scholarly journals Induction of Daptomycin Tolerance in Enterococcus faecalis by Fatty Acid Combinations

2020 ◽  
Vol 86 (20) ◽  
Author(s):  
William Brewer ◽  
Johnathan Harrison ◽  
Holly E. Saito ◽  
Elizabeth M. Fozo
Keyword(s):  
Fatty Acids ◽  
Antibiotic Resistance ◽  
Fatty Acid ◽  
Stearic Acid ◽  
Enterococcus Faecalis ◽  
Cell Envelope ◽  
Negative Effects ◽  
Gram Positive ◽  
Bacterial Populations ◽  
Content Type

ABSTRACT Enterococcus faecalis is a Gram-positive bacterium that normally exists as an intestinal commensal in humans but is also a leading cause of nosocomial infections. Previous work noted that growth supplementation with serum induced tolerance to membrane-damaging agents, including the antibiotic daptomycin. Specific fatty acids found within serum could independently provide tolerance to daptomycin (protective fatty acids), yet some fatty acids found in serum did not and had negative effects on enterococcal physiology (nonprotective fatty acids). Here, we measured a wide array of physiological responses after supplementation with combinations of protective and nonprotective fatty acids to better understand how serum induces daptomycin tolerance. When cells were supplemented with either nonprotective fatty acid, palmitic acid, or stearic acid, there were marked defects in growth and morphology, but these defects were rescued upon supplementation with either protective fatty acid, oleic acid, or linoleic acid. Membrane fluidity decreased with growth in either palmitic or stearic acid alone but returned to basal levels when a protective fatty acid was supplied. Daptomycin tolerance could be induced if a protective fatty acid was provided with a nonprotective fatty acid, and some specific combinations protected as well as serum supplementation. While cell envelope charge has been associated with tolerance to daptomycin in other Gram-positive bacteria, we concluded that it does not correlate with the fatty acid-induced protection we observed. Based on these observations, we conclude that daptomycin tolerance by serum is driven by specific, protective fatty acids found within the fluid. IMPORTANCE With an increasing prevalence of antibiotic resistance in the clinic, we strive to understand more about microbial defensive mechanisms. A nongenetic tolerance to the antibiotic daptomycin was discovered in Enterococcus faecalis that results in the increased survival of bacterial populations after treatment with the drug. This tolerance mechanism likely synergizes with antibiotic resistance in the clinic. Given that this tolerance phenotype is induced by incorporation of fatty acids present in the host, it can be assumed that infections by this organism require a higher dose of antibiotic for successful eradication. The mixture of fatty acids in human fluids is quite diverse, with little understanding between the interplay of fatty acid combinations and the tolerance phenotype we observe. It is crucial to understand the effects of fatty acid combinations on E. faecalis physiology if we are to suppress the tolerance physiology in the clinic.

scholarly journals Exogenous Fatty Acids Protect Enterococcus faecalis from Daptomycin-Induced Membrane Stress Independently of the Response Regulator LiaR

2016 ◽  
Vol 82 (14) ◽  
pp. 4410-4420 ◽  
Author(s):  
John R. Harp ◽  
Holly E. Saito ◽  
Allen K. Bourdon ◽  
Jinnethe Reyes ◽  
Cesar A. Arias ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Enterococcus Faecalis ◽  
Stress Responses ◽  
Nosocomial Infections ◽  
Response Regulator ◽  
Membrane Damage ◽  
Membrane Stress ◽  
Content Type ◽  
Induced Membrane

ABSTRACTEnterococcus faecalisis a commensal bacterium of the gastrointestinal tract that can cause nosocomial infections in immunocompromised humans. The hallmarks of this organism are its ability to survive in a variety of stressful habitats and, in particular, its ability to withstand membrane damage. One strategy used byE. faecalisto protect itself from membrane-damaging agents, including the antibiotic daptomycin, involves incorporation of exogenous fatty acids from bile or serum into the cell membrane. Additionally, the response regulator LiaR (a member of the LiaFSR [lipid II-interacting antibiotic response regulator and sensor] system associated with cell envelope stress responses) is required for the basal level of resistanceE. faecalishas to daptomycin-induced membrane damage. This study aimed to determine if membrane fatty acid changes could provide protection against membrane stressors in a LiaR-deficient strain ofE. faecalis. We noted that despite the loss of LiaR, the organism readily incorporated exogenous fatty acids into its membrane, and indeed growth in the presence of exogenous fatty acids increased the survival of LiaR-deficient cells when challenged with a variety of membrane stressors, including daptomycin. Combined, our results suggest thatE. faecaliscan utilize both LiaR-dependent and -independent mechanisms to protect itself from membrane damage.IMPORTANCEEnterococcus faecalisis responsible for a significant number of nosocomial infections. Worse, many of the antibiotics used to treatE. faecalisinfection are no longer effective, as this organism has developed resistance to them. The drug daptomycin has been successfully used to treat some of these resistant strains; however, daptomycin-resistant isolates have been identified in hospitals. Many daptomycin-resistant isolates are found to harbor mutations in the genetic locusliaFSR, which is involved in membrane stress responses. Another mechanism shown to increase tolerance to daptomycin involves the incorporation of exogenous fatty acids from host fluids like serum or bile. This improved tolerance was found to be independent ofliaFSRand suggests that there are additional ways to impact sensitivity to daptomycin. Thus, further studies are needed to understand how host fatty acid sources can influence antibiotic susceptibility.

scholarly journals The Two Functional Enoyl-Acyl Carrier Protein Reductases of Enterococcus faecalis Do Not Mediate Triclosan Resistance

mBio ◽  
2013 ◽  
Vol 4 (5) ◽  
Author(s):  
Lei Zhu ◽  
Hongkai Bi ◽  
Jincheng Ma ◽  
Zhe Hu ◽  
Wenbin Zhang ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Enterococcus Faecalis ◽  
Fatty Acid Synthesis ◽  
Acyl Carrier Protein ◽  
Acid Synthesis ◽  
Carrier Protein ◽  
Content Type ◽  
Enoyl Acp Reductase ◽  
Triclosan Resistance

ABSTRACTEnoyl-acyl carrier protein (enoyl-ACP) reductase catalyzes the last step of the elongation cycle in the synthesis of bacterial fatty acids. TheEnterococcus faecalisgenome contains two genes annotated as enoyl-ACP reductases, a FabI-type enoyl-ACP reductase and a FabK-type enoyl-ACP reductase. We report that expression of either of the two proteins restores growth of anEscherichia colifabItemperature-sensitive mutant strain under nonpermissive conditions.In vitroassays demonstrated that both proteins support fatty acid synthesis and are active with substrates of all fatty acid chain lengths. Although expression ofE. faecalis fabKconfers toE. colihigh levels of resistance to the antimicrobial triclosan, deletion offabKfrom theE. faecalisgenome showed that FabK does not play a detectable role in the inherent triclosan resistance ofE. faecalis. Indeed, FabK seems to play only a minor role in modulating fatty acid composition. Strains carrying a deletion offabKgrow normally without fatty acid supplementation, whereasfabIdeletion mutants make only traces of fatty acids and are unsaturated fatty acid auxotrophs.IMPORTANCEThe finding that exogenous fatty acids support growth ofE. faecalisstrains defective in fatty acid synthesis indicates that inhibitors of fatty acid synthesis are ineffective in counteringE. faecalisinfections because host serum fatty acids support growth of the bacterium.

scholarly journals Incorporation of Exogenous Fatty Acids Protects Enterococcus faecalis from Membrane-Damaging Agents

2014 ◽  
Vol 80 (20) ◽  
pp. 6527-6538 ◽  
Author(s):  
Holly E. Saito ◽  
John R. Harp ◽  
Elizabeth M. Fozo
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Enterococcus Faecalis ◽  
Stress Adaptation ◽  
Membrane Composition ◽  
Content Type ◽  
Wide Range ◽  
Low Levels ◽  
Single Fatty Acid ◽  
Composition Changes

ABSTRACTEnterococcus faecalisis a commensal bacterium of the mammalian intestine that can persist in soil and aquatic systems and can be a nosocomial pathogen to humans. It employs multiple stress adaptation strategies in order to survive such a wide range of environments. Within this study, we sought to elucidate whether membrane fatty acid composition changes are an important component for stress adaptation. We noted thatE. faecalisOG1RF was capable of changing its membrane composition depending upon growth phase and temperature. The organism also readily incorporated fatty acids from bile, serum, and medium supplemented with individual fatty acids, often dramatically changing the membrane composition such that a single fatty acid was predominant. Growth in either low levels of bile or specific individual fatty acids was found to protect the organism from membrane challenges such as high bile exposure. In particular, we observed that when grown in low levels of bile, serum, or the host-derived fatty acids oleic acid and linoleic acid,E. faecaliswas better able to survive the antibiotic daptomycin. Interestingly, the degree of membrane saturation did not appear to be important for protection from the stressors examined here; instead, it appears that a specific fatty acid or combination of fatty acids is critical for stress resistance.

scholarly journals Identification of the Lyso-Form N-Acyl Intramolecular Transferase in Low-GC Firmicutes

2017 ◽  
Vol 199 (11) ◽  
Author(s):  
Krista M. Armbruster ◽  
Timothy C. Meredith
Keyword(s):  
Fatty Acid ◽  
Cell Envelope ◽  
Transmembrane Protein ◽  
Physiological Role ◽  
Gram Positive ◽  
Gram Negative ◽  
Content Type ◽  
Gram Positive Bacteria ◽  
Unknown Gene ◽  
Gram Negative Organisms

ABSTRACT Bacterial lipoproteins are embedded in the cell membrane of both Gram-positive and Gram-negative bacteria, where they serve numerous functions central to cell envelope physiology. Lipoproteins are tethered to the membrane by an N-acyl-S-(mono/di)-acyl-glyceryl-cysteine anchor that is variously acylated depending on the genus. In several low-GC, Gram-positive firmicutes, a monoacyl-glyceryl-cysteine with an N-terminal fatty acid (known as the lyso form) has been reported, though how it is formed is unknown. Here, through an intergenic complementation rescue assay in Escherichia coli, we report the identification of a common orthologous transmembrane protein in both Enterococcus faecalis and Bacillus cereus that is capable of forming lyso-form lipoproteins. When deleted from the native host, lipoproteins remain diacylated with a free N terminus, as maturation to the N-acylated lyso form is abolished. Evidence is presented suggesting that the previously unknown gene product functions through a novel intramolecular transacylation mechanism, transferring a fatty acid from the diacylglycerol moiety to the α-amino group of the lipidated cysteine. As such, the discovered gene has been named lipoprotein intramolecular transacylase (lit), to differentiate it from the gene for the intermolecular N-acyltransferase (lnt) involved in triacyl lipoprotein biosynthesis in Gram-negative organisms. IMPORTANCE This study identifies a new enzyme, conserved among low-GC, Gram-positive bacteria, that is involved in bacterial lipoprotein biosynthesis and synthesizes lyso-form lipoproteins. Its discovery is an essential first step in determining the physiological role of N-terminal lipoprotein acylation in Gram-positive bacteria and how these modifications impact bacterial cell envelope function.

scholarly journals Multiple Roles for Enterococcus faecalis Glycosyltransferases in Biofilm-Associated Antibiotic Resistance, Cell Envelope Integrity, and Conjugative Transfer

2015 ◽  
Vol 59 (7) ◽  
pp. 4094-4105 ◽  
Author(s):  
Jennifer L. Dale ◽  
Julian Cagnazzo ◽  
Chi Q. Phan ◽  
Aaron M. T. Barnes ◽  
Gary M. Dunny
Keyword(s):  
Antibiotic Resistance ◽  
Enterococcus Faecalis ◽  
Cell Envelope ◽  
Opportunistic Pathogen ◽  
Growth Conditions ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Conjugative Transfer ◽  
Content Type ◽  
Resistance Determinants

ABSTRACTThe emergence of multidrug-resistant bacteria and the limited availability of new antibiotics are of increasing clinical concern. A compounding factor is the ability of microorganisms to form biofilms (communities of cells encased in a protective extracellular matrix) that are intrinsically resistant to antibiotics.Enterococcus faecalisis an opportunistic pathogen that readily forms biofilms and also has the propensity to acquire resistance determinants via horizontal gene transfer. There is intense interest in the genetic basis for intrinsic and acquired antibiotic resistance inE. faecalis, since clinical isolates exhibiting resistance to multiple antibiotics are not uncommon. We performed a genetic screen using a library of transposon (Tn) mutants to identifyE. faecalisbiofilm-associated antibiotic resistance determinants. Five Tn mutants formed wild-type biofilms in the absence of antibiotics but produced decreased biofilm biomass in the presence of antibiotic concentrations that were subinhibitory to the parent strain. Genetic determinants responsible for biofilm-associated antibiotic resistance include components of the quorum-sensing system (fsrA,fsrC, andgelE) and two glycosyltransferase (GTF) genes (epaIandepaOX). We also found that the GTFs play additional roles inE. faecalisresistance to detergent and bile salts, maintenance of cell envelope integrity, determination of cell shape, polysaccharide composition, and conjugative transfer of the pheromone-inducible plasmid pCF10. TheepaOXgene is located in a variable extended region of the enterococcal polysaccharide antigen (epa) locus. These data illustrate the importance of GTFs inE. faecalisadaptation to diverse growth conditions and suggest new targets for antimicrobial design.

Chlorella vulgaris as a Source of Essential Fatty Acids and Micronutrients: A Brief Commentary

2017 ◽  
Vol 10 (1) ◽  
pp. 92-99 ◽  
Author(s):  
Hércules Rezende Freitas
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Culture Medium ◽  
Essential Fatty Acids ◽  
Total Lipid Content ◽  
Dietary Quality ◽  
Negative Effects ◽  
Algae Biomass ◽  
Dietary Consumption ◽  
Acid Consumption

Polyunsaturated fatty acids (PUFAs) comprise about 35-40% of the total lipid content from green algaeChlorella, reaching up to 24% linoleic acid and 27% α-linolenic acid inC. vulgaris. Also, microalgae nutrient composition may be modulated by changes in the culture medium, increasing fatty acid and microelement concentrations in the algae biomass. PUFAs, such as α-linolenic (n-3) and linoleic (n-6) acids, as well as its derivatives, are considered essential for dietary consumption, and their ability to regulate body chemistry has been recently explored in depth. A balanced fatty acid consumption is shown to counteract the negative effects of western diets, such as chronic inflammation and glucose intolerance. In this brief commentary, technological and practical uses ofC. vulgarisare explored as means to improve dietary quality and, ultimately, human health.

Depot fatty acids of Aberdeen Angus and Friesian cattle reared on hay and barley diets

1977 ◽  
Vol 89 (3) ◽  
pp. 575-582 ◽  
Author(s):  
W. M. F. Leat
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Stearic Acid ◽  
Acid Composition ◽  
Visual Inspection ◽  
Subcutaneous Fat ◽  
Octadecenoic Acid ◽  
Hexadecenoic Acid ◽  
Friesian Cattle

SummaryAberdeen Angus and Friesian cattle were reared from 4 months of age to slaughter weight at 18–24 months on either high-barley or high-hay diets. Samples of subcutaneous fat were taken by biopsy at 3 monthly intervals, and the degree of fatness of each animal was estimated ultrasonically prior to slaughter, and by visual inspection of the carcasses.The barley-fed animals gained weight more rapidly, and fattened more quickly than the hay-fed animals with the Angus being fatter than the Friesian at the same age. The percentage stearic acid (C18:0) in subcutaneous fat decreased with age and was replaced by octadecenoic acid (C18:l) and hexadecenoic acid (C16:l), these changes being more rapid in barley-fed than in hay-fed animals. At the same degree of fatness the depot fats of the Friesians were more unsaturated than those of the Angus, and in both breeds the fatter the animal the more unsaturated was its depot fat.In the hay-fed cattle the percentage C16:0 in subcutaneous fat increased during the last half of the experiment and at slaughter the percentage C16:0 was significantly higher, and C18:l significantly lower, in all depot fats compared with those of the barley-fed animals.It is concluded that the fatty acid composition of bovine depot fats is modulated by the degree of fattening, and can be affected by diet.

scholarly journals CRISPR-Cas and Restriction-Modification Act Additively against Conjugative Antibiotic Resistance Plasmid Transfer in Enterococcus faecalis

mSphere ◽  
2016 ◽  
Vol 1 (3) ◽  
Author(s):  
Valerie J. Price ◽  
Wenwen Huo ◽  
Ardalan Sharifi ◽  
Kelli L. Palmer
Keyword(s):  
Antibiotic Resistance ◽  
High Risk ◽  
Enterococcus Faecalis ◽  
Resistance Genes ◽  
Treatment Options ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Content Type ◽  
New Genes ◽  
Restriction Modification

ABSTRACT Enterococcus faecalis is a bacterium that normally inhabits the gastrointestinal tracts of humans and other animals. Although these bacteria are members of our native gut flora, they can cause life-threatening infections in hospitalized patients. Antibiotic resistance genes appear to be readily shared among high-risk E. faecalis strains, and multidrug resistance in these bacteria limits treatment options for infections. Here, we find that CRISPR-Cas and restriction-modification systems, which function as adaptive and innate immune systems in bacteria, significantly impact the spread of antibiotic resistance genes in E. faecalis populations. The loss of these systems in high-risk E. faecalis suggests that they are immunocompromised, a tradeoff that allows them to readily acquire new genes and adapt to new antibiotics. Enterococcus faecalis is an opportunistic pathogen and a leading cause of nosocomial infections. Conjugative pheromone-responsive plasmids are narrow-host-range mobile genetic elements (MGEs) that are rapid disseminators of antibiotic resistance in the faecalis species. Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification confer acquired and innate immunity, respectively, against MGE acquisition in bacteria. Most multidrug-resistant E. faecalis isolates lack CRISPR-Cas and possess an orphan locus lacking cas genes, CRISPR2, that is of unknown function. Little is known about restriction-modification defense in E. faecalis. Here, we explore the hypothesis that multidrug-resistant E. faecalis strains are immunocompromised. We assessed MGE acquisition by E. faecalis T11, a strain closely related to the multidrug-resistant hospital isolate V583 but which lacks the ~620 kb of horizontally acquired genome content that characterizes V583. T11 possesses the E. faecalis CRISPR3-cas locus and a predicted restriction-modification system, neither of which occurs in V583. We demonstrate that CRISPR-Cas and restriction-modification together confer a 4-log reduction in acquisition of the pheromone-responsive plasmid pAM714 in biofilm matings. Additionally, we show that the orphan CRISPR2 locus is functional for genome defense against another pheromone-responsive plasmid, pCF10, only in the presence of cas9 derived from the E. faecalis CRISPR1-cas locus, which most multidrug-resistant E. faecalis isolates lack. Overall, our work demonstrated that the loss of only two loci led to a dramatic reduction in genome defense against a clinically relevant MGE, highlighting the critical importance of the E. faecalis accessory genome in modulating horizontal gene transfer. Our results rationalize the development of antimicrobial strategies that capitalize upon the immunocompromised status of multidrug-resistant E. faecalis. IMPORTANCE Enterococcus faecalis is a bacterium that normally inhabits the gastrointestinal tracts of humans and other animals. Although these bacteria are members of our native gut flora, they can cause life-threatening infections in hospitalized patients. Antibiotic resistance genes appear to be readily shared among high-risk E. faecalis strains, and multidrug resistance in these bacteria limits treatment options for infections. Here, we find that CRISPR-Cas and restriction-modification systems, which function as adaptive and innate immune systems in bacteria, significantly impact the spread of antibiotic resistance genes in E. faecalis populations. The loss of these systems in high-risk E. faecalis suggests that they are immunocompromised, a tradeoff that allows them to readily acquire new genes and adapt to new antibiotics.

Long-chain n-3 polyunsaturated fatty acids contained in liver and muscle tissues of Okinawan long-spine porcupinefish (Diodon holocanthus Linnaeus 1758)

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Yutaka Tashiro
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acids ◽  
Ryukyu Islands ◽  
Okinawa Island ◽  
Content Type ◽  
Muscle Tissues ◽  
The Ryukyu Islands ◽  
Total Fatty Acids ◽  
Long Chain

Purpose This study aimed to analyze the lipid content and fatty acid composition in the liver and muscle of a porcupinefish species inhabiting waters around the Ryukyu Islands to investigate their potential as a source of long-chain n-3 polyunsaturated fatty acids (LC-PUFAs). Design/methodology/approach Porcupinefish were collected along the Okinawa Island coast. The composition of fatty acids and cholesterol in both liver and muscle were analyzed using a gas chromatograph mass spectrometer. Findings The liver of Okinawan long-spine porcupinefish was rich in lipids whose content correlated to the proportion of liver/body weight. Fatty acid compositions in their liver and muscles were similar to each other. LC-PUFAs occupied 44% of total fatty acids, with docosahexaenoic acid (DHA) being the dominant (42%), whereas eicosapentaenoic acid occupied 2.4%. The liver contained 1,690 mg of cholesterol and 14.8 g of DHA per 100 g, whose proportion decreased in summer compared to other seasons (p = 0.036). Originality/value The liver of Okinawan long-spine porcupinefish, which has not yet been commercially used although its non-toxicity is claimed, can be an excellent source of LC-PUFAs, especially DHA, accentuating its potential in food supplements’ production.

scholarly journals Fatty acid chemistry of Atrichum undulatum and Hypnum andoi

2012 ◽  
Vol 66 (2) ◽  
pp. 207-209 ◽  
Author(s):  
Boris Pejin ◽  
Ljubodrag Vujisic ◽  
Marko Sabovljevic ◽  
Vele Tesevic ◽  
Vlatka Vajs
Keyword(s):  
Fatty Acids ◽  
Gas Chromatography ◽  
Fatty Acid ◽  
Linoleic Acid ◽  
Stearic Acid ◽  
Palmitic Acid ◽  
Linolenic Acid ◽  
Essential Fatty Acids ◽  
Behenic Acid ◽  
Moss Species

The fatty acid composition of the moss species Atrichum undulatum (Hedw.) P. Beauv. (Polytrichaceae) and Hypnum andoi A.J.E. Sm. (Hypnaceae) collected in winter time were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) as a contribution to their chemistry. Eight fatty acids were identified in the chloroform/methanol extract 1:1 of A. undulatum (linoleic acid 26.80%, palmitic acid 22.17%, ?-linolenic acid 20.50%, oleic acid 18.49%, arachidonic acid 6.21%, stearic acid 3.34%, cis-5,8,11,14,17-eicosapentaenoic acid 1.52% and behenic acid 1.01%), while six fatty acids were found in the same type of extract of H. andoi (palmitic acid 63.48%, erucic acid 12.38%, stearic acid 8.08%, behenic acid 6.26%, lignoceric acid 5.16% and arachidic acid 4.64%). According to this study, the moss A. undulatum can be considered as a good source of both essential fatty acids for humans (linoleic acid and ?-linolenic acid) during the winter.

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