scholarly journals Effect of Four Essential Oils on Cells Release Membrane and Biofilm Formation of Clinical Bacterial Isolated From Oral Infection

2019 ◽  
Vol 9 (6) ◽  
pp. 28-35
Author(s):  
Dimitri Tchami Ngongang ◽  
Ascension Maximilienne Nyegue ◽  
Del Florence Esther Ndedi ◽  
Xavier François Etoa
Keyword(s):  
Essential Oils ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Oral Bacteria ◽  
Oral Diseases ◽  
Extracellular Medium ◽  
Bacterial Form ◽  
Membrane Methods ◽  
Interesting Alternative

Background: Bacterial biofilms forming are current resistant bacterial form to the treatment of oral diseases that colonizes in the gingival and sub-gingival regions of the mouth. The present study aims to screen the anti-biofilm potential and evaluate the effect of four essential oils on cells release membrane. Methods: Seven type isolate bacteria obtained during previous work were screen to select those who had ability to form biofilm using Congo Red Agar method, tube method and crystal violet method. The inhibitory parameter of biofilm forming was determine using microtiter plate method. The effect of essential oil on cell membrane release of each selected bacterial was put in evidence by measuring cellular material that absorb at 260 nm and 280 nm after 0 min, 30 min and 60 min of exposure and confirm by measuring DNA, RNA and proteins release by treated cells on extracellular medium using Nanodrop 1000 spectrophotometer. Results: The crystal violet method shows twelve (12) strong, five (05) moderate and five (05) weak biofilm forming bacteria. The anti-biofilm activity against the oral bacteria who shown that most of essentials oils have activity on different biofilm formation and the MICs ranged from 0.31 mg/mL to 1.25 mg/mL. Concentration of intracellular material released in extracellular medium ranged from 186,56 ± 2,35 ng/µL to 766,6 ± 2,84 ng/µL for DNA, 158,06 ± 1,87 ng/µL to 628,53 ± 2,05 ng/µL for RNA and 695,9 ± 2,11ng/µL to 1125,23 ± 2,15 ng/µL for proteins. Conclusion: This study demonstrates that the selected EOs have a significant anti-biofilm activity, acting on the cell surface and causing the disruption of the bacterial membrane. These EOs are interesting alternative to conventional antimicrobials for the control of oral microorganisms. Keywords: Anti-biofilm activity, Biofilm, oral diseases

scholarly journals Effect of S-PRG Eluate on Biofilm Formation and Enzyme Activity of Oral Bacteria

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Masahiro Yoneda ◽  
Nao Suzuki ◽  
Yosuke Masuo ◽  
Akie Fujimoto ◽  
Kosaku Iha ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Composite Resin ◽  
Fusobacterium Nucleatum ◽  
Microtiter Plate ◽  
Oral Bacteria ◽  
Gelatin Film ◽  
Glass Ionomer ◽  
Microtiter Plate Assay ◽  
Substrate Hydrolysis ◽  
Adherence Ability

Recently, the antibacterial activity of a composite resin containing prereacted glass ionomer (S-PRG) filler was revealed. We examined the effect of an S-PRG eluate on various biologic activities ofStreptococcus mutansandPorphyromonas gingivalis. Adherence ability ofS. mutanswas evaluated by microtiter plate assay; protease and gelatinase activities ofP. gingivaliswere examined by synthetic substrate hydrolysis and gelatin film spot assay, respectively. Coaggregation ofP. gingivaliswithFusobacterium nucleatumwas also examined. S-PRG eluate was found to suppress streptococcal adherence. S-PRG eluate inhibited the protease and gelatinase activities ofP. gingivalisand the coaggregation betweenP. gingivalisandF. nucleatum. These results indicate that S-PRG eluate suppresses streptococcal adherence and inhibits the protease and coaggregation activities ofP. gingivalis. These findings may prompt research into novel strategies for preventing caries and periodontitis.

scholarly journals Microtiter Plate Assay for Assessment of Listeria monocytogenes Biofilm Formation

2002 ◽  
Vol 68 (6) ◽  
pp. 2950-2958 ◽  
Author(s):  
D. Djordjevic ◽  
M. Wiedmann ◽  
L. A. McLandsborough
Keyword(s):  
Listeria Monocytogenes ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Cellular Growth ◽  
Epifluorescence Microscopy ◽  
Simple Method ◽  
Microtiter Plate Assay ◽  
Plate Assay ◽  
Biofilm Production

ABSTRACT Listeria monocytogenes has the ability to form biofilms on food-processing surfaces, potentially leading to food product contamination. The objective of this research was to standardize a polyvinyl chloride (PVC) microtiter plate assay to compare the ability of L. monocytogenes strains to form biofilms. A total of 31 coded L. monocytogenes strains were grown in defined medium (modified Welshimer's broth) at 32°C for 20 and 40 h in PVC microtiter plate wells. Biofilm formation was indirectly assessed by staining with 1% crystal violet and measuring crystal violet absorbance, using destaining solution. Cellular growth rates and final cell densities did not correlate with biofilm formation, indicating that differences in biofilm formation under the same environmental conditions were not due to growth rate differences. The mean biofilm production of lineage I strains was significantly greater than that observed for lineage II and lineage III strains. The results from the standardized microtiter plate biofilm assay were also compared to biofilm formation on PVC and stainless steel as assayed by quantitative epifluorescence microscopy. Results showed similar trends for the microscopic and microtiter plate assays, indicating that the PVC microtiter plate assay can be used as a rapid, simple method to screen for differences in biofilm production between strains or growth conditions prior to performing labor-intensive microscopic analyses.

scholarly journals Adaptation of Congo Red Agar Method and Microtiter Plate Assay to Study Biofilm Formation in Streptomyces

2021 ◽  
Vol 18 (1) ◽  
pp. 113-123
Author(s):  
Rabha EL othmany ◽  
Hafida Zahir ◽  
Chorouk Zanane ◽  
Doha Mazigh ◽  
Mostafa Ellouali ◽  
...  
Keyword(s):  
Crystal Violet ◽  
Congo Red ◽  
Biofilm Formation ◽  
Staining Method ◽  
Microtiter Plate ◽  
Alcoholic Solution ◽  
Bioactive Molecules ◽  
Microtiter Plate Assay ◽  
Agar Method ◽  
Quantitative And Qualitative Methods

Streptomyces has many advantages for exploration in biotechnological applications because of their ability to elaborate a multitude of bioactive molecules and secondary metabolites. Despite the importance of this genus in biotechnology, biofilm formation in Streptomyces is under-investigated. The objective of this research is to adapt two assays for the assessment of biofilm formation in Streptomyces. In the present investigation, we assess and follow biofilm formation in eight Streptomyces strains using quantitative and qualitative methods. The quantitative study based on a staining of the retained biomass in the microtiter plate with crystal violet “5%” and destaining using ethanol/acetone mixture, the concentration of crystal violet in the alcoholic solution reflect the intensity of the attached biofilm. On the other hand, the qualitative one consists of using modified freeman’s method a modified congo red agar method based on the color of colonies. Quantification of biomass by crystal violet staining method confirmed that Streptomyces bellus A43 and Streptomyces bellus A61 are biofilm-forming and this ability increase with the period of incubation. Our results showed that sixStreptomyces strains arenon-slime producing/non-biofilm forming. Two Streptomyces strains are slime producing/biofilm forming; this character vanishes at five days. Further research on genes responsible for biofilm formation in Streptomyces is highly recommended for better understanding of the phenomenon.

scholarly journals Anti-biofilm property of essential oils from Cymbopogon sp. against pathogenic bacteria in single-culture and co-culture

2020 ◽  
Vol 42 ◽  
pp. e1
Author(s):  
Alessandra Farias Millezi ◽  
Vanessa Schuh ◽  
Janaina Schuh ◽  
Taciara Penno do Amaral
Keyword(s):  
Essential Oils ◽  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Microtiter Plate ◽  
Bacterial Cells ◽  
Viable Cells ◽  
Single Culture ◽  
Cymbopogon Flexuosus ◽  
Colony Forming Units ◽  
Cymbopogon Martinii

In this study, we evaluated whether the essential oils (EOs) from Cymbopogon flexuosus and Cymbopogon martinii can prevent production of biofilms either in single or combined culture of Staphylococcus aureus and Pseudomonas aeruginosa. Biofilm formation was assessed by microtiter-plate test with further quantification of viable cells and biofilm biomass. The evaluated EOs at 0.78 % significantly (P 0.05) reduced only the viable cells of S. aureus that inhabited biofilm. However, in single-and co-culture assays, both oils significantly (P 0.05) decreased the amount of biofilm biomass. Biofilm reductions between 52-83% and 60-93% were achieved for the treatments with EOs from C. flexuosus and C. martinii, respectively. Although the biomass reductions of simgle and co-cultivated biofilms were significant, the same was not true for viable cells, except for S. aureus. Considering that the remaining colony forming units can reconstitute the EPS matrix, studies with higher concentrations than those used in this research are suggested in order to obtain greater logarithmic reductions of viable bacterial cells.

Inhibition and Inactivation of Salmonella Typhimurium Biofilms from Polystyrene and Stainless Steel Surfaces by Essential Oils and Phenolic Constituent Carvacrol†

2013 ◽  
Vol 76 (2) ◽  
pp. 205-212 ◽  
Author(s):  
KAMLESH A. SONI ◽  
ADEMOLA OLADUNJOYE ◽  
RAMAKRISHNA NANNAPANENI ◽  
M. WES SCHILLING ◽  
JUAN L. SILVA ◽  
...  
Keyword(s):  
Stainless Steel ◽  
Essential Oils ◽  
Salmonella Typhimurium ◽  
Exposure Time ◽  
Crystal Violet ◽  
Biofilm Formation ◽  
Cell Mass ◽  
Minimum Concentration ◽  
Steel Surfaces ◽  
Phenolic Constituent

Persistence of Salmonella biofilms within food processing environments is an important source of Salmonella contamination in the food chain. In this study, essential oils of thyme and oregano and their antimicrobial phenolic constituent carvacrol were evaluated for their ability to inhibit biofilm formation and inactivate preformed Salmonella biofilms. A crystal violet staining assay and CFU measurements were utilized to quantify biofilm cell mass, with evaluating factors such as strain variation, essential oil type, their concentrations, exposure time, as well as biofilm formation surface. Of the three Salmonella strains, Salmonella Typhimurium ATCC 23564 and Salmonella Typhimurium ATCC 19585 produced stronger biofilms than Salmonella Typhimurium ATCC 14028. Biofilm formation by different Salmonella strains was 1.5- to 2-fold higher at 22°C than at 30 or 37°C. The presence of nonbiocidal concentrations of thyme oil, oregano oil, and phenolic carvacrol at 0.006 to 0.012% suppressed Salmonella spp. biofilm formation 2- to 4-fold, but could not completely eliminate biofilm formation. There was high correlation in terms of biofilm inactivation, as determined by the crystal violet–stained optical density (at a 562-nm wavelength) readings and the viable CFU counts. Reduction of biofilm cell mass was dependent on antimicrobial concentration. A minimum concentration of 0.05 to 0.1% of these antimicrobial agents was needed to reduce a 7-log CFU biofilm mass to a nondetectable level on both polystyrene and stainless steel surfaces within 1 h of exposure time.

Investigate of the Ability of Cronobacter sakazakii Isolated from Clinical Samples of Children Under Two Years to Induce Swimming, Swarming and Biofilm

2018 ◽  
pp. 123
Author(s):  
Luma Abdal Hady Zwein ◽  
Tharieyt Abdulrahman Motlag ◽  
Mohamed Mousa
Keyword(s):  
Cerebrospinal Fluid ◽  
Congo Red ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Clinical Samples ◽  
Cronobacter Sakazakii ◽  
Yellow Color ◽  
Golden Yellow ◽  
Vitek 2 ◽  
Biochemical Examination

      The study included 200 samples were collected   from   children  under two   years included (50 samples from each of Cerebrospinal fluid, Blood, Stool and Urine) from, Central Children Hospital and Children's Protections Educational Hospital. Isolates bacterial were obtained cultural, microscopic and biochemical examination and diagnosed to the species by using vitek2 system. The results showed there were contamination in 6.5% of clinical samples. The diagnosed colonies which gave pink color on the MacConkey agar , golden yellow color on the Trypton Soy agar and green color on the Birillent Enterobacter sakazakii agar and gave  a probability of 99% in the vitek 2 and were identified as Cronobacter sakazakii. The identification revealed of thirteen isolates: 6(46.16%) isolated from Cerebrospinal fluid samples, 7(53.84%) isolated from blood samples and not isolated bacteria from stool and urine samples. The results of the investigation of some virulence factors showed that all bacteria isolates were able to swimming with a diameter ranging (1-9 mm) and swarming with a diameter ranging (1-40 mm) and their  ability to biofilm formation  by using three methods. The results show the ability  of  isolates to form biofilm by using  Congo red media  methods where it is 12 (92.30 %) out of 13 isolated bacteria belonging to C. sakazakii  able to form biofilm on the Congo red media  which is 3 (23.07%) were  strong production  biofilm ,   8 (61.53%)  were intermediate  production  biofilm and  1 (7.69% ) were weak  biofilm formation , while the 1 (7.69%)  unable to form biofilm.  Tubes method were all isolates were able to form biofilm, it were found that 3 (23.07%)  isolates strong, and 8 (61.53%) intermediate  and 2( 15.38%)  weak biofilm formation. Microtiter plate method  gave 5 (38.46 %) isolates strong, 6 (46.15%) intermediate and 1 (7.69%) weak biofilm formation.  

Rebalance the oral microbiota as efficacy tool in endocrine, metabolic, and immune disorders

2020 ◽  
Vol 20 ◽  
Author(s):  
Ciro Gargiulo Isacco ◽  
Andrea Ballini ◽  
Danila De Vito ◽  
Kieu Cao Diem Nguyen ◽  
Stefania Cantore ◽  
...  
Keyword(s):  
Systemic Disease ◽  
Current Treatment ◽  
Oral Bacteria ◽  
The Body ◽  
Oral Microbiota ◽  
Oral Diseases ◽  
Philosophical Approach ◽  
Treatment And Prevention ◽  
Cardio Vascular Disease ◽  
Cardio Vascular

: The current treatment and prevention of oral disorders follow a very sectoral control and procedures considering mouth and its structures as system completely independent from the rest of the body. The main therapeutic approach is carried out on just to keep the levels of oral bacteria and hygiene in an acceptable range compatible with one-way vision of oral-mouth health completely separated from a systemic microbial homeostasis (eubiosis vs dysbiosis). This can negatively impact on the diagnosis of more complex systemic disease and its progression. Dysbiosis is consequence of oral and gut microbiota unbalance with consequences, as reported in current literature, in cardio vascular disease, diabetes mellitus, rheumatoid arthritis, and Alzheimer’s disease. Likewise, there is the need to highlight and develop a novel philosophical approach in the treatments for oral diseases that will necessarily involve non-conventional approaches.

Essential oils from Artemisia species inhibit biofilm formation and the virulence of Escherichia coli EPEC 2348/69

Biofouling ◽  
2021 ◽  
pp. 1-11
Author(s):  
Ahmed Mathlouthi ◽  
Nabil Saadaoui ◽  
Eugenia Pennacchietti ◽  
Daniela De Biase ◽  
Mossadok Ben-Attia
Keyword(s):  
Escherichia Coli ◽  
Essential Oils ◽  
Biofilm Formation ◽  
Artemisia Species

scholarly journals Virtual Screening and Biological Evaluation of Anti-Biofilm Agents Targeting LuxS in the Quorum Sensing System

2021 ◽  
Vol 16 (6) ◽  
pp. 1934578X2110196
Author(s):  
Zheng Liu ◽  
Lihua Zhang ◽  
Jincai Wang ◽  
Yanping Li ◽  
Yiqun Chang ◽  
...  
Keyword(s):  
Dental Caries ◽  
Quorum Sensing ◽  
Biofilm Formation ◽  
Binding Mode ◽  
Biological Evaluation ◽  
Oral Diseases ◽  
Traditional Use ◽  
New Approach ◽  
Sensing System ◽  
Plaque Biofilm

Biofilm formation is considered as a crucial factor in various oral diseases, such as dental caries. The quorum sensing (QS) signaling system was proved to have a crucial role in the microbial dental plaque biofilm formation of Streptococcus mutans ( S. mutans). LuxS was critical to regulating the QS system and survival of the bacterium, and, therefore, compounds which target LuxS may be a potential therapy for dental caries. The binding activities of 37,170 natural compounds to LuxS were virtually screened in this study. Baicalein and paeonol were chosen for further research of the binding mode and ΔG values with LuxS. Both baicalein and paeonol inhibited the biofilm formation without influence on the growth of S. mutans. Baicalein also distinctly reduced the production of both rhamnolipids and acids. The results provide us with a new approach to combat dental caries instead of the traditional use of antibacterial chemicals.

Modulatory Effect of Glycated Collagen on Oral Streptococcal Nanoadhesion

2020 ◽  
Vol 100 (1) ◽  
pp. 82-89
Author(s):  
C.M.A.P. Schuh ◽  
B. Benso ◽  
P.A. Naulin ◽  
N.P. Barrera ◽  
L. Bozec ◽  
...  
Keyword(s):  
Dental Caries ◽  
Biofilm Formation ◽  
Type I Collagen ◽  
Type I ◽  
Oral Streptococci ◽  
Oral Diseases ◽  
Collagen Crosslinking ◽  
Initial Adhesion ◽  
Collagen Glycation ◽  
The Impact

Biofilm-mediated oral diseases such as dental caries and periodontal disease remain highly prevalent in populations worldwide. Biofilm formation initiates with the attachment of primary colonizers onto surfaces, and in the context of caries, the adhesion of oral streptococci to dentinal collagen is crucial for biofilm progression. It is known that dentinal collagen suffers from glucose-associated crosslinking as a function of aging or disease; however, the effect of collagen crosslinking on the early adhesion and subsequent biofilm formation of relevant oral streptococci remains unknown. Therefore, the aim of this work was to determine the impact of collagen glycation on the initial adhesion of primary colonizers such as Streptococcus mutans UA159 and Streptococcus sanguinis SK 36, as well as its effect on the early stages of streptococcal biofilm formation in vitro. Type I collagen matrices were crosslinked with either glucose or methylglyoxal. Atomic force microscopy nanocharacterization revealed morphologic and mechanical changes within the collagen matrix as a function of crosslinking, such as a significantly increased elastic modulus in crosslinked fibrils. Increased nanoadhesion forces were observed for S. mutans on crosslinked collagen surfaces as compared with the control, and retraction curves obtained for both streptococcal strains demonstrated nanoscale unbinding behavior consistent with bacterial adhesin-substrate coupling. Overall, glucose-crosslinked substrates specifically promoted the initial adhesion, biofilm formation, and insoluble extracellular polysaccharide production of S. mutans, while methylglyoxal treatment reduced biofilm formation for both strains. Changes in the adhesion behavior and biofilm formation of oral streptococci as a function of collagen glycation could help explain the biofilm dysbiosis seen in older people and patients with diabetes. Further studies are necessary to determine the influence of collagen crosslinking on the balance between acidogenic and nonacidogenic streptococci to aid in the development of novel preventive and therapeutic treatment against dental caries in these patients.

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