USA; School of Environmental Science and Engineering, Southern University of Science and Technology, Shenzhen China; Liu, Yuanyuan [Pacific Northwest National Laboratory, Richland WA USA; School of Earth Science and Engineering, Nanjing University, Nanjing China; Bailey, Vanessa L. [Pacific Northwest National Laboratory, Richland WA USA

2017-11-01

Biofilms are critical locations for biogeochemical reactions in the subsurface environment. The occurrence and distribution of biofilms at microscale as well as their impacts on macroscopic biogeochemical reaction rates are still poorly understood. This paper investigated the formation and distributions of biofilms in heterogeneous sediments using multiscale models, and evaluated the effects of biofilm heterogeneity on local and macroscopic biogeochemical reaction rates. Sediment pore structures derived from X-ray computed tomography were used to simulate the microscale flow dynamics and biofilm distribution in the sediment column. The response of biofilm formation and distribution to the variations in hydraulic and chemical properties was first examined. One representative biofilm distribution was then utilized to evaluate its effects on macroscopic reaction rates using nitrate reduction as an example. The results revealed that microorganisms primarily grew on the surfaces of grains and aggregates near preferential flow paths where both electron donor and acceptor were readily accessible, leading to the heterogeneous distribution of biofilms in the sediments. The heterogeneous biofilm distribution decreased the macroscopic rate of biogeochemical reactions as compared with those in homogeneous cases. Operationally considering the heterogeneous biofilm distribution in macroscopic reactive transport models such as using dual porosity domain concept can significantly improve the prediction of biogeochemical reaction rates. Overall, this study provided important insights into the biofilm formation and distribution in soils and sediments as well as their impacts on the macroscopic manifestation of reaction rates.

Plasticity of Candida albicans Biofilms

Science.gov (United States)

Daniels, Karla J.

2016-01-01

SUMMARY Candida albicans, the most pervasive fungal pathogen that colonizes humans, forms biofilms that are architecturally complex. They consist of a basal yeast cell polylayer and an upper region of hyphae encapsulated in extracellular matrix. However, biofilms formed in vitro vary as a result of the different conditions employed in models, the methods used to assess biofilm formation, strain differences, and, in a most dramatic fashion, the configuration of the mating type locus (MTL). Therefore, integrating data from different studies can lead to problems of interpretation if such variability is not taken into account. Here we review the conditions and factors that cause biofilm variation, with the goal of engendering awareness that more attention must be paid to the strains employed, the methods used to assess biofilm development, every aspect of the model employed, and the configuration of the MTL locus. We end by posing a set of questions that may be asked in comparing the results of different studies and developing protocols for new ones. This review should engender the notion that not all biofilms are created equal. PMID:27250770

Growing and analyzing biofilms in flow chambers

DEFF Research Database (Denmark)

Tolker-Nielsen, Tim; Sternberg, Claus

2011-01-01

This unit describes the setup of flow chamber systems for the study of microbial biofilms, and methods for the analysis of structural biofilm formation. Use of flow chambers allows direct microscopic investigation of biofilm formation. The biofilms in flow chambers develop under hydrodynamic......, and disassembly and cleaning of the system. In addition, embedding and fluorescent in situ hybridization of flow chamber-grown biofilms are addressed....

Microbial composition of biofilms associated with lithifying rubble of Acropora palmata branches.

Science.gov (United States)

Beltrán, Yislem; Cerqueda-García, Daniel; Taş, Neslihan; Thomé, Patricia E; Iglesias-Prieto, Roberto; Falcón, Luisa I

2016-01-01

Coral reefs are among the most productive ecosystems on the planet, but are rapidly declining due to global-warming-mediated changes in the oceans. Particularly for the Caribbean region, Acropora sp. stony corals have lost ∼80% of their original coverage, resulting in vast extensions of dead coral rubble. We analyzed the microbial composition of biofilms that colonize and lithify dead Acropora palmata rubble in the Mexican Caribbean and identified the microbial assemblages that can persist under scenarios of global change, including high temperature and low pH. Lithifying biofilms have a mineral composition that includes aragonite and magnesium calcite (16 mole% MgCO(3)) and calcite, while the mineral phase corresponding to coral skeleton is basically aragonite. Microbial composition of the lithifying biofilms are different in comparison to surrounding biotopes, including a microbial mat, water column, sediments and live A. palmata microbiome. Significant shifts in biofilm composition were detected in samples incubated in mesocosms. The combined effect of low pH and increased temperature showed a strong effect after two-week incubations for biofilm composition. Findings suggest that lithifying biofilms could remain as a secondary structure on reef rubble possibly impacting the functional role of coral reefs. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Biodegradation and attenuation of steroidal hormones and alkylphenols by stream biofilms and sediments

Science.gov (United States)

Writer, Jeffrey; Barber, Larry B.; Ryan, Joseph N.; Bradley, Paul M.

2011-01-01

Biodegradation of select endocrine-disrupting compounds (17β-estradiol, estrone, 17α-ethynylestradiol, 4-nonylphenol, 4-nonylphenolmonoexthoylate, and 4-nonylphenoldiethoxylate) was evaluated in stream biofilm, sediment, and water matrices collected from locations upstream and downstream from a wastewater treatment plant effluent discharge. Both biologically mediated transformation to intermediate metabolites and biologically mediated mineralization were evaluated in separate time interval experiments. Initial time intervals (0–7 d) evaluated biodegradation by the microbial community dominant at the time of sampling. Later time intervals (70 and 185 d) evaluated the biodegradation potential as the microbial community adapted to the absence of outside energy sources. The sediment matrix was more effective than the biofilm and water matrices at biodegrading 4-nonylphenol and 17β-estradiol. Biodegradation by the sediment matrix of 17α-ethynylestradiol occurred at later time intervals (70 and 185 d) and was not observed in the biofilm or water matrices. Stream biofilms play an important role in the attenuation of endocrine-disrupting compounds in surface waters due to both biodegradation and sorption processes. Because sorption to stream biofilms and bed sediments occurs on a faster temporal scale (185 d), these compounds can accumulate in stream biofilms and sediments.

Bacterial Biofilms in Jones Tubes.

Science.gov (United States)

Ahn, Eric S; Hauck, Matthew J; Kirk Harris, Jonathan; Robertson, Charles E; Dailey, Roger A

To investigate the presence and microbiology of bacterial biofilms on Jones tubes (JTs) by direct visualization with scanning electron microscopy and polymerase chain reaction (PCR) of representative JTs, and to correlate these findings with inflammation and/or infection related to the JT. In this study, prospective case series were performed. JTs were recovered from consecutive patients presenting to clinic for routine cleaning or recurrent irritation/infection. Four tubes were processed for scanning electron microscopy alone to visualize evidence of biofilms. Two tubes underwent PCR alone for bacterial quantification. One tube was divided in half and sent for scanning electron microscopy and PCR. Symptoms related to the JTs were recorded at the time of recovery. Seven tubes were obtained. Five underwent SEM, and 3 out of 5 showed evidence of biofilms (60%). Two of the 3 biofilms demonstrated cocci and the third revealed rods. Three tubes underwent PCR. The predominant bacteria identified were Pseudomonadales (39%), Pseudomonas (16%), and Staphylococcus (14%). Three of the 7 patients (43%) reported irritation and discharge at presentation. Two symptomatic patients, whose tubes were imaged only, revealed biofilms. The third symptomatic patient's tube underwent PCR only, showing predominantly Staphylococcus (56%) and Haemophilus (36%) species. Two of the 4 asymptomatic patients also showed biofilms. All symptomatic patients improved rapidly after tube exchange and steroid antibiotic drops. Bacterial biofilms were variably present on JTs, and did not always correlate with patients' symptoms. Nevertheless, routine JT cleaning is recommended to treat and possibly prevent inflammation caused by biofilms.

Impact of Hydrodynamics on Oral Biofilm Strength

NARCIS (Netherlands)

Paramonova, E.; Kalmykowa, O. J.; van der Mei, H. C.; Busscher, H. J.; Sharma, P. K.

2009-01-01

Mechanical removal of oral biofilms is ubiquitously accepted as the best way to prevent caries and periodontal diseases. Removal effectiveness strongly depends on biofilm strength. To investigate the influence of hydrodynamics on oral biofilm strength, we grew single- and multi-species biofilms of

A novel approach for harnessing biofilm communities in moving bed biofilm reactors for industrial wastewater treatment

Directory of Open Access Journals (Sweden)

Joe A. Lemire

2015-10-01

Full Text Available Moving bed biofilm reactors (MBBRs are an effective biotechnology for treating industrial wastewater. Biomass retention on moving bed biofilm reactor (MBBR carriers (biofilm support materials, allows for the ease-of-operation and high treatment capacity of MBBR systems. Optimization of MBBR systems has largely focused on aspects of carrier design, while little attention has been paid to enhancing strategies for harnessing microbial biomass. Previously, our research group demonstrated that mixed-species biofilms can be harvested from an industrial wastewater inoculum [oil sands process water (OSPW] using the Calgary Biofilm Device (CBD. Moreover, the resultant biofilm communities had the capacity to degrade organic toxins (naphthenic acids—NAs that are found in OSPW. Therefore, we hypothesized that harnessing microbial communities from industrial wastewater, as biofilms, on MBBR carriers may be an effective method to bioremediate industrial wastewater.Here, we detail our methodology adapting the workflow employed for using the CBD, to generate inoculant carriers to seed an MBBR.In this study, OSPW-derived biofilm communities were successfully grown, and their efficacy evaluated, on commercially available MBBR carriers affixed within a modified CBD system. The resultant biofilms demonstrated the capacity to transfer biomass to recipient carriers within a scaled MBBR. Moreover, MBBR systems inoculated in this manner were fully active 2 days post-inoculation, and readily degraded a select population of NAs. Together, these findings suggest that harnessing microbial communities on carriers affixed within a modified CBD system may represent a facile and rapid method for obtaining functional inoculants for use in wastewater MBBR treatment systems.

Multidisciplinary management of subgingival crown-root fracture of an immature permanent maxillary central incisor

Directory of Open Access Journals (Sweden)

Samir Zahedpasha

2012-01-01

Full Text Available This case report describes the multidisciplinary management of subgingival horizontal crown-root fracture of an immature permanent maxillary central incisor in a 10-year-old boy. After removal of the fractured fragment, pulpotomy was performed within 48 h from the injury to promote apexogenesis. The tooth was orthodontically extruded until the fracture line was located above the alveolar bone level. Frenectomy, supracrestal fiberotomy, and crown lengthening were performed after adequate stabilization of the extruded tooth for 5 months. Finally, the tooth was restored with composite resin by using the acid etch technique. This report highlights that a multidisciplinary treatment approach with strict cooperation among specialists to manage a complicated crown-root fracture can save and restore a traumatized immature permanent tooth.

Cell death in Pseudomonas aeruginosa biofilm development

DEFF Research Database (Denmark)

Webb, J.S.; Thompson, L.S.; James, S.

2003-01-01

Bacteria growing in biofilms often develop multicellular, three-dimensional structures known as microcolonies. Complex differentiation within biofilms of Pseudomonas aeruginosa occurs, leading to the creation of voids inside microcolonies and to the dispersal of cells from within these voids....... However, key developmental processes regulating these events are poorly understood. A normal component of multicellular development is cell death. Here we report that a repeatable pattern of cell death and lysis occurs in biofilms of P. aeruginosa during the normal course of development. Cell death...... occurred with temporal and spatial organization within biofilms, inside microcolonies, when the biofilms were allowed to develop in continuous-culture flow cells. A subpopulation of viable cells was always observed in these regions. During the onset of biofilm killing and during biofilm development...

Modelling the growth of a methanotrophic biofilm

DEFF Research Database (Denmark)

Arcangeli, J.-P.; Arvin, E.

1999-01-01

This article discusses the growth of methanotrophic biofilms. Several independent biofilm growths scenarios involving different inocula were examined. Biofilm growth, substrate removal and product formation were monitored throughout the experiments. Based on the oxygen consumption it was concluded...... that heterotrophs and nitrifiers co-existed with methanotrophs in the biofilm. Heterotrophic biomass grew on soluble polymers formed by the hydrolysis of dead biomass entrapped in the biofilm. Nitrifier populations developed because of the presence of ammonia in the mineral medium. Based on these experimental...... was performed on this model. It indicated that the most influential parameters were those related to the biofilm (i.e. density; solid-volume fraction; thickness). This suggests that in order to improve the model, further research regarding the biofilm structure and composition is needed....

Biofilm biodiversity presented by fluorescent in situ hybridisation

Directory of Open Access Journals (Sweden)

Wolf Mirela

2017-01-01

Full Text Available Numerous microorganisms may be present in the water distribution system. This is associated with the imperfection of purification processes, or secondary water pollution. Not only it results in the deterioration of water quality parameters, but it also increases threat of epidemiological problems. The water that is biologically unstable creates ideal conditions for colonization of the microorganisms to the inner surface of pipelines which may form biofilm. The key issue, enabling prevention and control of the impact of the development of biofilms, is to assess the biodiversity of microbiocenosis. In order to obtain comprehensive characteristics of microorganisms communities on a particular substrate, it is necessary to combine several techniques. Further analysis using molecular biology methods are usually after traditional methods of assessing the microbiological quality of water. Standard methods do not reflect the actual species composition, because they are targeted at the bacteria that can be isolated and cultured in the laboratory. Conventional methods are capable of detecting less than 10% of the organisms in the sample. In order to study the biodiversity of organisms inhabiting a biofilm (apart from the conventional methods analyses of the diversity of nucleic acids should be used. The first method could be the polymerase chain reaction (PCR and denaturing gradient gel electrophoresis (DGGE. Another way may be fluorescence in situ hybridization, which allows to detect determined DNA sequence using specially labeled oligonucleotide probes. Visualization of the material is performed using a fluorescence microscope. The main purpose of this article is to present rapid and precise identification groups of microorganisms in their natural habitat in biofilm using fluorescent in situ hybridization method (FISH . FISH method can be successfully used to visualize these microorganisms, which show difficulties in culturing, as well as to provide

Microbial biofilms in water-mixed metalworking fluids; Mikrobielle Biofilme in wassergemischten Kuehlschmierstoffen

Energy Technology Data Exchange (ETDEWEB)

Koch, Thomas [Wisura GmbH, Bremen (Germany)

2013-05-15

The microbial load of water-miscible metalworking fluids (MWF) as well as the hygienic aspects and the cost-related impact on the production process due to the activity of microbes is in the focus of many scientific investigations and documented in the related publications. The majority of this research work is focused on the microbiology of the water body, i.e. with the microbial life in the liquid coolant. The habitat biofilm, i.e. the three-dimensional growth of bacteria and fungi on surfaces of the coolant systems has been scarcely considered. Based on the scientific findings made in the recent years studying biofilms it can be concluded, that the relevant microbial processes for the depletion of the MWF and its recontamination takes predominantly places in biofilms. This paper gives an overview of the structure, the formation and the life in biofilms and represents their relevance in MWF systems. (orig.)

Colloidal silver: a novel treatment for Staphylococcus aureus biofilms?

Science.gov (United States)

Goggin, Rachel; Jardeleza, Camille; Wormald, Peter-John; Vreugde, Sarah

2014-03-01

Colloidal silver is an alternative medicine consisting of silver particles suspended in water. After using this solution as a nasal spray, the symptoms of a previously recalcitrant Staphylococcus aureus (S. aureus)-infected chronic rhinosinusitis patient were observed to have improved markedly. The aim of this study was to determine whether colloidal silver has any direct bactericidal effects on these biofilms in vitro. S. aureus biofilms were grown from the ATCC 25923 reference strain on Minimum Biofilm Eradication Concentration (MBEC) device pegs, and treated with colloidal silver. Concentrations tested ranged from 10 to 150 μL colloidal silver diluted to 200 μL with sterile water in 50 μL cerebrospinal fluid (CSF) broth. Control pegs were exposed to equivalent volumes of CSF broth and sterile water. The sample size was 4 biomass values per treatment or control group. Confocal scanning laser microscopy and COMSTAT software were used to quantify biofilms 24 hours after treatment. Significant differences from control were found for all concentrations tested bar the lowest of 10 μL colloidal silver in 200 μL. At 20 μL colloidal silver, the reduction in biomass was 98.9% (mean difference between control and treatment = -4.0317 μm(3) /μm(2) , p colloidal silver (mean differences = -4.0681 and -4.0675μm(3) /μm(2) , respectively, p Colloidal silver directly attenuates in vitro S. aureus biofilms. © 2014 ARS-AAOA, LLC.

Casein Phosphopeptide-Amorphous Calcium Phosphate Reduces Streptococcus mutans Biofilm Development on Glass Ionomer Cement and Disrupts Established Biofilms.

Directory of Open Access Journals (Sweden)

Stuart G Dashper

Full Text Available Glass ionomer cements (GIC are dental restorative materials that are suitable for modification to help prevent dental plaque (biofilm formation. The aim of this study was to determine the effects of incorporating casein phosphopeptide-amorphous calcium phosphate (CPP-ACP into a GIC on the colonisation and establishment of Streptococcus mutans biofilms and the effects of aqueous CPP-ACP on established S mutans biofilms. S. mutans biofilms were either established in flow cells before a single ten min exposure to 1% w/v CPP-ACP treatment or cultured in static wells or flow cells with either GIC or GIC containing 3% w/w CPP-ACP as the substratum. The biofilms were then visualised using confocal laser scanning microscopy after BacLight LIVE/DEAD staining. A significant decrease in biovolume and average thickness of S. mutans biofilms was observed in both static and flow cell assays when 3% CPP-ACP was incorporated into the GIC substratum. A single ten min treatment with aqueous 1% CPP-ACP resulted in a 58% decrease in biofilm biomass and thickness of established S. mutans biofilms grown in a flow cell. The treatment also significantly altered the structure of these biofilms compared with controls. The incorporation of 3% CPP-ACP into GIC significantly reduced S. mutans biofilm development indicating another potential anticariogenic mechanism of this material. Additionally aqueous CPP-ACP disrupted established S. mutans biofilms. The use of CPP-ACP containing GIC combined with regular CPP-ACP treatment may lower S. mutans challenge.

Effects of probiotic fermented milk on biofilms, oral microbiota, and enamel

OpenAIRE

Lodi, Carolina Simonetti; Oliveira, Lidiane Viana; Brighenti, Fernanda Lourenção [UNESP; Delbem, Alberto Carlos Botazzo; Martinhon, Cleide Cristina Rodrigues

2015-01-01

The aim of this study was to evaluate in vitro and in vivo the effects of 2 brands of probiotic fermented milk on biofilms, oral microbiota, and enamel. For the in situ experiment, ten volunteers wore palatine devices containing four blocks of bovine dental enamel over 3 phases, during which 20% sucrose solution, Yakult® (Treatment A), and Batavito® (Treatment B) were dropped on the enamel blocks. Salivary microbial counts were obtained and biofilm samples were analyzed after each phase. For ...

The biofilm-positive Staphylococcus epidermidis isolates in raw materials, foodstuffs and on contact surfaces in processing plants.

Science.gov (United States)

Schlegelová, J; Babák, V; Holasová, M; Dendis, M

2008-01-01

Isolates from the "farm to fork" samples (182 isolates from 2779 samples) were examined genotypically (icaAB genes) and phenotypically (in vitro biofilm formation, typical growth on Congo red agar; CRA) with the aim to assess the risk of penetration of virulent strains of Staphylococcus epidermidis into the food chain. The contamination of meat and milk products was significantly higher in comparison with raw materials. Contamination of contact surfaces in the meat-processing plants was significantly lower than that of contact surfaces in the dairy plants. The ica genes (which precondition the biofilm formation) were concurrently detected in 20 isolates that also showed a typical growth on CRA. Two ica operon-negative isolates produced biofilm in vitro but perhaps by an ica-independent mechanism. The surfaces in the dairy plants and the milk products were more frequently contaminated with ica operon-positive strains (2.3 and 1.2 % samples) than the other sample types (0-0.6 % samples).

Anti-Biofilm Compounds Derived from Marine Sponges

Directory of Open Access Journals (Sweden)

Christian Melander

2011-10-01

Full Text Available Bacterial biofilms are surface-attached communities of microorganisms that are protected by an extracellular matrix of biomolecules. In the biofilm state, bacteria are significantly more resistant to external assault, including attack by antibiotics. In their native environment, bacterial biofilms underpin costly biofouling that wreaks havoc on shipping, utilities, and offshore industry. Within a host environment, they are insensitive to antiseptics and basic host immune responses. It is estimated that up to 80% of all microbial infections are biofilm-based. Biofilm infections of indwelling medical devices are of particular concern, since once the device is colonized, infection is almost impossible to eliminate. Given the prominence of biofilms in infectious diseases, there is a notable effort towards developing small, synthetically available molecules that will modulate bacterial biofilm development and maintenance. Here, we highlight the development of small molecules that inhibit and/or disperse bacterial biofilms specifically through non-microbicidal mechanisms. Importantly, we discuss several sets of compounds derived from marine sponges that we are developing in our labs to address the persistent biofilm problem. We will discuss: discovery/synthesis of natural products and their analogues—including our marine sponge-derived compounds and initial adjuvant activity and toxicological screening of our novel anti-biofilm compounds.

Focus on the physics of biofilms

International Nuclear Information System (INIS)

Lecuyer, Sigolene; Stocker, Roman; Rusconi, Roberto

2015-01-01

Bacteria are the smallest and most abundant form of life. They have traditionally been considered as primarily planktonic organisms, swimming or floating in a liquid medium, and this view has shaped many of the approaches to microbial processes, including for example the design of most antibiotics. However, over the last few decades it has become clear that many bacteria often adopt a sessile, surface-associated lifestyle, forming complex multicellular communities called biofilms. Bacterial biofilms are found in a vast range of environments and have major consequences on human health and industrial processes, from biofouling of surfaces to the spread of diseases. Although the study of biofilms has been biologists’ territory for a long time, a multitude of phenomena in the formation and development of biofilms hinges on physical processes. We are pleased to present a collection of research papers that discuss some of the latest developments in many of the areas to which physicists can contribute a deeper understanding of biofilms, both experimentally and theoretically. The topics covered range from the influence of physical environmental parameters on cell attachment and subsequent biofilm growth, to the use of local probes and imaging techniques to investigate biofilm structure, to the development of biofilms in complex environments and the modeling of colony morphogenesis. The results presented contribute to addressing some of the major challenges in microbiology today, including the prevention of surface contamination, the optimization of biofilm disruption methods and the effectiveness of antibiotic treatments. (editorial)

Impact of periodontal therapy on the subgingival microbiota of severe periodontitis: comparison between good responders and individuals with refractory periodontitis using the human oral microbe identification microarray.

Science.gov (United States)

Colombo, Ana Paula V; Bennet, Susan; Cotton, Sean L; Goodson, J Max; Kent, Ralph; Haffajee, Anne D; Socransky, Sigmund S; Hasturk, Hatice; Van Dyke, Thomas E; Dewhirst, Floyd E; Paster, Bruce J

2012-10-01

This study compares the changes to the subgingival microbiota of individuals with "refractory" periodontitis (RP) or treatable periodontitis (good responders [GR]) before and after periodontal therapy by using the Human Oral Microbe Identification Microarray (HOMIM) analysis. Individuals with chronic periodontitis were classified as RP (n = 17) based on mean attachment loss (AL) and/or >3 sites with AL ≥2.5 mm after scaling and root planing, surgery, and systemically administered amoxicillin and metronidazole or as GR (n = 30) based on mean attachment gain and no sites with AL ≥2.5 mm after treatment. Subgingival plaque samples were taken at baseline and 15 months after treatment and analyzed for the presence of 300 species by HOMIM analysis. Significant differences in taxa before and post-therapy were sought using the Wilcoxon test. The majority of species evaluated decreased in prevalence in both groups after treatment; however, only a small subset of organisms was significantly affected. Species that increased or persisted in high frequency in RP but were significantly reduced in GR included Bacteroidetes sp., Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella spp., Tannerella forsythia, Dialister spp., Selenomonas spp., Catonella morbi, Eubacterium spp., Filifactor alocis, Parvimonas micra, Peptostreptococcus sp. OT113, Fusobacterium sp. OT203, Pseudoramibacter alactolyticus, Streptococcus intermedius or Streptococcus constellatus, and Shuttlesworthia satelles. In contrast, Capnocytophaga sputigena, Cardiobacterium hominis, Gemella haemolysans, Haemophilus parainfluenzae, Kingella oralis, Lautropia mirabilis, Neisseria elongata, Rothia dentocariosa, Streptococcus australis, and Veillonella spp. were more associated with therapeutic success. Persistence of putative and novel periodontal pathogens, as well as low prevalence of beneficial species was associated with chronic refractory periodontitis.

Silver-Palladium Surfaces Inhibit Biofilm Formation

DEFF Research Database (Denmark)

Chiang, Wen-Chi; Schroll, Casper; Hilbert, Lisbeth Rischel

2009-01-01

Undesired biofilm formation is a major concern in many areas. In the present study, we investigated biofilm-inhibiting properties of a silver-palladium surface that kills bacteria by generating microelectric fields and electrochemical redox processes. For evaluation of the biofilm inhibition...... efficacy and study of the biofilm inhibition mechanism, the silver-sensitive Escherichia coli J53 and the silver-resistant E. coli J53[pMG101] strains were used as model organisms, and batch and flow chamber setups were used as model systems. In the case of the silver-sensitive strain, the silver......-palladium surfaces killed the bacteria and prevented biofilm formation under conditions of low or high bacterial load. In the case of the silver-resistant strain, the silver-palladium surfaces killed surface-associated bacteria and prevented biofilm formation under conditions of low bacterial load, whereas under...

Pattern formation in Pseudomonas aeruginosa biofilms

DEFF Research Database (Denmark)

Parsek, Matthew R.; Tolker-Nielsen, Tim

2008-01-01

Bacteria are capable of forming elaborate multicellular communities called biofilms. Pattern formation in biofilms depends on cell proliferation and cellular migration in response to the available nutrients and other external cues, as well as on self-generated intercellular signal molecules...... and the production of an extracellular matrix that serves as a structural 'scaffolding' for the biofilm cells. Pattern formation in biofilms allows cells to position themselves favorably within nutrient gradients and enables buildup and maintenance of physiologically distinct subpopulations, which facilitates...... survival of one or more subpopulations upon environmental insult, and therefore plays an important role in the innate tolerance displayed by biofilms toward adverse conditions....

Evaluation of combinations of putative anti-biofilm agents and antibiotics to eradicate biofilms of Staphylococcus aureus and Pseudomonas aeruginosa.

Science.gov (United States)

Belfield, Katherine; Bayston, Roger; Hajduk, Nadzieja; Levell, Georgia; Birchall, John P; Daniel, Matija

2017-09-01

To evaluate potential anti-biofilm agents for their ability to enhance the activity of antibiotics for local treatment of localized biofilm infections. Staphylococcus aureus and Pseudomonas aeruginosa in vitro biofilm models were developed. The putative antibiotic enhancers N-acetylcysteine, acetylsalicylic acid, sodium salicylate, recombinant human deoxyribonuclease I, dispersin B, hydrogen peroxide and Johnson's Baby Shampoo (JBS) were tested for their anti-biofilm activity alone and their ability to enhance the activity of antibiotics for 7 or 14 days, against 5 day old biofilms. The antibiotic enhancers were paired with rifampicin and clindamycin against S. aureus and gentamicin and ciprofloxacin against P. aeruginosa. Isolates from biofilms that were not eradicated were tested for antibiotic resistance. Antibiotic levels 10× MIC and 100× MIC significantly reduced biofilm, but did not consistently eradicate it. Antibiotics at 100× MIC with 10% JBS for 14 days was the only treatment to eradicate both staphylococcal and pseudomonal biofilms. Recombinant human deoxyribonuclease I significantly reduced staphylococcal biofilm. Emergence of resistance of surviving isolates was minimal and was often associated with the small colony variant phenotype. JBS enhanced the activity of antibiotics and several other promising anti-biofilm agents were identified. Antibiotics with 10% JBS eradicated biofilms produced by both organisms. Such combinations might be useful in local treatment of localized biofilm infections. © The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Killing of Serratia marcescens biofilms with chloramphenicol.

Science.gov (United States)

Ray, Christopher; Shenoy, Anukul T; Orihuela, Carlos J; González-Juarbe, Norberto

2017-03-29

Serratia marcescens is a Gram-negative bacterium with proven resistance to multiple antibiotics and causative of catheter-associated infections. Bacterial colonization of catheters mainly involves the formation of biofilm. The objectives of this study were to explore the susceptibility of S. marcescens biofilms to high doses of common antibiotics and non-antimicrobial agents. Biofilms formed by a clinical isolate of S. marcescens were treated with ceftriaxone, kanamycin, gentamicin, and chloramphenicol at doses corresponding to 10, 100 and 1000 times their planktonic minimum inhibitory concentration. In addition, biofilms were also treated with chemical compounds such as polysorbate-80 and ursolic acid. S. marcescens demonstrated susceptibility to ceftriaxone, kanamycin, gentamicin, and chloramphenicol in its planktonic form, however, only chloramphenicol reduced both biofilm biomass and biofilm viability. Polysorbate-80 and ursolic acid had minimal to no effect on either planktonic and biofilm grown S. marcescens. Our results suggest that supratherapeutic doses of chloramphenicol can be used effectively against established S. marcescens biofilms.

Biofilm behavior on sulfonated poly(ether-ether-ketone) (sPEEK)

Energy Technology Data Exchange (ETDEWEB)

Montero, Juan F.D. [Center for Research on Dental Implants (CEPID), School of Dentistry (ODT), Federal University of Santa Catarina - UFSC, Florianópolis, SC 88040-900 (Brazil); Tajiri, Henrique A.; Barra, Guilherme M.O.; Fredel, Márcio C. [Department of Mechanical Engineering (EMC), Federal University of Santa Catarina (UFSC), Florianópolis, SC 88040-900 (Brazil); Benfatti, Cesar A.M.; Magini, Ricardo S. [Center for Research on Dental Implants (CEPID), School of Dentistry (ODT), Federal University of Santa Catarina - UFSC, Florianópolis, SC 88040-900 (Brazil); Pimenta, Andréa L. [Integrated Laboratories Technologies (InteLAB), Dept. Chemical Engineering (EQA), Federal University of Santa Catarina - UFSC, Florianópolis, SC 88040-970 (Brazil); Department of Biologie, Université de Cergy Pontoise, 2, Av. Adolphe Chauvin, 95302 Cergy Pontoise (France); Souza, Júlio C.M., E-mail: julio.c.m.souza@ufsc.br [Center for Research on Dental Implants (CEPID), School of Dentistry (ODT), Federal University of Santa Catarina - UFSC, Florianópolis, SC 88040-900 (Brazil); Center for Microelectromechanical Systems (CMEMS), Dept. Mechanical Engineering (DEM), Campus Azurém, 4800-058 Guimarães (Portugal)

2017-01-01

Poly(ether-ether-ketone) (PEEK) has also shown to be very attractive for incorporating therapeutic compounds thanks to a sulfonation process which modifies the material structure resulting in a sulfonated-PEEK (sPEEK). Concerning biomedical applications, the objective of this work was to evaluate the influence of different sulfonation degree of sPEEK on the biofilm growth. PEEK samples were functionalized by using sulphuric acid (98%) and then dissolved into dimethyl-sulfoxide. A dip coating technique was used to synthesize sPEEK thin films. The sulfonation degree of the materials was analyzed by FT-IR, H NMR, TG and IEC. The surfaces were characterized by scanning electron microscopy, profilometry and contact angle analyses. Subsequently, the biofilm formation on sulfonated-PEEK based on Streptococcus mutans and Enterococcus faecalis was measured by spectrophotometry, colony forming units (CFU mL{sup −1}) and SEM. Results obtained from thermal and chemical analyses showed an intensification in sulfonation degree for sPEEK at 2 and 2.5 h. The E. faecalis or S. mutans biofilm growth revealed statistically significant differences (p < 0.05) between 2 and 3 h sulfonation groups. A significant decrease (p < 0.05) in CFU mL{sup −1} was recorded for S. mutans or E. faecalis biofilm grown on 2.5 or 3 h sPEEK. Regarding the thermal-chemical and microbiologic analyses, the sulfonation degree of sPEEK ranging from 2 up to 3 h was successful capable to decrease the biofilm growth. That revealed an alternative strategy to embed anti-biofilm and therapeutic compounds into PEEK avoiding infections in biomedical applications. - Highlights: • PEEK can be dissolved to incorporate therapeutic compounds. • High sulfonation degree on sPEEK affected the biofilm growth. • The sulfonation degree must be controlled to maintain the properties of sPEEK.

Biofilm behavior on sulfonated poly(ether-ether-ketone) (sPEEK)

International Nuclear Information System (INIS)

Montero, Juan F.D.; Tajiri, Henrique A.; Barra, Guilherme M.O.; Fredel, Márcio C.; Benfatti, Cesar A.M.; Magini, Ricardo S.; Pimenta, Andréa L.; Souza, Júlio C.M.

2017-01-01

Poly(ether-ether-ketone) (PEEK) has also shown to be very attractive for incorporating therapeutic compounds thanks to a sulfonation process which modifies the material structure resulting in a sulfonated-PEEK (sPEEK). Concerning biomedical applications, the objective of this work was to evaluate the influence of different sulfonation degree of sPEEK on the biofilm growth. PEEK samples were functionalized by using sulphuric acid (98%) and then dissolved into dimethyl-sulfoxide. A dip coating technique was used to synthesize sPEEK thin films. The sulfonation degree of the materials was analyzed by FT-IR, H NMR, TG and IEC. The surfaces were characterized by scanning electron microscopy, profilometry and contact angle analyses. Subsequently, the biofilm formation on sulfonated-PEEK based on Streptococcus mutans and Enterococcus faecalis was measured by spectrophotometry, colony forming units (CFU mL −1 ) and SEM. Results obtained from thermal and chemical analyses showed an intensification in sulfonation degree for sPEEK at 2 and 2.5 h. The E. faecalis or S. mutans biofilm growth revealed statistically significant differences (p < 0.05) between 2 and 3 h sulfonation groups. A significant decrease (p < 0.05) in CFU mL −1 was recorded for S. mutans or E. faecalis biofilm grown on 2.5 or 3 h sPEEK. Regarding the thermal-chemical and microbiologic analyses, the sulfonation degree of sPEEK ranging from 2 up to 3 h was successful capable to decrease the biofilm growth. That revealed an alternative strategy to embed anti-biofilm and therapeutic compounds into PEEK avoiding infections in biomedical applications. - Highlights: • PEEK can be dissolved to incorporate therapeutic compounds. • High sulfonation degree on sPEEK affected the biofilm growth. • The sulfonation degree must be controlled to maintain the properties of sPEEK.

Chitosanase purified from bacterial isolate Bacillus licheniformis of ruined vegetables displays broad spectrum biofilm inhibition.

Science.gov (United States)

Muslim, Sahira Nsayef; Al-Kadmy, Israa M S; Hussein, Nadheema Hammood; Mohammed Ali, Alaa Naseer; Taha, Buthainah Mohammed; Aziz, Sarah Naji; Kheraif, Abdulaziz Abdullah Al; Divakar, Darshan Devang; Ramakrishnaiah, Ravikumar

2016-11-01

A number of bacterial species produces chitosanases which has variety of applications because of its high biodegradability, non-toxicity and antimicrobial assets. In the present study chitosanase is purified from new bacterial species Bacillus licheniformis from spoiled vegetable. This novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples has the ability to produce the chitosanase enzyme when grown on chitosan substrate. Study also examined its antibiofilm properties against diverse bacterial species with biofilm forming ability. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria [biofilm producers] tested in this study in congo red agar and microtiter plate's methods. Highly antibiofilm activity of chitosanase was recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation upto 22 and 29%, respectively compared with [100] % of control. Biofilm formation has multiple role including ability to enhance resistance and self-protection from external stress. This chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug resistant pathogen-associated infections, especially in situation where biofilms are involved. Copyright © 2016 Elsevier Ltd. All rights reserved.

Prevalence of antibiotic resistant coliform bacteria, Enterococcus spp. and Staphylococcus spp. in wastewater sewerage biofilm.

Science.gov (United States)

Lépesová, Kristína; Kraková, Lucia; Pangallo, Domenico; Medveďová, Alžbeta; Olejníková, Petra; Mackuľak, Tomáš; Tichý, Jozef; Grabic, Roman; Birošová, Lucia

2018-03-28

Urban wastewater contains different micropollutants and high number of different microorganisms. Some bacteria in wastewater can attach to the surfaces and form biofilm, which gives bacteria advantage in fight against environmental stress. This work is focused on bacterial community analysis in biofilms isolated from influent and effluent sewerage of wastewater treatment plant in Bratislava. Biofilm microbiota detection was performed by culture-independent and culture-dependent approaches. Composition of bacterial strains was detected by denaturing gradient gel electrophoresis fingerprinting coupled with the construction of 16S rRNA clone libraries. The biofilm collected at the inlet point was characterized primarily by the presence of Pseudomonas sp., Acinetobacter sp. and Janthinobacterium sp. clones, while in the biofilm isolated at outflow of wastewater treatment plant members of Pseudomonas genus were largely detected. Beside this analysis prevalence of antibiotics and resistant coliforms, Enterococcus spp. and Staphylococcus spp. in sewerage was studied. In influent wastewater were dominant antibiotics like azithromycin, clarithromycin and ciprofloxacin. Removal efficiency of these antibiotics notably azithromycin and clarithromycin were 30% in most cases. The highest number of resistant bacteria with predominance of coliforms was detected in sample of effluent biofilm. Multidrug resistant strains in effluent biofilm showed very good ability to form biofilm. Copyright © 2018. Published by Elsevier Ltd.

Microbial community composition and dynamics of moving bed biofilm reactor systems treating municipal sewage.

Science.gov (United States)

Biswas, Kristi; Turner, Susan J

2012-02-01

Moving bed biofilm reactor (MBBR) systems are increasingly used for municipal and industrial wastewater treatment, yet in contrast to activated sludge (AS) systems, little is known about their constituent microbial communities. This study investigated the community composition of two municipal MBBR wastewater treatment plants (WWTPs) in Wellington, New Zealand. Monthly samples comprising biofilm and suspended biomass were collected over a 12-month period. Bacterial and archaeal community composition was determined using a full-cycle community approach, including analysis of 16S rRNA gene libraries, fluorescence in situ hybridization (FISH) and automated ribosomal intergenic spacer analysis (ARISA). Differences in microbial community structure and abundance were observed between the two WWTPs and between biofilm and suspended biomass. Biofilms from both plants were dominated by Clostridia and sulfate-reducing members of the Deltaproteobacteria (SRBs). FISH analyses indicated morphological differences in the Deltaproteobacteria detected at the two plants and also revealed distinctive clustering between SRBs and members of the Methanosarcinales, which were the only Archaea detected and were present in low abundance (<5%). Biovolume estimates of the SRBs were higher in biofilm samples from one of the WWTPs which receives both domestic and industrial waste and is influenced by seawater infiltration. The suspended communities from both plants were diverse and dominated by aerobic members of the Gammaproteobacteria and Betaproteobacteria. This study represents the first detailed analysis of microbial communities in full-scale MBBR systems and indicates that this process selects for distinctive biofilm and planktonic communities, both of which differ from those found in conventional AS systems.

Species sorting during biofilm assembly by artificial substrates deployed in a cold seep system

KAUST Repository

Zhang, Wei Peng

2014-10-17

Studies focusing on biofilm assembly in deep-sea environments are rarely conducted. To examine the effects of substrate type on microbial community assembly, biofilms were developed on different substrates for different durations at two locations in the Red Sea: in a brine pool and in nearby bottom water (NBW) adjacent to the Thuwal cold seep II. The composition of the microbial communities in 51 biofilms and water samples were revealed by classification of pyrosequenced 16S rRNA gene amplicons. Together with the microscopic characteristics of the biofilms, the results indicate a stronger selection effect by the substrates on the microbial assembly in the brine pool compared with the NBW. Moreover, the selection effect by substrate type was stronger in the early stages compared with the later stages of the biofilm development. These results are consistent with the hypotheses proposed in the framework of species sorting theory, which states that the power of species sorting during microbial community assembly is dictated by habitat conditions, duration and the structure of the source community. Therefore, the results of this study shed light on the control strategy underlying biofilm-associated marine fouling and provide supporting evidence for ecological theories important for understanding the formation of deep-sea biofilms.

Species sorting during biofilm assembly by artificial substrates deployed in a cold seep system

KAUST Repository

Zhang, Wei Peng; Wang, Yong; Tian, Ren Mao; Bougouffa, Salim; Yang, Bo; Cao, Hui Luo; Zhang, Gen; Wong, Yue Him; Xu, Wei; Batang, Zenon B.; Al-Suwailem, Abdulaziz M.; Zhang, Xixiang; Qian, Pei-Yuan

2014-01-01

Studies focusing on biofilm assembly in deep-sea environments are rarely conducted. To examine the effects of substrate type on microbial community assembly, biofilms were developed on different substrates for different durations at two locations in the Red Sea: in a brine pool and in nearby bottom water (NBW) adjacent to the Thuwal cold seep II. The composition of the microbial communities in 51 biofilms and water samples were revealed by classification of pyrosequenced 16S rRNA gene amplicons. Together with the microscopic characteristics of the biofilms, the results indicate a stronger selection effect by the substrates on the microbial assembly in the brine pool compared with the NBW. Moreover, the selection effect by substrate type was stronger in the early stages compared with the later stages of the biofilm development. These results are consistent with the hypotheses proposed in the framework of species sorting theory, which states that the power of species sorting during microbial community assembly is dictated by habitat conditions, duration and the structure of the source community. Therefore, the results of this study shed light on the control strategy underlying biofilm-associated marine fouling and provide supporting evidence for ecological theories important for understanding the formation of deep-sea biofilms.

Microbiële biofilms in tandheelkunde

NARCIS (Netherlands)

Krom, B.P.

2015-01-01

Aangehechte gemeenschappen van micro-organismen, ook wel biofilms genoemd, zijn altijd en overal aanwezig. Hoewel biofilms een slechte naam hebben, zijn ze meestal natuurlijk, gezond en zelfs gewenst. In de tandartspraktijk komen zowel gezonde (orale biofilms) als ongezonde (bijv. in de waterleiding

Microbiële biofilms in tandheelkunde

NARCIS (Netherlands)

Krom, B.P.

2015-01-01

Aangehechte gemeenschappen van micro-organismen, ook wel biofilms genoemd, zijn altijd en overal aanwezig. Hoewel biofilms een slechte naam hebben, zijn ze meestal natuurlijk, gezond en zelfs gewenst. In de mondzorgpraktijk komen zowel gezonde (orale biofilms) als ongezonde (bijv. in de waterleiding

Microbial biofilms: biosurfactants as antibiofilm agents.

Science.gov (United States)

Banat, Ibrahim M; De Rienzo, Mayri A Díaz; Quinn, Gerry A

2014-12-01

Current microbial inhibition strategies based on planktonic bacterial physiology have been known to have limited efficacy on the growth of biofilm communities. This problem can be exacerbated by the emergence of increasingly resistant clinical strains. All aspects of biofilm measurement, monitoring, dispersal, control, and inhibition are becoming issues of increasing importance. Biosurfactants have merited renewed interest in both clinical and hygienic sectors due to their potential to disperse microbial biofilms in addition to many other advantages. The dispersal properties of biosurfactants have been shown to rival those of conventional inhibitory agents against bacterial and yeast biofilms. This makes them suitable candidates for use in new generations of microbial dispersal agents and for use as adjuvants for existing microbial suppression or eradication strategies. In this review, we explore aspects of biofilm characteristics and examine the contribution of biologically derived surface-active agents (biosurfactants) to the disruption or inhibition of microbial biofilms.

Biofilm inhibitors that target amyloid proteins.

Science.gov (United States)

Romero, Diego; Sanabria-Valentín, Edgardo; Vlamakis, Hera; Kolter, Roberto

2013-01-24

Bacteria establish stable communities, known as biofilms, that are resistant to antimicrobials. Biofilm robustness is due to the presence of an extracellular matrix, which for several species-among them Bacillus subtilis-includes amyloid-like protein fibers. In this work, we show that B. subtilis biofilms can be a simple and reliable tool for screening of molecules with antiamyloid activity. We identified two molecules, AA-861 and parthenolide, which efficiently inhibited biofilms by preventing the formation of amyloid-like fibers. Parthenolide also disrupted pre-established biofilms. These molecules also impeded the formation of biofilms of other bacterial species that secrete amyloid proteins, such as Bacillus cereus and Escherichia coli. Furthermore, the identified molecules decreased the conversion of the yeast protein New1 to the prion state in a heterologous host, indicating the broad range of activity of the molecules. Copyright © 2013 Elsevier Ltd. All rights reserved.

Material modeling of biofilm mechanical properties.

Science.gov (United States)

Laspidou, C S; Spyrou, L A; Aravas, N; Rittmann, B E

2014-05-01

A biofilm material model and a procedure for numerical integration are developed in this article. They enable calculation of a composite Young's modulus that varies in the biofilm and evolves with deformation. The biofilm-material model makes it possible to introduce a modeling example, produced by the Unified Multi-Component Cellular Automaton model, into the general-purpose finite-element code ABAQUS. Compressive, tensile, and shear loads are imposed, and the way the biofilm mechanical properties evolve is assessed. Results show that the local values of Young's modulus increase under compressive loading, since compression results in the voids "closing," thus making the material stiffer. For the opposite reason, biofilm stiffness decreases when tensile loads are imposed. Furthermore, the biofilm is more compliant in shear than in compression or tension due to the how the elastic shear modulus relates to Young's modulus. Copyright © 2014 Elsevier Inc. All rights reserved.

Pseudomonas aeruginosa Biofilm, a Programmed Bacterial Life for Fitness.

Science.gov (United States)

Lee, Keehoon; Yoon, Sang Sun

2017-06-28

A biofilm is a community of microbes that typically inhabit on surfaces and are encased in an extracellular matrix. Biofilms display very dissimilar characteristics to their planktonic counterparts. Biofilms are ubiquitous in the environment and influence our lives tremendously in both positive and negative ways. Pseudomonas aeruginosa is a bacterium known to produce robust biofilms. P. aeruginosa biofilms cause severe problems in immunocompromised patients, including those with cystic fibrosis or wound infection. Moreover, the unique biofilm properties further complicate the eradication of the biofilm infection, leading to the development of chronic infections. In this review, we discuss the history of biofilm research and general characteristics of bacterial biofilms. Then, distinct features pertaining to each stage of P. aeruginosa biofilm development are highlighted. Furthermore, infections caused by biofilms on their own or in association with other bacterial species ( i.e. , multispecies biofilms) are discussed in detail.

Oral hygiene frequency and presence of visible biofilm in the primary dentition Freqüência de higiene bucal e presença de biofilme visível na dentição decídua

Directory of Open Access Journals (Sweden)

Ana Paula Pires dos Santos

2007-03-01

Full Text Available The purpose of this study was to associate oral hygiene frequency and presence of visible biofilm in the primary dentition. The sample consisted of 90 children, aged up to 4 years old, outpatients of the University Hospital of the Rio de Janeiro State University. The examinations were carried out in a dental office by a single trained examiner who was aided by an assistant. The parents answered a structured questionnaire about oral hygiene methods and frequency. Two biofilm indices, one simplified (BF1 and the other conventional (BF2, were used. BF1 classifies biofilm as absent, thin or thick, in anterior and/or posterior teeth, and provides a score for the patient, whereas BF2 classifies biofilm as absent or present, provides scores for three surfaces of each tooth and the final score is the percentage of tooth surfaces with biofilm. More than half of the parents (51 - 56.7% reported they cleaned their child's teeth at least twice a day, while 7 (7.8% had never cleaned their child's teeth. BF1 revealed that 12.2% (11 of the children had no visible biofilm, 37.8% (34 had thin biofilm in anterior and/or posterior teeth, 27.8% (25 had thick biofilm in anterior or posterior teeth and 22.2% (20 had thick biofilm in both anterior and posterior teeth. BF2 revealed a mean value of 21.8% (s.d. 16.5. No statistically significant correlations were found between oral hygiene frequency and the two biofilm indices (p > 0.05, indicating that oral hygiene frequency was not associated to oral hygiene quality in the evaluated sample.O objetivo deste estudo foi associar freqüência de higiene bucal e presença de biofilme visível na dentição decídua. A amostra constou de 90 crianças com idade até 4 anos, cadastradas no Hospital Universitário da Universidade do Estado do Rio de Janeiro. Os exames foram realizados em consultório odontológico por um único examinador treinado, auxiliado por um anotador. Os pais responderam a um questionário estruturado

Enhancing the formation and shear resistance of nitrifying biofilms on membranes by surface modification

DEFF Research Database (Denmark)

Lackner, Susanne; Holmberg, Maria; Terada, Akihiko

2009-01-01

Polypropylene (PP) membranes and polyethylene (PE) surfaces were modified to enhance formation and shear resistance of nitrifying biofilms for wastewater treatment applications. A combination of plasma polymerization and wet chemistry was employed to ultimately introduce poly(ethyleneglycol) (PEG......) chains with two different functional groups (-PEG-NH2 and -PEG-CH3). Biofilm growth experiments using a mixed nitrifying bacterial culture revealed that the specific combination of PEG chains with amino groups resulted in most biofilm formation on both PP and PE samples. Detachment experiments showed...... structure might be possible explanations of the superiority of the -PEG-NH2 modification. The success of the-PEG-NH2 modification was independent of the original surface and might, therefore, be used in wastewater treatment bioreactors to improve reactor performance by making biofilm formation more stable...

Molecular Basis for Saccharomyces cerevisiae Biofilm Development

DEFF Research Database (Denmark)

Andersen, Kaj Scherz

In this study, I sought to identify genes regulating the global molecular program for development of sessile multicellular communities, also known as biofilm, of the eukaryotic microorganism, Saccharomyces cerevisiae (yeast). Yeast biofilm has a clinical interest, as biofilms can cause chronic...... infections in humans. Biofilm is also interesting from an evolutionary standpoint, as an example of primitive multicellularity. By using a genome-wide screen of yeast deletion mutants, I show that 71 genes are essential for biofilm formation. Two-thirds of these genes are required for transcription of FLO11......, but only a small subset is previously described as regulators of FLO11. These results reveal that the regulation of biofilm formation and FLO11 is even more complex than what has previously been described. I find that the molecular program for biofilm formation shares many essential components with two...

The biofilm-specific antibiotic resistance gene ndvB is important for expression of ethanol oxidation genes in Pseudomonas aeruginosa biofilms.

Science.gov (United States)

Beaudoin, Trevor; Zhang, Li; Hinz, Aaron J; Parr, Christopher J; Mah, Thien-Fah

2012-06-01

Bacteria growing in biofilms are responsible for a large number of persistent infections and are often more resistant to antibiotics than are free-floating bacteria. In a previous study, we identified a Pseudomonas aeruginosa gene, ndvB, which is important for the formation of periplasmic glucans. We established that these glucans function in biofilm-specific antibiotic resistance by sequestering antibiotic molecules away from their cellular targets. In this study, we investigate another function of ndvB in biofilm-specific antibiotic resistance. DNA microarray analysis identified 24 genes that were responsive to the presence of ndvB. A subset of 20 genes, including 8 ethanol oxidation genes (ercS', erbR, exaA, exaB, eraR, pqqB, pqqC, and pqqE), was highly expressed in wild-type biofilm cells but not in ΔndvB biofilms, while 4 genes displayed the reciprocal expression pattern. Using quantitative real-time PCR, we confirmed the ndvB-dependent expression of the ethanol oxidation genes and additionally demonstrated that these genes were more highly expressed in biofilms than in planktonic cultures. Expression of erbR in ΔndvB biofilms was restored after the treatment of the biofilm with periplasmic extracts derived from wild-type biofilm cells. Inactivation of ethanol oxidation genes increased the sensitivity of biofilms to tobramycin. Together, these results reveal that ndvB affects the expression of multiple genes in biofilms and that ethanol oxidation genes are linked to biofilm-specific antibiotic resistance.

Biofilm extracellular polysaccharides degradation during starvation and enamel demineralization.

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Bárbara Emanoele Costa Oliveira

Full Text Available This study was conducted to evaluate if extracellular polysaccharides (EPS are used by Streptococcus mutans (Sm biofilm during night starvation, contributing to enamel demineralization increasing occurred during daily sugar exposure. Sm biofilms were formed during 5 days on bovine enamel slabs of known surface hardness (SH. The biofilms were exposed to sucrose 10% or glucose + fructose 10.5% (carbohydrates that differ on EPS formation, 8x/day but were maintained in starvation during the night. Biofilm samples were harvested during two moments, on the end of the 4th day and in the morning of the 5th day, conditions of sugar abundance and starvation, respectively. The slabs were also collected to evaluate the percentage of surface hardness loss (%SHL. The biofilms were analyzed for EPS soluble and insoluble and intracellular polysaccharides (IPS, viable bacteria (CFU, biofilm architecture and biomass. pH, calcium and acid concentration were determined in the culture medium. The data were analyzed by two-way ANOVA followed by Tukey's test or Student's t-test. The effect of the factor carbohydrate treatment for polysaccharide analysis was significant (p 0.05. Larger amounts of soluble and insoluble EPS and IPS were formed in the sucrose group when compared to glucose + fructose group (p < 0.05, but they were not metabolized during starvation time (S-EPS, p = 0.93; I-EPS, p = 0.11; and IPS = 0.96. Greater enamel %SHL was also found for the sucrose group (p < 0.05 but the demineralization did not increase during starvation (p = 0.09. In conclusion, the findings suggest that EPS metabolization by S. mutans during night starvation do not contribute to increase enamel demineralization occurred during the daily abundance of sugar.

Susceptibility of Staphylococcus aureus biofilms to reactive discharge gases.

Science.gov (United States)

Traba, Christian; Liang, Jun F

2011-08-01

Formation of bacterial biofilms at solid-liquid interfaces creates numerous problems in both industrial and biomedical sciences. In this study, the susceptibility of Staphylococcus aureus biofilms to discharge gas generated from plasma was tested. It was found that despite distinct chemical/physical properties, discharge gases from oxygen, nitrogen, and argon demonstrated very potent and almost the same anti-biofilm activity. The bacterial cells in S. aureus biofilms were killed (>99.9%) by discharge gas within minutes of exposure. Under optimal experimental conditions, no bacteria and biofilm re-growth from discharge gas treated biofilms was found. Further studies revealed that the anti-biofilm activity of the discharge gas occurred by two distinct mechanisms: (1) killing bacteria in biofilms by causing severe cell membrane damage, and (2) damaging the extracellular polymeric matrix in the architecture of the biofilm to release biofilm from the surface of the solid substratum. Information gathered from this study provides an insight into the anti-biofilm mechanisms of plasma and confirms the applications of discharge gas in the treatment of biofilms and biofilm related bacterial infections.

Sunlight-exposed biofilm microbial communities are naturally resistant to chernobyl ionizing-radiation levels.

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Marie Ragon

Full Text Available BACKGROUND: The Chernobyl accident represents a long-term experiment on the effects of exposure to ionizing radiation at the ecosystem level. Though studies of these effects on plants and animals are abundant, the study of how Chernobyl radiation levels affect prokaryotic and eukaryotic microbial communities is practically non-existent, except for a few reports on human pathogens or soil microorganisms. Environments enduring extreme desiccation and UV radiation, such as sunlight exposed biofilms could in principle select for organisms highly resistant to ionizing radiation as well. METHODOLOGY/PRINCIPAL FINDINGS: To test this hypothesis, we explored the diversity of microorganisms belonging to the three domains of life by cultivation-independent approaches in biofilms developing on concrete walls or pillars in the Chernobyl area exposed to different levels of radiation, and we compared them with a similar biofilm from a non-irradiated site in Northern Ireland. Actinobacteria, Alphaproteobacteria, Bacteroidetes, Acidobacteria and Deinococcales were the most consistently detected bacterial groups, whereas green algae (Chlorophyta and ascomycete fungi (Ascomycota dominated within the eukaryotes. Close relatives to the most radio-resistant organisms known, including Rubrobacter species, Deinococcales and melanized ascomycete fungi were always detected. The diversity of bacteria and eukaryotes found in the most highly irradiated samples was comparable to that of less irradiated Chernobyl sites and Northern Ireland. However, the study of mutation frequencies in non-coding ITS regions versus SSU rRNA genes in members of a same actinobacterial operational taxonomic unit (OTU present in Chernobyl samples and Northern Ireland showed a positive correlation between increased radiation and mutation rates. CONCLUSIONS/SIGNIFICANCE: Our results show that biofilm microbial communities in the most irradiated samples are comparable to non-irradiated samples in

Sunlight-exposed biofilm microbial communities are naturally resistant to chernobyl ionizing-radiation levels.

Science.gov (United States)

Ragon, Marie; Restoux, Gwendal; Moreira, David; Møller, Anders Pape; López-García, Purificación

2011-01-01

The Chernobyl accident represents a long-term experiment on the effects of exposure to ionizing radiation at the ecosystem level. Though studies of these effects on plants and animals are abundant, the study of how Chernobyl radiation levels affect prokaryotic and eukaryotic microbial communities is practically non-existent, except for a few reports on human pathogens or soil microorganisms. Environments enduring extreme desiccation and UV radiation, such as sunlight exposed biofilms could in principle select for organisms highly resistant to ionizing radiation as well. To test this hypothesis, we explored the diversity of microorganisms belonging to the three domains of life by cultivation-independent approaches in biofilms developing on concrete walls or pillars in the Chernobyl area exposed to different levels of radiation, and we compared them with a similar biofilm from a non-irradiated site in Northern Ireland. Actinobacteria, Alphaproteobacteria, Bacteroidetes, Acidobacteria and Deinococcales were the most consistently detected bacterial groups, whereas green algae (Chlorophyta) and ascomycete fungi (Ascomycota) dominated within the eukaryotes. Close relatives to the most radio-resistant organisms known, including Rubrobacter species, Deinococcales and melanized ascomycete fungi were always detected. The diversity of bacteria and eukaryotes found in the most highly irradiated samples was comparable to that of less irradiated Chernobyl sites and Northern Ireland. However, the study of mutation frequencies in non-coding ITS regions versus SSU rRNA genes in members of a same actinobacterial operational taxonomic unit (OTU) present in Chernobyl samples and Northern Ireland showed a positive correlation between increased radiation and mutation rates. Our results show that biofilm microbial communities in the most irradiated samples are comparable to non-irradiated samples in terms of general diversity patterns, despite increased mutation levels at the single

Sunlight-Exposed Biofilm Microbial Communities Are Naturally Resistant to Chernobyl Ionizing-Radiation Levels

Science.gov (United States)

Ragon, Marie; Restoux, Gwendal; Moreira, David; Møller, Anders Pape; López-García, Purificación

2011-01-01

Background The Chernobyl accident represents a long-term experiment on the effects of exposure to ionizing radiation at the ecosystem level. Though studies of these effects on plants and animals are abundant, the study of how Chernobyl radiation levels affect prokaryotic and eukaryotic microbial communities is practically non-existent, except for a few reports on human pathogens or soil microorganisms. Environments enduring extreme desiccation and UV radiation, such as sunlight exposed biofilms could in principle select for organisms highly resistant to ionizing radiation as well. Methodology/Principal Findings To test this hypothesis, we explored the diversity of microorganisms belonging to the three domains of life by cultivation-independent approaches in biofilms developing on concrete walls or pillars in the Chernobyl area exposed to different levels of radiation, and we compared them with a similar biofilm from a non-irradiated site in Northern Ireland. Actinobacteria, Alphaproteobacteria, Bacteroidetes, Acidobacteria and Deinococcales were the most consistently detected bacterial groups, whereas green algae (Chlorophyta) and ascomycete fungi (Ascomycota) dominated within the eukaryotes. Close relatives to the most radio-resistant organisms known, including Rubrobacter species, Deinococcales and melanized ascomycete fungi were always detected. The diversity of bacteria and eukaryotes found in the most highly irradiated samples was comparable to that of less irradiated Chernobyl sites and Northern Ireland. However, the study of mutation frequencies in non-coding ITS regions versus SSU rRNA genes in members of a same actinobacterial operational taxonomic unit (OTU) present in Chernobyl samples and Northern Ireland showed a positive correlation between increased radiation and mutation rates. Conclusions/Significance Our results show that biofilm microbial communities in the most irradiated samples are comparable to non-irradiated samples in terms of general

Red and Green Fluorescence from Oral Biofilms.

Science.gov (United States)

Volgenant, Catherine M C; Hoogenkamp, Michel A; Krom, Bastiaan P; Janus, Marleen M; Ten Cate, Jacob M; de Soet, Johannes J; Crielaard, Wim; van der Veen, Monique H

2016-01-01

Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

Red and Green Fluorescence from Oral Biofilms.

Directory of Open Access Journals (Sweden)

Catherine M C Volgenant

Full Text Available Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation as compared to the sucrose grown biofilms (cariogenic simulation. Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

Candida albicans biofilms and MMA surface treatment influence the adhesion of soft denture liners to PMMA resin

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Martinna de Mendonça e Bertolini

2014-01-01

Full Text Available The effect of Candida albicans biofilms and methyl methacrylate (MMA pretreatment on the bond strength between soft denture liners and polymethyl methacrylate (PMMA resin was analyzed. Specimens were prepared and randomly divided with respect to PMMA pretreatment, soft liner type (silicone-based or PMMA-based, and presence or absence of a C. albicans biofilm. Samples were composed of a soft denture liner bonded between two PMMA bars. Specimens (n = 10 were incubated to produce a C. albicans biofilm or stored in sterile PBS for 12 days. The tensile bond strength test was performed and failure type was determined using a stereomicroscope. Surface roughness (SR and scanning electron microscopy (SEM analysis were performed on denture liners (n = 8. Highest bond strength was observed in samples containing a silicone-based soft liner and stored in PBS, regardless of pretreatment (p < 0.01. Silicone-based specimens mostly underwent adhesive failures, while samples containing PMMA-based liners predominantly underwent cohesive failures. The silicone-based specimens SR decreased after 12 days of biofilm accumulation or PBS storage, while the SR of PMMA-based soft liners increased (p < 0.01. The PMMA-based soft liners surfaces presented sharp valleys and depressions, while silicone-based specimens surfaces exhibited more gentle features. In vitro exposure to C. albicans biofilms reduced the adhesion of denture liners to PMMA resin, and MMA pretreatment is recommended during relining procedures.

Crenarchaeal biofilm formation under extreme conditions.

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Andrea Koerdt

Full Text Available BACKGROUND: Biofilm formation has been studied in much detail for a variety of bacterial species, as it plays a major role in the pathogenicity of bacteria. However, only limited information is available for the development of archaeal communities that are frequently found in many natural environments. METHODOLOGY: We have analyzed biofilm formation in three closely related hyperthermophilic crenarchaeotes: Sulfolobus acidocaldarius, S. solfataricus and S. tokodaii. We established a microtitre plate assay adapted to high temperatures to determine how pH and temperature influence biofilm formation in these organisms. Biofilm analysis by confocal laser scanning microscopy demonstrated that the three strains form very different communities ranging from simple carpet-like structures in S. solfataricus to high density tower-like structures in S. acidocaldarius in static systems. Lectin staining indicated that all three strains produced extracellular polysaccharides containing glucose, galactose, mannose and N-acetylglucosamine once biofilm formation was initiated. While flagella mutants had no phenotype in two days old static biofilms of S. solfataricus, a UV-induced pili deletion mutant showed decreased attachment of cells. CONCLUSION: The study gives first insights into formation and development of crenarchaeal biofilms in extreme environments.

Characterization of the internal ion environment of biofilms based on charge density and shape of ion.

Science.gov (United States)

Kurniawan, Andi; Tsuchiya, Yuki; Eda, Shima; Morisaki, Hisao

2015-12-01

Biofilm polymers contain both electrically positively and negatively charged sites. These charged sites enable the biofilm to trap and retain ions leading to an important role of biofilm such as nutrient recycling and pollutant purification. Much work has focused on the ion-exchange capacity of biofilms, and they are known to adsorb ions through an exchange mechanism between the ions in solution and the ions adsorbed to the charged sites on the biofilm polymer. However, recent studies suggest that the adsorption/desorption behavior of ions in a biofilm cannot be explained solely by this ion exchange mechanism. To examine the possibility that a substantial amount of ions are held in the interstitial region of the biofilm polymer by an electrostatic interaction, intact biofilms formed in a natural environment were immersed in distilled water and ion desorption was investigated. All of the detected ion species were released from the biofilms over a short period of time, and very few ions were subsequently released over more time, indicating that the interstitial region of biofilm polymers is another ion reserve. The extent of ion retention in the interstitial region of biofilms for each ion can be determined largely by charge density, |Z|/r, where |Z| is the ion valence as absolute value and r is the ion radius. The higher |Z|/r value an ion has, the stronger it is retained in the interstitial region of biofilms. Ion shape is also a key determinant of ion retention. Spherical and non-spherical ions have different correlations between the condensation ratio and |Z|/r. The generality of these findings were assured by various biofilm samples. Thus, the internal regions of biofilms exchange ions dynamically with the outside environment. Copyright © 2015 Elsevier B.V. All rights reserved.

[Biofilms in otolaryngology].

Science.gov (United States)

Mena Viveros, Nicolás

2014-01-01

According to the National Institute of Health of the USA, «more than 60% of all microbial infections are caused by biofilms».'This can surprise us, but it is enough to consider that common infections like those of the genito-urinary tract, infections produced by catheters, middle ear infections in children, the formation of dental plaque and gingivitis are caused by biofilms, for this statement to seem more realistic. At present this is one of the subjects of great interest within medicine, particularly in otolaryngology. Bacteria have traditionally been considered to be in a free state without evident organization, partly perhaps by the ease of studying them in this form. Nevertheless, the reality is that, in nature, the great majority of these germs form complex colonies adhered to surfaces, colonies that have received the name of biofilms. These biofilms are more common than previously thought and almost all of the people have been in contact with them in the form of infections in the teeth or humid, slippery areas. New treatments that can eradicate them are currently being investigated. Copyright © 2012 Elsevier España, S.L. All rights reserved.

Bulk water phase and biofilm growth in drinking water at low nutrient conditions.