Decolonization of Human Anterior Nares of Staphylococcus aureus with Use of a Glycerol Monolaurate Nonaqueous Gel

ABSTRACT Staphylococcus aureus is a highly significant infection problem in health care centers, particularly after surgery. It has been shown that nearly 80% of S. aureus infections following surgery are the same as those in the anterior nares of patients, suggesting that the anterior nares is the source of the infection strain. This has led to the use of mupirocin ointment being applied nasally to reduce infections; mupirocin resistance is being observed. This study was undertaken to determine whether gel composed of 5% glycerol monolaurate solubilized in a glycol-based, nonaqueous gel (5% GML gel) could be used as an alternative. In our study, 40 healthy human volunteers swabbed their anterior nares for 3 days with the 5% GML gel. Prior to swabbing and 8 to 12 h after swabbing, S. aureus and coagulase-negative staphylococcal CFU per milliliter were determined by plating the swabs on mannitol salt agar. Fourteen of the volunteers had S. aureus in their nares prior to 5% GML gel treatment, most persons with the organisms present in both nares; five had pure cultures of S. aureus. All participants without pure culture of S. aureus were cocolonized with S. aureus and coagulase-negative staphylococci. Five of the S. aureus strains produced the superantigens commonly associated with toxic shock syndrome, though none of the participants became ill. For both S. aureus and coagulase-negative staphylococci, the 5% GML gel treatment resulted in a 3-log-unit reduction in microorganisms. For S. aureus, the reduction persisted for 2 or 3 days. IMPORTANCE In this microflora study, we show that a 5% glycerol monolaurate nonaqueous gel is safe for use in the anterior nares. The gel was effective in reducing Staphylococcus aureus nasally, a highly significant hospital-associated pathogen. The gel may be a useful alternative or additive to mupirocin ointment for nasal use prior to surgery, noting that 80% of hospital-associated S. aureus infections are due to the same organism found in the nose. This gel also kills all enveloped viruses tested and should be considered for studies to reduce infection and transmission of coronaviruses and influenza viruses.

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Case Study: Misdiagnosis of Nonhemolytic Staphylococcus aureus Isolates from Cases of Bovine Mastitis as Coagulase-Negative Staphylococci

Animals ◽

10.3390/ani11020252 ◽

2021 ◽

Vol 11 (2) ◽

pp. 252

Author(s):

Valerie E. Ryman ◽

Felicia M. Kautz ◽

Steve C. Nickerson

Keyword(s):

Staphylococcus Aureus ◽

Bovine Mastitis ◽

Dairy Farm ◽

Blood Agar ◽

Coagulase Negative Staphylococci ◽

Mannitol Salt Agar ◽

Lactating Cows ◽

Presumptive Identification ◽

Microbiological Techniques

Staphylococcus aureus is one of the most concerning mastitis-causing pathogens in dairy cattle. Using basic microbiological techniques, S. aureus is typically identified by colony characteristics and hemolysis on blood agar where isolates without hemolysis are typically considered to be coagulase-negative staphylococci (CNS) isolates. Herein, we present a decade-long case study where suspected S. aureus isolates from one Georgia dairy farm were further tested to confirm presumptive identification. Presumptive identification of bacterial growth from 222 mammary secretions from bred Holstein heifers and lactating cows was conducted at the time of collection. Presumptive identification of S. aureus on blood agar was based on observation of colony morphology, color, and presence or absence of a broad zone of incomplete hemolysis and a smaller zone of complete hemolysis at 48 h. Those without hemolysis were presumptively characterized as CNS. All isolates were further plated on mannitol salt agar and a coagulase test was performed. A positive for both of these tests together was deemed to be S. aureus. A selection of isolates was tested using API® Staph to biochemically confirm S. aureus identification. Data showed that 63.96% of isolates presumed to be CNS isolates were identified as S. aureus, 9.46% of isolates presumed to be CNS isolates were identified as coagulase-positive staphylococci (CPS) species (but not S. aureus), and 26.58% of samples that were presumed to be CNS isolates were identified correctly.

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Activity of Debio1452, a FabI Inhibitor with Potent Activity against Staphylococcus aureus and Coagulase-Negative Staphylococcus spp., Including Multidrug-Resistant Strains

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.05119-14 ◽

2015 ◽

Vol 59 (5) ◽

pp. 2583-2587 ◽

Cited By ~ 20

Author(s):

Robert K. Flamm ◽

Paul R. Rhomberg ◽

Nachum Kaplan ◽

Ronald N. Jones ◽

David J. Farrell

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Agents ◽

Fatty Acid Biosynthesis ◽

Multidrug Resistant ◽

Coagulase Negative Staphylococcus ◽

Significant Activity ◽

Coagulase Negative Staphylococci ◽

Content Type ◽

Mrsa Strains ◽

Human Infections

ABSTRACTStaphylococcus aureusand coagulase-negative staphylococci (CoNS) are responsible for a wide variety of human infections. The investigational antibacterial Debio1450 (previously AFN-1720), a prodrug of Debio1452 (previously AFN-1252), specifically targets staphylococci without significant activity against other Gram-positive or Gram-negative species. Debio1452 inhibits FabI, an enzyme critical to fatty acid biosynthesis in staphylococci. The activity of Debio1452 against CoNS, methicillin-susceptibleS. aureus(MSSA), and methicillin-resistantS. aureus(MRSA), including significant clones, was determined. A globally diverse collection of 574 patient isolates from 35 countries was tested that included CoNS (6 species, 103 strains), MSSA (154 strains), MRSA (163 strains), and molecularly characterized strains (includingspa-typed MRSA clones; 154 strains). The isolates were tested for susceptibility by CLSI broth microdilution methods against Debio1452 and 10 comparators. The susceptibility rates for the comparators were determined using CLSI and EUCAST breakpoint criteria. AllS. aureusand CoNS strains were inhibited by Debio1452 concentrations of ≤0.12 and ≤0.5 μg/ml, respectively. The MIC50s for MSSA, MRSA, and molecularly characterized MRSA strains were 0.004 μg/ml, and the MIC90s ranged from 0.008 to 0.03 μg/ml. The MICs were higher for the CoNS isolates (MIC50/90, 0.015/0.12 μg/ml). AmongS. aureusstrains, resistance was common for erythromycin (61.6%), levofloxacin (49.0%), clindamycin (27.6%), tetracycline (15.7%), and trimethoprim-sulfamethoxazole (7.0%). Debio1452 demonstrated potent activity against MSSA, MRSA, and CoNS. Debio1452 showed significantly greater activity overall (MIC50, 0.004 μg/ml) than the other agents tested against these staphylococcal species, which included dominant MRSA clones and strains resistant to currently utilized antimicrobial agents.

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Biosynthesis of the Unique Wall Teichoic Acid of Staphylococcus aureus Lineage ST395

mBio ◽

10.1128/mbio.00869-14 ◽

2014 ◽

Vol 5 (2) ◽

Cited By ~ 25

Author(s):

Volker Winstel ◽

Patricia Sanchez-Carballo ◽

Otto Holst ◽

Guoqing Xia ◽

Andreas Peschel

Keyword(s):

Staphylococcus Aureus ◽

Cell Wall ◽

Gene Transfer ◽

Horizontal Gene Transfer ◽

Biosynthesis Pathway ◽

Glycerol Phosphate ◽

Coagulase Negative Staphylococci ◽

Content Type ◽

Gram Positive Bacteria ◽

Wall Teichoic Acids

ABSTRACT The major clonal lineages of the human pathogen Staphylococcus aureus produce cell wall-anchored anionic poly-ribitol-phosphate (RboP) wall teichoic acids (WTA) substituted with d-Alanine and N-acetyl-d-glucosamine. The phylogenetically isolated S. aureus ST395 lineage has recently been found to produce a unique poly-glycerol-phosphate (GroP) WTA glycosylated with N-acetyl-d-galactosamine (GalNAc). ST395 clones bear putative WTA biosynthesis genes on a novel genetic element probably acquired from coagulase-negative staphylococci (CoNS). We elucidated the ST395 WTA biosynthesis pathway and identified three novel WTA biosynthetic genes, including those encoding an α-O-GalNAc transferase TagN, a nucleotide sugar epimerase TagV probably required for generation of the activated sugar donor substrate for TagN, and an unusually short GroP WTA polymerase TagF. By using a panel of mutants derived from ST395, the GalNAc residues carried by GroP WTA were found to be required for infection by the ST395-specific bacteriophage Φ187 and to play a crucial role in horizontal gene transfer of S. aureus pathogenicity islands (SaPIs). Notably, ectopic expression of ST395 WTA biosynthesis genes rendered normal S. aureus susceptible to Φ187 and enabled Φ187-mediated SaPI transfer from ST395 to regular S. aureus. We provide evidence that exchange of WTA genes and their combination in variable, mosaic-like gene clusters have shaped the evolution of staphylococci and their capacities to undergo horizontal gene transfer events. IMPORTANCE The structural highly diverse wall teichoic acids (WTA) are cell wall-anchored glycopolymers produced by most Gram-positive bacteria. While most of the dominant Staphylococcus aureus lineages produce poly-ribitol-phosphate WTA, the recently described ST395 lineage produces a distinct poly-glycerol-phosphate WTA type resembling the WTA backbone of coagulase-negative staphylococci (CoNS). Here, we analyzed the ST395 WTA biosynthesis pathway and found new types of WTA biosynthesis genes along with an evolutionary link between ST395 and CoNS, from which the ST395 WTA genes probably originate. The elucidation of ST395 WTA biosynthesis will help to understand how Gram-positive bacteria produce highly variable WTA types and elucidate functional consequences of WTA variation.

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Uncoupling of Pro- and Anti-Inflammatory Properties of Staphylococcus aureus

Infection and Immunity ◽

10.1128/iai.02832-14 ◽

2015 ◽

Vol 83 (4) ◽

pp. 1587-1597 ◽

Cited By ~ 18

Author(s):

Adam G. Peres ◽

Camille Stegen ◽

Junbin Li ◽

An Qi Xu ◽

Benoit Levast ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Inflammatory Responses ◽

Proinflammatory Response ◽

Healthy Human ◽

Content Type ◽

Cell Immunity ◽

Extracellular Signal ◽

Severe Infections ◽

Toll Like Receptor 2 ◽

Anti Inflammatory

Staphylococcus aureusis a Gram-positive bacterium that is carried by a quarter of the healthy human population and that can cause severe infections. This pathobiosis has been linked to a balance between Toll-like receptor 2 (TLR2)-dependent pro- and anti-inflammatory responses. The relationship between these two types of responses is unknown. Analysis of 16 nasal isolates ofS. aureusshowed heterogeneity in their capacity to induce pro- and anti-inflammatory responses, suggesting that these two responses are independent of each other. Uncoupling of these responses was corroborated by selective signaling through phosphoinositol 3-kinase (PI3K)-Akt-mTOR and extracellular signal-regulated kinase (ERK) for the anti-inflammatory response and through p38 for the proinflammatory response. Uncoupling was also observed at the level of phagocytosis and phagosomal processing ofS. aureus, which were required solely for the proinflammatory response. Importantly, the anti-inflammatory properties of anS. aureusisolate correlated with its ability to modulate T cell immunity. Our results suggest the presence of anti-inflammatory TLR2 ligands in the staphylococcal cell wall, whose identification may provide templates for novel immunomodulatory drugs.

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Novel Type XII Staphylococcal Cassette ChromosomemecHarboring a New Cassette Chromosome Recombinase, CcrC2

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01692-15 ◽

2015 ◽

Vol 59 (12) ◽

pp. 7597-7601 ◽

Cited By ~ 58

Author(s):

Zhaowei Wu ◽

Fan Li ◽

Dongliang Liu ◽

Huping Xue ◽

Xin Zhao

Keyword(s):

United States ◽

Staphylococcus Aureus ◽

Methicillin Resistance ◽

The United States ◽

Gene Complex ◽

Coagulase Negative Staphylococci ◽

Typing Method ◽

Content Type ◽

Staphylococcal Cassette Chromosome ◽

Sequence Identities

ABSTRACTExcision and integration of staphylococcal cassette chromosomemec(SCCmec) are mediated by cassette chromosome recombinases (Ccr), which play a crucial role in the worldwide spread of methicillin resistance in staphylococci. We report a novelccrgene,ccrC2, in the SCCmecof aStaphylococcus aureusisolate, BA01611, which showed 62.6% to 69.4% sequence identities to all publishedccrC1sequences. A further survey found that theccrC2gene was mainly located among coagulase-negative staphylococci (CoNS) and could be found in staphylococcal isolates from China, the United States, France, and Germany. Theccrgene complex harboring theccrC2gene was designated a type 9 complex, and the SCCmecof BA01611 was considered a novel type and was designated type XII (9C2). This novel SCCmecelement in BA01611 was flanked by a pseudo-SCC element (ΨSCCBA01611) carrying a truncatedccrA1gene. Both individual SCC elements and a composite SCC were excised from the chromosome based on detection of extrachromosomal circular intermediates. We advocate inclusion of the ccrC2gene and type 9ccrgene complex during revision of the SCCmectyping method.

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Rapid Microarray-Based Identification of DifferentmecAAlleles in Staphylococci

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00574-12 ◽

2012 ◽

Vol 56 (11) ◽

pp. 5547-5554 ◽

Cited By ~ 40

Author(s):

Stefan Monecke ◽

Elke Müller ◽

Stefan Schwarz ◽

Helmut Hotzel ◽

Ralf Ehricht

Keyword(s):

Staphylococcus Aureus ◽

Amino Acid Sequences ◽

Animal Origin ◽

Coagulase Negative Staphylococci ◽

Content Type ◽

Diagnostic Assays ◽

Rapid Discrimination ◽

Unique Amino Acid ◽

Full Length Gene

ABSTRACTTo screen isolates and to identifymecAalleles, publishedmecAsequences were analyzed, and a microarray for the rapid discrimination ofmecAalleles was designed. A GenBank analysis yielded 135 full-length gene sequences annotated asmecA. These sequences clustered into 32 different alleles corresponding to 28 unique amino acid sequences and to 15 distinct hybridization patterns on this microarray. A collection of 78 clinical and veterinary isolates ofStaphylococcusspp. was characterized using this assay. Nine of the 15 expected patterns, as well as one as-yet-unknown pattern, were identified. These patterns were detected in various epidemic methicillin-resistantStaphylococcus aureusstrains, inS. pseudintermedius, and in coagulase-negative species such asS. epidermidis,S. fleurettii, orS. haemolyticus. There was no correlation between the differentmecAhybridization patterns and the SCCmectype. Determination of MICs showed thatmecAalleles corresponding to only four of these nine patterns were associated with β-lactam resistance. ThemecAalleles that did not confer β-lactam resistance were largely restricted to coagulase-negative staphylococci of animal origin, such asS. sciuriandS. vitulinus. Because of the diversity of sequences and the different impact on β-lactam susceptibility, the existence of differentmecAalleles needs to be taken into account when designing diagnostic assays for the detection ofmecA.

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In VitroActivity against Staphylococcus aureus of a Novel Antimicrobial Agent, PRF-119, a Recombinant Chimeric Bacteriophage Endolysin

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00217-11 ◽

2011 ◽

Vol 55 (9) ◽

pp. 4416-4419 ◽

Cited By ~ 29

Author(s):

Evgeny A. Idelevich ◽

Christof von Eiff ◽

Alexander W. Friedrich ◽

Domenico Iannelli ◽

Guoqing Xia ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Agent ◽

The Novel ◽

Good Activity ◽

Coagulase Negative Staphylococci ◽

Methicillin Resistant ◽

Content Type ◽

Microdilution Method ◽

Antistaphylococcal Activity ◽

Bacteriophage Endolysin

ABSTRACTAntistaphylococcal activity of the novel chimeric endolysin PRF-119 was evaluated with the microdilution method. The MIC50and MIC90of 398 methicillin-susceptibleStaphylococcus aureusisolates were 0.098 μg/ml and 0.391 μg/ml, respectively (range, 0.024 to 0.780 μg/ml). Both the MIC50and MIC90values of 776 methicillin-resistantS. aureusisolates were 0.391 μg/ml (range, 0.024 to 1.563 μg/ml). All 192 clinical isolates of coagulase-negative staphylococci exhibited MIC values of >50 μg/ml. In conclusion, PRF-119 exhibited very good activity specifically againstS. aureus.

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Staphylococcus aureus: Prevalence among normal healthy individuals

Nepal Journal of Medical Sciences ◽

10.3126/njms.v2i2.8968 ◽

2013 ◽

Vol 2 (2) ◽

pp. 160-164

Author(s):

D Finney ◽

R Ranganathan ◽

AKS Khan ◽

J Shanmugam

Keyword(s):

Staphylococcus Aureus ◽

Nasal Swab ◽

Throat Swab ◽

Research Centre ◽

Carrier Status ◽

Healthy Individuals ◽

Coagulase Negative Staphylococci ◽

College Hospital ◽

Mannitol Salt Agar ◽

Hospital Infection Control

Background: To determine the prevalence of Staphylococcus aureus among normal healthy individuals in relation to age, gender and site of isolation. Methods: A cross sectional study was conducted among healthy students and staff volunteers between the age group 15-65 years from Gulf Medical University (GMU) and Gulf Medical College Hospital and Research Centre (GMCH&RC), Ajman. Aseptically collected nasal and throat swabs were processed for direct Gram stain Microscopy and cultured on appropriate media. Based on the growth on Mannitol Salt Agar and tube coagulase test Staphylococcus aureus and Coagulase negative Staphylococci (CoNS) were grouped. Results: Of the 127 voulnteers screened 67 were from GMU and 60 from GMCH&RC. From 49/127 (38.5%) volunteers 124 Staphylococci isolates were isolated. Of which 62 (50%) were Staphylococcus aureus and 62 (50%) were CoNS. Among the 62 Staphylococcus aureus 35 (56.45%) were from nasal swab and 27 (43.54%) were from throat swab. Of the 62 CoNS 44 (70.96%) were from nasal swab, 4 (6%) from throat swab and 14 (22.58%) were from both nasal and throat swabs. Conclusion: The study revealed the asymptomatic inhabitation of Staphyloccus aureus in the nose and throat of healthy individuals. This should be seriously looked into since this in future may lead to carrier status. This alarms the hospital infection control committee to screen students and staff on regular basis to minimize the carrier status to protect the community. Nepal Journal of Medical Sciences | Volume 02 | Number 02 | July-December 2013 | Page 160-164 DOI: http://dx.doi.org/10.3126/njms.v2i2.8968

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Dissemination of the Samecfr-Carrying Plasmid among Methicillin-Resistant Staphylococcus aureus and Coagulase-Negative Staphylococcal Isolates in China

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.04580-14 ◽

2015 ◽

Vol 59 (6) ◽

pp. 3669-3671 ◽

Cited By ~ 17

Author(s):

Jia Chang Cai ◽

Yan Yan Hu ◽

Hong Wei Zhou ◽

Gong-Xiang Chen ◽

Rong Zhang

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistant Staphylococcus Aureus ◽

Mobile Element ◽

Sequence Type ◽

Coagulase Negative Staphylococci ◽

Methicillin Resistant ◽

Content Type ◽

Staphylococcal Species ◽

Complete Sequencing ◽

Staphylococcal Cassette Chromosome

ABSTRACTSixcfr-harboring methicillin-resistantStaphylococcus aureus(MRSA) isolates, which belonged to the same clone of sequence type 5 (ST5)-staphylococcal cassette chromosomemecelement II (SCCmecII)-spat311, were investigated in this study. Complete sequencing of acfr-carrying plasmid, pLRSA417, revealed an 8,487-bp fragment containing a Tn4001-like transposon,cfr,orf1, and ISEnfa4. This segment, first identified in an animal plasmid, pSS-01, was observed in several plasmids from clinical coagulase-negative staphylococci in China, suggesting that thecfrgene, which might originate from livestock, was located in the same mobile element and disseminated among different clinical staphylococcal species.

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Heparin Mimics Extracellular DNA in Binding to Cell Surface-Localized Proteins and Promoting Staphylococcus aureus Biofilm Formation

mSphere ◽

10.1128/msphere.00135-17 ◽

2017 ◽

Vol 2 (3) ◽

Cited By ~ 15

Author(s):

Surabhi Mishra ◽

Alexander R. Horswill

Keyword(s):

Staphylococcus Aureus ◽

Biofilm Formation ◽

Binding Proteins ◽

Extracellular Dna ◽

Enhancement Effect ◽

Heparin Binding ◽

Coagulase Negative Staphylococci ◽

Content Type ◽

Catheter Lock ◽

Main Component

ABSTRACT Staphylococcus aureus and coagulase-negative staphylococci (CoNS) are the leading causes of catheter implant infections. Identifying the factors that stimulate catheter infection and the mechanism involved is important for preventing such infections. Heparin, the main component of catheter lock solutions, has been shown previously to stimulate S. aureus biofilm formation through an unknown pathway. This work identifies multiple heparin-binding proteins in S. aureus, and it reveals a potential mechanism through which heparin enhances biofilm capacity. Understanding the details of the heparin enhancement effect could guide future use of appropriate lock solutions for catheter implants. Staphylococcus aureus is a leading cause of catheter-related bloodstream infections. Biofilms form on these implants and are held together by a matrix composed of proteins, polysaccharides, and extracellular DNA (eDNA). Heparin is a sulfated glycosaminoglycan that is routinely used in central venous catheters to prevent thrombosis, but it has been shown to stimulate S. aureus biofilm formation through an unknown mechanism. Data presented here reveal that heparin enhances biofilm capacity in many S. aureus and coagulase-negative staphylococcal strains, and it is incorporated into the USA300 methicillin-resistant S. aureus (MRSA) biofilm matrix. The S. aureus USA300 biofilms containing heparin are sensitive to proteinase K treatment, which suggests that proteins have an important structural role during heparin incorporation. Multiple heparin-binding proteins were identified by proteomics of the secreted and cell wall fractions. Proteins known to contribute to biofilm were identified, and some proteins were reported to have the ability to bind eDNA, such as the major autolysin (Atl) and the immunodominant surface protein B (IsaB). Mutants defective in IsaB showed a moderate decrease in biofilm capacity in the presence of heparin. Our findings suggested that heparin is substituting for eDNA during S. aureus biofilm development. To test this model, eDNA content was increased in biofilms through inactivation of nuclease activity, and the heparin enhancement effect was attenuated. Collectively, these data support the hypothesis that S. aureus can incorporate heparin into the matrix and enhance biofilm capacity by taking advantage of existing eDNA-binding proteins. IMPORTANCE Staphylococcus aureus and coagulase-negative staphylococci (CoNS) are the leading causes of catheter implant infections. Identifying the factors that stimulate catheter infection and the mechanism involved is important for preventing such infections. Heparin, the main component of catheter lock solutions, has been shown previously to stimulate S. aureus biofilm formation through an unknown pathway. This work identifies multiple heparin-binding proteins in S. aureus, and it reveals a potential mechanism through which heparin enhances biofilm capacity. Understanding the details of the heparin enhancement effect could guide future use of appropriate lock solutions for catheter implants.

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