scholarly journals Differential Efficacy of Two Dental Implant Decontamination Techniques in Reducing Microbial Biofilm and Re-Growth onto Titanium Disks In Vitro

2019 ◽  
Vol 9 (15) ◽  
pp. 3191 ◽  
Author(s):  
Aida Meto ◽  
Enrico Conserva ◽  
Francesco Liccardi ◽  
Bruna Colombari ◽  
Ugo Consolo ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Real Time ◽  
Dental Implant ◽  
Clinical Implication ◽  
Nickel Titanium ◽  
Microbial Load ◽  
Clinical Progression ◽  
Missing Teeth ◽  
Biofilm Removal

Dental implants are crucial therapeutic devices for successful substitution of missing teeth. Failure cases are mainly pathogen-associated events, allowing clinical progression toward peri-mucositis or peri-implantitis. The aim of this study was to compare the performance of two mechanical decontamination systems, Nickel-Titanium brush (Brush) and Air-Polishing system with 40 µm bicarbonate powder (BIC-40), by means of a novel bioluminescence-based model that measures microbial load in real time. Briefly, 30 disks were contaminated using the bioluminescent Pseudomonas aeruginosa strain (BLI-P. aeruginosa), treated with Brush (30 s rounds, for 90 s) or BIC-40 (30 s, at 5 mm distance) procedure, and then assessed for microbial load, particularly, biofilm removal and re-growth. Our results showed that Brush and BIC-40 treatment reduced microbial load of about 1 and more than 3 logs, respectively. Furthermore, microbial re-growth onto Brush-treated disks rapidly occurred, while BIC-40-treated disks were slowly recolonized, reaching levels of microbial load consistently below those observed with the controls. In conclusion, we provide evidence on the good performance of BIC-40 as titanium device-decontamination system, the clinical implication for such findings will be discussed.

scholarly journals Pseudomonas aeruginosa as a Potential Contaminant of Packed Fresh-Cut Lettuce in a Controlled Atmosphere. The Role of Phenotypes muc+/muc–

2020 ◽  
Vol 11 (2) ◽  
pp. 8716-8724
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Real Time ◽  
Foodborne Pathogens ◽  
Rrna Gene ◽  
Modified Atmosphere ◽  
Fresh Cut ◽  
Time Specific ◽  
Sanitation Systems ◽  
Biofilm Development

In order to shed light on contamination risks along the ready-to-eat chain of fresh commodities by emerging foodborne pathogens, we investigated the biofilm development in vitro of two Pseudomonas aeruginosa strains on fresh-cut lettuce (Lactuca sativa L. var. Iceberg). The experiment was performed employing a floating bioreactor system where modified atmosphere package conditions were mimicked, and fresh-cut lettuce disks of 2 cm2 were put into contact with a 106 CFU/mL of a phenotypic mucoid P. aeruginosa phenotype (muc+) or a non-mucoid one (muc-). Following a simulated 2-day refrigerated-shelf quantitative Real-Time PCR, designed on a target gene region of the 16S rRNA gene, defined the different muc phenotypes behavior on biofilm in lettuce phyllo-plane. Between the two strains, a development difference of nearly 1.0 log CFU/cm2 occurred, with the muc+ phenotype being the most settled and adherent. This result clearly showed a distinct contamination risk according to P. aeruginosa phenotype and the need to develop real-time, specific, fast, and easy to use detection protocols along with specific sanitation systems for modified atmosphere package ready-to-eat commodities.

scholarly journals Real-time monitoring of Pseudomonas aeruginosa biofilm formation on endotracheal tubes in vitro

2018 ◽  
Vol 18 (1) ◽  
Author(s):  
Eva Pericolini ◽  
Bruna Colombari ◽  
Gianmarco Ferretti ◽  
Ramona Iseppi ◽  
Andrea Ardizzoni ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Real Time ◽  
Biofilm Formation ◽  
Real Time Monitoring ◽  
Endotracheal Tubes

scholarly journals Influence of biofilm growth age, media and antibiotics exposure time on Staphylococcus aureus and Pseudomonas aeruginosa biofilm removal in vitro

2020 ◽  
Author(s):  
Xiaofeng Chen ◽  
Yijuan Xu ◽  
Heinz Winkler ◽  
Trine Rolighed Thomsen
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Treatment Duration ◽  
Time Dependent ◽  
Growth Media ◽  
Biofilm Growth ◽  
Biofilm Removal ◽  
Mueller Hinton ◽  
Experimental Parameters

Abstract Biofilm is known to be tolerant towards antibiotics and difficult to eradicate. Numerous studies have reported Minimum Biofilm Eradication Concentration (MBEC) values of antibiotics for many known biofilm pathogens. However, the experimental parameters applied in these studies differ considerably, and often the rationale behind the experimental design are not well described. This makes it difficult to compare the findings. To demonstrate the importance of experimental parameters, we investigated the influence of biofilm growth age, antibiotic treatment duration and growth media on biofilm eradication in this study. The commonly used biofilm model Calgary biofilm device was used to grow 24 h and 72 h biofilms of Staphylococcus aureus and Pseudomonas aeruginosa , which were treated with time-dependent vancomycin and concentration-dependent tobramycin, respectively. Two common bacteriological growth media Tryptic Soy Broth (TSB) and Cation-adjusted Mueller Hinton Broth (CaMHB) were tested. We found for both species that biofilms were more difficult to kill in TSB than in CaMHB. Furthermore, young biofilms (24 h) were easier to eradicate than old biofilms (72 h). In agreement with vancomycin being time-dependent, extension of the vancomycin exposure increased killing of S. aureus biofilms. Tobramycin treatment of 24 h P. aeruginosa biofilms was found concentration-dependent and time-independent, however, increasing killing was indicated for 72 h P. aeruginosa biofilms. This study demonstrated biofilm removal efficacy was influenced by media, biofilm age and antibiotics treatment duration. It is therefore necessary to taking these parameters into consideration when designing experiments.

scholarly journals Influence of biofilm growth age, media, antibiotic concentration and exposure time on Staphylococcus aureus and Pseudomonas aeruginosa biofilm removal in vitro

2020 ◽  
Author(s):  
Xiaofeng Chen ◽  
Trine Rolighed Thomsen ◽  
Heinz Winkler ◽  
Yijuan Xu
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Treatment Duration ◽  
Time Dependent ◽  
Growth Media ◽  
Antibiotic Concentration ◽  
Biofilm Growth ◽  
Biofilm Removal ◽  
Experimental Parameters

Abstract Background: Biofilm is known to be tolerant towards antibiotics and difficult to eradicate. Numerous studies have reported Minimum Biofilm Eradication Concentration (MBEC) values of antibiotics for many known biofilm pathogens. However, the experimental parameters applied in these studies differ considerably, and often the rationale behind the experimental design are not well described. This makes it difficult to compare the findings. To demonstrate the importance of experimental parameters, we investigated the influence of biofilm growth age, antibiotic concentration and treatment duration, and growth media on biofilm eradication. Additionally, OSTEOmycinTM, a clinically used antibiotic containing allograft bone product, was tested for antibiofilm efficacy. Results: The commonly used Calgary biofilm device was used to grow 24 h and 72 h biofilms of Staphylococcus aureus and Pseudomonas aeruginosa, which were treated with time-dependent vancomycin (up to 3000 mg/L) and concentration-dependent tobramycin (up to 80 mg/L), respectively. Two common bacteriological growth media Tryptic Soy Broth (TSB) and Cation-adjusted Mueller Hinton Broth (CaMHB) were tested. We found for both species that biofilms were more difficult to kill in TSB than in CaMHB. Furthermore, young biofilms (24 h) were easier to eradicate than old biofilms (72 h). In agreement with vancomycin being time-dependent, extension of the vancomycin exposure increased killing of S. aureus biofilms. Tobramycin treatment of 24 h P. aeruginosa biofilms was found concentration-dependent and time-independent, however, increasing killing was indicated for 72 h P. aeruginosa biofilms. Treatment with tobramycin containing OSTEOmycin TTM removed 72 h and 168 h P. aeruginosa biofilms after one day treatment, while few 72h S. aureus biofilms survived after two days treatment with vancomycin containing OSTEOmycin VTM. Conclusions: This study demonstrated biofilm removal efficacy was influenced by media, biofilm age and antibiotic concentration and treatment duration. It is therefore necessary to taking these parameters into consideration when designing experiments. The results of OSTEOmycin products indicated that simple in vitro biofilm test could be used for initial screening of antibiofilm products. For clinical application, a more clinically relevant biofilm model for the specific biofilm infection in question should be developed to guide the amount of antibiotics used for local antibiofilm treatment.

scholarly journals Influence of biofilm growth age, media, antibiotic concentration and exposure time on Staphylococcus aureus and Pseudomonas aeruginosa biofilm removal in vitro

2020 ◽  
Author(s):  
Xiaofeng Chen ◽  
Trine Rolighed Thomsen ◽  
Heinz Winkler ◽  
Yijuan Xu
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Treatment Duration ◽  
Time Dependent ◽  
Growth Media ◽  
Antibiotic Concentration ◽  
Biofilm Growth ◽  
Biofilm Removal ◽  
Experimental Parameters

Abstract Background: Biofilm is known to be tolerant towards antibiotics and difficult to eradicate. Numerous studies have reported minimum biofilm eradication concentration (MBEC) values of antibiotics for many known biofilm pathogens. However, the experimental parameters applied in these studies differ considerably, and often the rationale behind the experimental design are not well described. This makes it difficult to compare the findings. To demonstrate the importance of experimental parameters, we investigated the influence of biofilm growth age, antibiotic concentration and treatment duration, and growth media on biofilm eradication. Additionally, OSTEOmycinTM, a clinically used antibiotic containing allograft bone product, was tested for antibiofilm efficacy. Results: The commonly used Calgary biofilm device was used to grow 24 h and 72 h biofilms of Staphylococcus aureus and Pseudomonas aeruginosa, which were treated with time-dependent vancomycin (up to 3000 mg L-1) and concentration-dependent tobramycin (up to 80 mg L-1), respectively. Two common bacteriological growth media tryptic soy broth (TSB) and cation-adjusted Mueller Hinton broth (CaMHB) were tested. We found for both species that biofilms were more difficult to kill in TSB than in CaMHB. Furthermore, young biofilms (24 h) were easier to eradicate than old biofilms (72 h). In agreement with vancomycin being time-dependent, extension of the vancomycin exposure increased killing of S. aureus biofilms. Tobramycin treatment of 24 h P. aeruginosa biofilms was found concentration-dependent and time-independent, however, increasing killing was indicated for 72 h P. aeruginosa biofilms. Treatment with tobramycin containing OSTEOmycin TTM removed 72 h and 168 h P. aeruginosa biofilms after one day treatment, while few 72h S. aureus biofilms survived after two days treatment with vancomycin containing OSTEOmycin VTM. Conclusions: This study demonstrated biofilm removal efficacy was influenced by media, biofilm age and antibiotic concentration and treatment duration. It is therefore necessary to taking these parameters into consideration when designing experiments. The results of OSTEOmycinTM products indicated that simple in vitro biofilm test could be used for initial screening of antibiofilm products. For clinical application, a more clinically relevant biofilm model for the specific biofilm infection in question should be developed to guide the amount of antibiotics used for local antibiofilm treatment.

scholarly journals Development of In Vitro resistance to fluoroquinolones in Pseudomonas aeruginosa

2020 ◽  
Author(s):  
Lei Zhao ◽  
Shiqi Wang ◽  
Xiaobing Li ◽  
Xiaojing He ◽  
Lingyan Jian
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Real Time ◽  
Real Time Pcr ◽  
Sanger Sequencing ◽  
Target Genes ◽  
Efflux Pump ◽  
Relative Quantitation ◽  
Evolutionary Trajectories ◽  
Quantitation Method

Abstract Background: The objectives of this study were to investigate the dynamics of different resistant mechanisms in P.aeruginosa populations that have evolved under fluoroquinolone pressure, and any interactions between these mechanisms in the evolutionary trajectories. Methods: In this study, bacteria of the strain ATCC27853 were selected under different concentrations of levofloxacin and ciprofloxacin for six parallel lineages. The four target genes in the quinolone-resistance determining region were amplified and then Sanger sequencing was used to find the mutations. The expression of four efflux pump proteins were evaluated by real-time PCR, using the relative quantitation method, and the ATCC27853 was selected as a control. Results: we found that the P.aeruginosa was killed by ciprofloxacin earlier than levofloxacin. We found five different mutations in three subunits of QRDRs in our study; gyrA was the main mutated gene for conferring resistance to fluoroquinolone. A greater number of mutations appeared at 4mg/L for levofloxacin and at 2mg/L for ciprofloxacin. The main efflux pump that was expressed was MexCD-OprJ, and the first over expressed was evident at 0.5mg/L for levofloxacin and 0.25mg/L for ciprofloxacin. Conclusions: The mutation of gyrA83 and overexpression of MexCD-OprJ were the main mechanisms that conferred resistance of P.aeruginosa to levofloxacin and ciprofloxacin. Ciprofloxacin had a stronger ability to kill the bacteria, while may render bacteria more susceptible to resistance.

scholarly journals Pseudomonas aeruginosa Induces Membrane Blebs in Epithelial Cells, Which Are Utilized as a Niche for Intracellular Replication and Motility

2008 ◽  
Vol 76 (5) ◽  
pp. 1992-2001 ◽  
Author(s):  
Annette A. Angus ◽  
Amanda Ackerman Lee ◽  
Danielle K. Augustin ◽  
Ellen J. Lee ◽  
David J. Evans ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Epithelial Cells ◽  
Real Time ◽  
Cell Types ◽  
Intracellular Survival ◽  
Bacterial Survival ◽  
Intracellular Replication ◽  
Membrane Blebs ◽  
Cell Niche

ABSTRACT Pseudomonas aeruginosa is known to invade epithelial cells during infection and in vitro. However, little is known of bacterial or epithelial factors modulating P. aeruginosa intracellular survival or replication after invasion, except that it requires a complete lipopolysaccharide core. In this study, real-time video microscopy revealed that invasive P. aeruginosa isolates induced the formation of membrane blebs in multiple epithelial cell types and that these were then exploited for intracellular replication and rapid real-time motility. Further studies revealed that the type three secretion system (T3SS) of P. aeruginosa was required for blebbing. Mutants lacking either the entire T3SS or specific T3SS components were instead localized to intracellular perinuclear vacuoles. Most T3SS mutants that trafficked to perinuclear vacuoles gradually lost intracellular viability, and vacuoles containing those bacteria were labeled by the late endosomal marker lysosome-associated marker protein 3 (LAMP-3). Interestingly, mutants deficient only in the T3SS translocon structure survived and replicated within the vacuoles that did not label with LAMP-3. Taken together, these data suggest two novel roles of the P. aeruginosa T3SS in enabling bacterial intracellular survival: translocon-dependent formation of membrane blebs, which form a host cell niche for bacterial growth and motility, and effector-dependent bacterial survival and replication within intracellular perinuclear vacuoles.

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