scholarly journals Potential of Lactobacillus casei shirota’s strain against the biofilm-forming of Salmonella Spp

2019 ◽  
Vol 8 (2) ◽  
pp. 54-63
Author(s):  
Annisa Rizka Fauziah ◽  
Meiska Bahar ◽  
Aprilla Ayu Wulandari
Keyword(s):  
Biofilm Formation ◽  
Lactobacillus Casei ◽  
Microtiter Plate ◽  
Assay Method ◽  
Antibiofilm Activity ◽  
Salmonella Spp ◽  
Microplate Reader ◽  
Competitive Adhesion ◽  
Concentration Series

Biofilm of Salmonella spp. is formed through the expression of biofilm genes associated with proteins (bapA) regulated by curli synthesis genes (csg) which carry out adhesion, colonization, maturation, and dispersion on the surface of the intestinal epithelium. This study aimed to determine the antibiofilm activity of Lactobacillus casei Shirota’S strain (LcS) as an inhibitor of Salmonella spp. biofilm formation in vitro. This research was a true experimental study using Microtiter Plate 96 wells Biofilm Assay method. The sample used was the suspension of Salmonella spp. The treatment was in the form of adding a LcS suspension with a concentration series of 10-1;10-2; 10-3;10-4; and 10-5. Biofilm measurements were carried out using a microplate reader and obtained quantitative data in the form of Optical Density at a wavelength of 595nm. The results of this study showed that LcS suspension has antibiofilm activity ranging from 10-5 concentrations with a percentage of 36.58% (p<0.05). The results of exometabolism LcS can reduce Salmonella growth. Exopolysaccharide (EPS) and sortase-dependent proteins (SrtA) of LcS form barriers as competitive adhesion in inhibiting pathogenic biofilm formation.

scholarly journals The Anti-biofilm Activity of Nanometric Zinc doped Bioactive Glass against Putative Periodontal Pathogens: An in vitro Study

2018 ◽  
Vol 4 (1) ◽  
pp. 95-107
Author(s):  
Nasrin Esfahanizadeh ◽  
Mohammad Reza Nourani ◽  
Abbas Bahador ◽  
Nasrin Akhondi ◽  
Mostafa Montazeri
Keyword(s):  
Biofilm Formation ◽  
Sol Gel ◽  
Antimicrobial Properties ◽  
In Vitro Study ◽  
Inhibitory Effect ◽  
Antibiofilm Activity ◽  
Periodontal Pathogens ◽  
Microplate Reader ◽  
45S5 Bioglass

Abstract Colonization of periodontal pathogens on the surgical sites is one of the primary reasons for the failure of regenerative periodontal therapies. Bioactive glasses (BGs) owing to their favorable structural and antimicrobial properties have been proposed as promising materials for the reconstruction of periodontal and peri-implant bone defects. This study aimed to investigate the antibiofilm activity of zinc-doped BG (Zn/BG) compared with 45S5 Bioglass® (BG®) on putative periodontal pathogens. In this in vitro experimental study, the nano BG doped with 5-mol% zinc and BG® were synthesized by sol-gel method. Mono-species biofilms of Aggregatibacter actinomycetemcomitans (A. a), Porphyromonas gingivalis (P. g), and Prevotella intermedia (P. i)were prepared separately in a well-containing microplate. After 48 hours of exposure to generated materials at 37°C, the anti-biofilm potential of the samples was studied by measuring the optical density (OD) at 570nm wavelengths with a microplate reader. Two-way ANOVA then analyzed the results. Both Zn/BG and BG® significantly reduced the biofilm formation ability of all examined strains after 48 hours of incubation (P=0.0001). Moreover, the anti-biofilm activity of Zn/BG was significantly stronger than BG® (P=0.0001), which resulted in the formation of a weak biofilm (OD<1) compared with a moderately adhered biofilm observed with BG® (1<OD<2). Zn/BG showed a significant inhibitory effect on the biofilm formation of all examined periodontal pathogens. Given the enhanced regenerative and anti-biofilm properties of this novel biomaterial, further investigations are required for its implementation in clinical situations.

scholarly journals In vitro Antibiofilm Activity of Pomegranate (Punica granatum) Juice on Oral Pathogens

2019 ◽  
Vol 1 (2) ◽  
pp. 49
Author(s):  
Jemima Pramadita ◽  
Armelia Sari Widyarman
Keyword(s):  
Biofilm Formation ◽  
Brain Heart Infusion ◽  
Punica Granatum ◽  
Pomegranate Juice ◽  
Bacterial Suspension ◽  
Antibiofilm Activity ◽  
Microplate Reader ◽  
Level Of Significance ◽  
Negative Controls

Introduction: Pomegranate (Punica granatum) fruit contains valuable ingredients, such as ellagitannins and flavonoids, that have many potential effects, including antibacterial, antifungal, and anti-inflammatory functions. Objectives: The aim of this study was to investigate the effects of pomegranate fruit juice on F. nucleatum and S. sanguinis monospecies and multispecies biofilm formation in vitro. Methods: Pomegranate juice was obtained using a juicer and diluted using a brain heart infusion (BHI) broth into five different concentrations. The biofilm assay was performed as follows: F. nucleatum and S. sanguinis were cultured separately in the BHI broth for 48 hours at 37°C in an anaerobic atmosphere. A 200 mL bacterial suspension (107 CFU/mL) was distributed into a 96-well plate and incubated for 24 hours to form  a biofilm. Subsequently, pomegranate juice was added to the biofilm well and observed after 1 hours, 3 hours, 6 hours, and 24 hours. The biofilm mass was measured using a microplate reader (490 nm) after crystal violet staining. Chlorhexidine (0.2%) and the biofilms without treatment were used as the positive and negative controls, respectively. The data were statistically analyzed using one-way analysis of variance, with p<0.05 as the level of significance. Result: There was a significant biofilm reduction after treatment with pomegranate juice for all the concentrations and incubation times (p<0.05). The effective concentrations to inhibit the biofilm monospecies F. nucleatum and S. sanguinis and the multispecies were 6.25% (OD 0.148±0.019), 50% (OD 0.211±0.026), and 6.25% (OD 0.024±0.209), respectively. Conclusion: Pomegranate juice inhibits F. nucleatum and S. sanguinis biofilm formation as a monospecies and a multispecies. Future studies are needed to observe the mechanism of this active substance.

scholarly journals In vitro Antibiofilm Activity of Pomegranate (Punica granatum) Juice on Oral Pathogens

2019 ◽  
Vol 2 (1) ◽  
pp. 15
Author(s):  
Jemima Pramadita ◽  
Armelia Sari Widyarman
Keyword(s):  
Biofilm Formation ◽  
Brain Heart Infusion ◽  
Punica Granatum ◽  
Pomegranate Juice ◽  
Bacterial Suspension ◽  
Antibiofilm Activity ◽  
Microplate Reader ◽  
Level Of Significance ◽  
Negative Controls

  Introduction: Pomegranate (Punica granatum) fruit contains valuable ingredients, such as ellagitannins and flavonoids, that have many potential effects, including antibacterial, antifungal, and anti-inflammatory functions.   Objectives: The aim of this study was to investigate the effects of pomegranate fruit juice on F. nucleatum and S. sanguinis monospecies and multispecies biofilm formation in vitro.   Methods: Pomegranate juice was obtained using a juicer and diluted using a brain heart infusion (BHI) broth into five different concentrations. The biofilm assay was performed as follows: F. nucleatum and S. sanguinis were cultured separately in the BHI broth for 48 hours at 37°C in an anaerobic atmosphere. A 200 mL bacterial suspension (107 CFU/mL) was distributed into a 96-well plate and incubated for 24 hours to form  a biofilm. Subsequently, pomegranate juice was added to the biofilm well and observed after 1 hours, 3 hours, 6 hours, and 24 hours. The biofilm mass was measured using a microplate reader (490 nm) after crystal violet staining. Chlorhexidine (0.2%) and the biofilms without treatment were used as the positive and negative controls, respectively. The data were statistically analyzed using one-way analysis of variance, with p<0.05 as the level of significance.   Result: There was a significant biofilm reduction after treatment with pomegranate juice for all the concentrations and incubation times (p<0.05). The effective concentrations to inhibit the biofilm monospecies F. nucleatum and S. sanguinis and the multispecies were 6.25% (OD 0.148±0.019), 50% (OD 0.211±0.026), and 6.25% (OD 0.024±0.209), respectively.   Conclusion: Pomegranate juice inhibits F. nucleatum and S. sanguinis biofilm formation as a monospecies and a multispecies. Future studies are needed to observe the mechanism of this active substance.

scholarly journals Antibiofilm and antivirulence potential of silver nanoparticles against multidrug-resistant Acinetobacter baumannii

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Helal F. Hetta ◽  
Israa M. S. Al-Kadmy ◽  
Saba Saadoon Khazaal ◽  
Suhad Abbas ◽  
Ahmed Suhail ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Antibacterial Activity ◽  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Diffusion Method ◽  
Resistance Pattern ◽  
Infection Model ◽  
The Impact

AbstractWe aimed to isolate Acinetobacter baumannii (A. baumannii) from wound infections, determine their resistance and virulence profile, and assess the impact of Silver nanoparticles (AgNPs) on the bacterial growth, virulence and biofilm-related gene expression. AgNPs were synthesized and characterized using TEM, XRD and FTIR spectroscopy. A. baumannii (n = 200) were isolated and identified. Resistance pattern was determined and virulence genes (afa/draBC, cnf1, cnf2, csgA, cvaC, fimH, fyuA, ibeA, iutA, kpsMT II, PAI, papC, PapG II, III, sfa/focDE and traT) were screened using PCR. Biofilm formation was evaluated using Microtiter plate method. Then, the antimicrobial activity of AgNPs was evaluated by the well-diffusion method, growth kinetics and MIC determination. Inhibition of biofilm formation and the ability to disperse biofilms in exposure to AgNPs were evaluated. The effect of AgNPs on the expression of virulence and biofilm-related genes (bap, OmpA, abaI, csuA/B, A1S_2091, A1S_1510, A1S_0690, A1S_0114) were estimated using QRT-PCR. In vitro infection model for analyzing the antibacterial activity of AgNPs was done using a co-culture infection model of A. baumannii with human fibroblast skin cell line HFF-1 or Vero cell lines. A. baumannii had high level of resistance to antibiotics. Most of the isolates harbored the fimH, afa/draBC, cnf1, csgA and cnf2, and the majority of A. baumannii produced strong biofilms. AgNPs inhibited the growth of A. baumannii efficiently with MIC ranging from 4 to 25 µg/ml. A. baumannii showed a reduced growth rate in the presence of AgNPs. The inhibitory activity and the anti-biofilm activity of AgNPs were more pronounced against the weak biofilm producers. Moreover, AgNPs decreased the expression of kpsMII , afa/draBC,bap, OmpA, and csuA/B genes. The in vitro infection model revealed a significant antibacterial activity of AgNPs against extracellular and intracellular A. baumannii. AgNPs highly interrupted bacterial multiplication and biofilm formation. AgNPs downregulated the transcription level of important virulence and biofilm-related genes. Our findings provide an additional step towards understanding the mechanisms by which sliver nanoparticles interfere with the microbial spread and persistence.

Evaluation of the Role of Staphylococci in the Pathomechanism of Conjunctivitis

2021 ◽  
Author(s):  
Ewa Jasińska ◽  
Agnieszka Bogut ◽  
Agnieszka Magryś ◽  
Alina Olender
Keyword(s):  
Antibiotic Resistance ◽  
Epithelial Cells ◽  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Microtiter Plate ◽  
Host Cells ◽  
Diffusion Method ◽  
Microtiter Plate Assay ◽  
Low Prevalence

Abstract Purpose: Determination of the association between ica genes and phenotypic biofilm formation in staphylococcal isolates involved in conjunctivitis, their antibiotic resistance as well as detection of selected virulence characteristics: adhesion to epithelial cells and in vitro cytotoxicity.Methods: The study included 26 Staphylococcus aureus (SA) and 26 Staphylococcus epidermidis (SE) isolates. The presence of icaAD genes and ica operon was determined by the PCR assay. Phenotypic biofilm formation was verified using the microtiter plate assay. Antibiotic resistance was performed using the disc diffusion method. Staphylococcal ability to attach to host cells was assessed by flow cytometry. Cytotoxicity on epithelial cells was evaluated by LDH assay.Results: The ica genes were detected in 26.9% of SE and in 42.3% of SA isolates. Only 15.3% of isolates (SE) were positive for both the icaAD and the ica operon. Phenotypically, 19.2% of SE isolates were strong biofilm producers, among which three were both icaAD- and ica operon-positive. 26.9% of SA isolates were strong biofilm producers. Methicillin resistance (MR) was detected in 34.6% of SE and 26.9% of SA isolates. 75% of MR isolates were multidrug resistant. SA isolates adhered to host cells more extensively than SE. SA isolates released higher level of LDH than SE.Conclusions: Adherence abilities were commonly observed in staphylococci associated with conjunctivitis. However, low prevalence of isolates positive for a complete and functional ica locus and low prevalence of strong biofilm producers was detected. SA adhered to a greater extent to eukaryotic cells than SE and were more cytotoxic.

scholarly journals Effect of Silver Nanoparticles on Biofilm Formation and EPS Production of Multidrug-Resistant Klebsiella pneumoniae

2020 ◽  
Vol 2020 ◽  
pp. 1-9 ◽  
Author(s):  
Muhammad Hussnain Siddique ◽  
Bilal Aslam ◽  
Muhammad Imran ◽  
Asma Ashraf ◽  
Habibullah Nadeem ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Microtiter Plate ◽  
Cellular Protein ◽  
Antibiofilm Activity ◽  
Biofilm Inhibition ◽  
Microtiter Plate Assay ◽  
Protein Leakage ◽  
Hela Cell Lines

Antibiotic resistance against present antibiotics is rising at an alarming rate with need for discovery of advanced methods to treat infections caused by resistant pathogens. Silver nanoparticles are known to exhibit satisfactory antibacterial and antibiofilm activity against different pathogens. In the present study, the AgNPs were synthesized chemically and characterized by UV-Visible spectroscopy, scanning electron microscopy, and X-ray diffraction. Antibacterial activity against MDR K. pneumoniae strains was evaluated by agar diffusion and broth microdilution assay. Cellular protein leakage was determined by the Bradford assay. The effect of AgNPs on production on extracellular polymeric substances was evaluated. Biofilm formation was assessed by tube method qualitatively and quantitatively by the microtiter plate assay. The cytotoxic potential of AgNPs on HeLa cell lines was also determined. AgNPs exhibited an MIC of 62.5 and 125 μg/ml, while their MBC is 250 and 500 μg/ml. The production of extracellular polymeric substance decreased after AgNP treatment while cellular protein leakage increased due to higher rates of cellular membrane disruption by AgNPs. The percentage biofilm inhibition was evaluated to be 64% for K. pneumoniae strain MF953600 and 86% for MF953599 at AgNP concentration of 100 μg/ml. AgNPs were evaluated to be minimally cytotoxic and safe at concentrations of 15-120 μg/ml. The data evaluated by this study provided evidence of AgNPs being safe antibacterial and antibiofilm compounds against MDR K. pneumoniae.

scholarly journals Antimicrobial and Antibiofilm Activity against Staphylococcus aureus of Opuntia ficus-indica (L.) Mill. Cladode Polyphenolic Extracts

Antioxidants ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 117 ◽  
Author(s):  
Federica Blando ◽  
Rossella Russo ◽  
Carmine Negro ◽  
Luigi De Bellis ◽  
Stefania Frassinetti
Keyword(s):  
Staphylococcus Aureus ◽  
Antioxidant Activity ◽  
Antioxidant Capacity ◽  
Biofilm Formation ◽  
Ex Vivo ◽  
Trolox Equivalent Antioxidant Capacity ◽  
Total Phenolic ◽  
Antibiofilm Activity ◽  
Opuntia Ficus Indica

Plant extracts are a rich source of natural compounds with antimicrobial properties, which are able to prevent, at some extent, the growth of foodborne pathogens. The aim of this study was to investigate the potential of polyphenolic extracts from cladodes of Opuntia ficus-indica (L.) Mill. to inhibit the growth of some enterobacteria and the biofilm formation by Staphylococcus aureus. Opuntia ficus-indica cladodes at two stages of development were analysed for total phenolic content and antioxidant activity by Oxygen Radical Absorbance Capacity (ORAC) and Trolox equivalent antioxidant capacity (TEAC) (in vitro assays) and by cellular antioxidant activity in red blood cells (CAA-RBC) (ex vivo assay). The Liquid Chromatography Time-of-Flight Mass Spectrometry (LC/MS–TOF) analysis of the polyphenolic extracts revealed high levels of piscidic acid, eucomic acid, isorhamnetin derivatives and rutin, particularly in the immature cladode extracts. Opuntia cladodes extracts showed a remarkable antioxidant activity (in vitro and ex vivo), a selective inhibition of the growth of Gram-positive bacteria, and an inhibition of Staphylococcus aureus biofilm formation. Our results suggest and confirm that Opuntia ficus-indica cladode extracts could be employed as functional food, due to the high polyphenolic content and antioxidant capacity, and used as natural additive for food process control and food safety.

scholarly journals Peptide Mix from Olivancillaria hiatula Interferes with Cell-to-Cell Communication in Pseudomonas aeruginosa

2019 ◽  
Vol 2019 ◽  
pp. 1-12
Author(s):  
Edward Ntim Gasu ◽  
Hubert Senanu Ahor ◽  
Lawrence Sheringham Borquaye
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Biofilm Formation ◽  
Antimicrobial Agents ◽  
Cell Communication ◽  
Microtiter Plate ◽  
Antibiofilm Activity ◽  
Dependent Manner ◽  
Biofilm Inhibition ◽  
Swarming Motility

Bacteria in biofilms are encased in an extracellular polymeric matrix that limits exposure of microbial cells to lethal doses of antimicrobial agents, leading to resistance. In Pseudomonas aeruginosa, biofilm formation is regulated by cell-to-cell communication, called quorum sensing. Quorum sensing facilitates a variety of bacterial physiological functions such as swarming motility and protease, pyoverdine, and pyocyanin productions. Peptide mix from the marine mollusc, Olivancillaria hiatula, has been studied for its antibiofilm activity against Pseudomonas aeruginosa. Microscopy and microtiter plate-based assays were used to evaluate biofilm inhibitory activities. Effect of the peptide mix on quorum sensing-mediated processes was also evaluated. Peptide mix proved to be a good antibiofilm agent, requiring less than 39 μg/mL to inhibit 50% biofilm formation. Micrographs obtained confirmed biofilm inhibition at 1/2 MIC whereas 2.5 mg/mL was required to degrade preformed biofilm. There was a marked attenuation in quorum sensing-mediated phenotypes as well. At 1/2 MIC of peptide, the expression of pyocyanin, pyoverdine, and protease was inhibited by 60%, 72%, and 54%, respectively. Additionally, swarming motility was repressed by peptide in a dose-dependent manner. These results suggest that the peptide mix from Olivancillaria hiatula probably inhibits biofilm formation by interfering with cell-to-cell communication in Pseudomonas aeruginosa.

scholarly journals Biofilm Formation and Interaction with the Surfaces of Gallstones by Salmonella spp.

2002 ◽  
Vol 70 (5) ◽  
pp. 2640-2649 ◽  
Author(s):  
A. M. Prouty ◽  
W. H. Schwesinger ◽  
J. S. Gunn
Keyword(s):  
Salmonella Enterica ◽  
Biofilm Formation ◽  
Asymptomatic Carrier ◽  
Carrier State ◽  
Luria Bertani ◽  
Salmonella Spp ◽  
Human Gallbladder ◽  
Luria Bertani Broth ◽  
High Concentrations

ABSTRACT Salmonellae can exist in an asymptomatic carrier state in the human gallbladder. Individuals with gallstones are more likely to become typhoid carriers, and antibiotic treatments are often ineffectual against Salmonella enterica serovar Typhi in carriers with gallstones. Therefore, we hypothesized that Salmonella spp. form biofilms on the surfaces of gallstones, where the bacteria are protected from high concentrations of bile and antibiotics. A number of methods were utilized to examine biofilm formation on human gallstones and glass coverslips in vitro, including confocal, light, and scanning electron microscopy. In our assays, salmonellae formed full biofilms on the surfaces of gallstones within 14 days and appeared to excrete an exopolysaccharide layer that bound them to the surfaces and to other bacteria. Efficient biofilm formation on gallstones was dependent upon the presence of bile, as a biofilm did not form on gallstones within 14 days in Luria-Bertani broth alone. The biofilms formed by a Salmonella enterica serovar Typhi Vi antigen mutant, as well as strains with mutations in genes that eliminate production of four different fimbriae, were indistinguishable from the biofilms formed by the parents. Mutants with an incomplete O-antigen, mutants that were nonmotile, and mutants deficient in quorum sensing were unable to develop complete biofilms. In addition, there appeared to be selectivity in salmonella binding to the gallstone surface that did not depend on the topology or surface architecture. These studies should aid in the understanding of the Salmonella carrier state, an important but underresearched area of typhoid fever pathogenesis. If the basis of carrier development can be understood, it may be possible to identify effective strategies to prevent or treat this chronic infection.

scholarly journals In Vitro Antibacterial and Antibiofilm Activity of Lippia alba Essential Oil, Citral, and Carvone against Staphylococcus aureus

2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Emanuela Mesquita Porfírio ◽  
Hider Machado Melo ◽  
Antônio Matheus Gomes Pereira ◽  
Theodora Thays Arruda Cavalcante ◽  
Geovany Amorim Gomes ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Essential Oil ◽  
Microtiter Plate ◽  
Antibiofilm Activity ◽  
Bacterial Cells ◽  
Lippia Alba ◽  
Microdilution Method ◽  
Aerial Parts ◽  
Clinical Interest

In vitro antimicrobial and antibiofilm activities of the Lippia alba essential oil and its major components (citral and carvone) against Staphylococcus aureus were investigated. Essential oils (LA1EO, LA2EO, and LA3EO) were extracted from the aerial parts of three L. alba specimens by hydrodistillation and analyzed by gas chromatography coupled to a mass spectrometer. Minimum Inhibitory Concentrations (MIC) and Minimum Bacterial Concentration (MBC) were determined by the microdilution method. For the antibiofilm assays, the biomass formation in the biofilm was evaluated by the microtiter-plate technique with the crystal violet (CV) assay and the viability of the bacterial cells was analyzed. All oils and their major components presented antibacterial activity, and the lowest MIC and MBC values were 0.5 mg mL−1 when LA1EO and citral were used. Potential inhibition (100%) of S. aureus biofilm formation at the concentration of 0.5 mg mL−1 of all EOs was observed. However, the elimination of biofilm cells was confirmed at concentrations of 1 mg mL−1, 2 mg mL−1, 2 mg mL−1, and 0.5 mg mL−1 for LA1EO, LA2EO, LA3EO, and citral, respectively. The results obtained in the present research point to the promising antibacterial and antibiofilm potential of L. alba EOs against S. aureus, a species of recognized clinical interest.

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