Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 125

Search results for: Streptococcus mutans

125 Molecular Basis of Anti-Biofilm and Anti-Adherence Activity of Syzygium aromaticum on Streptococcus mutans: In Vitro and in Vivo Study

Authors: Mohd Adil, Rosina Khan, Asad U. Khan, Vasantha Rupasinghe HP

Abstract:

The study examined the effects of Syzygium aromaticum extracts on the virulence properties of Streptococcus mutans. The activity of glucosyltransferases in the presence of crude and diethylether fraction was reduced to 80% at concentration 78.12μg/ml and 39.06μg/ml respectively. The glycolytic pH drop by S. mutans cells was also disrupted by these extracts without affecting the bacterial viability. Microscopic analysis revealed morphological changes of the S. mutans biofilms, indicating that these plant extracts at sub-MICs could significantly affect the ability of S. mutans to form biofilm with distorted extracellular matrix. Furthermore, with the help of quantitative RT-PCR, the expression of different genes involved in adherence, quorum sensing, in the presence of these extracts were down regulated. The crude and active fractions were found effective in preventing caries development in rats. The data showed that S. aromaticum holds promise as a naturally occurring source of compounds that may prevent biofilm-related oral diseases.

Keywords: biofilm, quorum sensing, Streptococcus mutans, Syzygium aromaticum extract

Procedia PDF Downloads 233
124 Oral Microflora and the Risk of Dental Caries in Portuguese Children

Authors: Sara Sousa, Veronique Gomes, Nélio Veiga, Maria José Correia

Abstract:

Objectives: To assess the presence or absence of Streptococcus mutans, Streptococcus gordonii and Streptococcus salivarius in the oral biofilm of children in an elementary school of Viseu, Portugal, and verify the relationship between Streptococcus gordonii and Streptococcus salivarius and the absence of dental caries. Methods: A cross-sectional study was designed with a final sample of 40 children aged 6-11 years old. Oral examination was accomplished with the identification of their oral health status and oral biofilm collection. Analysis of biological samples by molecular techniques of DNA isolation and identification of three Streptococci bacteria by Polimerase Chain Reaction (PCR) was made. Results: We identified Streptococcus salivarius and Streptococcus gordoni only in the lower interincisal region. These species were also present mainly in the first permanent non-decayed molars. On the contrary, Streptococcus mutans was found mostly in decayed first permanent molars. Conclusion: This preliminary study establishes a possible association between the absence of dental caries and the presence of Streptococcus gordonii and Streptococcus salivarius. Since these two species are described as alkali producers, it is suggested that their presence somehow confers protection against caries. These results support new dental caries prevention strategies based on oral biofilm modulation by enrichment with alkalinogenic species.

Keywords: dental caries, oral biofilm, Streptococcus gordonii, Streptococcus salivarius

Procedia PDF Downloads 211
123 Antimicrobial Effect of Toothpastes Containing Fluoride, Xylitol or Xylitol-Probiotic on Salivary Streptococcus mutans and Lactobacillus in Children

Authors: Eda Arat Maden, Ceyhan Altun, Bilal Ozmen, Feridun Basak

Abstract:

Purpose: The purpose of this study was to compare the antimicrobial effect of toothpastes containing fluoride, xylitol or xylitol-probiotic in vivo, Streptococcus mutans and Lactobacillus in 13-15 years old children. Method: The study consisted of 60 pediatric patients were randomly divided into 3 groups of 20 each. Group 1 received fluoride toothpaste (Colgate Max Fresh), group 2 used xylitol toothpaste (Xyliwhite) and group 3 used xylitol-probiotic toothpaste (PerioBiotic). Subjects were asked to use the allocated dentifrice two times a day, for 6 weeks. We performed tests on the samples of saliva in the beginning of the study and after 6 weeks’ duration following the use of toothpaste. Result and Conclusion: All of the participants of the study stated that they brushed their teeth well twice a day by using the toothpastes given to them for 6 weeks. Majority of the subjects had high counts of salivary mutans streptococci and Lactobacillus at baseline. When the number of cariogenic bacteria (S. mutans and Lactobacillus) at the start of the PerioBiotic Probiotic toothpaste usage are compared with the results measured after 6 weeks, an important decrease is observed in the S. mutans and Lactobacillus bacteria according to the CRT Tests. After the 6-week use of Probiotic toothpaste, the S. mutans (≥105) decreased to 20% from 75% in the group with S. mutans and Lactobacillus (≥105) decreased to 30% from 60% in the group with Lactobacillus. In addition, an important decrease was recorded in the participants with the S. mutans percentage (80% - 45%) and Lactobacillus (70% - 55%) after using the Colgate Max Fresh toothpaste for six weeks. On the other hand, it was determined with the Chi-square that there were not important changes between the Xyliwhite toothpaste group and the other groups with S. mutans (80% - 75%) and Lactobacillus (75% -65%). It was also determined after the comparison of the groups that the decrease in the S. mutans was higher than the group using PerioBiotic Probiotic toothpaste at a significant level, when compared with the Colgate Max Fresh toothpaste and Xyliwhite toothpaste. S. mutans were more sensitive to the antimicrobial activity of the PerioBiotic Probiotic toothpaste and to the Colgate Max Fresh toothpaste when compared with the Lactobacillus. In the light of the data obtained in this in vivo study, the use of probiotics ensure the balance between the bacterial flora in the oral cavity.

Keywords: lactobacillus, probiotic, Streptococcus mutans, toothpaste

Procedia PDF Downloads 186
122 Survey of the Effect of the Probiotic Bacterium Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT/PTEN, and MAPK Signaling Pathways at Co-Culture with KB Oral Cancer Cell Line and HUVEC Cells

Authors: Negar Zaheddoust, Negin Zaheddoust, Abbas Asoudeh-Fard

Abstract:

Probiotic bacteria have been employed as a novel and less side-effect strategy for anticancer therapy. Since the oral cavity is a host for probiotic and pathogen bacteria to colonize, more investigation is needed to evaluate the effectiveness of this novel adjunctive treatment for oral cancer. We considered Lactobacillus plantarum as a probiotic and Streptococcus mutans as a pathogen bacterium in our study. The aim of this study is to examine the effect of Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT / PTEN, and MAPK signaling pathway, which is involved in apoptosis or survival of oral cancer KB cells. On the other hand, to study the effects of these bacteria on normal cells, we used HUVEC cells. The KB and HUVEC cell lines were co-cultured with Lactobacillus plantarum and Streptococcus mutans isolated from traditional Iranian dairy and dental plaque, respectively. The growth-inhibitory effects of these two bacteria on KB and HUVEC cells were determined by (3-(4, 5-dimethylthiazolyl-2)-2,5diphenyltetrazolium bromide) MTT assay. MTT results demonstrated that the proliferation of KB cells was affected in a time, dose, and strain-dependent manner. In the following, the examination of induced apoptosis or necrosis in co-cultured KB cells with the best IC50 concentration of the Lactobacillus plantarum and Streptococcus mutans will be analyzed by FACS flow cytometry, and the changes in gene expression of Casp3, AKT / PTEN, MAPK genes will be evaluated using real-time polymerase chain reaction.

Keywords: cancer therapy, induced apoptosis, oral cancer, probiotics

Procedia PDF Downloads 108
121 Inhibition of Streptococcus Mutans Biofilm Development of Dental Caries In Vitro and In Vivo by Trachyspermum ammi Seeds: An Approach of Alternative Medicine

Authors: Mohd Adil, Rosina Khan, Danishuddin, Asad U. Khan

Abstract:

The aim of this study was to evaluate the influence of the crude and active solvent fraction of Trachyspermum ammi on S. mutans cariogenicity, effect on expression of genes involved in biofilm formation and caries development in rats. GC–MS was carried out to identify the major components present in the crude and the active fraction of T. ammi. The crude extract and the solvent fraction exhibiting least MIC were selected for further experiments. Scanning electron microscopy was carried out to observe the effect of the extracts on S. mutans biofilm. Comparative gene expression analysis was carried out for nine selected genes. 2-Isopropyl-5-methyl-phenol was found as major compound in crude and the active fraction. Binding site of this compound within the proteins involved in biofilm formation was mapped with the help of docking studies. Real-time RT-PCR analyses revealed significant suppression of the genes involved in biofilm formation. All the test groups showed reduction in caries (smooth surface as well as sulcal surface caries) in rats. Moreover, it also provides new insight to understand the mechanism influencing biofilm formation in S. mutans. Furthermore, the data suggest the putative cariostatic properties of T. Ammi and hence can be used as an alternative medicine to prevent caries infection.

Keywords: bio-film, Streptococcus mutans, dental caries, bio-informatic

Procedia PDF Downloads 385
120 4-Allylpyrocatechol Loaded Polymeric Micelles for Solubility Enhancing and Effects on Streptococcus mutans Biofilms

Authors: Siriporn Okonogi, Pimpak Phumat, Sakornrat Khongkhunthian

Abstract:

Piper betle has been extensively reported for various pharmacological effects including antimicrobial activity. 4-Allylpyrocatechol (AC) is a principle active compound found in P. betle. However, AC has a problem of solubility in water. The aims of the present study were to prepare AC loaded polymeric micelles for enhancing its water solubility and to evaluate its anti-biofilm activity against oral phathogenic bacteria. AC was loaded in polymeric micelles (PM) of Pluronic F127 by using thin film hydration method to obtain AC loaded PM (PMAC). The results revealed that AC in the form of PMAC possessed high water solubility. PMAC particles were characterized using a transmission electron microscope and photon correlation spectroscopy. Determination of entrapment efficiency (EE) and loading capacity (LC) of PMAC was done by using high-performance liquid chromatography. The highest EE (86.33 ± 14.27 %) and LC (19.25 ± 3.18 %) of PMAC were found when the weight ratio of polymer to AC was 4 to 1. At this ratio, the particles showed spherical in shape with the size of 38.83 ± 1.36 nm and polydispersity index of 0.28 ± 0.10. Zeta potential of the particles is negative with the value of 16.43 ± 0.55 mV. Crystal violet assay and confocal microscopy were applied to evaluate the effects of PMAC on Streptococcus mutans biofilms using chlorhexidine (CHX) as a positive control. PMAC contained 1.5 mg/mL AC could potentially inhibit (102.01 ± 9.18%) and significantly eradicate (85.05 ± 2.03 %) these biofilms (p < 0.05). Comparison with CHX, PMAC showed slightly similar biofilm inhibition but significantly stronger biofilm eradication (p < 0.05) than CHX. It is concluded that PMAC can enhance water solubility and anti-biofilm activity of AC.

Keywords: pluronic, polymeric micelles, solubility, 4-allylpyrocathecol, Streptococcus mutans, anti-biofilm

Procedia PDF Downloads 70
119 The Comparison of pH Saliva before and after Brushing Teeth Using Tooth Paste Containing Betel Leaf Extracts

Authors: Ika Anisyah, Nety Trisnawaty

Abstract:

Mechanical brushing can help control plaque and is the first step to control dental caries. The type of toothpaste used is one of the contributing factors in it since the benefits of toothpaste are to reduce plaque formation and strengthen the teeth against dental caries, clean and polish tooth surfaces, eliminate or reduce bad breath, give a fresh taste to the mouth and maintain gingival health. Betel leaf toothpaste has the ability to inhibit the Streptococcus mutans bacteria that can cause the increase of pH saliva. Betel leaf extracts can increase the pH saliva because betel leaf has an anti bacterial characteristic against Streptococcus mutans so that pH saliva increases. This study aims to see the difference between pH saliva before and after brushing teeth with toothpaste containing betel leaf extracts. This type of research is pre-experimental using One Group Pretest-Posttest Design. This study was conducted on 32 subjects taken randomly from the representatives of students aged 11-12 years old in SD Pesanggrahan 03. The result of statistic test using non parametric test showed a value of 0.000. The resulted value being smaller than 0.05 (p < 0.05) means there is a significant salivary pH difference before and after teeth brushing using toothpaste containing betel leaf. The conclusion of this study showed an increase in salivary pH after teeth brushing with toothpaste containing betel leaves extracts in children aged 11-12 years old.

Keywords: pH saliva, brushing teeth, tooth paste, betel leaves extracts

Procedia PDF Downloads 59
118 Binding of Avian Excreta-Derived Enteroccoci to a Streptococcocus mutans: Implications for Avian to Human Transmission

Authors: Richard K. Jolley, Jonathan A. Coffman

Abstract:

Since Enterococci has been implicated in oral disease, we hypothesized the transmission of avian Enterococci to humans via fecal-oral transmission facilitated by adherence to dental plaque. To demonstrate the capability of Enterococci to bind to a dental plaque we filtered avian excreta and incubated the filtrate on a sucrose-induced, Streptococcus mutans biofilm. The biofilm was washed several times with a detergent to remove bacteria binding non-specifically to the biofilm, DNA was isolated from the biofilm, 16S rDNA was amplified, sequenced by Ion Torrent DNA sequencing and analyzed with bioinformatics. Enterococci and other known bacterial pathogens were shown to adhere to the biofilm. Culturing the washed biofilm with Bile Esculin Azide (BEA) agar also confirmed the presence of Enterococci as verified with Sanger sequencing. The results suggest that Enteroccoci in avian excreta has the ability to adhere to human dental plaque and may be a mechanism of entry when humans encounter contaminated aerosols, water or food.

Keywords: Enterococci, avian excreta, dental plaque, NGS

Procedia PDF Downloads 83
117 Mothwash Formulation of Moringa Leaf (Moringa Oleifera) and Its Activity as an Antibacterial for Streptococcus Mutans

Authors: Amalia Dwi Berliyanti Amel

Abstract:

Streptococcus mutants bacteria are bacteria that are believed to be the cause of the growth of dental plaque which can further adversely affect dental caries if left unchecked. Previous research has shown that Moringa leaf extract can slow down the growth rate of this bacterium. This study aims to make the best formulation of mouthwash with the active ingredient of Moringa leaf extract based on its antibacterial and organoleptic test results. Nine mouthwash variations were carried out with two factors and three levels, namely a comparison of the concentration of sorbitol (A) with three levels namely 15% (A1), 20% (A2), and 25% (A3), and peppermint added (B) with three levels, namely 0.2% (B1), 0.25% (B2), and 0.3% (B3). The test parameters performed as the determination of the best mouthwash are based on physicochemical properties which include pH and viscosity as well as organoleptic test results which include color, viscosity, aroma, taste, sensation in the mouth, and general appearance. The results showed that the bright zone as a test for the antibacterial activity of Streptococcus mutants began to be seen at a concentration of 5%. Moringa leaf mouthwash formulation has a pH value between 6 - 7, with a control of 6. Whereas the mucosa leaf mouthwash vascularity produced between 1.1 - 1.7 cP with a control of 1.1 cP. Moringa leaf mouthwash and control have the same total number of microbes, namely 0 colonies / mL. Based on organoleptic tests performed with 20 panelists, it was shown that the best mouthwash formulation was formulation A1B3 with sorbitol composition 15% and peppermint 0.3%.

Keywords: antibasteria, formula, moringa leaf, mouthwash

Procedia PDF Downloads 46
116 Evaluation of Oral Biofilm Suppression by Carribean Herbal Extracts

Authors: Ravi Teja Chitturi Suryaprakash, Chandrashekhar Unakal, Haytham Al-Bayaty, Duraisamy Saravanakumar

Abstract:

Background and significance: Oral biofilm formation is a well-known causative factor for caries and periodontal diseases. Scientists over the years have been trying to find a solution against the formation of oral biofilms. Though several advances have been made to understand the microbial ecology and how the bio film survives, it is still an enigma to researchers to find a chemical product that not only can inhibit the formation of oral bio film but also not disturb the oral micro flora required for oral health and not to cause damage to the cells of the oral cavity. One such product that has never been investigated much are herbal preparations. Some of the microorganisms important in the formation of biofilm are Streptococcus mutans, Actinomyces naeslundi, Streptococuss oralis and Prevotella intermedia. The aim of this study was to study the antimicrobial property of some herbal extracts available in Trinidad and Tobago against these pathogens. The significance of this study is that identification of biologically effective plant extracts can result in indigenous development of mouth rinses and tooth pastes that the people can benefit from to not only develop effective but also a cheap solution. Methodology: The extracts from the leaves of Plectranthus ambonicus, Ocmium tenuiflorum, Azadirchata indica, Anacardium occidentale, Psidium guajava were prepared by dissolving them in water. The extracts from the roots of Curcuma longa were prepared similarly and the antimicrobial activity of these six plant extracts was determined by the agar well diffusion method using minimum inhibitory concentration (MIC) against Streptococcus mutans, Actinomyces naeslundi, Streptococuss oralis and Prevotella intermedia and compared with chlorhexidine. Results: The six plant extracts showed variable effect on the oral micro-organisms. Ocmium tenuiflorum (16.66 ± 0.44, 14 ± 0.58, 13.33 ± 0.88, 12.83 ± 0.60), Azadirchata indica (17.5 ± 0.28, 14.83 ± 0.17, 15 ± 0.58, 12.83 ± 0.6) and Curcuma longa (16.16 ± 0.44, 13.66 ± 0.88, 12.33 ± 0.88, 11.33 ± 0.67) were found to have highest inhibitory activity against all the four pathogens (Streptococcus mutans, Streptococuss oralis, Actinomyces naeslundi, and Prevotella intermedia) respectively. Conclusion: Although the extracts were not pure compounds we obtained antimicrobial results which determine that they are potent antimicrobial agents. Further derivation of pure compounds from these extracts could be lucrative as it might lead to the development of a cost effective and biologically safe medicine to act against oral biofilms. Acknowledgement: The authors would like to acknowledge the Campus Research and Publication Fund Committee, The University of the West Indies for funding this study and would also like to acknowledge Dr. Leonette Cox, Department of Chemistry, Faculty of Science and Technology, The University of the West Indies, St. Augustine Campus, Trinidad and Tobago for helping to prepare the plant extracts.

Keywords: agar well diffusion method, herbal extracts, minimum inhibitory concentration, oral biofilm forming microorganisms

Procedia PDF Downloads 92
115 Genotypic Identification of Oral Bacteria Using 16S rRNA in Children with and without Early Childhood Caries in Kelantan, Malaysia

Authors: Zuliani Mahmood, Thirumulu Ponnuraj Kannan, Yean Yean Chan, Salahddin A. Al-Hudhairy

Abstract:

Caries is the most common childhood disease which develops due to disturbances in the physiological equilibrium in the dental plaque resulting in demineralization of tooth structures. Plaque and dentine samples were collected from three different tooth surfaces representing caries progression (intact, over carious lesion and dentine) in children with early childhood caries (ECC, n=36). In caries free (CF) children, plaque samples were collected from sound tooth surfaces at baseline and after one year (n=12). The genomic DNA was extracted from all samples and subjected to 16S rRNA PCR amplification. The end products were cloned into pCR®2.1-TOPO® Vector. Five randomly selected positive clones collected from each surface were sent for sequencing. Identification of the bacterial clones was performed using BLAST against GenBank database. In the ECC group, the frequency of Lactobacillus sp. detected was significantly higher in the dentine surface (p = 0.031) than over the cavitated lesion. The highest frequency of bacteria detected in the intact surfaces was Fusobacterium nucleatum subsp. polymorphum (33.3%) while Streptococcus mutans was detected over the carious lesions and dentine surfaces at a frequency of 33.3% and 52.7% respectively. Fusobacterium nucleatum subsp. polymorphum was also found to be highest in the CF group (41.6%). Follow up at the end of one year showed that the frequency of Corynebacterium matruchotii detected was highest in those who remained caries free (16.6%), while Porphyromonas catoniae was highest in those who developed caries (25%). In conclusion, Streptococcus mutans and Porphyromonas catoniae are strongly associated with caries progression, while Lactobacillus sp. is restricted to deep carious lesions. Fusobacterium nucleatum subsp. polymorphum and Corynebacterium matruchotii may play a role in sustaining the healthy equilibrium in the dental plaque. These identified bacteria show promise as potential biomarkers in diagnosis which could help in the management of dental caries in children.

Keywords: early childhood caries, genotypic identification, oral bacteria, 16S rRNA

Procedia PDF Downloads 209
114 An Antibacterial Dental Restorative Containing 3,4-Dichlorocrotonolactone: Synthesis, Formulation and Evaluation

Authors: Dong Xie, Leah Howard, Yiming Weng

Abstract:

The objective of this study was to synthesize and characterize 5-acryloyloxy-3,4-dichlorocrotonolactone (a furanone derivative), use this derivative to modify a dental restorative, and study the effect of the derivative on the antibacterial activity and compressive strength of the formed restorative. In this study, a furanone derivative was synthesized, characterized, and used to formulate a dental restorative. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the formed restorative. The fabricated restorative specimens were photocured and conditioned in distilled water at 37oC for 24 h, followed by direct testing for CS or/and incubating with S. mutans for 48 h for antibacterial testing. The results show that the modified dental restorative showed a significant antibacterial activity without substantially decreasing the mechanical strengths. With addition of the antibacterial derivative up to 30%, the restorative kept its original CS nearly unchanged but showed a significant antibacterial activity with 68% reduction in the S. mutans viability. Furthermore, the antibacterial function of the modified restorative was not affected by human saliva. The aging study also indicates that the modified restorative may have a long-lasting antibacterial function. It is concluded that this experimental antibacterial restorative may potentially be developed into a clinically attractive dental filling restorative due to its high mechanical strength and antibacterial function.

Keywords: antibacterial, dental restorative, compressive strength, S. mutans viability

Procedia PDF Downloads 237
113 Antibacterial Effect of Silver Diamine Fluoride Incorporated in Fissure Sealants

Authors: Nélio Veiga, Paula Ferreira, Tiago Correia, Maria J. Correia, Carlos Pereira, Odete Amaral, Ilídio J. Correia

Abstract:

Introduction: The application of fissure sealants is considered to be an important primary prevention method used in dental medicine. However, the formation of microleakage gaps between tooth enamel and the fissure sealant applied is one of the most common reasons of dental caries development in teeth with fissure sealants. The association between various dental biomaterials may limit the major disadvantages and limitations of biomaterials functioning in a complementary manner. The present study consists in the incorporation of a cariostatic agent – silver diamine fluoride (SDF) – in a resin-based fissure sealant followed by the study of release kinetics by spectrophotometry analysis of the association between both biomaterials and assessment of the inhibitory effect on the growth of the reference bacterial strain Streptococcus mutans (S. mutans) in an in vitro study. Materials and Methods: An experimental in vitro study was designed consisting in the entrapment of SDF (Cariestop® 12% and 30%) into a commercially available fissure sealant (Fissurit®), by photopolymerization and photocrosslinking. The same sealant, without SDF was used as a negative control. The effect of the sealants on the growth of S. mutans was determined by the presence of bacterial inhibitory halos in the cultures at the end of the incubation period. In order to confirm the absence of bacteria in the surface of the materials, Scanning Electron Microscopy (SEM) characterization was performed. Also, to analyze the release profile of SDF along time, spectrophotometry technique was applied. Results: The obtained results indicate that the association of SDF to a resin-based fissure sealant may be able to increase the inhibition of S. mutans growth. However, no SDF release was noticed during the in vitro release studies and no statistical significant difference was verified when comparing the inhibitory halo sizes obtained for test and control group.  Conclusions: In this study, the entrapment of SDF in the resin-based fissure sealant did not potentiate the antibacterial effect of the fissure sealant or avoid the immediate development of dental caries. The development of more laboratorial research and, afterwards, long-term clinical data are necessary in order to verify if this association between these biomaterials is effective and can be considered for being used in oral health management. Also, other methodologies for associating cariostatic agents and sealant should be addressed.

Keywords: biomaterial, fissure sealant, primary prevention, silver diamine fluoride

Procedia PDF Downloads 179
112 Temporal Change in Bonding Strength and Antimicrobial Effect of a Zirconia after Nonthermal Atmospheric Pressure Plasma Treatment

Authors: Chan Park, Sang-Won Park, Kwi-Dug Yun, Hyun-Pil Lim

Abstract:

Purpose: Plasma treatment under various conditions has been studied to increase the bonding strength and surface sterilization of dental ceramic materials. We assessed the evolution of the shear bond strength (SBS) and antimicrobial effect of nonthermal atmospheric pressure plasma (NTAPP) treatment over time. Methods: Presintered zirconia specimens were manufactured as discs (diameter: 15 mm, height: 2 mm) after final sintering. The specimens then received a 30-min treatment with argon gas (Ar², 99.999%; 10 L/min) using an NTAPP device. Five post-treatment intervals were evaluated: control (no treatment), P0 (within 1 h), P1 (24 h), P2 (48 h), and P3 (72 h). This study investigated the surface characteristics, SBS of two different resin cement (RelyXTM U200 self-adhesive resin cement, Panavia F2.0 methacryloyloxydecyl dihydrogen phosphate (MDP)-based resin cement), and Streptococcus mutans biofilm formation. Results: The SBS of RelyXTM U200 increased significantly (p < 0.05) within 2 days following plasma treatment (P0, P1, P2). For Panavia F 2.0, a significant decrease (p < 0.05) was detected only in the group that had undergone cementation immediately after plasma treatment (P0). S. mutans adhesion decreased significantly (p < 0.05) within 2 days of plasma treatment (P0, P1, P2) compared to the control group. The P0 group displayed a lower biofilm thickness than the P1 and P2 groups (p < 0.05). Conclusions: After NTAPP treatment of zirconia, the effects on bonding strength and antimicrobial growth persist for a limited duration. The effect of NTAPP treatment on bonding strength depends on the resin cement.

Keywords: NTAPP, SBS, antimicrobial effect, zirconia

Procedia PDF Downloads 104
111 Antimicrobial and Antibiofilm Properties of Fatty Acids Against Streptococcus Mutans

Authors: A. Mulry, C. Kealey, D. B. Brady

Abstract:

Planktonic bacteria can form biofilms which are microbial aggregates embedded within a matrix of extracellular polymeric substances (EPS). They can be found attached to abiotic or biotic surfaces. Biofilms are responsible for oral diseases such as dental caries, gingivitis and the progression of periodontal disease. Biofilms can resist 500 to 1000 times the concentration of biocides and antibiotics used to kill planktonic bacteria. Biofilm development on oral surfaces involves four stages, initial attachment, early development, maturation and dispersal of planktonic cells. The Minimum Inhibitory Concentration (MIC) was determined using a range of saturated and unsaturated fatty acids using the resazurin assay, followed by serial dilution and spot plating on BHI agar plates to establish the Minimum Bactericidal Concentration (MBC). Log reduction of bacteria was also evaluated for each fatty acid. The Minimum Biofilm Inhibition Concentration (MBIC) was determined using crystal violet assay in 96 well plates on forming and pre-formed S. mutans biofilms using BHI supplemented with 1% sucrose. Saturated medium-chain fatty acids Octanoic (C8.0), Decanoic (C10.0) and Undecanoic acid (C11.0) do not display strong antibiofilm properties; however, Lauric (C12.0) and Myristic (C14.0) display moderate antibiofilm properties with 97.83% and 97.5% biofilm inhibition with 1000 µM respectively. Monounsaturated, Oleic acid (C18.1) and polyunsaturated large chain fatty acids, Linoleic acid (C18.2) display potent antibiofilm properties with biofilm inhibition of 99.73% at 125 µM and 100% at 65.5 µM, respectively. Long-chain polyunsaturated Omega-3 fatty acids α-Linoleic (C18.3), Eicosapentaenoic Acid (EPA) (C20.5), Docosahexaenoic Acid (DHA) (C22.6) have displayed strong antibiofilm efficacy from concentrations ranging from 31.25-250µg/ml. DHA is the most promising antibiofilm agent with an MBIC of 99.73% with 15.625µg/ml. This may be due to the presence of six double bonds and the structural orientation of the fatty acid. To conclude, fatty acids displaying the most antimicrobial activity appear to be medium or long-chain unsaturated fatty acids containing one or more double bonds. Most promising agents include Omega-3-fatty acids Linoleic, α-Linoleic, EPA and DHA, as well as Omega-9 fatty acid Oleic acid. These results indicate that fatty acids have the potential to be used as antimicrobials and antibiofilm agents against S. mutans. Future work involves further screening of the most potent fatty acids against a range of bacteria, including Gram-positive and Gram-negative oral pathogens. Future work will involve incorporating the most effective fatty acids onto dental implant devices to prevent biofilm formation.

Keywords: antibiofilm, biofilm, fatty acids, S. mutans

Procedia PDF Downloads 33
110 Effects of Gelatin on Characteristics and Dental Pathogen Inhibition by Silver Nanoparticles Synthesized from Ascorbic Acid

Authors: Siriporn Okonogi, Temsiri Suwan, Sakornrat Khongkhunthian, Jakkapan Sirithunyalug

Abstract:

In this study, silver nanoparticles (AgNPs) were prepared using ascorbic acid as a reducing agent and silver nitrate as a precursor. The effects of gelatin (G) on particle characteristics and dental pathogen inhibition were investigated. The spectra of AgNPs and G-AgNPs were compared using UV-Vis and Energy-dispersive X-ray (EDX) spectroscopy. The obtained AgNPs and G-AgNPs showed the maximum absorption at 410 and 430 nm, respectively, and EDX spectra of both systems confirmed Ag element. Scanning electron microscope showed that AgNPs and G-AgNPs were spherical in shape. Particles size, size distribution, and zeta potential were determined using dynamic light scattering approach. The size of AgNPs and G-AgNPs were 56 ± 2.4 and 67 ± 3.6 nm, respectively with a size distribution of 0.23 ± 0.03 and 0.19 ± 0.02, respectively. AgNPs and G-AgNPs exhibited negative zeta potential of 24.1 ± 2.7 mV and 32.7 ± 1.2 mV, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the obtained AgNPs and G-AgNPs against three strains of dental pathogenic bacteria; Streptococcus gordonii, Streptococcus mutans, and Staphylococcus aureus were determined using broth dilution method. AgNPs and G-AgNPs showed the strongest inhibition against S. gordonii with the MIC of 0.05 and 0.025 mg/mL, respectively and the MBC of 0.1 and 0.05 mg/mL, respectively. Cytotoxicity test of AgNPs and G-AgNPs on human breast cancer cells using MTT assay indicated that G-AgNPs (0.1 mg/mL) was significantly stronger toxic than AgNPs with the cell inhibition of 91.1 ± 5.4%. G-AgNPs showed significantly less aggregation after storage at room temperature for 90 days than G-AgNPs.

Keywords: antipathogenic activity, ascorbic acid, cytotoxicity, stability

Procedia PDF Downloads 72
109 Peculiarities of Microflora of Odontogenic Inflammatory Processes in the Central Kazakhstan Region

Authors: Aliya Tokbergenova, Maida Tusupbekova, Daulet Dzhangaliyev, Alena Lavrinenko

Abstract:

Background: Odontogenic phlegmons are ranked the first among pyoinflammatory processes in the frequency of hospitalization in maxillofacial surgery in the post-Soviet countries. The main role in etiology is played by obligate anaerobes and aerobes. According to numerous data, the structure of aerobic pathogens is dominated by staphylococci and gram-negative bacteria. Aim: The research aim is to study the microflora of the purulent discharge odontogenic inflammatory processes. Materials and methods: A total of 220 patients have been examined, of which 120 patients aged 25-59 years have been included in the research who did not have comorbidity hospitalized in the maxillofacial hospital in Karaganda (Kazakhstan) from January 2016 to July 2017. The bacteriological research has been carried out on the basis of the multiaccess laboratory of the KSMU, through the Matrix Assisted Laser Desorption/Ionization (MALDI) apparatus. The material sample was pus from the inflammation focus, taken during the operating period. Results: According to the research among 120 patients (100%), 15 patients (12.5%) have had microorganisms not grown. From 105 (87.5%) bacteriological results, it has been revealed the following 1) Streptococcus: 51 (42.5%): Streptococcus beta-haemolytic: 17 (14.2%), Streptococcus pneumoniae: 12 (10%), Streptococcus anginosus: 8 (6.6%), Streptococcus oralis: 8 (6.6%), Streptococcus constellatus: 6 (5.0%); 2) Staphylococci: 27 (22.5%): Staphylococci aureus: 14 (11.7%) and Staphylococci epidermidis: 13 (10.8%); 3) Pseudomonas aeruginosa: 12 (10%); 4) Neisseria: 11 (9.1%): Neisseria mucosa: 5 (4.1%) and Neisseria macacae: 6 (5.0%); 5) Klebsiella pneumoniae: 2 (1.7%); 6) Stenotrophomonas maltophilia: 2 (1.7%). 15 patients (12.5%) experienced complications in the form of 1) The dissemination of the process in 10 patients (8.4%). 2) Osteomyelitis in 3 (2.5%). 3) Mediastinitis in 1 (0.8%). 4) Sinusitis in 1 (0.8%). 15 patients (100%) were carried out repeated bacteriological examination, the following was revealed: 1) Streptococcus: 10 (66.7%): Streptococcus beta-haemolytic: 4 (26.7%), Streptococcus pneumoniae: 2 (13.3%), Streptococcus аnginosus: 2 (13.3%), Streptococcus oralis: 1 (6.7%), Streptococcus constellatus: 1 (6.7%); 2) Staphylococci: 4 (26.7%): Staphylococci aureus: 3 (20%) and Staphylococci epidermidis: 1 (6.7%); 3) Pseudomonas aeruginosa: 1 (6.7%). Conclusions: Thus, according to our research data, streptococci predominate in the odontogenic processes microflora in aerobic flora in the central Kazakhstan region, which refutes the leading role of staphylococci in the development of odontogenic inflammatory processes, thus creating prerequisites for studying new treatment approaches.

Keywords: maxillofacial surgery, microflora, odontogenic phlegmons, pyo-inflammatory

Procedia PDF Downloads 111
108 Evaluation of Chemical Compositions and Biological Activities of Five Essential Oils

Authors: G. Ozturk, B. Demirci

Abstract:

It is well known that essential oils used for therapeutic purposes for many years. In this study, five different Pharmacopoeia grade essential oils (Achillea millefolium L., Pimpinella anisum L., Matricaria recutita L., Eucalyptus globulus L., Salvia officinalis L.) which obtained from commercial sources were evaluated for chemical compositions, synergistic antimicrobial activities, and lipoxygenase enzyme inhibitions. Volatile components were determined by gas chromatography/flame ionization detector and gas chromatography/mass spectrometer, simultaneously. The potential antimicrobial activity of essential oils was tested against oral pathogenic standard strains such as Streptococcus mutans, Streptococcus sanguinis, Staphylococcus aureus, Corynebacterium striatum, Candida albicans and Candida krusei by broth microdilution methods. Ciprofloxacin and ketoconazole were used positive controls. It has been observed that the essential oils tested have average inhibitory antimicrobial activity against oral pathogens with a Minimum Inhibition Concentration of 20-0.625 mg/mL. The active essential oils have been combined with antibiotics and synergistic effects have been evaluated by Checkerboard method. ƩFIC values were determined. In combination with antibiotics M. recutita essential oil has been shown to have a synergistic effect against S. aureus in combination with tetracycline (ƩFIC 0.46). In addition, 5-LOX inhibitory activity was measured by modifying the spectrophotometric method developed by Baylac and Racine. As a result, 5-LOX % inhibition of S. officinalis, E. globulus and M. recutita were calculated as 34.0 ± 6.66, 72.7 ± 2.78 and 27.7 ± 0.60, respectively.

Keywords: antimicrobial activity, essential oils, synergistic activity, 5-lipoxygenase inhibition

Procedia PDF Downloads 40
107 Bioactivity of Local Isolated Probiotic to Inhibiting Important Bacterial Pathogens in Aquaculture

Authors: Abhichet Nobhiwong, Jiraporn Rojtinnakorn, Udomluk Sompong

Abstract:

Six probiotic strains isolated from Chiang Mai and Chiang Rai province, Thailand; CR1-2, CM3-4, CM5-2, CR7-8, CM10-5 and CM10-8 were used to study their morphology and inhibition activity on three pathogenic bacteria; Aeromonas sp., Streptococcus sp. and Flavobacterium sp. that isolated from infected Nile tilapia. The agar well diffusion technique was applied for 24 and 48 hours incubation. Interestingly, some probiotics showed good inhibition activity both 24 and 48 hours on each 3 bacterial pathogens. The capable inhibiting Aeromonas sp. were CR1-2 and CR5-2 with inhibition diameters of 13.0 mm and 11.2 mm, respectively. For Streptococcus sp., effective probiotics were CR10-2 with inhibition diameters of 10.7 mm. Whereas for Flavobacterium sp., effective probiotics were CR5-2 with inhibition diameter of 9.7 mm. It can be concluded that these probiotics have potentiality to develop as the pathogens biocontrol products. These will be support for safety and organic aquaculture that which the most worthy for people health.

Keywords: probiotics, Aeromanas sp., Streptococcus sp., Flavobacterium sp.

Procedia PDF Downloads 200
106 Use of Lactic Strains Isolated from Algerian Ewe's Milk in the Manufacture of a Natural Yogurt

Authors: Chougrani Fadela, Cheriguene Abderrahim

Abstract:

Fifty three strains of thermophilic and mesophilic lactic acid bacteria were isolated from the ewe’s milk. Identification reveals the presence of nineteen strains (36%) of Lactobacillus sp., seventeen strains (32%) of Lactococcus sp., nine strains (17%) of Streptococcus thermophilus and eight strains (15%) of Leuconostoc sp. The strains were characterized for their technological properties. A high diversity of properties among the studied strains was demonstrated. On the basis of technological characteristics, two strains (Lactobacillus bulgaricus and Streptococcus thermophilus) were screened with respect to their acid and flavour production for the preparation of a natural yogurt and compared to a commercial starter cultures. Sensorial analyses revealed that the product manufactured on the basis of the isolated strains have a cohesiveness and adhesiveness corresponding to standard products. The pH and the acidity recorded are also within accepted levels during all the period of conservation.

Keywords: Lactobacillus bulgaricus, Streptococcus thermophilus, yoghurt, cohesiveness, adhesiveness, Algerian ewe’s milk

Procedia PDF Downloads 253
105 Characterization of Genus Candida Yeasts Isolated from Oral Microbiota of Brazilian Schoolchildren with Different Caries Experience

Authors: D. S. V. Barbieri, R. R. Gomes, G. D. Santos, P. F. Herkert, M. Moreira, E. S. Trindade, V. A. Vicente

Abstract:

The importance of yeast infections has increased in recent decades. The monitoring of Candida yeasts has been relevant in the study of groups and populations. This research evaluated 31 Candida spp. isolates from oral microbiota of 12 Brazilian schoolchildren coinfected with Streptococcus mutans. The isolates were evaluated for their ability to form biofilm in vitro and molecularly characterized based on the sequencing of intergenic spacer regions ITS1-5,8S-ITS2 and variable domains of the large subunit (D1/D2) regions of the rDNA, as well as ABC system genotyping. The sequencing confirmed 26 lineages of Candida albicans, three Candida tropicalis, one Candida guillhermondii and one Candida glabrata. Genetic variability and differences on in biofilm formation were observed among Candida yeasts lineages. At least one Candida strain from each caries activity child was C.albicans genotype A or Candida non-albicans. C. tropicalis was associated with highest cavities rates. These results indicate that the presence of C. albicans genotype A or multi-colonization by non albicans species seem to be associates to the potentialization of caries risk.

Keywords: biofilm, Candida albicans, oral microbiota, caries

Procedia PDF Downloads 435
104 Streptococcus anginosus Infections; Clinical and Bacteriologic Characteristics: A 6-Year Retrospective Study of Adult Patients in Qatar

Authors: Adila Shaukat, Hussam Al Soub, Muna Al Maslamani, Abdullatif Al Khal

Abstract:

Background: The aim of this study was to assess clinical presentation and antimicrobial susceptibility of Streptococcus (S.) anginosus group infections in Hamad General Hospital, a tertiary care hospital in the state of Qatar, which is a multinational community. The S. anginosus group is a subgroup of viridans streptococci that consist of 3 different species: S. anginosus, S. constellatus, and S. intermedius. Although a part of the human bacteria flora, they have potential to cause suppurative infections. Method: We studied a total of 101 patients with S. anginosus group infections from January 2006 until March 2012 by reviewing medical records and identification of organisms by VITEK 2 and MALDI-TOF. Results: The most common sites of infection were skin and soft tissue, intra-abdominal, and bacteremia (28.7%, 24.8%, and 22.7%, respectively). Abscess formation was seen in approximately 30% of patients. Streptococcus constellatus was the most common isolated species (40%) followed by S. anginosus(30%) and S. intermedius(7%). In 23% of specimens, the species was unidentified. The most common type of specimen for organism isolation was blood followed by pus and tissue (50%, 22%, and 8%, respectively). Streptococcus constellatus was more frequently associated with abdominal and skin and soft tissue infections than the other 2 species, whereas S. anginosus was isolated more frequently from blood. All isolates were susceptible to penicillin, ceftriaxone, and vancomycin. Susceptibility to erythromycin and clindamycin was also good, reaching 91% and 95%, respectively. Forty percent of patients needed surgical drainage along with antibiotic therapy. Conclusions: Identification of S. anginosus group to species level is helpful in clinical practice because different species exhibit different pathogenic potentials.

Keywords: abscess, bacterial infection, bacteremia, Streptococcus anginosus

Procedia PDF Downloads 60
103 Effect of Capsule Storage on Viability of Lactobacillus bulgaricus and Streptococcus thermophilus in Yogurt Powder

Authors: Kanchana Sitlaothaworn

Abstract:

Yogurt capsule was made by mixing 14% w/v of reconstitution of skim milk with 2% FOS. The mixture was fermented by commercial yogurt starter comprising Lactobacillus bulgaricus and Streptococcus thermophilus. These yogurts were made as yogurt powder by freeze-dried. Yogurt powder was put into capsule then stored for 28 days at 4oc. 8ml of commercial yogurt was found to be the most suitable inoculum size in yogurt production. After freeze-dried, the viability of L. bulgaricus and S. thermophilus reduced from 109 to 107 cfu/g. The precence of sucrose cannot help to protect cell from ice crystal formation in freeze-dried process, high (20%) sucrose reduced L. bulgaricus and S. thermophilus growth during fermentation of yogurt. The addition of FOS had reduced slowly the viability of both L. bulgaricus and S. thermophilus similar to control (without FOS) during 28 days of capsule storage. The viable cell exhibited satisfactory viability level in capsule storage (6.7x106cfu/g) during 21 days at 4oC.

Keywords: yogurt capsule, Lactobacillus bulgaricus, Streptococcus thermophilus, freeze-drying, sucrose

Procedia PDF Downloads 252
102 Use of a New Multiplex Quantitative Polymerase Chain Reaction Based Assay for Simultaneous Detection of Neisseria Meningitidis, Escherichia Coli K1, Streptococcus agalactiae, and Streptococcus pneumoniae

Authors: Nastaran Hemmati, Farhad Nikkhahi, Amir Javadi, Sahar Eskandarion, Seyed Mahmuod Amin Marashi

Abstract:

Neisseria meningitidis, Escherichia coli K, Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complications. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections. The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains. The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours. The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.

Keywords: cerebrospinal fluid, meningitis, quantitative polymerase chain reaction, simultaneous detection, diagnosis testing

Procedia PDF Downloads 24
101 Studies on Some Aspects of Sub Clinical Mastitis in Cattle

Authors: Kavita Jaidiya, Anju Chahar, Chitra Jaidiya

Abstract:

The present study was conducted on 200 quarters from 50 apparently healthy cows. Samples are subjected to California Mastitis Test (CMT), cultural examination, and mPCR. Milk samples were also subjected to changes in composition Viz. fat, protein, and lactose. The prevalence of subclinical mastitis based on culture examination was 30(60/200), 36 (72/200), and 40 percent (93/200) based on CMT, culture examination, and mPCR on a quarterly basis. The prevalence of subclinical mastitis on animal basis was 40 (20/50), 46 (23/50), and 52 percent (26/50) based on CMT, Culture examination, and mPCR. The highest prevalence was observed in IVth parity on a quarterly basis and in Vth parity on cow basis. On culture examination, Staphylococcus aureus was the most prevalent organism (50.56%), followed by Streptococcus dysaglactiae (11.33%), E. coli (7.8 %), Staphylococcus agalactiae (13.48 %), Staphylococcus epidermidis (2.2 %), Streptococcus hyicus (6.94%), Streptococcus uberis (5.16%), Klebsiella pneumonia (6.74%). On isolation by bacterial mPCR, Staphylococcus spp. (42%) was the major pathogen. Organisms isolated in mixed infections are Streptococcus spp., Klebsiella pneumonia, E.coli and Pseudomonas aeruginous. The average mean value of fat, protein, and lactose content in subclinically affected milk samples were 3.40 ± 0.101, 3.009 ± 0.033, and 4.48 ± 0.03, and the mean value of fat, protein, and lactose content in normal milk were 4.13 ± 0.035, 3.39 ± 0.021, and 5.10 ± 0.016. The mean blood level of reduced glutathione in subclinical mastitis (30.44 ± 1.87 ng/ml) was lower than healthy cows (47.98 ± 4.04ng/ml). The concentration of malondialdehyde (10.026 ± 0.21mmol/L) in subclinical mastitis was significantly higher as compared to healthy group cows (2.19 ± 0.23mmol/L).

Keywords: cow, subclinical mastitis, mPCR, California Mastitis test

Procedia PDF Downloads 70
100 Isolation and Characterization of Lactic Acid Bacteria from Libyan Traditional Fermented Milk "Laban"

Authors: M. H. Nahaisi, N. M. Almaroum

Abstract:

Laban is a Libyan traditional fermented milk product. This lactic fermentation has been known in many cities of Libya long time ago as stable, nutritious, refreshing drink especially during the summer. 16 naturally fermented milk samples were collected from different cities located in North West of Libya. The average pH, titratable acidity, fat and total solids were 4.16, 0.73%, 1.54% and 8.12 % respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴ and 41 ×10³ cfu/ ml. respectively. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of one hundred forty two lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests have revealed that the most frequently Lactobacillus species was found to be Lactobacillus delbrueckii ssp. lactis (62.32%) followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis and Leuconostoc mesenteroides ssp. cremoris.

Keywords: traditional fermented milk, laban, lactococcus, streptococcus, mesophilic lactobacillus, thermophilic lactobacillus counts

Procedia PDF Downloads 285
99 High-Throughput Artificial Guide RNA Sequence Design for Type I, II and III CRISPR/Cas-Mediated Genome Editing

Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

Abstract:

A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.

Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles

Procedia PDF Downloads 176
98 Rapid and Cheap Test for Detection of Streptococcus pyogenes and Streptococcus pneumoniae with Antibiotic Resistance Identification

Authors: Marta Skwarecka, Patrycja Bloch, Rafal Walkusz, Oliwia Urbanowicz, Grzegorz Zielinski, Sabina Zoledowska, Dawid Nidzworski

Abstract:

Upper respiratory tract infections are one of the most common reasons for visiting a general doctor. Streptococci are the most common bacterial etiological factors in these infections. There are many different types of Streptococci and infections vary in severity from mild throat infections to pneumonia. For example, S. pyogenes mainly contributes to acute pharyngitis, palatine tonsils and scarlet fever, whereas S. Streptococcus pneumoniae is responsible for several invasive diseases like sepsis, meningitis or pneumonia with high mortality and dangerous complications. There are only a few diagnostic tests designed for detection Streptococci from the infected throat of patients. However, they are mostly based on lateral flow techniques, and they are not used as a standard due to their low sensitivity. The diagnostic standard is to culture patients throat swab on semi selective media in order to multiply pure etiological agent of infection and subsequently to perform antibiogram, which takes several days from the patients visit in the clinic. Therefore, the aim of our studies is to develop and implement to the market a Point of Care device for the rapid identification of Streptococcus pyogenes and Streptococcus pneumoniae with simultaneous identification of antibiotic resistance genes. In the course of our research, we successfully selected genes for to-species identification of Streptococci and genes encoding antibiotic resistance proteins. We have developed a reaction to amplify these genes, which allows detecting the presence of S. pyogenes or S. pneumoniae followed by testing their resistance to erythromycin, chloramphenicol and tetracycline. What is more, the detection of β-lactamase-encoding genes that could protect Streptococci against antibiotics from the ampicillin group, which are widely used in the treatment of this type of infection is also developed. The test is carried out directly from the patients' swab, and the results are available after 20 to 30 minutes after sample subjection, which could be performed during the medical visit.

Keywords: antibiotic resistance, Streptococci, respiratory infections, diagnostic test

Procedia PDF Downloads 42
97 Antibacterial Activity and Cytotoxicity of Silver Nanoparticles Synthesized by Moringa oleifera Extract as Reducing Agent

Authors: Temsiri Suwan, Penpicha Wanachantararak, Sakornrat Khongkhunthian, Siriporn Okonogi

Abstract:

In the present study, silver nanoparticles (AgNPs) were synthesized by green synthesis approach using Moringa oleifera aqueous extract (ME) as a reducing agent and silver nitrate as a precursor. The obtained AgNPs were characterized using UV-Vis spectroscopy (UV-Vis), dynamic light scattering (DLS), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray diffractometry (XRD). The results from UV-Vis revealed that the maximum absorption of AgNPs was at 430 nm and the EDX spectrum confirmed Ag element. The results from DLS indicated that the amount of ME played an important role in particle size, size distribution, and zeta potential of the obtained AgNPs. The smallest size (62.4 ± 1.8 nm) with narrow distribution (0.18 ± 0.02) of AgNPs was obtained after using 1% w/v of ME. This system gave high negative zeta potential of -36.5 ± 2.8 mV. SEM results indicated that the obtained AgNPs were spherical in shape. Antibacterial activity using dilution method revealed that the minimum inhibitory and minimum bactericidal concentrations of the obtained AgNPs against Streptococcus mutans were 0.025 and 0.1 mg/mL, respectively. Cytotoxicity test of AgNPs on adenocarcinomic human alveolar basal epithelial cells (A549) indicated that the particles impacted against A549 cells. The percentage of cell growth inhibition was 87.5 ± 3.6 % when only 0.1 mg/mL AgNPs was used. These results suggest that ME is the potential reducing agent for green synthesis of AgNPs.

Keywords: antibacterial activity, Moringa oleifera extract, reducing agent, silver nanoparticles

Procedia PDF Downloads 50
96 A Furaneol-Containing Glass-Ionomer Cement for Enhanced Antibacterial Activity

Authors: Dong Xie, Yuling Xu, Leah Howard

Abstract:

Secondary caries is found to be one of the main reasons to the restoration failure of dental restoratives. To prevent secondary caries formation, dental restoratives ought to be made antibacterial. In this study, a natural fruit component furaneol was tethered onto polyacid, the formed polyacid was used to formulate the light-curable glass-ionomer cements, and then the effect of this new antibacterial compound on compressive strength (CS) and antibacterial activity of the formed cement was evaluated. Fuji II LC glass powders were used as fillers. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the formed cement. The experimental cement showed a significant antibacterial activity, accompanying with an initial CS reduction. Increasing the compound loading significantly decreased the S. mutans viability from 5 to 81% and also reduced the initial CS of the formed cements from 4 to 58%. The cement loading with 7% antibacterial polymer showed 168 MPa, 7.8 GPa, 243 MPa, 46 MPa, and 57 MPa in yield strength, modulus, CS, diametral tensile strength and flexural strength, respectively, as compared to 141, 6.9, 236, 42 and 53 for Fuji II LC. The cement also showed an antibacterial function to other bacteria. No human saliva effect was noticed. It is concluded that the experimental cement may potentially be developed to a permanent antibacterial cement.

Keywords: antibacterial, dental materials, strength, cell viability

Procedia PDF Downloads 237