Enterococcal Species Associated with Slovak Raw Goat Milk, Their Safety and Susceptibility to Lantibiotics and Durancin ED26E/7

Goat milk has become a popular item of human consumption due to its originality. Enterococci are ubiquitous bacteria, and they can also be found in traditional dairy products. This study focuses on the safety of enterococci from Slovak raw goat milk and on their susceptibility to lantibiotic bacteriocins and durancin ED26E/7, which has not previously been studied. Biofilm formation ability in enterococci, virulence factor genes, enzyme production and antibiotic profile were investigated. Samples of raw goat milk (53) were collected from 283 goats in Slovakia. MALDI-TOF mass spectrometry identified three enterococcal species: Enterococcus faecium, E. hirae and E. mundtii, with dominant occurrence of the species E. faecium. Low-grade biofilm formation ability (0.1 ≤ A570 < 1.0) was found in four strains of E. faecium.Gelatinase, hyaluronidase, aggregation substance and enterococcal surface protein genes were absent in these enterococci. Gene efaAfm (adhesin) was detected in five E. faecium strains. However, it was not detected in biofilm-forming strains. Enterococci detected in Slovak raw goat milk were found not to have pathogenic potential; four strains even produced high amounts of useful β-galactosidase. The strains were susceptible to lantibiotic bacteriocin treatment and to durancin ED26E/7 as well, which represents original information in dairy production.

Planktonic Interference and Biofilm Alliance between Aggregation Substance and Endocarditis- and Biofilm-Associated Pili inEnterococcus faecalis

ABSTRACTLike many bacteria,Enterococcus faecalisencodes a number of adhesins involved in colonization or infection of different niches. Two well-studiedE. faecalisadhesins, aggregation substance (AS) and endocarditis- and biofilm-associated pili (Ebp), both contribute to biofilm formation on abiotic surfaces and in endocarditis, suggesting that they may be expressed at the same time. Because different regulatory pathways have been reported for AS and Ebp, here, we examined if they are coexpressed on the same cells and what is the functional impact of coexpression on individual cells and within a population. We found that while Ebp are only expressed on a subset of cells, when Ebp and AS are expressed on the same cells, pili interfere with AS-mediated clumping and impede AS-mediated conjugative plasmid transfer during planktonic growth. However, when the population density increases, horizontal gene transfer rates normalize and are no longer affected by pilus expression. Instead, at higher cell densities during biofilm formation, Ebp and AS differentially contribute to biofilm development and structure, synergizing to promote maximal biofilm formation.IMPORTANCEMost bacteria express multiple adhesins that contribute to surface attachment and colonization. However, the network and relationships between the various adhesins of a single bacterial species are less well understood. Here, we examined two well-characterized adhesins inEnterococcus faecalis, aggregation substance and endocarditis- and biofilm-associated pili, and found that they exhibit distinct functional contributions depending on the growth stage of the bacterial community. Pili interfere with aggregation substance-mediated clumping and plasmid transfer under planktonic conditions, whereas the two adhesins structurally complement one another during biofilm development. This study advances our understanding of howE. faecalis, a ubiquitous member of the human gut microbiome and an opportunistic pathogen, uses multiple surface structures to evolve and thrive.

Planktonic interference and biofilm alliance between aggregation substance and endocarditis and biofilm associated pili inEnterococcus faecalis

ABSTRACTLike many bacteria,Enterococcus faecalisencodes a number of adhesins involved in colonization or infection of different niches. Two well-studiedE. faecalisadhesins, aggregation substance (AS) and endocarditis and biofilm-associated pili (Ebp), both contribute to biofilm formation on abiotic surfaces and in endocarditis, suggesting they may be expressed at the same time. Because different regulatory pathways have been reported for AS and Ebp, here we examined if they are co-expressed on the same cells and the functional impact of co-expression on individual cells and within a population. We found that while Ebp are only expressed on a subset of cells, when Ebp and AS are expressed on the same cells, pili interfere with AS-mediated clumping and impede AS-mediated conjugative plasmid transfer during planktonic growth. However, when the population density increases, horizontal gene transfer rates normalize and are no longer affected by pilus expression. Instead, at higher cell densities during biofilm formation, Ebp and AS differentially contribute to biofilm development and structure, synergizing to promote maximal biofilm formation.

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.

ENTEROCOCCUS FAECALIS CAN SURVIVE EXTREME CHALLENGES - OVERVIEW

Enterococcus faecalis is a micro-organism that can survive extreme challenges. Its pathogenicity ranges from life-threatening diseases in compromised individuals to less severe conditions, systemic diseases such as endocarditis, brain abscesses, and septicaemia to infection of obturated root canals with chronic apical periodontitis. This article highlights some of the virulence factors of E. faecalis that may be related to endodontic infections and the periradicular inflammatory response. The most-cited virulence factors are aggregation substance, surface adhesins, sex pheromones, lipoteichoic acid, extracellular superoxide production, the lytic enzymes gelatinase and hyaluronidase, and the toxin cytolysin. Each of them may be associated with various stages of an endodontic infection as well as with periapical inflammation. While some products of the bacterium may be directly linked to damage of the periradicular tissues, a large part of the tissue damage is probably mediated by the host response to the bacterium and its products.

Complex Nature of Enterococcal Pheromone-Responsive Plasmids

Pheromone-responsive plasmids constitute a unique group of approximately 20 plasmids identified, as yet, only among enterococcal species. Several of their representatives, e.g. pAD1, pCF10, pPD1 and pAM373 have been extensively studied. These plasmids possess a sophisticated conjugation mechanism based on response to sex pheromones--small peptides produced by plasmid-free recipient cells. Detailed analysis of regulation and function of the pheromone response process revealed its great complexity and dual role--in plasmid conjugation and modulation of enterococcal virulence. Among other functional modules identified in pheromone plasmids, the stabilization/partition systems play a crucial role in stable maintenance of the plasmid molecule in host bacteria. Among them, the par locus of pAD1 is one of the exceptional RNA addiction systems. Pheromone-responsive plasmids contribute also to enterococcal phenotype being an important vehicle of antibiotic resistance in this genus. Both types of acquired vancomycin resistance determinants, vanA and vanB, as well many other resistant phenotypes, were found to be located on these plasmids. They also encode two basic agents of enterococcal virulence, i.e. aggregation substance (AS) and cytolysin. AS participates in mating-pair formation during conjugation but can also facilitate the adherence ofenterococci to human tissues during infection. The second protein, cytolysin, displays hemolytic activity and helps to invade eukaryotic cells. There are still many aspects of the nature of pheromone plasmids that remain unclear and more detailed studies are needed to understand their uniqueness and complexity.