Effects of sarang semut (Myrmecodia Pendens Merr. & Perry) extracts on Enterococcus faecalis sensitivity

Background: Enterococcus faecalis (E. faecalis) is a gram positive oral pathogen that reported at the main agent infection of endodontic treatment. Its activities are influenced by the virulence factors facilitating the interaction process between agents with host cells. Like aggregation substance, cytolysin, extracellular superoxide, gelatinase, hyaluronidase, sex pheromones, and surface adhesions molecules. Plant extracts are reported as the material antibacterial as well as E. faecalis in pathogenesis of endodontic infections. Purpose: Purpose of this study was to analyse of sarang semut extracts (Myrmecodia Pendens Merr. & Perry) towards sensitivity of E. faecalis. Method: This research used the methanol extract of sarang semut, E. faecalis ATCC 29212, and fosfomycin also chlorhexidine as the positive controls. Whereas, Bradford protein method was measured the concentration of the surface protein of E. faecalis and active component of the sarang semut extract. Result: Generally, the sarang semut extract possessed low sensitivity toward E. faecalis (≤ 13 mm), but on the concentrations of 100 µg/ml and 75 µg/ml better than inhibition of other concentrations, round 10.6-11.6 (mm). Specifically, on 100 µg/ml has indicator the minimal bactericidal concentration (MBC) on E. faecalis. Whereas minimal inhibition concentration (MIC) on the concentration of 3,125 µg/ml. Conclusion: Based on MBC and MIC assay, the extract of sarang semut has potential effects to adherence growth of E. faecalis, mainly on the highest concentration 100 µg/ml also MIC on 3,125 µg/ ml.

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Antibodies to a Surface-Exposed, N-terminal Domain of Aggregation Substance Are Not Protective in the Rabbit Model of Enterococcus faecalis Infective Endocarditis

Infection and Immunity ◽

10.1128/iai.69.5.3305-3314.2001 ◽

2001 ◽

Vol 69 (5) ◽

pp. 3305-3314 ◽

Cited By ~ 27

Author(s):

John K. McCormick ◽

Helmut Hirt ◽

Christopher M. Waters ◽

Timothy J. Tripp ◽

Gary M. Dunny ◽

...

Keyword(s):

Infective Endocarditis ◽

Enterococcus Faecalis ◽

Rabbit Model ◽

Surface Protein ◽

Conjugative Plasmid ◽

Aggregation Substance ◽

Exposed Region ◽

Correct Sequence

ABSTRACT The aggregation substance (AS) surface protein fromEnterococcus faecalis has been implicated as an important virulence factor for the development of infective endocarditis. To evaluate the role of antibodies specific for Asc10 (the AS protein from the conjugative plasmid pCF10) in protective immunity to infective endocarditis, an N-terminal region of Asc10 lacking the signal peptide and predicted to be surface exposed (amino acids 44 to 331; AS44–331) was cloned with a C-terminal histidine tag translational fusion and expressed fromEscherichia coli. N-terminal amino acid sequencing of the purified protein revealed the correct sequence, and rabbit polyclonal antisera raised against AS44–331 reacted specifically to Asc10 expressed from E. faecalis OG1SSp, but not to other proteins as judged by Western blot analysis. Using these antisera, flow cytometry analysis demonstrated that antibodies to AS44–331 bound to a surface-exposed region of Asc10. Furthermore, antibodies specific for AS44–331were opsonic for E. faecalis expressing Asc10 in vitro but not for cells that did not express Asc10. New Zealand White rabbits immunized with AS44–331 were challenged intravenously withE. faecalis cells constitutively expressing Asc10 in the rabbit model of experimental endocarditis. Highly immune animals did not show significant differences in clearance of organisms from the blood or spleen or in formation of vegetations on the aortic valve, in comparison with nonimmune animals. Although in vivo expression of Asc10 was demonstrated by immunohistochemistry, these experiments provide evidence that immunity to Asc10 does not play a role in protection from experimental infective endocarditis due toE. faecalis and may have important implications for the development of immunological approaches to combat enterococcal endocarditis.

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Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected in Sardinia (Italy)

Journal of Medical Microbiology ◽

10.1099/jmm.0.05038-0 ◽

2003 ◽

Vol 52 (6) ◽

pp. 491-498 ◽

Cited By ~ 109

Author(s):

I. Duprè ◽

S. Zanetti ◽

A. M. Schito ◽

G. Fadda ◽

L. A. Sechi

Keyword(s):

Enterococcus Faecalis ◽

Enterococcus Faecium ◽

Virulence Genes ◽

Protein Gene ◽

Surface Protein ◽

Virulence Determinants ◽

Colonization Factor ◽

Aggregation Substance ◽

Epithelial Cell Lines ◽

Surface Protein Gene

Enterococci are widely distributed in the environment; within the human body, they are normal commensals of the oral cavity, gastrointestinal tract and vagina. In recent years, enterococci have become one of the most frequent causes of acquired nosocomial infections worldwide. The molecular mechanism of virulence of these bacteria is still not completely understood. The aims of this work were to characterize phenotypically 47 isolates of Enterococcus faecalis and Enterococcus faecium collected in Sardinia (Italy) by their abilities to adhere to different epithelial cell lines (Vero and Caco-2 cells) and to associate their phenotypes with the presence of known virulence genes detected within their genomes by PCR. The following genes were amplified: AS (aggregation substance), esp (surface protein gene), ace (accessory colonization factor), efaA (E. faecalis endocarditis antigen) and gelE (gelatinase). The virulence genes were detected in E. faecalis isolates only, with the exception of esp, which was found in both species. The phenotypic and genotypic results were also compared with the susceptibility of isolates to various antibiotics.

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The Antibacterial Effect of Soursop Leaf Extract on Staphylococcus aureus ATCC® 25923tm (In Vitro)

Journal of Biomimetics Biomaterials and Biomedical Engineering ◽

10.4028/www.scientific.net/jbbbe.48.119 ◽

2020 ◽

Vol 48 ◽

pp. 119-125

Author(s):

Minasari Nasution ◽

Sri Amelia ◽

Khairiyani Asri Hasibuan

Keyword(s):

Staphylococcus Aureus ◽

Leaf Extract ◽

Antibacterial Effect ◽

Control Group ◽

Dilution Method ◽

Minimal Bactericidal Concentration ◽

Minimal Inhibition Concentration ◽

Mannitol Salt Agar ◽

Inhibition Concentration ◽

Significant Difference

Purpose: This study was to determine the Minimal Inhibition Concentration (MIC) and Minimal Bactericidal Concentration (MBC) soursop leaf extract antibacterial activity against bacteria Staphylococcus aureus ATCC® 25923TM at a concentration of 100, 90, 80, 70, 60 and 50%. Method: This study is an experimental laboratory with design Post Test Only Control Group Design by using soursop leaf extract at a concentration of 100, 90, 80, 70, 60 and 50% respectively. Each test is repeated four times to determine the average Minimal Inhibition Concentration (MIC) and Minimal Bactericidal Concentration (MBC). Soursop leaf extract is made by the dilution method using a medium Nutrient Broth (NB) and sub-cultured on Mannitol Salt Agar (MSA). The number of colonies of Staphylococcus aureus ATCC® 25923TM are counted using a manual calculation in the media Mueller Hinton Agar (MHA). Test Kruskkal Wallis there are differences in the antibacterial effect that was significant (p <0.05), soursop leaf extract against Staphylococcus aureus ATCC® 25923TM from each treatment group. Results: The minimum inhibitory concentration was 90% and the minimum bactericidal concentration is 100% committed. The number of colonies of Staphylococcus aureus bacteria ATCC® 25923TM at 100% concentration 0 CFU/mL 90% 234.50 CFU/ml and at concentrations of 80, 70, 60 and 50% could not count the number of colonies due to > 300 CFU/mL. As for the diameter of inhibitory zone at 100% concentration 10,625 mm, 90% 8,875 mm, 6,750 mm 80% and 70%, 60%, 50% reporting no inhibition zone diameter. Conclusion: The p-value of Kruskkal Wallis is p<0,05 which shows that there is a significant difference between the inhibition of soursop leaf extract with a concentration of 100, 90, 80, 70, 60 and 50% on growth of Staphylococcus aureus ATCC® 25923TM. Thus, soursop leaf extract has antibacterial effects against the growth of Staphylococcus aureus ATCC® 25923TM.

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Adhesion to Bile Drain Materials and Physicochemical Surface Properties of Enterococcus faecalis Strains Grown in the Presence of Bile

Applied and Environmental Microbiology ◽

10.1128/aem.68.8.3855-3858.2002 ◽

2002 ◽

Vol 68 (8) ◽

pp. 3855-3858 ◽

Cited By ~ 20

Author(s):

Karola Waar ◽

Henny C. van der Mei ◽

Hermie J. M. Harmsen ◽

John E. Degener ◽

Henk J. Busscher

Keyword(s):

Surface Properties ◽

Enterococcus Faecalis ◽

Photoelectron Spectroscopy ◽

Surface Protein ◽

Lower Surface ◽

Contact Angles ◽

Surface Proteins ◽

Water Contact ◽

Zeta Potentials ◽

Aggregation Substance

ABSTRACT The aim of this study is to determine whether growth in the presence of bile influences the surface properties and adhesion to hydrophobic bile drain materials of Enterococcus faecalis strains expressing aggregation substance (Agg) or enterococcal surface protein (Esp), two surface proteins that are associated with infections. After growth in the presence of bile, the strains were generally more hydrophobic by water contact angles and the zeta potentials were more negative than when the strains were grown in the absence of bile. Nitrogen was found in lower surface concentrations upon growth in the presence of bile, whereas higher surface concentrations of oxygen were measured by X-ray photoelectron spectroscopy. Moreover, an up to twofold-higher number of bacteria adhered after growth in bile for E. faecalis not expressing Agg or Esp and E. faecalis with Esp on its surface. E. faecalis expressing Agg did not adhere in higher numbers after growth in bile, possibly because they mainly adhere through positive cooperativity and less through direct interactions with a substratum surface. Since adhesion of bacteria is the first step in biomaterial-centered infection, it can be concluded that growth in bile increases the virulence of E. faecalis.

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Rottlerin Inhibits Chlamydial Intracellular Growth and Blocks Chlamydial Acquisition of Sphingolipids from Host Cells

Applied and Environmental Microbiology ◽

10.1128/aem.02151-07 ◽

2007 ◽

Vol 74 (4) ◽

pp. 1243-1249 ◽

Cited By ~ 11

Author(s):

Pooja Shivshankar ◽

Lei Lei ◽

Jie Wang ◽

Guangming Zhong

Keyword(s):

Host Cell ◽

Chlamydial Infection ◽

Host Factors ◽

Host Cells ◽

Potential Mechanism ◽

Infection Cycle ◽

Intracellular Growth ◽

Minimal Inhibition Concentration ◽

Middle Stage ◽

Inhibition Concentration

ABSTRACT We report that rottlerin, a plant-derived compound known to inhibit various mammalian kinases, profoundly inhibited chlamydial growth in cell culture with a minimal inhibition concentration of 1 μM. The inhibition was effective even when rottlerin was added as late as the middle stage of chlamydial infection cycle, against multiple Chlamydia species, and in different host cell lines. Pretreatment of host cells with rottlerin prior to infection also blocked chlamydial growth, suggesting that rottlerin targets host factors. Moreover, rottlerin did not alter the chlamydial infection rate and did not directly target chlamydial protein synthesis and secretion. The rottlerin-mediated inhibition of chlamydial replication and inclusion expansion correlated well with the rottlerin-induced blockade of host cell sphingolipid trafficking from the Golgi apparatus into chlamydial inclusions. These studies not only allowed us to identify a novel antimicrobial activity for rottlerin but also allowed us to uncover a potential mechanism for rottlerin inhibition of chlamydial growth.

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Inducible Expression of Enterococcus faecalis Aggregation Substance Surface Protein Facilitates Bacterial Internalization by Cultured Enterocytes

Infection and Immunity ◽

10.1128/iai.68.12.7190-7194.2000 ◽

2000 ◽

Vol 68 (12) ◽

pp. 7190-7194 ◽

Cited By ~ 41

Author(s):

Carol L. Wells ◽

Elizabeth A. Moore ◽

Julie A. Hoag ◽

Helmut Hirt ◽

Gary M. Dunny ◽

...

Keyword(s):

Lactococcus Lactis ◽

Enterococcus Faecalis ◽

Surface Protein ◽

Inducible Expression ◽

Free Medium ◽

Heterologous Host ◽

Aggregation Substance ◽

Ht 29

ABSTRACT Aggregation substance (AS) is an Enterococcus faecalissurface protein that may contribute to virulence. Using a recently described system for controlled expression of AS in E. faecalis and the heterologous host Lactococcus lactis, experiments were designed to assess the effect of AS on bacterial internalization by HT-29 and Caco-2 enterocytes. AS expression was associated with increased internalization of E. faecalis by HT-29 enterocytes and of L. lactis by HT-29 and Caco-2 enterocytes. Compared to enterocytes cultivated under standard conditions, either cultivation in hypoxia or 1-h pretreatment of enterocytes with calcium-free medium resulted in increased internalization of both E. faecalis and L. lactis (with and without AS expression). Also, AS expression augmented these increases when E. faecalis was incubated with pretreated HT-29 enterocytes and when L. lactis was incubated with pretreated Caco-2 and HT-29 enterocytes. These data indicated that AS might facilitate E. faecalisinternalization by cultured enterocytes.

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Analysis of Functional Domains of theEnterococcus faecalis Pheromone-Induced Surface Protein Aggregation Substance

Journal of Bacteriology ◽

10.1128/jb.183.19.5659-5667.2001 ◽

2001 ◽

Vol 183 (19) ◽

pp. 5659-5667 ◽

Cited By ~ 46

Author(s):

C. M. Waters ◽

G. M. Dunny

Keyword(s):

Cell Surface ◽

High Efficiency ◽

Cell Surface Hydrophobicity ◽

Surface Protein ◽

Cell Aggregation ◽

Surface Hydrophobicity ◽

Host Cells ◽

Functional Domains ◽

Aggregation Substance ◽

C Terminus

ABSTRACT Pheromone-inducible aggregation substance (AS) proteins ofEnterococcus faecalis are essential for high-efficiency conjugation of the sex pheromone plasmids and also serve as virulence factors during host infection. A number of different functions have been attributed to AS in addition to bacterial cell aggregation, including adhesion to host cells, adhesion to fibrin, increased cell surface hydrophobicity, resistance to killing by polymorphonuclear leukocytes and macrophages, and increased vegetation size in an experimental endocarditis model. Relatively little information is available regarding the structure-activity relationship of AS. To identify functional domains, a library of 23 nonpolar 31-amino-acid insertions was constructed in Asc10, the AS encoded by the plasmid pCF10, using the transposons TnlacZ/in and TnphoA/in. Analysis of these insertions revealed a domain necessary for donor-recipient aggregation that extends further into the amino terminus of the protein than previously reported. In addition, insertions in the C terminus of the protein also reduced aggregation. As expected, the ability to aggregate correlates with efficient plasmid transfer. The results also indicated that an increase in cell surface hydrophobicity resulting from AS expression is not sufficient to mediate bacterial aggregation.

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Aggregation Substance Promotes Adherence, Phagocytosis, and Intracellular Survival of Enterococcus faecalis within Human Macrophages and Suppresses Respiratory Burst

Infection and Immunity ◽

10.1128/iai.68.9.4900-4906.2000 ◽

2000 ◽

Vol 68 (9) ◽

pp. 4900-4906 ◽

Cited By ~ 118

Author(s):

Sigurd D. Süßmuth ◽

Albrecht Muscholl-Silberhorn ◽

Reinhard Wirth ◽

Milorad Susa ◽

Reinhard Marre ◽

...

Keyword(s):

Enterococcus Faecalis ◽

Respiratory Burst ◽

Superoxide Anion ◽

Surface Protein ◽

Human Monocyte ◽

Human Macrophages ◽

Aggregation Substance ◽

Type 3 ◽

Inhibition Studies ◽

Macrophage Killing

ABSTRACT The aggregation substance (AS) of Enterococcus faecalis, encoded on sex pheromone plasmids, is a surface-bound glycoprotein that mediates aggregation between bacteria thereby facilitating plasmid transfer. Sequencing of the pAD1-encoded Asa1 revealed that this surface protein contains two RGD motifs which are known to ligate integrins. Therefore, we investigated the influence of AS on the interaction of E. faecalis with human monocyte-derived macrophages which constitutively express β2 integrins (e.g., CD18). AS was found to cause a greater-than-fivefold increase in enterococcal adherence to macrophages and a greater-than-sevenfold increase in phagocytosis. Adherence was mediated by an interaction between the RGD motif and the integrin CD11b/CD18 (complement receptor type 3) as demonstrated by inhibition studies with monoclonal antibodies and RGD peptide. AS-bearing enterococci were significantly more resistant to macrophage killing during the first 3 h postinfection, probably due to inhibition of the respiratory burst as indicated by reduced concentrations of superoxide anion.

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The Aggregation Domain of Aggregation Substance, Not the RGD Motifs, Is Critical for Efficient Internalization by HT-29 Enterocytes

Infection and Immunity ◽

10.1128/iai.71.10.5682-5689.2003 ◽

2003 ◽

Vol 71 (10) ◽

pp. 5682-5689 ◽

Cited By ~ 24

Author(s):

Christopher M. Waters ◽

Carol L. Wells ◽

Gary M. Dunny

Keyword(s):

Enterococcus Faecalis ◽

Surface Protein ◽

Cell Types ◽

Functional Domain ◽

Aggregation Substance ◽

Bacterial Aggregation ◽

Bacterial Aggregates ◽

Different Cell Types ◽

Ht 29 ◽

Dependent Mechanism

ABSTRACT Aggregation substance (AS), a surface protein encoded on the pheromone-inducible plasmids of Enterococcus faecalis, has been shown to increase adherence and internalization into a number of different cell types, presumably through integrin binding mediated by the N-terminal RGD motif of AS. Here, defined mutations constructed in Asc10, the AS encoded by the plasmid pCF10, are analyzed for their ability to promote increased internalization levels into HT-29 enterocytes. The results clearly show that the previously identified Asc10 functional domain, not the RGD motifs, is critical for Asc10-directed internalization of E. faecalis into HT-29 enterocytes. Also, expression of Asc10 in the nonaggregating E. faecalis strain INY3000 is unable to mediate HT-29 internalization. However, Asc10-expressing E. faecalis cells are not internalized as bacterial aggregates, suggesting bacterial aggregation is not a prerequisite for HT-29 internalization. These data show that Asc10 directs internalization of E. faecalis into HT-29 enterocytes through a non-RGD-dependent mechanism.

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