Name: experimental endocarditis model of methicillin resistant staphylococcus aureus (mrsa) in rat
Uploaded: 2012-06-04T12:00:00
Description: Watch this Scientific Journal Video about Experimental Endocarditis Model of Methicillin Resistant Staphylococcus aureus MRSA in Rat at JoVE.com

This work was supported by the U.S. National Institutes of Health [grant R01AI-39108 to A.S.B.] and the American Heart Association [grants SDG 0630219N and AID 09GRNT2180065 to Y.Q.X.].

1. Preparing the MRSA Strains for Infection
<ol>
	<li>Inoculate a loopful of MRSA culture from a stock tube at -80 &deg;C to a sheep blood Trypticase Soy agar (TSA) plate (see Table of specific reagents and equipment), and incubate at 37 &deg;C for overnight.</li>
	<li>Check the purity of the culture on the blood agar plate (similar colony phenotypes) to make sure there is no contamination.</li>
	<li>Pick one colony from the sheep blood TSA plate and inoculate the colony into 5 ml Trypticase Soy broth (TSB) in a 15 ml ...

<ol>
	<li>Petti, C. A., Fowler, V. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Staphylococcus+aureus+bacteremia+and+endocarditis.">Staphylococcus aureus bacteremia and endocarditis.</a> Cardiol. Clin. 21, 219-233 (2003).</li><li>Bashore, T. M., Cabell, C., Fowler, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Update+on+infective+endocarditis.">Update on infective endocarditis.</a> Curr. Probl. Cardiol. 31, 274-352 (2006).</li><li>Fowler, V. 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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+outcomes+of+patients+receiving+daptomycin+for+the+treatment+of+Staphylococcus+aureus+infections+and+assessment+of+clinical+factors+for+daptomycin+failure:+a+retrospective+cohort+study+utilizing+the+Cubicin+Outcomes+Registry+and+Experience.">Clinical outcomes of patients receiving daptomycin for the treatment of Staphylococcus aureus infections and assessment of clinical factors for daptomycin failure: a retrospective cohort study utilizing the Cubicin Outcomes Registry and Experience.</a> Clin. Ther. 31, 1936-1945 (2009).</li><li>Xiong, Y. Q., Willard, J., Kadurugamuwa, J. L., Yu, J., Francis, K. P., Bayer, A. 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The rat endocarditis is an important and well-characterized animal model for in vivo studies in the pathogenesis and antimicrobial agents in the treatment of bacterial infections 9-11. In addition, rat endocarditis model represents a composite of acute and sub-acute infections, and closely mimics human counterpart and human native valve endocarditis. Moreover, accompanied by an in dwelling catheter, it represents a classic biofilm-associated infection, a common and difficult issue in clinical se...

<table border="1">
<tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Blood agar plate, 5% sheep blood in Trypticase Soy agar(TSA)</td>
 <td>Hardy Diagnostics</td>
 <td>A10BX</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Trypticase Soy broth (TSB)</td>
 <td>Becton Dickinson</td>
 <td>211825</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Shaking incubator</td>
 <td>Labnet</td>
 <td>I5311-DS</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Polyethylene tubing </td>
 <td>Becton Dickinson</td>
 <td>427401</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Catheter introducer </td>
 <td>Becton Dickinson</td>
 <td>6999</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Isofluorance</td>
 <td>Western Medical Suppy, Inc</td>
 <td>2147</td>
 <td>Follow safety and handling information</td>
 </tr>
 <tr>
 <td>Surgical Instruments</td>
 <td>Fine Science Tools and Biomedical Research Instruments, Inc.</td>
 <td>&nbsp;</td>
 <td>Find the instruments from the two companies for your needs</td>
 </tr>
 </tbody>
</table>

experimental endocarditis model of methicillin resistant staphylococcus aureus (mrsa) in rat

Endovascular infections, including endocarditis, are life-threatening infectious syndromes1-3. Staphylococcus aureus is the most common world-wide cause of such syndromes with unacceptably high morbidity and mortality even with appropriate antimicrobial agent treatments4-6. The increase in infections due to methicillin-resistant S. aureus (MRSA), the high rates of vancomycin clinical treatment failures and growing problems of linezolid and daptomycin resistance have all further complicated the management of patients with such infections, and led to high healthcare costs7, 8. In addition, it should be emphasized that most recent studies with antibiotic treatment outcomes have been based in clinical settings, and thus might well be influenced by host factors varying from patient-to-patient. Therefore, a relevant animal model of endovascular infection in which host factors are similar from animal-to-animal is more crucial to investigate microbial pathogenesis, as well as the efficacy of novel antimicrobial agents. Endocarditis in rat is a well-established experimental animal model that closely approximates human native valve endocarditis. This model has been used to examine the role of particular staphylococcal virulence factors and the efficacy of antibiotic treatment regimens for staphylococcal endocarditis. In this report, we describe the experimental endocarditis model due to MRSA that could be used to investigate bacterial pathogenesis and response to antibiotic treatment.

Watch this Scientific Journal Video about Experimental Endocarditis Model of Methicillin Resistant Staphylococcus aureus MRSA in Rat at JoVE.com

Experimental Endocarditis Model of Methicillin Resistant Staphylococcus aureus MRSA in Rat

Experimental rat endocarditis model due to methicillin-resistant S. aureus.

Experimental Endocarditis Model of Methicillin Resistant Staphylococcus aureus (MRSA) in Rat

Endovascular infections, including endocarditis, are life-threatening infectious syndromes1-3. Staphylococcus aureus is the most common world-wide cause ...

Research

JoVE Journal

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The Use of Drip Flow and Rotating Disk Reactors for Staphylococcus aureus Biofilm Analysis

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Subcutaneous Infection of Methicillin Resistant Staphylococcus Aureus (MRSA)

Subcutaneous Infection of Methicillin Resistant Staphylococcus Aureus MRSA

MRSA is a worldwide threat to public health, and MRSA skin and soft-tissue infections now account for more than half of all soft-tissue infections in the ...

Overcoming Unresponsiveness in Experimental Autoimmune Encephalomyelitis (EAE) Resistant Mouse Strains by Adoptive Transfer and Antigenic Challenge

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Staphylococcus aureus Growth using Human Hemoglobin as an Iron Source

Staphylococcus aureus Growth using Human Hemoglobin as an Iron Source

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Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus

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Staphylococcal Cassette Chromosome mec (SCCmec) typing  is a very important molecular tool for understanding the epidemiology and  clonal strain ...

Studying Interactions of Staphylococcus aureus with Neutrophils by Flow Cytometry and Time Lapse Microscopy

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We present methods to study the effect of phenol soluble modulins (PSMs)  and other toxins produced and secreted by Staphylococcus  aureus on neutrophils. ...

Targeting Biofilm Associated Staphylococcus aureus Using Resazurin Based Drug-susceptibility Assay

Targeting Biofilm Associated Staphylococcus aureus Using Resazurin Based Drug-susceptibility Assay

Most pathogenic bacteria are able to form biofilms during infection, but due to the difficulty of manipulating and assessing biofilms, the vast majority ...

Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR (RS-PCR)

Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR RS-PCR

The ribosomal spacer PCR (RS-PCR) is a highly resolving and robust genotyping method for S. aureus that allows a high throughput at moderate costs and is, ...

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus

One important feature of the major opportunistic human pathogen Staphylococcus aureus is its extraordinary ability to rapidly acquire resistance to ...

A Mouse Model to Assess Innate Immune Response to Staphylococcus aureus Infection

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Staphylococcus aureus (S. aureus)&#160;infections, including methicillin resistant stains, are an enormous burden on the healthcare system. With incidence ...