Chambers, H F
Strains of staphylococci resistant to methicillin were identified immediately after introduction of this drug. Methicillin-resistant strains have unusual properties, the most notable of which is extreme variability in expression of the resistance trait. The conditions associated with this heterogeneous expression of resistance are described. Methicillin resistance is associated with production of a unique penicillin-binding protein (PBP), 2a, which is bound and inactivated only at high concentrations of beta-lactam antibiotics. PBP2a appears to be encoded by the mec determinant, which also is unique to methicillin-resistant strains. The relationships between PBP2a and expression of resistance and implications for the mechanism of resistance are discussed. The heterogeneous expression of methicillin resistance by staphylococci poses problems in the detection of resistant strains. Experience with several susceptibility test methods is reviewed and guidelines for performance of these tests are given. Treatment of infections caused by methicillin-resistant staphylococci is discussed. Vancomycin is the treatment of choice. Alternatives have been few because methicillin-resistant strains often are resistant to multiple antibiotics in addition to beta-lactam antibiotics. New agents which are active against methicillin-resistant staphylococci are becoming available, and their potential role in treatment is discussed. Images PMID:3069195
Srisuwan, Sutthirat; Tongtawe, Pongsri; Srimanote, Potjanee; Voravuthikunchai, Supayang Piyawan
Background The increasing resistance of Staphylococcus aureus to conventional antibiotics poses a major health problem. Moreover, S. aureus can survive within phagocytes, thus evading some antibiotics and the innate immune response. Rhodomyrtone, a bioactive compound from the leaves of Rhodomyrtus tomentosa, possesses potent antibacterial activity against methicillin-resistant S. aureus (MRSA). This study was to investigate the immunomodulatory effects of rhodomyrtone on THP-1 monocytes in response to MRSA. Methods THP-1 monocytes were stimulated with heat-killed MRSA, followed by treatment with rhodomyrtone. The cell pellets were prepared to detect pro-inflammatory molecules using real-time PCR. The supernatants were collected to assess nitric oxide production using Griess assay. Assays for phagocytosis and bacterial killing by THP-1 monocytes were performed to determine if they were affected by rhodomyrtone. Results Expression of pro-inflammatory molecules including IL-1β, TNF-α, IL-6, and iNOS was enhanced in THP-1 monocytes stimulated with high doses of heat-killed MRSA (108 to 109 cfu/ml). In contrast, monocytes stimulated with MRSA at lower doses (106 to 107 cfu/ml) did not induce the expression of these cytokines. However, rhodomyrtone significantly increased the expression of pro-inflammatory mediators, IL-6 and iNOS in monocytes stimulated with heat-killed MRSA at low doses, and displayed some anti-inflammatory activity by reducing TNF-α expression in monocytes stimulated with heat-killed MRSA at high doses. Treatment with rhodomyrtone also significantly up-regulated the expression of the key pattern recognition receptors, TLR2 and CD14, in THP-1 monocytes stimulated with heat-killed MRSA at 106 to 109 cfu/ml, while heat-killed MRSA alone did not induce the expression of these molecules. The ability of rhodomyrtone to eliminate MRSA from the monocytes was observed within 24 h after treatment. Conclusion Rhodomyrtone enhanced the expression of pattern
Akilandeswari, K; Ruckmani, K
Methicillin-resistant Staphylococcus aureus (MRSA) infections are easily spread among infected patients, where resistance has dramatically increased resulted in serious health issues. Therefore, there is a need to develop alternative natural or combination drug therapies. Apigenin (AP) is a natural poly phenolic flavonoid has been found to possess many beneficial biological actions. The aim of this study was to investigate the anti-MRSA efficacy and synergistic effect of apigenin (AP) and in combination with ampicillin (AM) and ceftriaxone (CEF). The antibacterial activity of apigenin was assessed by the broth macro dilution, checkerboard micro dilution method and time-kill assay. The mode of action was studied by outer and inner membrane permeabilisation assays, scanning electron microscopy and transmission electron microscopy. The minimum inhibitory concentration (MIC) of apigenin against gram positive and gram negative strain ranged from 32.5 to 62.5µg/ml. In checkerboard method apigenin markedly reduced the MIC of the antibiotics ampicillin 800 µg/ml shifted to 107 µg/ml (AM+AP) and ceftriaxone 58 µg/ml shifted to 2.6 µg/ml (CEF+AP) against MRSA. The synergistic activity of ampicillin and ceftriaxone plus apigenin combinations with FIC indices (CI) between 0.18-0.47. The modulation of methicillin-resistance by apigenin significantly enhanced the activities of ampicillin and ceftriaxone. The result of time-kill assays of the two drug combinations AM +AP and CEF+AP against MRSA showed significant inhibitory effect and reduced the colony count by approximately 99% after 8 h The results for outer membrane (OM) and inner membrane (IM) permeabilization showed that ampicillin and ceftriaxone in combination with apigenin damaged MRSA cytoplasmic membrane and caused subsequent leakage of intracellular constituents. Electron microscopy clearly showed that the above said combination also caused marked morphological damage of cell wall, cell shape and plasma
Haque, N; Bari, M S; Bilkis, L; Haque, N; Haque, S; Sultana, S
Staphylococcus aureus strains those are resistant to methicillin are referred to as Methicillin resistant Staphylococcus aureus. These express mecA gene to produce altered penicillin binding protein. At present Methicillin resistant Staphylococcus aureus has been increasing as a serious nosocomial and community pathogen having the property of multi drug resistant. Humans are the natural reservoir for Staphylococcus aureus and asymptomatic colonization is far more common than infection. Many hospitals of different country of the world including Bangladesh are struggling with increasing number of this versatile pathogen. Early and specific diagnosis is important to ensure a favourable outcome. In this paper we attempted to explore history, prevalence, transmission, risk factors, pathogenicity, laboratory diagnosis, prevention and control of Methicillin resistant Staphylococcus aureus as a critical review to provide some new upgrade regarding this super bug.
Medeiros Barreto, Humberto; Cerqueira Fontinele, Filipe; Pereira de Oliveira, Aldeídia; Arcanjo, Daniel Dias Rufino; Cavalcanti dos Santos, Bernadete Helena; de Abreu, Aislan Pereira Lira; Douglas Melo Coutinho, Henrique; Alves Carvalho da Silva, Romezio; Oliveira de Sousa, Taciana; Freire de Medeiros, Maria das Graças; Lopes Citó, Antonia Maria das Graças; Dantas Lopes, José Arimateia
The Lippia origanoides H.B.K. ethanol extract (LOEE) and hexane (LOHEX), dichloromethane (LODCM), and ethyl acetate (LOEA) fractions were tested for their antimicrobial activity alone or in combination with antibiotics against a methicillin resistant Staphylococcus aureus (MRSA) strain. The natural products did not show antimicrobial activity against multidrug resistant strain at the clinically significant concentrations tested. However, a modulatory effect in the antibacterial activity of the neomycin and amikacin was verified when LOEE, LOHEX and LODCM were added to the growth medium at subinhibitory concentrations. A similar modulation was found when the natural products were changed for chlorpromazine, an inhibitor of bacterial efflux pumps, suggesting the involvement of resistance mediated by efflux system in the MRSA tested. The fractions LOHEX and LODCM showed a modulatory activity bigger than their majority compounds (carvacrol, thymol, and naringenin), indicating that this activity is not due to their majority compounds only, but it is probably due to a synergism between their chemical components. These results indicate that L. origanoides H.B.K. can be a source of phytochemicals able to modify the phenotype of resistance to aminoglycosides in MRSA. PMID:24683545
van Duijkeren, Engeline; Catry, Boudewijn; Greko, Christina; Moreno, Miguel A; Pomba, M Constança; Pyörälä, Satu; Ruzauskas, Modestas; Sanders, Pascal; Threlfall, E John; Torren-Edo, Jordi; Törneke, Karolina
Staphylococcus pseudintermedius is an important opportunistic pathogen of companion animals, especially dogs. Since 2006 there has been a significant emergence of methicillin-resistant S. pseudintermedius (MRSP) mainly due to clonal spread. This article reviews research on MRSP with a focus on occurrence, methods used for identification, risk factors for colonization and infection, zoonotic potential and control options. Potential areas for future research are also discussed.
Ippolito, Giuseppe; Leone, Sebastiano; Lauria, Francesco N; Nicastri, Emanuele; Wenzel, Richard P
Over the last decade, methicillin-resistant Staphylococcus aureus (MRSA) strains have emerged as serious pathogens in the nosocomial and community setting. Hospitalization costs associated with MRSA infections are substantially greater than those associated with methicillin-sensitive S. aureus (MSSA) infections, and MRSA has wider economic effects that involve indirect costs to the patient and to society. In addition, there is some evidence suggesting that MRSA infections increase morbidity and the risk of mortality. Glycopeptides are the backbone antibiotics for the treatment of MRSA infections. However, several recent reports have highlighted the limitations of vancomycin, and its role in the management of serious infections is now being reconsidered. Several new antimicrobials demonstrate in vitro activity against MRSA and other Gram-positive bacteria. Data from large surveys indicate that linezolid, daptomycin, and tigecycline are almost universally active against MRSA. This review will briefly discuss the epidemiology, costs, outcome, and therapeutic options for the management of MRSA infections.
Panesso, Diana; Planet, Paul J; Diaz, Lorena; Hugonnet, Jean-Emmanuel; Tran, Truc T; Narechania, Apurva; Munita, Jose M; Rincon, Sandra; Carvajal, Lina P; Reyes, Jinnethe; Londoño, Alejandra; Smith, Hannah; Sebra, Robert; Deikus, Gintaras; Weinstock, George M; Murray, Barbara E; Rossi, Flavia; Arthur, Michel; Arias, Cesar A
We report characterization of a methicillin-susceptible, vancomycin-resistant bloodstream isolate of Staphylococcus aureus recovered from a patient in Brazil. Emergence of vancomycin resistance in methicillin-susceptible S. aureus would indicate that this resistance trait might be poised to disseminate more rapidly among S. aureus and represents a major public health threat.
van Duijkeren, E; Box, A T A; Heck, M E O C; Wannet, W J B; Fluit, A C
Staphylococci isolated from animals (n=311) were screened for methicillin resistance by oxacillin agar screening. Oxacillin-resistant strains were tested for the presence of the mecA gene by PCR. Isolates were identified by standard techniques and 16S rDNA analysis, and their antimicrobial susceptibilities were tested using an agar diffusion method. MecA-positive strains were further analyzed using pulsed-field gel electrophoresis (PFGE). From 11 multidrug-resistant staphylococci, 6 were mecA-positive: 2 methicillin-resistant Staphylococcus aureus (MRSA) and 4 Staphylococcus haemolyticus. Screening of 300 staphylococci (100 S. aureus, 100 S. intermedius and 100 coagulase-negative staphylococci (CNS)) randomly chosen from the strain collection of the Veterinary Microbiological Diagnostic Center yielded five oxacillin-resistant coagulase-negative staphylococci, four of which were mecA-positive. PFGE showed that all mecA-positive staphylococci isolated from animals had distinct patterns. However, one MRSA isolated from a flank fistula of a dog showed homology to a human epidemic MRSA cluster, suggesting that transfer of MRSA between humans and dogs might occur.
Salaheen, Serajus; Peng, Mengfei; Joo, Jungsoo; Teramoto, Hironori; Biswas, Debabrata
The therapeutic roles of phenolic blueberry (Vaccinium corymbosum) and blackberry (Rubus fruticosus) pomace (commercial byproduct) extracts (BPE) and their mechanism of actions were evaluated against methicillin resistant Staphylococcus aureus (MRSA). Five major phenolic acids of BPE, e.g., protocatechuic, p. coumaric, vanillic, caffeic, and gallic acids, as well as crude BPE completely inhibited the growth of vegetative MRSA in vitro while BPE+methicillin significantly reduced MRSA biofilm formation on plastic surface. In addition, BPE restored the effectiveness of methicillin against MRSA by down-regulating the expression of methicillin resistance (mecA) and efflux pump (norA, norB, norC, mdeA, sdrM, and sepA) genes. Antibiogram with broth microdilution method showed that MIC of methicillin reduced from 512 μg/mL to 4 μg/mL when combined with only 200 μg Gallic Acid Equivalent (GAE)/mL of BPE. Significant reduction in MRSA adherence to and invasion into human skin keratinocyte Hek001 cells were also noticed in the presence of BPE. BPE induced anti-apoptosis and anti-autophagy pathways through overexpression of Bcl-2 gene and down-regulation of TRADD and Bax genes (inducers of apoptosis pathway) in Hek001 cells. In summary, novel and sustainable prophylactic therapy can be developed with BPE in combination with currently available antibiotics, especially methicillin, against skin and soft tissue infections with MRSA. PMID:28270804
Rehm, Susan J; Tice, Alan
The evolution of methicillin-resistant and vancomycin-resistant Staphylococcus aureus has demanded serious review of antimicrobial use and development of new agents and revised approaches to prevent and overcome drug resistance. Depending on local conditions and patient risk factors, empirical therapy of suspected S. aureus infection may require coverage of drug-resistant organisms with newer agents and novel antibiotic combinations. The question of treatment with inappropriate antibiotics raises grave concerns with regard to methicillin-resistant S. aureus selection, overgrowth, and increased virulence. Several strategies to reduce the nosocomial burden of resistance are suggested, including shortened hospital stays and outpatient parenteral antimicrobial therapy of the most serious infections.
Domínguez, M A; Liñares, J; Martín, R
Methicillin-resistant Staphylococcus aureus (MRSA) strains are among the most common nosocomial pathogens. The most significant mechanism of resistance to methicillin in this-species is the acquisition of a genetic determinant (mecA gene). However, resistance seems to have a more complex molecular basis, since additional chromosomal material is involved in such resistance. Besides, overproduction of penicillinase and/or alterations in the PBPs can contribute to the formation of resistance phenotypes. Genetic and environmental factors leading to MRSA are reviewed.
Börjesson, Stefan; Landén, Annica; Bergström, Martin; Andersson, Ulrika Grönlund
Staphylococcus pseudintermedius is an opportunistic pathogen that is one of the most frequent causes of infections in dogs. In Europe, there are increasing reports of methicillin-resistant S. pseudintermedius (MRSP), and in Sweden, MRSP has also been more frequently isolated during recent years. However, there is limited knowledge regarding the epidemiology and genetic relationship among the Swedish isolates. This study therefore investigated the genetic relationship of MRSP isolated from companion animals in Sweden. In the study, MRSP isolates taken in the period January 2008-June 2010 from a total of 226 dogs and cats were characterized by spa typing, pulsed-field gel electrophoresis, and antimicrobial susceptibility testing. In addition, the geographical distribution of the isolates based on year of isolation and genetic typing was determined using a geographical information system. One multiresistant clonal lineage dominated among Swedish MRSP isolates, corresponding to the European winning lineage ST71-J-t02-SCCmec II-III. Furthermore, the geographical dissemination of MRSP corresponded to areas with high dog densities, centered on the three major cities in Sweden where the largest animal hospitals are situated.
Mattana, C.M.; Satorres, S.E.; Sosa, A.; Fusco, M.; Alcará, L.E.
Antibacterial activity of organic and aqueous extracts of Acacia aroma was evaluated against methicillin-resistant Staphylococcus aureus (MRSA), methicillin sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus epidermidis. Inhibition of bacterial growth was determined using agar diffusion and bioautographic methods. Among all assayed organic extracts only ethanolic and ethyl acetate extracts presented highest activities against all tested Staphylococcus strains with minimal inhibitory concentration (MIC) values ranging from 2.5 to 10 mg/ml and from 2.5 to 5 mg/ml respectively. The aqueous extracts show little antibacterial activity against Staphylococcus strains. The bioautography assay demonstrated well-defined growth inhibition zones against S. aureus in correspondence with flavonoids and saponins. A. aroma would be an interesting topic for further study and possibly for an alternative treatment for skin infections. PMID:24031532
Goodacre, R; Rooney, P J; Kell, D B
Curie-point pyrolysis mass spectra were obtained from 15 methicillin-resistant and 22 methicillin-susceptible Staphylococcus aureus strains. Cluster analysis showed that the major source of variation between the pyrolysis mass spectra resulted from the phage group of the bacteria, not their resistance or susceptibility to methicillin. By contrast, artificial neural networks could be trained to recognize those aspects of the pyrolysis mass spectra that differentiated methicillin-resistant from methicillin-sensitive strains. The trained neural network could then use pyrolysis mass spectral data to assess whether an unknown strain was resistant to methicillin. These results give the first demonstration that the combination of pyrolysis mass spectrometry with neural networks can provide a very rapid and accurate antibiotic susceptibility testing technique.
Ma, Xiao Xue; Galiana, Antonio; Pedreira, Walter; Mowszowicz, Martin; Christophersen, Inés; Machiavello, Silvia; Lope, Liliana; Benaderet, Sara; Buela, Fernanda; Vicentino, Walter; Albini, María; Bertaux, Olivier; Constenla, Irene; Bagnulo, Homero; Llosa, Luis; Ito, Teruyo
A novel, methicillin-resistant Staphylococcus aureus clone (Uruguay clone) with a non–multidrug-resistant phenotype caused a large outbreak, including 7 deaths, in Montevideo, Uruguay. The clone was distinct from the highly virulent community clone represented by strain MW2, although both clones carried Panton-Valentine leukocidin gene and cna gene. PMID:15963301
DiCicco, Matthew; Weese, Scott; Neethirajan, Suresh; Rousseau, Joyce; Singh, Ameet
The effectiveness of fosfomycin was examined across 31 methicillin-resistant Staphylococcus pseudintermedius (MRSP) strains by agar dilution. Prevalence of the fosfomycin-resistance determinant gene, fosB, was assessed by PCR analysis. Results found that 84% of isolates were fosfomycin-susceptible. Interestingly, 87% of isolates possessed fosB, indicating no association between this putative staphylococci resistance gene and phenotypic resistance. Further evaluation of fosfomycin as a potential treatment of MRSP in dogs is warranted.
Methicillin-Resistant Staphylococcus aureus (MRSA) Infections in the Department of Defense (DOD): Annual Summary 2013 NMCPHC-EDC-TR-44...December 2014 EpiData Center Department Abstract Methicillin-resistant Staphylococcus aureus (MRSA) infections are a national concern for public...Navy (DON) beneficiary populations. This report provides a summary of Methicillin-resistant Staphylococcus aureus (MRSA) incidence and prevalence
Gilani, Mehreen; Usman, Javaid; Latif, Mahwish; Munir, Tehmina; Gill, Maria Mushtaq; Anjum, Rabia; Babar, Nazish
The objective of our study was to determine the frequency of methicillin resistance in coagulase negative Staphylococcus (CoNS) and to determine its in-vitro antimicrobial susceptibility to various other routinely used antibiotics. It was a cross sectional study conducted at the department of Microbiology, Army Medical College, Rawalpindi, Pakistan from June 2011 to May 2012. The organisms were identified on the basis of colony morphology, Gram staining, catalase, DNAase and slide/tube coagulase tests. The organisms were considered to be methicillin resistant when the diameter of zone of inhibition was less than 25mm around 30μg cefoxitin disc. Antibiotic sensitivity was determined using the Modified Kirby-Bauer disc diffusion method. From a total of 337 CoNS, 201 were methicillin resistant and were included in the study. All were resistant to Penicillin, followed by Erythromycin (93•1%), Ciprofloxacin (77%), Co-trimoxazole (74•8%), Gentamicin (68•3%), Clindamycin (51•06%), Tetracycline (44•6%), Fusidic acid (40%), Rifampicin (39•5%), Chloramphenicol (19•3%), Linezolid (2%), Minocycline (1•1%), and Vancomycin (0%). More than half of CoNS were methicillin resistant. Vancomycin is the only drug to which all of the MRCoNS were sensitive, with more than 98% of the isolates being sensitive to Linezolid and Minocycline.
Full-Genome Sequencing Identifies in the Genetic Background Several Determinants That Modulate the Resistance Phenotype in Methicillin-Resistant Staphylococcus aureus Strains Carrying the Novel mecC Gene.
Milheiriço, Catarina; de Lencastre, Hermínia; Tomasz, Alexander
Most methicillin-resistant Staphylococcus aureus (MRSA) strains are resistant to beta-lactam antibiotics due to the presence of the mecA gene, encoding an extra penicillin-binding protein (PBP2A) that has low affinity for virtually all beta-lactam antibiotics. Recently, a new resistance determinant-the mecC gene-was identified in S. aureus isolates recovered from humans and dairy cattle. Although having typically low MICs to beta-lactam antibiotics, MRSA strains with the mecC determinant are also capable of expressing high levels of oxacillin resistance when in an optimal genetic background. In order to test the impact of extensive beta-lactam selection on the emergence of mecC-carrying strains with high levels of antibiotic resistance, we exposed the prototype mecC-carrying MRSA strain, LGA251, to increasing concentrations of oxacillin. LGA251 was able to rapidly adapt to high concentrations of oxacillin in growth medium. In such laboratory mutants with increased levels of oxacillin resistance, we identified mutations in genes with no relationship to the mecC regulatory system, indicating that the genetic background plays an important role in the establishment of the levels of oxacillin resistance. Our data also indicate that the stringent stress response plays a critical role in the beta-lactam antibiotic resistance phenotype of MRSA strains carrying the mecC determinant.
Moodley, Arshnee; Damborg, Peter; Nielsen, Søren Saxmose
Staphylococcus pseudintermedius is a commensal and a common opportunistic pathogen causing mainly infections of the integumentary system in dogs. The recent emergence of multidrug-resistant S. pseudintermedius isolates, in particular methicillin-resistant strains (MRSP) is a threat to small animal health and highlights the need for antimicrobial resistance surveillance to detect trends and potentially perform timeous interventions. We systematically reviewed 202 published articles to investigate temporal changes in antimicrobial resistance in clinical and commensal S. pseudintermedius isolated from dogs in 27 countries between 1980 and 2013. Resistance to the most common antimicrobials tested for in published studies and important for the treatment of staphylococcal infections in dogs were assessed separately for methicillin resistant (MRSP) and methicillin susceptible (MSSP) isolates. Stratified by MSSP and MRSP, no significant increases in antimicrobial resistance were observed over time, except for the penicillinase-labile penicillins (penicillin and ampicillin) among MSSP. However, in recent years, a few studies have reported higher-level of resistance to amikacin, gentamicin and enrofloxacin amongst MSSP. The review highlights inconsistencies between studies as a result of several factors, for example the use of different antimicrobial susceptibility testing methods and interpretation criteria. We recommend that data on susceptibility in important companion animal pathogens are collected and presented in a more harmonized way to allow more precise comparison of susceptibility patterns between studies. One way to accomplish this would be through systematic surveillance either at the country-level or at a larger scale across countries e.g. EU level.
de Carvalho, Milton Jorge; Pimenta, Fabiana Cristina; Hayashida, Miyeko; Gir, Elucir; da Silva, Adriana Maria; Barbosa, Caio Parente; da Silva Canini, Silvia Rita Marin; Santiago, Silvana
INTRODUCTION: S. aureus is one of the main agents of nosocomial infection and is sometimes difficult to treat with currently available active antimicrobials. PURPOSE: To analyze the prevalence of methicillin-susceptible S.aureus (MSSA) and methicillin-resistant S. aureus (MRSA) as well as the MRSA antimicrobial susceptibility profile isolated in the saliva of health professionals at a large public education hospital. MATERIALS AND METHODS: The project was approved by the research and ethics committee of the institution under study. Three samples of saliva from 340 health professionals were collected. The saliva analysis used to identify S. aureus was based on mannitol fermentation tests, catalase production, coagulase, DNAse, and lecithinase. In order to detect MRSA, samples were submitted to the disk diffusion test and the oxacillin agar screening test. In order to identify the minimum inhibitory concentration, the Etest® technique was used. RESULTS: The prevalence of MSSA was 43.5% (148/340), and MRSA was 4.1% (14/340). MRSA detected by the diffusion disk test, was 100% resistant to penicillin and oxacillin, 92.9% resistant to erythromycin, 57.1% resistant to clindamycin, 42.9% resistant to ciprofloxacin and 57.1% resistant to cefoxetin. CONCLUSION: This subject is important for both the education of health professionals and for preventative measures. Standard and contact-precautions should be employed in professional practice. PMID:19488585
Lau, W. Y.; Teoh-Chan, C. H.; Fan, S. T.; Lau, K. F.
Five hundred strains of methicillin-resistant Staphylococcus aureus were tested against various anti-staphylococcal agents. Vancomycin, fusidic acid and fosfomycin were found to be the most effective. Only 1 strain out of 500 was resistant to fosfomycin. Three patients with methicillin-resistant Staphylococcus aureus septicaemia were successfully treated by fosfomycin. We conclude that fosfomycin could be the drug of choice for methicillin-resistant Staphylococcus aureus infection. PMID:3637200
Vitko, Nicholas P.; Richardson, Anthony R.
Staphylococcus aureus is an important bacterial pathogen in the hospital and community settings, especially Staphylococcus aureus clones that exhibit methicillin-resistance (MRSA). Many strains of S. aureus are utilized in the laboratory, underscoring the genetic differences inherent in clinical isolates. S. aureus grows quickly at 37°C with aeration in rich media (e.g. BHI) and exhibits a preference for glycolytic carbon sources. Furthermore, S. aureus has a gold pigmentation, exhibits β-hemolysis, and is catalase and coagulase positive. The four basic laboratory protocols presented in this unit describe how to culture S. aureus on liquid and solid media, how to identify S. aureus strains as methicillin resistant, and how to generate a freezer stock of S. aureus for long-term storage. PMID:23408135
Geria, Aanand N; Schwartz, Robert A
Impetigo is a bacterial infection of the superficial epidermis most commonly seen in infants and children. It is clinically characterized by crusted erosions or ulcers that may arise as a primary infection in which bacterial invasion occurs through minor breaks in the cutaneous surface or a secondary infection of a preexisting dermatosis or infestation. Impetigo occurs in 2 forms: bullous and nonbullous. Staphylococcus aureus currently is the most common overall cause of impetigo, but Streptococcus pyogenes remains an important cause in developing nations. Community-acquired methicillin-resistant S aureus (CA-MRSA) poses a challenge because of its enhanced virulence and increasing prevalence in children. For limited uncomplicated impetigo, either topical mupirocin or fusidic acid is as effective if not more effective than systemic antibiotics. For extensive or complicated impetigo, systemic antibiotics may be warranted, but beta-lactam antibiotics should be avoided if methicillin-resistant S aureus (MRSA) is suspected.
Methicillin-resistant Staphylococcus aureus (MRSA) is a significant cause of health care-associated infections. Vancomycin remains an acceptable treatment option. There has been a welcome increase in the number of agents available for the treatment of MRSA infection. These drugs have certain differentiating attributes and may offer some advantages over vancomycin, but they also have significant limitations. These agents provide some alternative when no other options are available. PMID:28032484
Bogiel, Tomasz; Mikucka, Agnieszka; Deptuła, Aleksander; Gospodarek, Eugenia
Many identification and relatedness studies methods had been commonly used for epidemiological studies in microbiological laboratories. Apart from phenotypic methods, genotypic are also often used. The aim of this study was to compare, obtained by PFGE chromosomal DNA patterns of methicillin-resistant S. epidermidis strains isolated from clinical material. 46 methicillin-resistant S. epidermidis strains were included in this study. Most of them were isolated from wound swabs (65.2%) and catheters (19.6%) from different surgical clinics (76.1%). To identify strains and receive biochemical profiles, ID 32 Staph tests and GPI cards of Vitek 1 system were used. Pulsed-field gel electrophoresis and Tenover et al. interpretation were used to compare chromosomal DNA patterns of examined strains. 44 and 42 PFGE patterns of chromosomal DNA were received, using visual interpretation classifying two pairs of strains as the same, two pairs as closely related and three pairs as probably related. Strains classified as identical and similar in visual evaluation were indistinguishable in Molecular Analyst DST interpretation, probably due to tolerance in bands location pattern. Strains probably related in visual interpretation represent at least 96% similarity in Molecular Analyst DST but different susceptibility and biochemical profiles obtained by ID 32 Staph and Vitek 1. PFGE analysis had foremost capacity to distinguish methicillin-resistant S. epidermidis strains using visual interpretation and Molecular Analyst DST (Bio-Rad) program and seems to be useful method in epidemiological studies. Strains with the same PFGE pattern, had different susceptibility and biochemical profiles.
Hamdad, F; Donda, F; Lefebvre, J F; Laurans, G; Biendo, M; Thomas, D; Canarelli, B; Rousseau, F; Eb, F
Eighty-five atypical isolates of Staphylococcus aureus divided into 73 aminoglycoside-susceptible methicillinresistant (AS-MRSA) and 12 kanamycin-tobramycin-resistant methicillin-susceptible (KTR-MSSA) were phenotypically and genotypically examined for methicillin resistance. Among these tests, the diffusion method using the oxacillin and cefoxitin disks on Mueller-Hinton agar with and without NaCl, the incubation at 35 degrees C or 30 degrees C for 24 or 48 hr, respectively, and the determination of oxacillin MICs by E-test were performed. We also examined the presence of the mecA gene by PCR and its product PBP 2a by the Slidex MRSA Detection test after induction by cefoxitin disk. All of the AS-MRSA strains (100%) were detected by the cefoxitin disk in all conditions and by the oxacillin disk on Mueller-Hinton agar with 2% of NaCl at 35 degrees C. Without NaCl, the sensitivity fell to 97.2% by oxacillin disk. The oxacillin MICs for these isolates ranged from 2 to 128 mg/L. The mecA gene determinant and its product PBP 2a were detected in all AS-MRSA strains. All KTR-MSSA strains were phenotypically methicillin-susceptible and oxacillin MICs were below or borderline of breakpoint (< or =2 mg/L). The mecA gene determinant and its product were detected in one strain. Pulsed-field gel electrophoresis (PFGE) was applied and revealed the presence of two major patterns A (36.9%) and B (46.2%) in AS-MRSA isolates and seven patterns in the KTR-MSSA strains.
Rota, Ada; Milani, Chiara; Drigo, Ilenia; Drigo, Michele; Corrò, Michela
The overuse of antimicrobials can select resistant bacteria strains; staphylococci have the ability to become resistant to all beta-lactam antimicrobials and are a significant concern in human medicine and a growing issue for veterinary medicine. Because antimicrobials are sometimes incorrectly used in breeding kennels, the objective of the work was to assess the occurrence of methicillin-resistant coagulase-positive staphylococci in breeding dogs. The research was carried out in 13 kennels that were allotted to three categories according to the intensity of antimicrobial use. Vaginal and milk swabs were taken from 87 healthy bitches around parturition and also from multiple organs of 27 of their pups that died within the first 2 weeks. Standard bacteriological examinations were carried out and coagulase-positive staphylococci were identified. All the coagulase-positive staphylococci resulted to be Staphylococcus pseudintermedius. Susceptibility to oxacillin and the presence of the mecA gene were tested. Nine out of 89 strains (six isolated from the bitches' milk and three from dead puppies, all belonging to kennels characterized by an excessive use of antimicrobials) were multidrug-resistant, methicillin-resistant and mecA positive. Our results confirm that excessive use of antimicrobials entails the risk of selecting resistant staphylococci strains. Our data also indicate that the bacterial flora of healthy dogs belonging to specific populations may act as a reservoir of resistance genes.
Sasaki, Takashi; Kikuchi, Ken; Tanaka, Yoshikazu; Takahashi, Namiko; Kamata, Shinichi; Hiramatsu, Keiichi
We surveyed methicillin-resistant coagulase-positive staphylococcus (MRCPS) strains from 57 (26 inpatient and 31 outpatient) dogs and 20 veterinary staff in a veterinary teaching hospital. From the staff, three MRCPS strains were isolated, and two were methicillin-resistant Staphylococcus aureus (MRSA). In contrast, 18 MRCPS strains were detected in both inpatient (12 of 26 [46.2%]) and outpatient (6 of 31 [19.4%]) dogs. Among them, only one strain was MRSA. Using direct sequencing of sodA and hsp60 genes, the 18 MRCPS strains other than MRSA from a staff and 17 dogs, were finally identified as Staphylococcus pseudintermedius, a novel species of Staphylococcus from a cat. All of the methicillin-resistant S. pseudintermedius (MRSP) strains were multidrug resistant to erythromycin, clindamycin, trimethoprim-sulfamethoxazole, and levofloxacin. Most of the MRSP strains showed high-level resistance to oxacillin (>/=128 mug/ml, 15 of 18 [83.3%]), and 10 of 15 (66.7%) high-level oxacillin-resistant MRSP strains carried type III SCCmec. DNA fingerprinting of MRSP strains by pulsed-field gel electrophoresis yielded eight clusters: clone A with four subtypes, clone B with four subtypes, clone C with three subtypes, and five other different single clones. MRSP strains from the staff and some inpatient and outpatient dogs shared three major clones (clones A, B, and C), but the strains of the other five different clusters were distributed independently among inpatient or outpatient dogs. This genetic diversity suggested that the MRSP strains were not only acquired in this veterinary teaching hospital but also acquired in primary veterinary clinics in the community. To our knowledge, this is the first report of MRSP in dogs and humans in a veterinary institution.
Cheong, I; Tan, S C; Wong, Y H; Zainudin, B M; Rahman, M Z
Between August 1990 to November 1991, 905 of 2583 (35.4%) isolates of Staphylococcus aureus were found to be methicillin-resistant in a general hospital in Malaysia. A detailed study of 539 of these isolates showed a high prevalence of methicillin resistant Staphylococcus aureus (MRSA) in the surgical/orthopaedic wards, paediatric wards and the special care unit. The yield of MRSA was highest from wounds/ulcers/skin swabs accounting for 64.2 per cent followed by 6.9 per cent in blood cultures. Vancomycin remains the drug of choice with no resistance detected. The resistance to ciprofloxacin was 6.7 per cent, rifampicin 4.5 per cent and fusidic acid 2.0 per cent. Most isolates were resistant to aminoglycosides. In view of the high prevalence of MRSA in this hospital, the authorities must introduce more effective measures to control its spread as a nosocomial pathogen. Otherwise it may seriously disrupt the efficient delivery of health care services in the country.
Argudín, M Angeles; Butaye, Patrick
The use of metals as feed supplement has been recognized as a potential driver for co-selection of methicillin-resistant Staphylococcus aureus in pigs. However, the prevalence of these determinants in methicillin-resistant coagulase-negative staphylococci (MRCoNS) is largely unknown. In this study, a collection of 130 MRCoNS from pigs and veal calves were investigated for the presence of metal-resistance genes (czrC, copB, cadD, arsA) associated to SCCmec. Near half of the isolates carried metal resistance genes (czrC 5.4%, copB 38.5%, cadD 7.7%, arsA 26.2%) regardless of their SCCmec type. The increased use of metals in livestock animals, especially zinc in pigs in several European countries may co-select for methicillin-resistance in several staphylococcal species.
Qin, Li; Da, Fei; Tan, Daniel C. S.; Nguyen, Thuan H.; Fu, Chih-Lung; Tan, Vee Y.; Sturdevant, Daniel E.
Bacterial sepsis is a major killer in hospitalized patients. Coagulase-negative staphylococci (CNS) with the leading species Staphylococcus epidermidis are the most frequent causes of nosocomial sepsis, with most infectious isolates being methicillin-resistant. However, which bacterial factors underlie the pathogenesis of CNS sepsis is unknown. While it has been commonly believed that invariant structures on the surface of CNS trigger sepsis by causing an over-reaction of the immune system, we show here that sepsis caused by methicillin-resistant S. epidermidis is to a large extent mediated by the methicillin resistance island-encoded peptide toxin, PSM-mec. PSM-mec contributed to bacterial survival in whole human blood and resistance to neutrophil-mediated killing, and caused significantly increased mortality and cytokine expression in a mouse sepsis model. Furthermore, we show that the PSM-mec peptide itself, rather than the regulatory RNA in which its gene is embedded, is responsible for the observed virulence phenotype. This finding is of particular importance given the contrasting roles of the psm-mec locus that have been reported in S. aureus strains, inasmuch as our findings suggest that the psm-mec locus may exert effects in the background of S. aureus strains that differ from its original role in the CNS environment due to originally “unintended” interferences. Notably, while toxins have never been clearly implied in CNS infections, our tissue culture and mouse infection model data indicate that an important type of infection caused by the predominant CNS species is mediated to a large extent by a toxin. These findings suggest that CNS infections may be amenable to virulence-targeted drug development approaches. PMID:28151994
Fu, Xiu-jun; Fang, Yong; Yao, Min
Nowadays methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common multidrug resistant bacteria both in hospitals and in the community. In the last two decades, there has been growing concern about the increasing resistance to MRSA of the most potent antibiotic glycopeptides. MRSA infection poses a serious problem for physicians and their patients. Photosensitizer-mediated antimicrobial photodynamic therapy (PDT) appears to be a promising and innovative approach for treating multidrug resistant infection. In spite of encouraging reports of the use of antimicrobial PDT to inactivate MRSA in large in vitro studies, there are only few in vivo studies. Therefore, applying PDT in the clinic for MRSA infection is still a long way off. PMID:23555074
Price, Lance B.; Stegger, Marc; Hasman, Henrik; Aziz, Maliha; Larsen, Jesper; Andersen, Paal Skytt; Pearson, Talima; Waters, Andrew E.; Foster, Jeffrey T.; Schupp, James; Gillece, John; Driebe, Elizabeth; Liu, Cindy M.; Springer, Burkhard; Zdovc, Irena; Battisti, Antonio; Franco, Alessia; Żmudzki, Jacek; Schwarz, Stefan; Butaye, Patrick; Jouy, Eric; Pomba, Constanca; Porrero, M. Concepción; Ruimy, Raymond; Smith, Tara C.; Robinson, D. Ashley; Weese, J. Scott; Arriola, Carmen Sofia; Yu, Fangyou; Laurent, Frederic; Keim, Paul; Skov, Robert; Aarestrup, Frank M.
ABSTRACT Since its discovery in the early 2000s, methicillin-resistant Staphylococcus aureus (MRSA) clonal complex 398 (CC398) has become a rapidly emerging cause of human infections, most often associated with livestock exposure. We applied whole-genome sequence typing to characterize a diverse collection of CC398 isolates (n = 89), including MRSA and methicillin-susceptible S. aureus (MSSA) from animals and humans spanning 19 countries and four continents. We identified 4,238 single nucleotide polymorphisms (SNPs) among the 89 core genomes. Minimal homoplasy (consistency index = 0.9591) was detected among parsimony-informative SNPs, allowing for the generation of a highly accurate phylogenetic reconstruction of the CC398 clonal lineage. Phylogenetic analyses revealed that MSSA from humans formed the most ancestral clades. The most derived lineages were composed predominantly of livestock-associated MRSA possessing three different staphylococcal cassette chromosome mec element (SCCmec) types (IV, V, and VII-like) including nine subtypes. The human-associated isolates from the basal clades carried phages encoding human innate immune modulators that were largely missing among the livestock-associated isolates. Our results strongly suggest that livestock-associated MRSA CC398 originated in humans as MSSA. The lineage appears to have undergone a rapid radiation in conjunction with the jump from humans to livestock, where it subsequently acquired tetracycline and methicillin resistance. Further analyses are required to estimate the number of independent genetic events leading to the methicillin-resistant sublineages, but the diversity of SCCmec subtypes is suggestive of strong and diverse antimicrobial selection associated with food animal production. PMID:22354957
Green, Bart N.; Johnson, Claire D.; Egan, Jonathon Todd; Rosenthal, Michael; Griffith, Erin A.; Evans, Marion Willard
Objective Methicillin-resistant Staphylococcus aureus (MRSA) is associated with difficult-to-treat infections and high levels of morbidity. Manual practitioners work in environments where MRSA is a common acquired infection. The purpose of this review is to provide a practical overview of MRSA as it applies to the manual therapy professions (eg, physical and occupational therapy, athletic training, chiropractic, osteopathy, massage, sports medicine) and to discuss how to identify and prevent MRSA infections in manual therapy work environments. Methods PubMed and CINAHL were searched from the beginning of their respective indexing years through June 2011 using the search terms MRSA, methicillin-resistant Staphylococcus aureus, and Staphylococcus aureus. Texts and authoritative Web sites were also reviewed. Pertinent articles from the authors' libraries were included if they were not already identified in the literature search. Articles were included if they were applicable to ambulatory health care environments in which manual therapists work or if the content of the article related to the clinical management of MRSA. Results Following information extraction, 95 citations were included in this review, to include 76 peer-reviewed journal articles, 16 government Web sites, and 3 textbooks. Information was organized into 10 clinically relevant categories for presentation. Information was organized into the following clinically relevant categories: microbiology, development of MRSA, risk factors for infection, clinical presentation, diagnostic tests, screening tests, reporting, treatment, prevention for patients and athletes, and prevention for health care workers. Conclusion Methicillin-resistant S aureus is a health risk in the community and to patients and athletes treated by manual therapists. Manual practitioners can play an essential role in recognizing MRSA infections and helping to control its transmission in the health care environment and the community
Miletić, Maja; Vuković, Dragana; Živanović, Irena; Dakić, Ivana; Soldatović, Ivan; Maletić, Dejan; Lazović, Saša; Malović, Gordana; Petrović, Zoran; Puač, Nevena
In numerous recent papers plasma chemistry of non equilibrium plasma sources operating at atmospheric pressure has been linked to plasma medical effects including sterilization. In this paper we present a study of the effectiveness of an atmospheric pressure plasma source, known as plasma needle, in inhibition of the growth of biofilm produced by methicillin resistant Staphylococcus aureus (MRSA). Even at the lowest powers the biofilms formed by inoculi of MRSA of 104 and 105 CFU have been strongly affected by plasma and growth in biofilms was inhibited. The eradication of the already formed biofilm was not achieved and it is required to go to more effective sources.
Wang, Yucheng; Dai, Tianhong; Gu, Ying
Background: With the increasing emergence of multidrug-resistant (MDR) bacterial strains, there is a pressing need for the development of alternative treatment for infections. Antimicrobial blue light (aBL) has provided a simple and effective approach. Methods: We first investigated the effectiveness of aBL (415 nm) inactivation of USA300 LAClux (a communityacquired Methicillin-resistant Staphylococcus aureus strain) both in the planktonic and biofilm forms. The survival of the bacteria in suspensions was determined by serial dilution and that of the biofilm-embedded bacteria was determined by bioluminescence quantification. Using a mouse model of thermal burn infected with USA300 LAClux, we further assessed the effectiveness of aBL for treating localized infections. Bioluminescence imaging was performed to monitor in real time bacterial viability in vivo. Results: In vitro study showed that, for the planktonic counterpart of the bacteria or the 24-h-old biofilms, an irradiance of 55 mW/cm2 for 60 min resulted in a 4.61 log10 or 2.56 log10 inactivation, respectively. In vivo study using infected mouse burns demonstrated that a 2.56-log10 inactivation was achieved after 100-mW/cm2 irradiation for 62 min. Conclusions: aBL is a potential alternative approach for treating Methicillin-resistant Staphylococcus aureus infections.
Alex, Aniltta; Letizia, MariJo
Methicillin-resistant "Staphylococcus aureus" (MRSA) is a disease-causing organism that has been present in hospital settings since the 1960s. However, a genetically distinct strain of MRSA, called community-acquired methicillin-resistant "Staphylococcus aureus" (CA-MRSA), has emerged in recent years in community settings among healthy…
Atkinson Smith, Mary
In the specialty of orthopaedics, methicillin-resistant Staphylococcus aureus (MRSA) is a major contributor to infections of the soft tissues, surgical sites, and joints, in addition to increasing disability, mortality, and healthcare costs. Inappropriate prescribing and misuse of antibiotics have led to bacterial resistance and the rapid emergence of MRSA. It is imperative for healthcare providers and facilities to improve quality, promote safety, and decrease costs related to MRSA infections. The healthcare profession and society as a whole play an important role in minimizing the transmission of pathogens, reducing the incidence of MRSA infections, and decreasing the development of future antibiotic resistant pathogens. This article discusses the epidemiology of MRSA and describes evidence-based guidelines pertaining to the prevention, minimization, and treatment of MRSA-related infections. Specific application to orthopaedics are discussed in the context of patient risk factors, perioperative and postoperative prophylaxis, and current trends regarding education and reporting strategies.
Barbarin, Alexis M; Hu, Baofeng; Nachamkin, Irving; Levy, Michael Z
A recent paper published by Lowe and Romney in Emerging Infectious Diseases titled, Bed bugs as Vectors for Drug-Resistant Bacteria has sparked a renewed interest in bed bug vector potential. We followed a pyrethroid resistant strain of the human bed bug (Cimex lectularius, L.) fed either human blood or human blood with added methicillin resistant Staphylococcus aureus (MRSA) for 9 days post-feeding. Results indicated that while the bed bug midgut is a hospitable environment for MRSA, the bacteria does not survive longer than 9 days within the midgut. Additionally, MRSA is not amplified within the midgut of the bug as the infection was cleared within 9 days. Due to the weekly feeding behaviours of bed bugs, these results suggest that bed bug transmission of MRSA is highly unlikely.
Joffe, Daniel; Goulding, Fiona; Langelier, Ken; Magyar, Gabor; McCurdy, Les; Milstein, Moe; Nielsen, Kia; Villemaire, Stephanie
Pyoderma in dogs is most commonly caused by Staphylococcus spp., and significant emergence of methicillin resistance in staphylococcal pyoderma has been reported. This preliminary study of the prevalence of methicillin resistance in canine pyoderma cases in Canadian primary care veterinary practices revealed that methicillin-resistant Staphylococcus spp. were present in 12.1% of 149 staphylococcal positive skin culture cases. PMID:26483585
Joffe, Daniel; Goulding, Fiona; Langelier, Ken; Magyar, Gabor; McCurdy, Les; Milstein, Moe; Nielsen, Kia; Villemaire, Stephanie
Pyoderma in dogs is most commonly caused by Staphylococcus spp., and significant emergence of methicillin resistance in staphylococcal pyoderma has been reported. This preliminary study of the prevalence of methicillin resistance in canine pyoderma cases in Canadian primary care veterinary practices revealed that methicillin-resistant Staphylococcus spp. were present in 12.1% of 149 staphylococcal positive skin culture cases.
Hensler, Mary E.; Jang, Kyoung Hwa; Thienphrapa, Wdee; Vuong, Lisa; Tran, Dan N.; Soubih, Evaristus; Lin, Leo; Haste, Nina M.; Cunningham, Mark L.; Kwan, Bryan P.; Shaw, Karen Joy; Fenical, William; Nizet, Victor
Anthracimycin is a recently discovered novel marine-derived compound with activity against Bacillus anthracis. We tested anthracimycin against an expanded panel of Staphylococcus aureus strains in vitro and in vivo. All strains of S. aureus tested, including methicillin-sensitive (MSSA), methicillin-resistant (MRSA), and vancomycin-resistant strains of S. aureus were sensitive to anthracimycin at minimum inhibitory concentrations (MIC) of < 0.25 mg/L. Although its post-antibiotic effects were minimal, anthracimycin exhibited potent and rapid bactericidal activity, with a > 4-log kill of USA300 MRSA within 3 hours at 5 times its MIC. At concentrations significantly below the MIC, anthracimycin slowed MRSA growth and potentiated the bactericidal activity of the human cathelicidin, LL-37. The bactericidal activity of anthracimycin was somewhat mitigated in the presence of 20% human serum, and the compound was minimally toxic to human cells, with an IC50 = 70 mg/L against human carcinoma cells. At concentrations near the MIC anthracimycin inhibited S. aureus nucleic acid synthesis as determined by optimized macromolecular synthesis methodology, with inhibition of DNA and RNA synthesis occurring in the absence of DNA intercalation. Anthracimycin at a single dose of 1 or 10 mg/kg was able to protect mice from MRSA-induced mortality in a murine peritonitis model of infection. Anthracimycin provides an interesting new scaffold for future development of a novel MRSA antibiotic. PMID:24736856
Weese, J S; Sweetman, K; Edson, H; Rousseau, J
Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is an increasingly important cause of opportunistic infections in dogs and cats. Treatment options are often limited because of the high level of antimicrobial resistance. Doxycycline resistance is common, but variable mechanisms of doxycycline resistance exist, with some conferring resistance to doxycycline but not minocycline. However, there has been limited study of minocycline susceptibility in clinical MRSP isolates nor has the association of susceptibility and clonal complex been clearly established. The objective of this study was to evaluate the susceptibility of MRSP to tetracycline, doxycycline and minocycline, to determine the prevalence of tet(M) and tet(K) and to evaluate the impact of strain on minocycline susceptibility. One hundred seven MRSP isolates from dogs from Canada and the US were included; 79 from clinical infections and 28 from colonization sites. Thirty-nine (36%) isolates were susceptible to tetracycline, 41 (38%) to doxycycline and 70 (65%) to minocycline. Two main dru types, dt9a and dt11a, were present. When tetracycline or doxycycline resistant, dru type dt9a and related strains predominantly harboured tet(K) and were susceptible to minocycline. In contrast, dt11a and related strains tended to harbour tet(M), which confers resistance to all three tetracyclines. Minocycline might be a treatment option for some MRSP infections, even those that are doxycycline resistant; however, interpretive breakpoints may need to be re-assessed. Study of the pharmacokinetics and clinical efficacy of minocycline in dogs and cats is warranted.
Springer, Burkhard; Orendi, Ulrike; Much, Peter; Höger, Gerda; Ruppitsch, Werner; Krziwanek, Karina; Metz-Gercek, Sigrid; Mittermayer, Helmut
Staphylococcus aureus is a major cause of infection in hospitals and the community. One third of the general population is colonized by the bacterium, constituting a risk factor for acquisition of infection with this pathogen. Worldwide, the increasing antibiotic resistance of S. aureus complicates treatment of infection and control measures. Soon after the introduction of methicillin, the first isolates resistant to this antibiotic were reported and named methicillin-resistant S. aureus (MRSA). During the past decade a major change in MRSA epidemiology has been observed: whereas in the past MRSA was almost exclusively regarded a hospital pathogen, the advent of community-acquired MRSA has led to infections in people without hospital-related risk factors. Recent evidence has also identified a link between colonization of livestock and MRSA carriage and infections in people who work with animals. Screening of pigs and pig farmers in the Netherlands revealed high prevalence of MRSA sequence type (ST) 398 and it has become clear that the emergence of ST398 is not just a Dutch problem, as reports on livestock colonization and human infections are appearing worldwide. In Austria, the ST398 lineage has been detected in dust samples from pig breeding facilities and in food samples. Since the first Austrian detection of this emerging lineage in 2006, 21 human isolates, partially associated with infections, have been observed. MRSA has to be regarded as a new emerging zoonotic agent and livestock may constitute a growing reservoir of the ST398 lineage. More information is needed so that control measures to reduce the impact of the emerging MRSA ST398 lineage on public health can be developed and implemented.
Modulation of mecA Gene Expression by Essential Oil from Salvia sclarea and Synergism with Oxacillin in Methicillin Resistant Staphylococcus epidermidis Carrying Different Types of Staphylococcal Chromosomal Cassette mec
Chovanová, Romana; Mikulášová, Mária; Vaverková, Štefánia
The essential oil (EO) from Salvia sclarea was shown to increase the susceptibility of methicillin resistant Staphylococcus epidermidis (MRSE) isolates to oxacillin. The purpose of this study was to investigate the effect of EO from S. sclarea on expression of mecA gene of MRSE carrying different types of staphylococcal chromosomal cassette (SCCmec) and to evaluate potential synergistic effect of EO with oxacillin. Using real-time PCR we found that EO alone inhibited the expression of the resistant genes mecA, mecR1, and mecI and blaZ, blaR1, and blaI. The use of the combination of EO with oxacillin resulted in significantly inhibited expression of mecA gene in all tested strains with different types of SCCmec. Using time-kill assay and checkerboard assay we confirmed synergistic effect of EO from S. sclarea and oxacillin in MRSE. PMID:26880926
Turabelidze, George; Lin, Mei; Wolkoff, Barbara; Dodson, Douglas; Gladbach, Stephen; Zhu, Bao-Ping
Methicillin-resistant Staphylococcus aureus (MRSA) infections outside the healthcare setting are an increasing concern. We conducted a case-control study to investigate an MRSA outbreak during 2002-2003 in a Missouri prison and focused on hygiene factors. Information on sociodemographic characteristics, medical history, and hygiene practices of study participants was collected by interview and medical record review. Logistic regression was used to evaluate MRSA infection in relation to hygiene factors individually and as a composite hygiene score; potential confounding factors were controlled. Selected MRSA isolates were analyzed by pulsed-field gel electrophoresis (PFGE). MRSA infection was significantly associated with a low composite hygiene score. Transmission among prison inmates appeared to be responsible for this outbreak. PFGE analysis showed that isolates were indistinguishable and associated with community-onset MRSA infections in other US prisons. Improving hygiene practices and environmental conditions may help prevent and interrupt future MRSA outbreaks in prison settings.
Sowash, Madeleine G.; Uhlemann, Anne-Catrin
Over the past decade, the emergence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has changed the landscape of S. aureus infections around the globe. Initially recognized for its ability to cause disease in young and healthy individuals without healthcare exposures as well as for its distinct genotype and phenotype, this original description no longer fully encompasses the diversity of CA-MRSA as it continues to expand its niche. Using four case studies, we highlight a wide range of the clinical presentations and challenges of CA-MRSA. Based on these cases we further explore the globally polygenetic background of CA-MRSA with a special emphasis on generally less characterized populations. PMID:24085688
De Lucia, M; Moodley, A; Latronico, F; Giordano, A; Caldin, M; Fondati, A; Guardabassi, L
The overall prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) was 2% (10/590) among 590 canine specimens submitted to an Italian veterinary diagnostic laboratory during a two-month period, and 21% (10/48) among Staphylococcus intermedius group (SIG) isolates. All methicillin-resistant strains exhibited additional resistance to fluoroquinolones, gentamicin, lincosamides, tetracyclines, and potentiated sulfonamides, belonged predominantly to spa type t02 and harboured SCCmec type II-III cassette.
Marples, R R; Reith, S
Methicillin-resistant Staphylococcus aureus (MRSA) were detected soon after the introduction of methicillin in 1960, and reports of their isolation increased up to 1971. Changes in antibiotic usage were associated with a fall and then, in the early 1980s, a further rise in the number of reports. This article reviews the various surveys that have been conducted to establish the frequency, distribution and strain varieties of MRSA. The first strain to be recognised as epidemic (ie, affecting more than one hospital), was defined by phage typing and antibiogram, confirmed with molecular typing, and designated EMRSA-1. It was first detected in 1981 and became progressively more widespread until it began to decline in 1987. Only three health regions reported this strain in the first quarter of 1991. EMRSA-2 has remained restricted to the South East and South West Thames regions. EMRSA-3 appeared in the South East Thames region in 1987 and has since spread, being reported from eight health regions in the first quarter of 1991. At least 11 other strains of MRSA affecting more than one hospital have been detected and ten endemic strains (restricted to single hospitals) have been identified. Imported strains of MRSA, often introduced following the repatriation of road traffic accident victims, may include strains with epidemic potential and local spread has followed importation in at least two incidents. Continued surveillance of epidemic MRSA strains and the search for simple and widely applicable markers, such as unusual antibiotic resistance patterns or biochemical features, are needed for the prompt application of control measures.
Kumari, Jyoti; Shenoy, Shalini M.; Baliga, Shrikala; Chakrapani, M.; Bhat, Gopalkrishna K.
Objectives: Healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen worldwide and its multidrug resistance is a major concern. This study aimed to determine the clinical characteristics and antibiotic susceptibility profile of healthcare-associated MRSA with emphasis on resistance to macrolide-lincosamide-streptogramin B (MLSB) phenotypes and vancomycin. Methods: This cross-sectional study was carried out between February 2014 and February 2015 across four tertiary care hospitals in Mangalore, South India. Healthcare-associated infections among 291 inpatients at these hospitals were identified according to the Centers for Disease Control and Prevention guidelines. Clinical specimens were collected based on infection type. S. aureus and MRSA isolates were identified and antibiotic susceptibility tests performed using the Kirby-Bauer disk diffusion method. The minimum inhibitory concentration of vancomycin was determined using the Agar dilution method and inducible clindamycin resistance was detected with a double-disk diffusion test (D-test). Results: Out of 291 healthcare-associated S. aureus cases, 88 were MRSA (30.2%). Of these, 54.6% were skin and soft tissue infections. All of the isolates were susceptible to teicoplanin and linezolid. Four MRSA isolates exhibited intermediate resistance to vancomycin (4.6%). Of the MRSA strains, 10 (11.4%) were constitutive MLSB phenotypes, 31 (35.2%) were inducible MLSB phenotypes and 14 (15.9%) were macrolide-streptogramin B phenotypes. Conclusion: Healthcare-associated MRSA multidrug resistance was alarmingly high. In routine antibiotic susceptibility testing, a D-test should always be performed if an isolate is resistant to erythromycin but susceptible to clindamycin. Determination of the minimum inhibitory concentration of vancomycin is necessary when treating patients with MRSA infections. PMID:27226908
Goodacre, Royston; Rooney, Paul J.; Kell, Douglas B.
FTIR spectra were obtained from 15 methicillin-resistant and 22 methicillin-susceptible Staphylococcus aureus strains using our DRASTIC approach. Cluster analysis showed that the major source of variation between the IR spectra was not due to their resistance or susceptibility to methicillin; indeed early studies suing pyrolysis mass spectrometry had shown that this unsupervised analysis gave information on the phage group of the bacteria. By contrast, artificial neural networks, based on a supervised learning, could be trained to recognize those aspects of the IR spectra which differentiated methicillin-resistant from methicillin- susceptible strains. These results give the first demonstration that the combination of FTIR with neural networks can provide a very rapid and accurate antibiotic susceptibility testing technique.
Wong, San S.; Smith, Peter R.; Ayaz, Asim; Sepkowitz, Douglas
We report a case of vertebral osteomyelitis due to methicillin-resistant Staphylococcus aureus (MRSA) with a slowly progressive, relatively asymptomatic course in a young woman with suspected hyperimmunoglobulin E syndrome (Job's syndrome). PMID:26839768
Tawil, Nancy; Mouawad, Flaviana; Lévesque, Simon; Sacher, Edward; Mandeville, Rosemonde; Meunier, Michel
Two hundred fifty Staphylococcus aureus clinical isolates were studied to determine their susceptibilities to β-lactam antibiotics. Among these isolates, 16 were methicillin-sensitive S. aureus (MSSA), 207 were methicillin-resistant S. aureus (MRSA) and 27 were borderline oxacillin-resistant S. aureus (BORSA). Currently, the reported mechanism of methicillin resistance in S. aureus is the production of a distinctive penicillin binding protein 2a (PBP2a), which exhibits low affinity toward β-lactams. A surface plasmon resonance biosensor was evaluated for its ability to identify MRSA and to distinguish these strains from MSSA and BORSA, by specifically detecting PBP2a. We found that the system permits label-free, real-time, specific detection of pathogens for concentrations as low as 10 colony forming units/milliliter (CFU/ml), in less than 20 min. This system promises to become a diagnostic tool for bacteria that cause major public concern in clinical settings.
Robinson, Joan L; Salvadori, Marina I
Uncomplicated skin abscesses in previously well children are typically managed with drainage alone. An increasing percentage of such abscesses are due to methicillin-resistant Staphylococcus aureus infections. Although definitive data are lacking, drainage alone appears to be a reasonable strategy for methicillin-resistant S aureus skin abscesses, with antibiotics reserved for infants younger than three months of age, or for children who are systemically unwell, have underlying medical problems or have significant surrounding cellulitis. PMID:22294871
Ciftci, Alper; Findik, Arzu; Onuk, Ertan Emek; Savasan, Serap
This study aimed to detect methicillin resistant and slime producing Staphylococcus aureus in cases of bovine mastitis. A triplex PCR was optimized targetting 16S rRNA, nuc and mecA genes for detection of Staphylococcus species, S. aureus and methicillin resistance, respectively. Furthermore, for detection of slime producing strains, a PCR assay targetting icaA and icaD genes was performed. In this study, 59 strains were detected as S. aureus by both conventional tests and PCR, and 13 of them were found to be methicillin resistant and 4 (30.7%) were positive for mecA gene. Although 22 of 59 (37.2%) S. aureus isolates were slime-producing in Congo Red Agar, in PCR analysis only 15 were positive for both icaA and icaD genes. Sixteen and 38 out of 59 strains were positive for icaA and icaD gene, respectively. Only 2 of 59 strains were positive for both methicillin resistance and slime producing, phenotypically, suggesting lack of correlation between methicillin resistance and slime production in these isolates. In conclusion, the optimized triplex PCR in this study was useful for rapid and reliable detection of methicillin resistant S. aureus. Furthermore, only PCR targetting icaA and icaD may not sufficient to detect slime production and further studies targetting other ica genes should be conducted for accurate evaluation of slime production characters of S. aureus strains. PMID:24031354
Crombé, Florence; Argudín, M. Angeles; Vanderhaeghen, Wannes; Hermans, Katleen; Haesebrouck, Freddy; Butaye, Patrick
From the mid-2000s on, numerous studies have shown that methicillin-resistant Staphylococcus aureus (MRSA), renowned as human pathogen, has a reservoir in pigs and other livestock. In Europe and North America, clonal complex (CC) 398 appears to be the predominant lineage involved. Especially worrisome is its capacity to contaminate humans in close contact with affected animals. Indeed, the typical multi-resistant phenotype of MRSA CC398 and its observed ability of easily acquiring genetic material suggests that MRSA CC398 strains with an increased virulence potential may emerge, for which few therapeutic options would remain. This questions the need to implement interventions to control the presence and spread of MRSA CC398 among pigs. MRSA CC398 shows a high but not fully understood transmission potential in the pig population and is able to persist within that population. Although direct contact is probably the main route for MRSA transmission between pigs, also environmental contamination, the presence of other livestock, the herd size, and farm management are factors that may be involved in the dissemination of MRSA CC398. The current review aims at summarizing the research that has so far been done on the transmission dynamics and risk factors for introduction and persistence of MRSA CC398 in farms. PMID:23518663
Chen, Yi-Wen; Wang, Hong; Hupert, Mateusz; Soper, Steven A
Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-acquired (HA-MRSA) infection worldwide. As a result, the rapid and specific detection of MRSA is crucial not only for early prevention of disease spread, but also for the effective treatment of these infections. We report here an integrated modular-based microfluidic system for MRSA identification, which can carry out the multi-step assay used for MRSA identification in a single disposable fluidic cartridge. The multi-step assay included PCR amplification of the mecA gene harboring methicillin resistance loci that can provide information on drug susceptibility, ligase detection reaction (LDR) to generate fluorescent ligation products appended with a zip-code complement that directs the ligation product to a particular address on a universal array containing zip-code probes and a universal DNA array, which consisted of a planar waveguide for evanescent excitation. The fluidic cartridge design was based on a modular format, in which certain steps of the molecular processing pipeline were poised on a module made from a thermoplastic. The cartridge was comprised of a module interconnected to a fluidic motherboard configured in a 3-dimensional network; the motherboard was made from polycarbonate, PC, and was used for PCR and LDR, while the module was made from poly(methylmethacrylate), PMMA, and contained an air-embedded waveguide serving as the support for the universal array. Fluid handling, thermal management and optical readout hardware were situated off-chip and configured into a small footprint instrument. In this work, the cartridge was used to carry out a multiplexed PCR/LDR coupled with the universal array allowed for simultaneous detection of five genes that encode for 16S ribosomal RNA (SG16S), protein A (spa), the femA protein of S. epidermidis (femA), the virulence factor of Panton-Valentine leukocidin (PVL) and the gene that confers methicillin resistance (mecA). Results
Tesch, W; Strässle, A; Berger-Bächi, B; O'Hara, D; Reynolds, P; Kayser, F H
A 6.2-kilobase chromosomal DNA fragment from a methicillin-resistant Staphylococcus epidermidis strain was cloned into Staphylococcus carnosus by using staphylococcal plasmid pCA44 as the vector. The recombinant plasmid obtained, pBBB21, conferred methicillin resistance on its host and was responsible for the synthesis of a low-affinity penicillin-binding protein (PBP), PBP 2'. PBP 2' determined by the S. epidermidis DNA and expressed as a membrane-bound PBP in S. carnosus reacted with monoclonal antibodies directed against PBP 2' of Staphylococcus aureus origin, and the cloned S. epidermidis DNA hybridized to the methicillin (mec)-specific DNA from S. aureus. These findings point to a common origin of the methicillin resistance determinant in staphylococci. Images PMID:2903715
Green, Christopher F; Elbe, Laura A; Neal, Tyler D; Lowe, John J; Gibbs, Shawn G
Antibiotic misuse and overuse in both the healthcare and agricultural fields have dramatically increased the prevalence of antibiotic resistance in human pathogens. Two strains of methicillin-resistant Staphylococcus aureus (MRSA) (ATCC 43330 and a wild-type) and 1 strain of methicillin-susceptible S. aureus (ATCC 25923) were challenged (9 runs in triplicate) in a preliminary study with ultraviolet germicidal irradiation (UVGI) doses ranging from 0.25 to 3.00 mJ/cm(2). The mean percent kill was calculated for each strain when compared with the control plates (no exposure to UVGI). Then, each strain was challenged (22 runs in triplicate) with UVGI doses of 2.00, 2.50, and 3.00 mJ/cm(2). The results suggest a difference between the doses required to disinfect surfaces with each strain. Assuming a standard error rate of α = 0.05, there was a significant difference in variance between the MRSA (ATCC 43330 and wild type) strains and the S. aureus (ATCC 25923) methicillin-susceptible strain.
Yasuda, T; Yoshimura, S; Katsuno, Y; Takada, H; Ito, M; Takahashi, M; Yahazaki, F; Iriyama, J; Ishigo, S; Asano, Y
Various disinfectants were compared in terms of the duration of bactericidal activity against methicillin-sensitive Staphylococcus aureus (MSSA), and methicillin-resistant Staphylococcus aureus (MRSA), among S. aureus isolated in our hospital. Strains of S. aureus which showed minimum inhibitory concentrations of cloxacillin of less than 1.56 micrograms/ml and of 3.13 micrograms/ml or higher were designated MSSA and MRSA respectively. There was no difference in sensitivity to disinfectants between MSSA and MRSA. There was a great variation in the duration of bactericidal activity of chlorhexidine gluconate against these species with the majority requiring contact times of between 2 minutes and over 20 minutes. All strains except for one strain of MRSA were killed within 20 seconds after disinfection with benzalkonium chloride. All strains were killed within 20 seconds after disinfection with alkyldiaminoethylglycine hydrochloride or povidone-iodine.
Pournajaf, Abazar; Ardebili, Abdollah; Goudarzi, Leyla; Khodabandeh, Mahmoud; Narimani, Tahmineh; Abbaszadeh, Hassan
Objective To evaluated the PCR for mecA gene compared with the conventional oxacillin disk diffusion method for methicillin-resistant Staphylococcus aureus (S. aureus) identification. Methods A total of 292 S. aureus strains were isolated from various clinical specimens obtained from hospitalized patients. Susceptibility test to several antimicrobial agents was performed by disk diffusion agar according to Clinical and Laboratory Standards Institute guidelines. The PCR amplification of the mecA gene was carried out in all the clinical isolates. Results Among antibiotics used in our study, penicillin showed the least anti-staphylococcal activity and vancomycin was the most effective. The rate of methicillin-resistant S. aureus prevalence determined by oxacillin disk diffusion method was 47.6%; whereas, 45.1% of S. aureus isolates were mecA- positive in the PCR assay. Conclusions This study is suggestive that the PCR for detection of mecA gene is a fast, accurate and valuable diagnostic tool, particularly in hospitals in areas where methicillin-resistant S. aureus is endemic. PMID:25183100
Cherian, Joel; Singh, Rahul; Varma, Muralidhar; Vidyasagar, Sudha; Mukhopadhyay, Chiranjay
Pyogenic liver abscesses are rare with an incidence of 0.5% to 0.8% and are mostly due to hepatobiliary causes (40% to 60%). Most are polymicrobial with less than 10% being caused by Staphylococcus aureus. Of these, few are caused by methicillin-resistant Staphylococcus aureus (MRSA) and fewer still by a community-acquired strain. Here we present a case study of a patient with a community-acquired MRSA liver abscess. The patient presented with fever since 1 month and tender hepatomegaly. Blood tests revealed elevated levels of alkaline phosphatase, C-reactive protein, erythrocyte sedimentation rate, and neutrophilic leukocytosis. Blood cultures were sterile. Ultrasound of the abdomen showed multiple abscesses, from which pus was drained and MRSA isolated. Computed tomography of the abdomen did not show any source of infection, and an amebic serology was negative. The patient was started on vancomycin for 2 weeks, following which he became afebrile and was discharged on oral linezolid for 4 more weeks. Normally a liver abscess is treated empirically with ceftriaxone for pyogenic liver abscess and metronidazole for amebic liver abscess. However, if the patient has risk factors for a Staphylococcal infection, it is imperative that antibiotics covering gram-positive organisms be added while waiting for culture reports. PMID:27540556
Soong, Grace; Paulino, Franklin; Wachtel, Sarah; Parker, Dane; Wickersham, Matthew; Zhang, Dongni; Brown, Armand; Lauren, Christine; Dowd, Margaret; West, Emily; Horst, Basil; Planet, Paul
ABSTRACT Skin is the most common site of Staphylococcus aureus infection. While most of these infections are self-limited, recurrent infections are common. Keratinocytes and recruited immune cells participate in skin defense against infection. We postulated that S. aureus is able to adapt to the milieu within human keratinocytes to avoid keratinocyte-mediated clearance. From a collection of S. aureus isolated from chronically infected patients with atopic dermatitis, we noted 22% had an agr mutant-like phenotype. Using several models of human skin infection, we demonstrate that toxin-deficient, agr mutants of methicillin-resistant S. aureus (MRSA) USA300 are able to persist within keratinocytes by stimulating autophagy and evading caspase-1 and inflammasome activation. MRSA infection induced keratinocyte autophagy, as evidenced by galectin-8 and LC3 accumulation. Autophagy promoted the degradation of inflammasome components and facilitated staphylococcal survival. The recovery of more than 58% agr or RNAIII mutants (P < 0.0001) of an inoculum of wild-type (WT) MRSA from within wortmannin-treated keratinocytes compared to control keratinocytes reflected the survival advantage for mutants no longer expressing agr-dependent toxins. Our results illustrate the dynamic interplay between S. aureus and keratinocytes that can result in the selection of mutants that have adapted specifically to evade keratinocyte-mediated clearance mechanisms. PMID:25900653
Okuda, Ken-ichi; Zendo, Takeshi; Sugimoto, Shinya; Iwase, Tadayuki; Tajima, Akiko; Yamada, Satomi; Sonomoto, Kenji; Mizunoe, Yoshimitsu
Control of biofilms formed by microbial pathogens is an important subject for medical researchers, since the development of biofilms on foreign-body surfaces often causes biofilm-associated infections in patients with indwelling medical devices. The present study examined the effects of different kinds of bacteriocins, which are ribosomally synthesized antimicrobial peptides produced by certain bacteria, on biofilms formed by a clinical isolate of methicillin-resistant Staphylococcus aureus (MRSA). The activities and modes of action of three bacteriocins with different structures (nisin A, lacticin Q, and nukacin ISK-1) were evaluated. Vancomycin, a glycopeptide antibiotic used in the treatment of MRSA infections, showed bactericidal activity against planktonic cells but not against biofilm cells. Among the tested bacteriocins, nisin A showed the highest bactericidal activity against both planktonic cells and biofilm cells. Lacticin Q also showed bactericidal activity against both planktonic cells and biofilm cells, but its activity against biofilm cells was significantly lower than that of nisin A. Nukacin ISK-1 showed bacteriostatic activity against planktonic cells and did not show bactericidal activity against biofilm cells. Mode-of-action studies indicated that pore formation leading to ATP efflux is important for the bactericidal activity against biofilm cells. Our results suggest that bacteriocins that form stable pores on biofilm cells are highly potent for the treatment of MRSA biofilm infections.
Chanchaithong, Pattrarat; Prapasarakul, Nuvee
This study aimed to detect inducible clindamycin (iCLI) resistance in Staphylococcus pseudintermedius isolated from dogs in Thailand using D-zone testing. Strains that were iCLI-resistant were characterized by molecular typing and antibiogram and were detected in 10/200 S. pseudintermedius isolates (5%) from 7/41 dogs (17%). All were methicillin-resistant S. pseudintermedius (MRSP) and demonstrated multidrug resistance. The iCLI-resistant MRSP contained erm(B) and had identical or closely related DNA fingerprint patterns by pulsed-field gel electrophoresis. All iCLI-resistant MRSP strains belonged to the same clonal complex 112 (sequence types 111 and 112) by multilocus sequence typing. To avoid misinterpretation of clindamycin susceptibility, D-zone testing is recommended to promote rational antimicrobial selection and limit the clonal expansion of multidrug resistant bacteria.
Oteo, Jesús; Belén Aracil, María
Multi-drug resistance in bacterial pathogens increases morbidity and mortality in infected patients and it is a threat to public health concern by their high capacity to spread. For both reasons, the rapid detection of multi-drug resistant bacteria is critical. Standard microbiological procedures require 48-72 h to provide the antimicrobial susceptibility results, thus there is emerging interest in the development of rapid detection techniques. In recent years, the use of selective and differential culture-based methods has widely spread. However, the capacity for detecting antibiotic resistance genes and their low turnaround times has made molecular methods a reference for diagnosis of multidrug resistance. This review focusses on the molecular methods for detecting some mechanisms of antibiotic resistance with a high clinical and epidemiological impact: a) Enzymatic resistance to broad spectrum β-lactam antibiotics in Enterobacteriaceae, mainly extended spectrum β-lactamases (ESBL) and carbapenemases; and b) methicillin resistance in Staphylococcus aureus.
Brief report Longitudinal characterization of Acinetobacter baumannii-calcoaceticus complex, Klebsiella pneumoniae , and methicillin-resistant...resistant Acinetobacter baumannii-calcoaceticus complex Klebsiella pneumoniae Methicillin-resistant Staphylococcus aureus MRSA Drug-resistant...Acinetobacter baumannii-calcoaceticus complex, Klebsiella pneumoniae , and methicillin- resistant Staphylococcus aureus colonize and infect combat casualties
Ganesan, Anuradha; Crawford, Katrina; Mende, Katrin; Murray, Clinton K; Lloyd, Bradley; Ellis, Michael; Tribble, David R; Weintrob, Amy C
A total of 102 methicillin-resistant Staphylococcus aureus (MRSA) isolates collected from 50 injured service members (June 2009 to December 2011) at U.S. military treatment facilities were analyzed for the conventional mecA gene and mecC homologue by using standard PCR-based methods. The prevalence of the mecC homologue was zero.
Paludi, D; Vergara, A; Festino, A R; Di Ciccio, P; Costanzo, C; Conter, M; Zanardi, E; Ghidini, S; Ianieri, A
There is increasing concern about the impact on public health of methicillin-resistant Staphylococcus aureus (MRSA) associated with animal food products. MRSA remains a serious problem because of the high incidence and multidrug resistance of the strains, even for strains isolated from foods, food environments and food handlers. The objectives of this study are: (i) to evaluate the susceptibility of S. aureus strains isolated from food, food handlers and food-processing environments to 14 antibiotics currently used in veterinary and human therapy; (ii) to assess the presence of the mecA gene. A total of 1007 samples were collected from food, food handlers, and environments and were analyzed for the presence of S. aureus. S. aureus was present in 165 of the 1007 samples. A total of 157 isolates were methicillin-susceptible S. aureus (MSSA) and 8 isolates were MRSA. In particular, out of 8 MRSA strains detected, 4 strains harboured the mecA gene. All MRSA strains were resistant to at least one of the tested antibiotics and 6 strains demonstrated multi-resistance. Considering the high level of resistances in S. aureus and the isolation of MRSA strains, the surveillance of antimicrobial resistance and the spreading of this pathogen is of crucial importance in the food production chain. These data are useful in improving background data on antimicrobial resistance of S. aureus isolated from food, processing environments and food handlers, supporting the prudent use of antibiotics and the development of international control programs.
The presence of antibiotic resistant bacteria and antibiotic resistance genes in rural sewage treatment plants are not well reported in the literature. The aim of the present study was to study the frequency occurrence of Methicillin Resistant Staphylococcus aureus (MRSA) in a rural sewage treatment plant. This study was conducted using raw sewage as well as treated sewage from a small town sewage treatment plant in rural southeast Louisiana of USA. Results showed the presence of MRSA consistently in both raw and treated sewage. The presence of mecA gene responsible for methicillin resistance was confirmed in the raw and treated sewage water samples.
Diederen, Bram; van Duijn, Inge; van Belkum, Alex; Willemse, Piet; van Keulen, Peter; Kluytmans, Jan
CHROMagar MRSA was evaluated for its ability to identify methicillin-resistant Staphylococcus aureus (MRSA). A well-defined collection consisting of 216 MRSA strains and 241 methicillin-susceptible Staphylococcus aureus isolates was used. The sensitivity of CHROMagar MRSA after 24 h of incubation was 95.4%, increasing to 100% after 48 h. The specificity was already 100% after 24 h. PMID:15815020
Liu, Xiaoyu; Zhang, Shijie
Increasing cases of infections caused by methicillin-resistant Staphylococcus aureus (MRSA) strains in healthy individuals have raised concerns worldwide. MRSA strains are resistant to almost the entire family of β-lactam antibiotics due to the acquisition of an extra penicillin-binding protein, PBP2a. Studies have shown that spoVG is involved in oxacillin resistance, while the regulatory mechanism remains elusive. In this study, we have found that SpoVG plays a positive role in oxacillin resistance through promoting cell wall synthesis and inhibiting cell wall degradation in MRSA strain N315. Deletion of spoVG in strain N315 led to a significant decrease in oxacillin resistance and a dramatic increase in Triton X-100-induced autolytic activity simultaneously. Real-time quantitative reverse transcription-PCR revealed that the expression of 8 genes related to cell wall metabolism or oxacillin resistance was altered in the spoVG mutant. Electrophoretic mobility shift assay indicated that SpoVG can directly bind to the putative promoter regions of lytN (murein hydrolase), femA, and lytSR (the two-component system). These findings suggest a molecular mechanism in which SpoVG modulates oxacillin resistance by regulating cell wall metabolism in MRSA. PMID:27001809
Costa, Dayane de Melo; Kipnis, André; Leão-Vasconcelos, Lara Stefânia Netto de Oliveira; Rocha-Vilefort, Larissa Oliveira; Telles, Sheila Araújo; André, Maria Cláudia Dantas Porfírio Borges; Tipple, Anaclara Ferreira Veiga; Lima, Ana Beatriz Mori; Ribeiro, Nádia Ferreira Gonçalves; Pereira, Mayara Regina; Prado-Palos, Marinésia Aparecida
The aim of the study was to analyze epidemiological and microbiological aspects of oral colonization by methicillin-resistant Staphylococcus of health care workers in a cancer hospital. Interview and saliva sampling were performed with 149 health care workers. Antimicrobial resistance was determined by disk diffusion and minimum inhibitory concentration. Polymerase Chain Reaction, Internal Transcribed Spacer-Polymerase Chain Reaction and Pulsed Field Gel Electrophoresis were performed for genotypic characterization of methicillin-resistant Staphylococcus. Risk factors were determined by logistic regression. Methicillin-resistant Staphylococcus colonization prevalence was 19.5%, denture wearing (p = 0.03), habit of nail biting (p = 0.04) and preparation and administration of antimicrobial (p = 0.04) were risk factors identified. All methicillin-resistant Staphylococcus were S. epidermidis, 94.4% of them had mecA gene. Closely related and indistinguishable methicillin-resistant S. epidermidis were detected. These results highlight that HCWs which have contact with patient at high risk for developing infections were identified as colonized by MRSE in the oral cavity, reinforcing this cavity as a reservoir of these bacteria and the risk to themselves and patients safety, because these microorganisms may be spread by coughing and talking.
Moon, Bo Youn; Youn, Jung-Ho; Shin, Sook; Hwang, Sun Young; Park, Yong Ho
Staphylococci were isolated from veterinary staff, hospitalized animals, and medical equipment from 2 major tertiary veterinary hospitals in South Korea to investigate antimicrobial resistance and genetic relatedness. The detection rate for staphylococci was 55.2% (111/201 samples), and 11 species were identified among the collected staphylococcal strains. The most prevalent species were Staphylococcus pseudintermedius (52/111, 46.8%), Staphylococcus epidermidis (21/111, 18.9%), and Staphylococcus aureus (19/111, 17.1%). The methicillin-resistance rates of staphylococci isolated from veterinary staff and medical equipment were higher than those from hospitalized animals. The genotype of methicillin-resistant S. aureus (MRSA) strains in the current study was sequence type (ST)72-SCCmec IVc-t324, which is similar to the genotype of prevalent MRSA strains in human beings and food animals in South Korea. Among the mecA-positive S. pseudintermedius isolates, SCCmec V was most prevalent in strains originating from both veterinary staff and hospitalized animals. SCCmec IVa was detected in methicillin-resistant S. epidermidis, whereas SCCmec IVc was found in other methicillin-resistant, coagulase-negative staphylococci. The SCCmec typing, antimicrobial susceptibility tests, and pulsed field gel electrophoresis results showed that methicillin-resistant staphylococci dissemination between hospitalized animals and veterinary staff is possible in South Korean veterinary hospitals.
Rose-Martel, Megan; Kulshreshtha, Garima; Ahferom Berhane, Nahom; Jodoin, Joelle; Hincke, Maxwell T.
Staphylococcus aureus, a human pathogen associated with many illnesses and post-surgical infections, can resist treatment due to the emergence of antibiotic-resistant strains and through biofilm formation. The current treatments for chronic biofilm infections are antibiotics and/or surgical removal of the contaminated medical device. Due to higher morbidity and mortality rates associated with overuse/misuse of antibiotics, alternate treatments are essential. This study reports the antibiofilm activity of avian erythrocyte histones against methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA). Fluorescence and scanning electron microscopy revealed membrane damage to bacteria in histone-treated biofilms. Histones and indolicidin (positive control) increased the expression of apsS and apsR, which are associated with the Antimicrobial Peptide (AMP) sensor/regulator system in S. aureus. The expression of dltB, and vraF, associated with AMP resistance mechanisms, were under histone inducible control in the biofilm-embedded bacterial cells. The time kill kinetics for histones against S. aureus revealed a rapid biocidal activity (<5 min). Purified erythrocyte-specific histone H5 possessed 3–4 fold enhanced antimicrobial activity against planktonic cells compared to the histone mixture (H1, H2A, H2B, H3, H4, H5). These results demonstrate the promise of histones and histone-like derivatives as novel antibiotics against pathogens in their planktonic and biofilm forms. PMID:28378802
Shahkarami, Fatemeh; Rashki, Ahmad; Rashki Ghalehnoo, Zahra
Background: Today, significant increase in the prevalence and emergence of methicillin-resistant Staphylococcus aureus (MRSA) is a serious public health concern and is likely to have a dramatic negative impact on many current medical practices. Therefore, identification of MRSA strains is important for both clinical and epidemiological implications. Objectives: The present study was carried out to determine the frequency of methicillin resistant; antibiotic susceptibility and plasmid profiles of S. aureus recovered from different types of clinical samples of patients in Zabol, Iran. Material and Methods: Clinical samples from 500 outpatient and hospitalized patients were tested for S. aureus. The susceptibility of 106 S. aureus to 11 antibiotics was evaluated by the disk diffusion method and Etest oxacillin strips. The presence of mecA gene was investigated by polymerase chain reaction (PCR). The plasmid profile patterns of all isolates were determined by a modified alkaline lysis method. Results: A total of 67 (63.20%) strains were found to be MRSA isolates. Most of MRSA isolates showed high level of resistance to ampicillin, erythromycin, nalidixic acid, penicillin, and tetracycline. Twenty-six percent of MRSA isolates showed high level of resistance to oxacillin (minimum inhibitory concentration [MIC] ≥ 256 μg/mL). mecA gene was detected among 62 MRSA isolates. Totally, 75 isolates of both strains harbored plasmid. Conclusions: Resistance to oxacillin and other antibiotics was high, and most of the isolates were found to be multi-drug resistance (MDR). Plasmid analysis of representative S. aureus isolates also demonstrates the presence of a wide range of plasmid sizes, with no consistent relationship between plasmid profiles and resistance phenotypes. Regular surveillance of hospital infections and monitoring of their antibiotic sensitivity patterns are required to reduce MRSA prevalence. High prevalence and multi-drug resistance of MRSA isolates in southeast
Fungwithaya, Punpichaya; Chanchaithong, Pattrarat; Phumthanakorn, Nathita; Prapasarakul, Nuvee
This study aimed to investigate the nasal carriage of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in dogs treated with oral cephalexin monohydrate. Ten dogs with superficial pyoderma were monitored longitudinally for carriage of MRSP for up to 1 year after treatment; the strains were typed and antibiograms were determined. Methicillin-susceptible S. pseudintermedius (MSSP) was recovered prior to treatment in all dogs and could be isolated after 12 months in 1 dog. Methicillin-resistant Staphylococcus pseudintermedius was detected within 1 week of treatment in all dogs, and 3 clones represented by ST45, ST112, and ST181 were consistently present for up to 12 months after treatment. All MRSP isolates were resistant to at least 7 common antimicrobials. Oral cephalexin monohydrate treatment selected for strains of multi-resistant MRSP, which were still present after 1 year.
Laarhoven, Laura M; de Heus, Phebe; van Luijn, Jeanine; Duim, Birgitta; Wagenaar, Jaap A; van Duijkeren, Engeline
Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is an emerging pathogen in dogs and has been found in Europe, Asia and North America. To date most studies are one-point prevalence studies and therefore little is known about the dynamics of MRSP in dogs and their surrounding. In this longitudinal study MRSP colonization in dogs and the transmission of MRSP to humans, contact animals and the environment was investigated. Sixteen dogs with a recent clinical MRSP infection were included. The index dogs, contact animals, owners and environments were sampled once a month for six months. Samples taken from the nose, perineum and infection site (if present) of the index cases and contact animals, and the nares of the owners were cultured using pre-enrichment. Index cases were found positive for prolonged periods of time, in two cases during all six samplings. In five of the 12 households that were sampled during six months, the index case was intermittently found MRSP-positive. Contact animals and the environment were also found MRSP-positive, most often in combination with a MRSP-positive index dog. In four households positive environmental samples were found while no animals or humans were MRSP-positive, indicating survival of MRSP in the environment for prolonged periods of time. Genotyping revealed that generally similar or indistinguishable MRSP isolates were found in patients, contact animals and environmental samples within the same household. Within two households, however, genetically distinct MRSP isolates were found. These results show that veterinarians should stay alert with (former) MRSP patients, even after repeated MRSP-negative cultures or after the disappearance of the clinical infection. There is a considerable risk of transmission of MRSP to animals in close contact with MRSP patients. Humans were rarely MRSP-positive and never tested MRSP-positive more than once suggesting occasional contamination or rapid elimination of colonization of
Schelli, Katie; Rutowski, Joshua; Roubidoux, Julia; Zhu, Jiangjiang
Recently, novel bioanalytical methods, such as NMR and mass spectrometry based metabolomics approaches, have started to show promise in providing rapid, sensitive and reproducible detection of Staphylococcus aureus antibiotic resistance. Here we performed a proof-of-concept study focused on the application of HPLC-MS/MS based targeted metabolic profiling for detecting and monitoring the bacterial metabolic profile changes in response to sub-lethal levels of methicillin exposure. One hundred seventy-seven targeted metabolites from over 20 metabolic pathways were specifically screened and one hundred and thirty metabolites from in vitro bacterial tests were confidently detected from both methicillin susceptible and methicillin resistant Staphylococcus aureus (MSSA and MRSA, respectively). The metabolic profiles can be used to distinguish the isogenic pairs of MSSA strains from MRSA strains, without or with sub-lethal levels of methicillin exposure. In addition, better separation between MSSA and MRSA strains can be achieved in the latter case using principal component analysis (PCA). Metabolite data from isogenic pairs of MSSA and MRSA strains were further compared without and with sub-lethal levels of methicillin exposure, with metabolic pathway analyses additionally performed. Both analyses suggested that the metabolic activities of MSSA strains were more susceptible to the perturbation of the sub-lethal levels of methicillin exposure compared to the MRSA strains.
Mikołajczyk, Dorota; Kaczmarek, Agnieszka; Budzytiska, Anna; Gospodarek, Eugenia
The aim of this study was to determine resistance to co-trimoxazole among 476 strains of methicillin-resistant Staphylococus aureus (MRSA) and 2137 strains of methicillin resistant coagulase-negative staphylococci (MRCNS) isolation during three years (2001-2003). Their susceptibility testing were performed by the disc-diffusion techniques according to recomendation of NCCLS (National Committe for Clinical Laboratory Standards). Co-trimoxazole resistance was demonstrated more frequently among MRCNS (52,8%, 51,4%, 63,9% in 2001, 2002, 2003), than among MRSA (23,4%, 19,5%, 16,8% adequately).
Riley, Matthew C.; Bemis, David A.
We report the first complete genome sequences of three predominant clones (ST68, ST71, and ST84) of methicillin-resistant Staphylococcus pseudintermedius in North America. All strains were isolated from canine infections and have different SCCmec elements and antibiotic resistance gene patterns. PMID:27795289
Riley, Matthew C; Perreten, Vincent; Bemis, David A; Kania, Stephen A
We report the first complete genome sequences of three predominant clones (ST68, ST71, and ST84) of methicillin-resistant Staphylococcus pseudintermedius in North America. All strains were isolated from canine infections and have different SCCmec elements and antibiotic resistance gene patterns.
Kumar, Vasudevan Anil; Nair, Nisha; Thachathodiyl, Rajesh; Nandakumar, Aswathy; Dinesh, Kavitha R; Thatcher, Eileen; Karim, Shamsul; Biswas, Raja
Though pericardial disease is common in patients with renal disease, purulent pericarditis is very rare. We report a fatal case of purulent pericarditis and sepsis due to methicillin-resistant Staphylococcus aureus in a 78-year-old male with systemic hypertension and renal disease along with the molecular characterization of its resistant mechanism. PMID:24701111
Bhattacharya, Susmita; Jain, Sonia; Chatterjee, Shiv Sekhar; Konar, Jayashree
Introduction Surgical Site Infection (SSI) is the most common healthcare associated infection that could be averted by antibiotics prophylaxis against the probable offending organisms. As Staphylococcus aureus has been playing a substantial role in the aetiology of SSIs, Methicillin Resistant Staphylococcus aureus (MRSA) happens to be a problem while dealing with the postoperative wound infection. Aim To determine the prevalence of SSI caused by MRSA and the antibiotic sensitivity pattern of MRSA. Materials and Methods A cross-sectional study was conducted at Nil Ratan Sircar Medical College, Kolkata, West Bengal from July 2009 to December 2012. A total of 19,359 surgical procedures were done of which 3003 culture positive SSIs have been documented. The clinical samples were collected from patients of both sexes and all ages suspected to be suffering from SSI from different specialities. Samples were processed according to CLSI, 2007 guidelines. The isolated strains of Staphylococcus aureus were screened for MRSA by detection of resistance to Cefoxitin disc (zone of inhibition was ≤21 mm) and slidex staph latex agglutination tests were done on cefoxitin resistant strains to spot phenotypic expression of mec A gene. Then PCR was performed for detection of mecA gene. Antibiotic sensitivity test was done following Kirby Bauer technique. Results In this 3½ year study, 1049 Staphylococcus aureus (34.93%) were reported from 3003 cases of SSI followed by Escherichia coli (20.34%), Klebsiella spp. (18.08%), Pseudomonas spp. (7.99%), Acinetobacter spp. (7.49%) respectively. Among the Staphylococcus aureus, 267 strains were derived as MRSA (25.45%). MRSA were isolated from 167 (62.54%) male patients and 100 (37.45%) female patients having surgical site infections. Inpatients and outpatients distribution of MRSA were 235 (88.01%) and 32 (11.98%) respectively. Majority of the MRSA cases were reported from Surgery (12.49%) and Orthopaedics (11.85%) departments in the age
Infection is a common complication of chronic wounds that delays healing. Community-acquired methicillin-resistant Staphylococcus aureus has emerged as a common pathogen and major impediment to healing affected chronic wounds. Community-acquired methicillin-resistant S aureus is virulent, highly communicable, and difficult to eradicate. Treatment options include incision and drainage, debridement, and systemic antimicrobials. Early aggressive wound management and appropriate antibiotic therapy are considered essential to successful treatment. Facility-specific protocols should be developed to minimize the spread of this organism to the general population, with particular attention focused on protecting patients burdened with chronic wounds. This article reviews current knowledge of community-acquired methicillin-resistant S aureus, focusing on its impact on persons with chronic wounds.
Kang, Min-Hee; Chae, Min-Joo; Yoon, Jang-Won; Lee, So-Young; Yoo, Jong-Hyun; Park, Hee-Myung
Resistance to fluoroquinolones and methicillin was determined for 49 ophthalmic isolates of Staphylococcus pseudintermedius from dogs with and without ophthalmic disease. Resistance was observed for ciprofloxacin (40.8%), ofloxacin (38.8%), enrofloxacin (38.8%), levofloxacin (34.7%), and moxifloxacin (4.1%). Eighteen isolates, 16 of which were resistant to oxacillin, were mecA-positive. Nine of the 16 oxacillin-resistant mecA-positive S. pseudintermedius isolates were resistant to more than one fluoroquinolone and 2 isolates were resistant to 5 fluoroquinolones. The frequency of mecA gene occurrence and fluoroquinolone resistance was twice as high among S. pseudintermedius isolates derived from dogs with ophthalmic disease compared with isolates for dogs without ophthalmic disease. The high prevalence of methicillin and fluoroquinolone resistance in S. pseudintermedius from dogs with ophthalmic disease is a concern.
Wang, Dengfeng; Wang, Zhicai; Yan, Zuoting; Wu, Jianyong; Ali, Tariq; Li, Jianjun; Lv, Yanli; Han, Bo
The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infection in dairy animals is of great concern for livestock and public health. The aim of present study was to detect new trends of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) towards antibiotic susceptibility, resistance genes and molecular typing by methods of disc diffusion, multiplex PCR assay and multilocus sequence typing (MLST). A total of 219 S. aureus strains were isolated from bovine mastitis cases from six provinces of China, including 34 MRSA strains. The results revealed that more than 70% isolated strains showed resistance to various antibiotics, and multiple-drugs resistance to more than five categories of antibiotics was found more common. The ermC was the most prevalent resistance gene, followed by other genes; however, ermA was the least frequently detected gene. Twenty-eight mecA-negative MRSA and six mecA-positive MRSA strains were detected, and in which three strains were ST97-MRSA-IV, others were ST965-MRSA-IV, ST6-MRSA-IV and ST9-MRSA-SCCmec-NT. The mecA-negative MRSA strains were found resistant to most of the antibiotics, and harbored aac(6')/aph(2''), aph(3')-III and tetM genes higher than MSSA strains. The resistance to most of the antibiotics was significantly higher in MRSA than in MSSA strains. The MLST profiles showed that these strains mainly belonged to CC5, CC398, CC121 and CC50 lineage, especially within ST97 and ST398, while some novel sequence types (ST2154, ST2165 and ST2166) were identified and deposited in the MLST database. This indicates that the resistance of S. aureus is becoming more complicated by changes in multi-drug resistance mechanism and appearance of mecA-negative MRSA isolates, and importantly, MRSA-IV strains in different MLST types are emerging.
Eftekhar, Fereshteh; Raei, Fereshteh
Staphylococcus epidermidis is the most important member of coagulase negative staphylococci responsible for community and hospital acquired infections. Most clinical isolates of S. epidermidis are resistant to methicillin making these infections difficult to treat. In this study, correlation of methicillin resistance phenotype was compared with methicillin resistance (mecA) gene carriage in 55 clinical isolates of S. epidermidis. Susceptibility was measured by disc diffusion using methicillin discs, and minimum inhibitory concentrations (MIC) were measured using broth microdilution. Methicillin resistance gene (MecA gene) carriage was detected by specific primers and PCR. Disc susceptibility results showed 90.9% resistance to methicillin. Considering a MIC of 4 µg/ml, 78.1% of the isolates were methicillin resistant, 76.36% of which carried the mecA gene. On the other hand, when a breakpoint of 0.5 µg/ml was used, 89.09% were methicillin resistant, of which 93.75% were mecA positive. There was a better correlation between MIC of 0.5 µg/ml with disc diffusion results and mecA gene carriage. The findings suggest that despite the usefulness of molecular methods for rapid diagnosis of virulence genes, gene carriage does not necessarily account for virulence phenotype. Ultimately, gene expression, which is controlled by the environment, would determine the outcome.
Bittencourt, Paloma Horejs; Pimentel, Carlos Sidney Silva; Bonfim, Bianca Sampaio; Marostica, Paulo José; Souza, Edna Lúcia
In Brazil the knowledge about methicillin-resistant Staphylococcus aureus infection in cystic fibrosis patients is scarce. This study aimed to determine the incidence of respiratory tract colonization and the identification rates after a standardized treatment. A retrospective cohort was performed highlighting the history of respiratory colonizations between January 2008 and June 2015. Patients under the age of 21 years with cystic fibrosis confirmed by sweat test or genetic study receiving care at the outpatient clinics of a Teaching Hospital were included. The treatment consisted of trimethoprim/sulfamethoxazole, rifampicin, nasal mupirocin and chlorhexidine 2%. The mean follow-up period was of 22.2 months and those with ≥3 negative cultures were considered free of methicillin-resistant Staphylococcus aureus. Forty-two patients were included. Methicillin-resistant Staphylococcus aureus was identified in six patients. Most patients had methicillin-sensitive S. aureus isolation prior to methicillin-resistant Staphylococcus aureus. Five children used the standardized treatment, none presented side effects. Only one child had a new isolation of methicillin-resistant Staphylococcus aureus during follow-up (after 20 months). The incidence of methicillin-resistant Staphylococcus aureus infection was high and occurred in young patients. The therapeutic regimen was effective, safe and being a good option to treat methicillin-resistant Staphylococcus aureus infection.
Jang, Yunho; Bae, Dong hwa; Cho, Jae-Keun; Bahk, Gyung Jin; Lim, Suk-Kyung; Lee, Young Ju
Staphylococci were isolated from dogs in animal hospitals, animal shelters, and the Daegu PET EXPO to investigate the characteristics of circulating methicillin-resistant Staphylococcal (MRS) strains in companion animals in Korea. A total of 36/157 isolates were classified as MRS, and subdivided as follows: 1 methicillin-resistant Staphylococcus aureus (MRSA), 4 methicillin-resistant Staphylococcus epidermidis, 2 methicillin-resistant Staphylococcus haemolyticus, and 29 MRS spp. Among the 36 MRS isolates tested, 100% were resistant to oxacillin and penicillin, and at least 50% were resistant to sulfamethoxazole/trimethoprim (69.4%), erythromycin (63.9%), tetracycline (58.3%), cefoxitin (55.6%), clindamycin (50.0%) or pirlimycin (50.0%). Additionally, 34/36 MRS isolates (94.4%) were mecA positive, 15 of which were further classified as SCCmec type V, 6 isolates as type I, 4 isolates as type IIIb, 1 isolate as type IVa, 1 isolate as type IV, with 7 isolates being non-classifiable. The results of multilocus sequence typing and spa typing for the one MRSA strain were ST 72 (1-4-1-8-4-4-3) and spa t148. Our results provide evidence that companion animals like dogs may be MRS carriers, and that continued surveillance of MRS in companion animals is required to prevent increased incidences in humans.
Wang, Pin-Jia; Xie, Cheng-Bin; Sun, Feng-Hui; Guo, Li-Juan; Dai, Min; Cheng, Xi; Ma, Yong-Xin
Staphylococcus epidermidis, especially methicillin-resistant strains, may be the source of surgical site infections and may be a reservoir of staphylococcal cassette chromosome mec (SCCmec) for S. aureus. The aim of this study was to investigate the prevalence of methicillin-resistant S. epidermidis (MRSE) on the abdominal skin of females before laparotomy and determine the molecular characteristics and antimicrobial susceptibility patterns of these isolates. MRSE was found in 54 of 157 isolates based on mecA gene detection, and there was no difference in icaA gene carriage rate between MRSE and methicillin-susceptible S. epidermidis (MSSE) isolates. Antimicrobial susceptibility profiles were determined by broth microdilution antimicrobial susceptibility testing according to the latest CLSI manuals. All MRSE isolates had unfavorable antimicrobial susceptibility patterns. Twenty-three MRSE strains (42.6%) were multi-drug resistant. SCCmec typing and pulsed field gel electrophoresis (PFGE) typing was performed. Thirty-nine (72.2%) had a single SCCmec type, whereas 1.9% had two types. Fourteen strains (25.9%) were non-typeable (NT). The most frequent MRSE genotype was SCCmec type IVa. High diversity with PFGE patterns was obtained for MRSE, and there were no isolates exhibiting identical pulsotype. The results confirm that methicillin-resistant strains are frequently present among S. epidermidis on the abdominal skin of females before laparotomy. Moreover, resistance profiles seem to have no association with the SCCmec types or PFGE types for most common antibiotics. PMID:27338374
Wang, Pin-Jia; Xie, Cheng-Bin; Sun, Feng-Hui; Guo, Li-Juan; Dai, Min; Cheng, Xi; Ma, Yong-Xin
Staphylococcus epidermidis, especially methicillin-resistant strains, may be the source of surgical site infections and may be a reservoir of staphylococcal cassette chromosome mec (SCCmec) for S. aureus. The aim of this study was to investigate the prevalence of methicillin-resistant S. epidermidis (MRSE) on the abdominal skin of females before laparotomy and determine the molecular characteristics and antimicrobial susceptibility patterns of these isolates. MRSE was found in 54 of 157 isolates based on mecA gene detection, and there was no difference in icaA gene carriage rate between MRSE and methicillin-susceptible S. epidermidis (MSSE) isolates. Antimicrobial susceptibility profiles were determined by broth microdilution antimicrobial susceptibility testing according to the latest CLSI manuals. All MRSE isolates had unfavorable antimicrobial susceptibility patterns. Twenty-three MRSE strains (42.6%) were multi-drug resistant. SCCmec typing and pulsed field gel electrophoresis (PFGE) typing was performed. Thirty-nine (72.2%) had a single SCCmec type, whereas 1.9% had two types. Fourteen strains (25.9%) were non-typeable (NT). The most frequent MRSE genotype was SCCmec type IVa. High diversity with PFGE patterns was obtained for MRSE, and there were no isolates exhibiting identical pulsotype. The results confirm that methicillin-resistant strains are frequently present among S. epidermidis on the abdominal skin of females before laparotomy. Moreover, resistance profiles seem to have no association with the SCCmec types or PFGE types for most common antibiotics.
Kizerwetter-Świda, Magdalena; Chrobak-Chmiel, Dorota; Rzewuska, Magdalena; Binek, Marian
We investigated in vitro activity of a novel veterinary fluoroquinolone, pradofloxacin, against methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolates and compared with other fluoroquinolones. A total of 38 MRSP isolates were subjected to agar disk diffusion tests for sensitivity to pradofloxacin, orbifloxacin, marbofloxacin, enrofloxacin, and ciprofloxacin. The minimal inhibitory concentration (MIC) values of pradofloxacin, ciprofloxacin, and enrofloxacin were determined. Mutations in the genes encoding DNA gyrase subunit A (GyrA) and topoisomerase IV (GrlA) proteins associated with fluoroquinolone resistance were studied by an analysis of partial sequences of the genes encoding these proteins. Two MRSP isolates were susceptible in disk diffusion and microdilution test to all fluoroquinolones tested, including pradofloxacin. Based on the results of the disk diffusion testing, 33 of 38 isolates showed resistance to pradofloxacin and 3 were intermediate, whereas, by pradofloxacin MIC testing, 35 isolates were classified as resistant and 1 as intermediate. Single alterations in GyrA and GrlA proteins were observed in the 35 resistant isolates and the 1 intermediate isolate (MIC results). These same 36 isolates were also resistant to the other tested fluoroquinolones. The results of the current study showed that MRSP isolates are usually resistant to all fluoroquinolones, including pradofloxacin. Therefore, in routine susceptibility testing to pradofloxacin by disk diffusion, the results should be carefully interpreted for MRSP isolates, especially those resistant to other fluoroquinolones and, in questionable cases, the pradofloxacin MIC should be determined to confirm the susceptibility testing results.
Choi, Eun-Jin; Kim, Hyung-Ip; Kim, Ji-Ae; Jun, Soo Youn; Kang, Sang Hyeon; Park, Dong June; Son, Seok-Jun; Kim, Younghoon; Shin, Ok Sarah
The emergence of antibiotic resistant strains such as methicillin-resistant Staphylococcus aureus (MRSA) reminds us an urgent need to develop a new immune-modulating agent for preventing S. aureus infection. In this study, we found that herbal medicines, honokiol and magnolol, caused a significant cellular immune modulatory effect during S. aureus infection. In mouse macrophages, these compounds drove upregulation of an antioxidant effect in response to S. aureus, resulting in a dampened total cellular reactive oxygen species (ROS) production and decreased production of inflammatory cytokines/chemokines, whereas honokiol induced increased types I and III interferon messenger RNA (mRNA) expression levels in response to MSSA infection. Moreover, the internalization of S. aureus by human alveolar epithelial cells was inhibited by these compounds. Furthermore, honokiol and magnolol treatment promoted a delay in killing during MSSA infection in Caenorhabditis elegans, suggesting antimicrobial function in vivo. In conclusion, honokiol and magnolol may be considered as attractive immune-modulating treatment for S. aureus infection.
Chah, Kennedy F; Gómez-Sanz, Elena; Nwanta, John A; Asadu, Brendan; Agbo, Ifeoma C; Lozano, Carmen; Zarazaga, Myriam; Torres, Carmen
The occurrence, resistance phenotype and molecular mechanisms of resistance of methicillin-resistant staphylococci from groin swabs of 109 clinically healthy dogs in Nsukka, Nigeria were investigated. The groin swab samples were cultured on mannitol salt agar supplemented with 10 μg of cloxacillin. Sixteen methicillin-resistant coagulase negative staphylococci (MRCoNS), all harbouring the mecA gene were isolated from 14 (12.8%) of the 109 dogs studied. The MRCoNS isolated were: S. sciuri subspecies rodentium, S. lentus, S. haemolyticus, and S. simulans with S. sciuri subspecies rodentium (62.5%) being the predominant species. Thirteen (81.3%) of the MRCoNS were resistant to tetracycline while 12 (75%) and 10 (62.5%) were resistant to kanamycin and trimthoprim-sulphamethoxazole respectively. None of the isolates was resistant to fusidic acid, linezolid and vancomycin. Thirteen (81.3%) of the MRCoNS were multi-drug resistance (MDR). Other antimicrobial genes detected were: blaZ, tet(K), tet(M), tet(L), erm(B), lnu(A), aacA-aphD, aphA3, str, dfr(G), cat pC221 , and cat pC223 . Methicillin-resistant staphylococci are common colonizers of healthy dogs in Nigeria with a major species detected being S. sciuri subsp. rodentium.
Starlander, Gustaf; Börjesson, Stefan; Grönlund-Andersson, Ulrika; Tellgren-Roth, Christian; Melhus, Asa
The dog-associated Staphylococcus pseudintermedius is a rare pathogen in humans. Here we describe a cluster of infections caused by the methicillin-resistant S. pseudintermedius clone ST71-J-t02-II-III. It involved four elderly patients at a tertiary hospital. Three patients had wound infections, and the strain had a tendency to cause bullous skin lesions.
An 8-year-old, neutered male Labrador retriever dog with generalized pruritis had a history of recurring atopic dermatitis and superficial pyoderma. Cocci and yeast were found on cytology and methicillin-resistant Staphylococcus pseudintermedius was cultured. A regimen of marbofloxacin, dexamethasone, ketoconazole, and cyclosporine in addition to bathing with 2% chlorhexidine shampoo resulted in marked improvement.
Nicolasora, Nelson P; Zacharek, Mark A; Malani, Anurag N
Staphylococcus aureus has long been recognized as a cause of acute bacterial parotitis. A case of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) parotitis is presented, highlighting the emergence of this increasingly important pathogen to cause a wide variety of infections. Also reviewed are the salient clinical and microbiologic features of this novel infection.
Strains of methicillin resistant Staphylococcus aureus (MRSA), a frequent human pathogen, may also be found in the flora of healthy persons and in the environments that they frequent. Strains of MRSA circulating in the community classified as USA 300 are now found not only in the community but also...
Carroll, Ronan K; Burda, Whittney N; Roberts, Jill C; Peak, Kealy K; Cannons, Andrew C; Shaw, Lindsey N
We present the draft genome sequence of methicillin-resistant Staphylococcus aureus strain CBD-635, from the USA100 lineage. This is a sepsis isolate obtained from Tampa General Hospital. This strain is spa type t003 and multilocus sequence typing (MLST) type ST5, and it has been used by our group in the study of novel antimicrobial chemotherapeutics.
Huijsdens, Xander W; Janssen, Maria; Renders, Nicole H M; Leenders, Alexander; van Wijk, Paul; van Santen Verheuvel, Marga G; van Driel, Jolanda Koel; Morroy, Gabriella
An outbreak of community-associated USA300 methicillin-resistant Staphylococcus aureus occurred in a beautician and 2 of her customers. Eight other persons, who were either infected (n = 5) or colonized (n = 3), were linked to this outbreak, including a family member, a household contact, and partners of customers.
Many, Patricia S.
Methicillin-resistant "Staphylococcus aureus" (MRSA) was once thought to be a bacterium causing infections in only hospitalized patients. However, a new strain of MRSA has emerged among healthy individuals who have not had any recent exposure to a hospital or to medical procedures. This new strain is known as "community-associated…
Phillips, M; Malloy, G; Nedunchezian, D; Lukrec, A; Howard, R G
Several antibiotics have disulfiram-like effects; we evaluated disulfiram for its antibiotic-like effects. Disulfiram inhibited the in vitro growth of methicillin-resistant Staphylococcus aureus, with an MIC of 1.33 micrograms/ml, but was not effective against members of the family Enterobacteriaceae or Pseudomonas species. PMID:2069390
An 8-year-old, neutered male Labrador retriever dog with generalized pruritis had a history of recurring atopic dermatitis and superficial pyoderma. Cocci and yeast were found on cytology and methicillin-resistant Staphylococcus pseudintermedius was cultured. A regimen of marbofloxacin, dexamethasone, ketoconazole, and cyclosporine in addition to bathing with 2% chlorhexidine shampoo resulted in marked improvement. PMID:25392557
Larsen, Jesper; Stegger, Marc; Andersen, Paal S.; Petersen, Andreas; Larsen, Anders R.; Westh, Henrik; Agersø, Yvonne; Fetsch, Alexandra; Kraushaar, Britta; Käsbohrer, Annemarie; Feβler, Andrea T.; Schwarz, Stefan; Cuny, Christiane; Witte, Wolfgang; Butaye, Patrick; Denis, Olivier; Haenni, Marisa; Madec, Jean-Yves; Jouy, Eric; Laurent, Frederic; Battisti, Antonio; Franco, Alessia; Alba, Patricia; Mammina, Caterina; Pantosti, Annalisa; Monaco, Monica; Wagenaar, Jaap A.; de Boer, Enne; van Duijkeren, Engeline; Heck, Max; Domínguez, Lucas; Torres, Carmen; Zarazaga, Myriam; Price, Lance B.; Skov, Robert L.
We investigated the evolution and epidemiology of a novel livestock-associated methicillin-resistant Staphylococcus aureus strain, which colonizes and infects urban-dwelling Danes even without a Danish animal reservoir. Genetic evidence suggests both poultry and human adaptation, with poultry meat implicated as a probable source. PMID:27655995
Wigmore, Sarah M.; Wareham, David W.
ABSTRACT Staphylococcus haemolyticus strain SW007 was isolated from a nasal swab taken from a healthy dog. The isolate is resistant to methicillin, mupirocin, macrolides, and sulfonamides. The SW007 draft genome is 2,325,410 bp and contains 2,277 coding sequences, including 60 tRNAs and nine complete rRNA-coding regions. PMID:28385855
Metz-Gercek, Sigrid; Mittermayer, Helmut
Methicillin-resistant Staphylococcus aureus (MRSA) clonal type ST398 is usually associated with animals. We examined 1,098 confirmed MRSA samples from human patients and found that 21 were MRSA ST398. Most (16) patients were farmers. Increasing prevalence from 1.3% (2006) to 2.5% (2008) shows emergence of MRSA ST398 in humans in Austria. PMID:19402964
Krziwanek, Karina; Metz-Gercek, Sigrid; Mittermayer, Helmut
Methicillin-resistant Staphylococcus aureus (MRSA) clonal type ST398 is usually associated with animals. We examined 1,098 confirmed MRSA samples from human patients and found that 21 were MRSA ST398. Most (16) patients were farmers. Increasing prevalence from 1.3% (2006) to 2.5% (2008) shows emergence of MRSA ST398 in humans in Austria.
Methicillin-resistant Staphylococcus aureus (MRSA) is a significant human pathogen and has also been associated with wounded or ill companion animals. Healthy animals may also harbor MRSA without presenting any symptoms, but little is known about the prevalence of MRSA among these animals. Therefo...
The main goal of this study was to determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA), particularly livestock-associated (LA)-MRSA in pigs and pork. Genotypic relatedness of isolates on-farm, at slaughter and retail was assessed. Paired nasal and peri-anal swab samples we...
Saddiq, Amna Ali; Mohamed, Azza Mostafa
The aim of this study was to explore the protective impact of aqueous extract of Saudi red propolis against rat lung damage induced by the pathogenic bacteria namely methicillin resistant Staphylococcus aureus (MRSA) ATCC 6538 strain. Infected rats were received a single intraperitoneal (i.p.) injection of bacterial suspension at a dose of 1 X 10(6) CFU / 100g body weight. Results showed that oral administration of an aqueous extract of propolis (50mg/100g body weight) daily for two weeks to infected rats simultaneously with bacterial infection, effectively ameliorated the alteration of oxidative stress biomarker, malondialdehyde (MDA), as well as the antioxidant markers, glutathione peroxidase (GPx) and superoxide dismutase (SOD), in lungs of infected rats compared with infected untreated ones. Also, the used propolis extract successfully modulated the alterations in proinflammatory mediators, tumor necrosis factor-α (TNF- α) and vascular endothelial growth factor (VEGF) in serum. In addition, the propolis extract successfully modulated the oxidative DNA damage and the apoptosis biomarker, caspase 3, in lungs of S aureus infected rats compared with infected untreated animals. The biochemical results were supported by histo-pathological observation of lung tissues. In conclusion, the beneficial prophylactic role of the aqueous extract of Saudi red propolis against lung damage induced by methicillin resistant S aureus may be related to the antioxidant, anti-inflammatory, immunomodulatory and antiapoptosis of its active constituents.
Varaldo, P E; Debbia, E; Schito, G C
The antistaphylococcal activity of rifapentine, a new rifamycin SV derivative, was evaluated in vitro and compared with that of rifampin. A total of 313 staphylococcal strains freshly isolated from clinical material and including representatives of all currently recognized Staphylococcus species of human origin were used. The susceptibility to methicillin of all the test strains was determined preliminarily. Despite minor differences with some species, the MICs of rifapentine were found to be substantially similar to those of rifampin. Methicillin-resistant strains of all species were most resistant to rifapentine and rifampin than were their methicillin-susceptible counterparts. For most strains tested, the MBCs of both rifamycins exceeded by twofold the respective MICs. Both the checkerboard dilution and time-kill methods were used to determine the interactions of rifapentine or rifampin with six different antibiotics: cefamandole, vancomycin, teicoplanin, gentamicin, erythromycin, and fusidic acid. No significant differences between the two rifamycins in the combinations were observed against either methicillin-susceptible or methicillin-resistant strains. Minor differences were noted depending on the second antibiotic tested or the staphylococcal species examined. Antagonism was never observed, and indifference was the prevalent response. Cases of synergism were observed occasionally with the checkerboard method and slightly more often with the time-kill method. PMID:3847272
Mandal, Santi M; Ghosh, Ananta K; Pati, Bikas R
Vancomycin-resistant Staphylococcus aureus (VRSA) and methicillin-resistant S aureus (MRSA) strains were examined in hospital effluents. Most S aureus strains are resistant to methicillin (MRSA), followed by tetracycline. Approximately 15% of MRSA strains are also resistant to vancomycin (VRSA). All VRSA strains developed a VanR/VanS-regulated 2-component system of VanA-type resistance in their genome. Results indicate that there is a possibility of developing resistance to aminoglycosides by VRSA strains in the near future.
Robinson, D. Ashley; Enright, Mark C.
Five major lineages of methicillin-resistant Staphylococcus aureus (MRSA) have evolved since the introduction of methicillin for the treatment of infections caused by penicillin-resistant S. aureus in 1959. The clones of these lineages are responsible for the vast majority of hospital-acquired MRSA disease globally. We have constructed high-resolution evolutionary models for each lineage using a parsimony approach with 15 partial gene sequences from 147 geographically diverse isolates. On the basis of these models, we infer that MRSA has emerged at least 20 times upon acquisition of the methicillin resistance determinant, which is carried on a mobile genetic element called the staphylococcal cassette chromosome mec (SCCmec). The acquisition of SCCmec by sensitive clones was four times more common than the replacement of one SCCmec with another. Notably, SCCmec type IV was found in twice as many clones as any other SCCmec type, and it is this SCCmec type which is commonly found in clones from patients with community-acquired MRSA disease. Our findings suggest that most clones of MRSA arise by the acquisition of SCCmec type IV by methicillin-sensitive isolates. PMID:14638503
Hammad, Ahmed M; Watanabe, Wataru; Fujii, Tomoko; Shimamoto, Tadashi
Staphylococci are not part of the normal fish microflora. The presence of staphylococci on fish is an indication of (a) post-harvest contamination due to poor personnel hygiene, or (b) disease in fish. The aim of this study was to determine the prevalence, molecular genetic characteristics, antibiotic resistance and virulence factors of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS) isolated from 200 samples of retail ready-to-eat raw fish (sashimi) collected from the Japanese prefecture of Hiroshima. We characterized 180 staphylococcal strains. A majority of the grocery stores surveyed (92%, 23/25) contained fish contaminated with Staphylococcus species. We recovered 175 S. aureus isolates from 174 (87%, 174/200) samples, with 170 isolates of MSSA. For the MRSA and MR-CoNS, 10 isolates were obtained from 10 samples (5%, 10/200) collected from 10 shops (40%, 10/25) belonging to four supermarket chains. SCCmec typing revealed the presence of a type IV.1 SCCmec cassette in S. warneri isolates, a type II.1 SCCmec cassette in S. haemolyticus isolates and a cassette in methicillin-resistant S. aureus (MRSA) isolates that could not be typed. Molecular typing of two MRSA isolates by spa sequencing and multilocus sequence typing (MLST) identified t1767 and ST8, respectively. Antibiotic resistance genes that confer resistance to aminoglycosides, tetracyclines, β-lactams, macrolides, lincosamides and streptogramin B (MLS(B)) antibiotics were detected. Genes encoding one or more of the following virulence factors: staphylococcal enterotoxins (seb, and sed), toxic shock syndrome toxin 1 (tst), exfoliative toxin (etaA) were detected in 14.2% (25/175) of S. aureus isolates. The accessory gene regulator (agr) typing of S. aureus isolates revealed that agr type 1 was most prevalent (96.5%, 169/175) followed by type 2 (2.2%, 4/175) and type 3 (1.1%, 2/175). None of
Atyah, M A S; Zamri-Saad, M; Siti-Zahrah, A
Swabs from the brain, eyes and kidneys of tilapia from 11 farms were collected for a period of 2 years. They were grown on blood agar before cultures of suspected Staphylococcus aureus were subjected to ABI STAPH Detection Kit and PCR for identification. They were then grown on oxacillin resistance screening agar base (ORSAB) and subjected to PCR using the MRSA 17 kb forward and reverse primers to identify the methicillin-resistant S. aureus (MRSA). A total of 559 isolates of Staphylococcus spp. were obtained, from which 198 (35%) isolates were identified as S. aureus. Of the 198 S. aureus isolated from tilapias, 98 (50%) were identified as methicillin-resistant S. aureus (MRSA). Since global spread of multi-drug-resistant bacteria has increased in the past decade, this new finding in fish should be of concern.
Millar, Eugene V.; Crawford, Katrina B.; Cui, Tianyuan; Lanier, Jeffrey B.; Tribble, David R.; Ellis, Michael W.
Chlorhexidine has been increasingly utilized in outpatient settings to control methicillin-resistant Staphylococcus aureus (MRSA) outbreaks and as a component of programs for MRSA decolonization and prevention of skin and soft-tissue infections (SSTIs). The objective of this study was to determine the prevalence of chlorhexidine resistance in clinical and colonizing MRSA isolates obtained in the context of a community-based cluster-randomized controlled trial for SSTI prevention, during which 10,030 soldiers were issued chlorhexidine for body washing. We obtained epidemiological data on study participants and performed molecular analysis of MRSA isolates, including PCR assays for determinants of chlorhexidine resistance and high-level mupirocin resistance and pulsed-field gel electrophoresis (PFGE). During the study period, May 2010 to January 2012, we identified 720 MRSA isolates, of which 615 (85.4%) were available for molecular analysis, i.e., 341 clinical and 274 colonizing isolates. Overall, only 10 (1.6%) of 615 isolates were chlorhexidine resistant, including three from the chlorhexidine group and seven from nonchlorhexidine groups (P > 0.99). Five (1.5%) of the 341 clinical isolates and five (1.8%) of the 274 colonizing isolates harbored chlorhexidine resistance genes, and four (40%) of the 10 possessed genetic determinants for mupirocin resistance. All chlorhexidine-resistant isolates were USA300. The overall prevalence of chlorhexidine resistance in MRSA isolates obtained from our study participants was low. We found no association between extended chlorhexidine use and the prevalence of chlorhexidine-resistant MRSA isolates; however, continued surveillance is warranted, as this agent continues to be utilized for infection control and prevention efforts. PMID:24841265
Banerjee, Ritu; Gretes, Michael; Basuino, Li; Strynadka, Natalie; Chambers, Henry F
Methicillin-resistant Staphylococcus aureus (MRSA) is resistant to beta-lactam antibiotics because it expresses penicillin-binding protein 2a (PBP2a), a low-affinity penicillin-binding protein. An investigational broad-spectrum cephalosporin, ceftobiprole (BPR), binds PBP2a with high affinity and is active against MRSA. We hypothesized that BPR resistance could be mediated by mutations in mecA, the gene encoding PBP2a. We selected BPR-resistant mutants by passage in high-volume broth cultures containing subinhibitory concentrations of BPR. We used strain COLnex (which lacks chromosomal mecA) transformed with pAW8 (a plasmid vector only), pYK20 (a plasmid carrying wild-type mecA), or pYK21 (a plasmid carrying a mutant mecA gene corresponding to five PBP2a mutations). All strains became resistant to BPR by day 9 of passaging, but MICs continued to increase until day 21. MICs increased 256-fold (from 1 to 256 microg/ml) for pAW8, 32-fold (from 4 to 128 microg/ml) for pYK20, and 8-fold (from 16 to 128 mug/ml) for pYK21. Strains carrying wild-type or mutant mecA developed six (pYK20 transformants) or four (pYK21 transformants) new mutations in mecA. The transformation of COLnex with a mecA mutant plasmid conferred BPR resistance, and the loss of mecA converted resistant strains into susceptible ones. Modeling studies predicted that several of the mecA mutations altered BPR binding; other mutations may have mediated resistance by influencing interactions with other proteins. Multiple mecA mutations were associated with BPR resistance in MRSA. BPR resistance also developed in the strain lacking mecA, suggesting a role for chromosomal genes.
Olowe, Olugbenga Adekunle; Kukoyi, Olayinka Oluwatoyin; Taiwo, Samuel Sunday; Ojurongbe, Olusola; Opaleye, Oluyinka Oladele; Bolaji, Oloyede Samuel; Adegoke, Abiodun Adebimpe; Makanjuola, Olufunmilola Bamidele; Ogbolu, David Olusoga; Alli, Oyebode Terry
Introduction The characteristics and antimicrobial resistance profiles of Staphylococcus aureus differs according to geographical regions and in relation to antibiotic usage. The aim of this study was to determine the biochemical characteristics of the prevalent S. aureus from Ekiti State, Nigeria, and to evaluate three commonly used disk diffusion methods (cefoxitin, oxacillin, and methicillin) for the detection of methicillin resistance in comparison with mecA gene detection by polymerase chain reaction. Materials and methods A total of 208 isolates of S. aureus recovered from clinical specimens were included in this study. Standard microbiological procedures were employed in isolating the strains. Susceptibility of each isolate to methicillin (5 μg), oxacillin (1 μg), and cefoxitin (30 μg) was carried out using the modified Kirby–Bauer/Clinical and Laboratory Standard Institute disk diffusion technique. They were also tested against panels of antibiotics including vancomycin. The conventional polymerase chain reaction method was used to detect the presence of the mecA gene. Results Phenotypic resistance to methicillin, oxacillin, and cefoxitin were 32.7%, 40.3%, and 46.5%, respectively. The mecA gene was detected in 40 isolates, giving a methicillin-resistant S. aureus (MRSA) prevalence of 19.2%. The S. aureus isolates were resistant to penicillin (82.7%) and tetracycline (65.4%), but largely susceptible to erythromycin (78.8% sensitive), pefloxacin (82.7%), and gentamicin (88.5%). When compared to the mecA gene as the gold standard for MRSA detection, methicillin, oxacillin, and cefoxitin gave sensitivity rates of 70%, 80%, and 100%, and specificity rates of 76.2%, 69.1%, and 78.5% respectively. Conclusion When compared with previous studies employing mecA polymerase chain reaction for MRSA detection, the prevalence of 19.2% reported in Ekiti State, Nigeria in this study is an indication of gradual rise in the prevalence of MRSA in Nigeria. A cefoxitin (30
Argudín, M Angeles; Lauzat, Birgit; Kraushaar, Britta; Alba, Patricia; Agerso, Yvonne; Cavaco, Lina; Butaye, Patrick; Porrero, M Concepción; Battisti, Antonio; Tenhagen, Bernd-Alois; Fetsch, Alexandra; Guerra, Beatriz
Livestock associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has emerged in animal production worldwide. Most LA-MRSA in Europe belong to the clonal complex (CC) 398. The reason for the LA-MRSA emergence is not fully understood. Besides antimicrobial agents used for therapy, other substances with antimicrobial activity applied in animal feed, including metal-containing compounds might contribute to their selection. Some of these genes have been found in various novel SCCmec cassettes. The aim of this study was to assess the occurrence of metal-resistance genes among a LA-S. aureus collection [n=554, including 542 MRSA and 12 methicillin-susceptible S. aureus (MSSA)] isolated from livestock and food thereof. Most LA-MRSA isolates (76%) carried at least one metal-resistance gene. Among the LA-MRSA CC398 isolates (n=456), 4.8%, 0.2%, 24.3% and 71.5% were positive for arsA (arsenic compounds), cadD (cadmium), copB (copper) and czrC (zinc/cadmium) resistance genes, respectively. In contrast, among the LA-MRSA non-CC398 isolates (n=86), 1.2%, 18.6% and 16.3% were positive for the cadD, copB and czrC genes, respectively, and none were positive for arsA. Of the LA-MRSA CC398 isolates, 72% carried one metal-resistance gene, and the remaining harboured two or more in different combinations. Differences between LA-MRSA CC398 and non-CC398 were statistically significant for arsA and czrC. The czrC gene was almost exclusively found (98%) in the presence of SCCmec V in both CC398 and non-CC398 LA-MRSA isolates from different sources. Regarding the LA-MSSA isolates (n=12), some (n=4) were also positive for metal-resistance genes. This study shows that genes potentially conferring metal-resistance are frequently present in LA-MRSA.
Horká, Marie; Tesařová, Marie; Karásek, Pavel; Růžička, Filip; Holá, Veronika; Sittová, Martina; Roth, Michal
Serious bloodstream infections are a significant complication in critically ill patients. The treatment of these infections has become more difficult because of the increasing prevalence of multiresistant strains, especially methicillin-resistant Staphylococcus aureus (MRSA). Rapid differentiation of low number of MRSA from methicillin-susceptible S. aureus (MSSA) cells (10(1)-10(2) cells mL(-1)) in blood is necessary for fast effective antibiotic therapy. Currently, three groups of techniques, phenotyping, genotyping, and mass spectrometry, are used for MRSA and MSSA strains differentiation. Most of these techniques are time-consuming. PCR and other molecular techniques allow the detection and differentiation between MSSA and MRSA directly from blood cultures. These methods alone are rapid and they have good reproducibility and repeatability. Potential disadvantages of the genotyping methods include their discrimination ability, technical complexity, financial costs, and difficult interpretation of the results. Recently, capillary electrophoresis (CZE) was successfully used to differentiate between the agar-cultivated MRSA and MSSA strains in fused silica capillaries etched with supercritical water and modified with (3-glycidyloxypropyl)trimethoxysilane. The possible use of CZE as a fast and low-cost method for distinguishing between the blood-incubated MRSA or MSSA cells has been tested in this manuscript. Our goal was to test low amounts of bacteria (∼10(2) cell mL(-1)) similar to those in clinical samples. The migration times of the purified blood-incubated cells and the agar-cultivated cells were different from each other. However, their isoelectric point was the same for all strains.
Sweeney, Debora; Shinabarger, Dean L; Arhin, Francis F; Belley, Adam; Moeck, Greg; Pillar, Chris M
Methicillin-resistant Staphylococcus aureus (MRSA) infections constitute a threat to the public health due to their prevalence and associated mortality and morbidity. Several agents have been recently approved to treat MRSA skin infections including lipoglycopeptides (dalbavancin, oritavancin, and telavancin), ceftaroline, and tedizolid. This study compared the MIC, minimum bactericidal concentration (MBC), and time-kill of these agents alongside daptomycin, linezolid, and vancomycin against MRSA (n=15); meropenem, cefazolin, and nafcillin were also included against methicillin-susceptible S. aureus (MSSA [n=12]). MIC and MBC testing was conducted in accordance with Clinical and Laboratory Standards Institute guidelines, and time-kills were evaluated at multiples of the MIC and the free-drug maximum plasma concentration (fCmax) at both standard and high inoculum densities for a subset of MRSA (n=2) and MSSA (n=2). MRSA and MSSA were highly susceptible to all agents, with the lipoglycopeptides having the most potent activity by MIC50/90. All agents excluding tedizolid and linezolid were bactericidal by MBC for MRSA and MSSA, though dalbavancin and telavancin exhibited strain-specific bactericidal activity for MRSA. All agents excluding tedizolid and linezolid were bactericidal by time-kill at their respective fCmax against MRSA and MSSA at standard inoculum density, though oritavancin exhibited the most rapid bactericidal activity. Oritavancin and daptomycin at their respective fCmax maintained similar kill curves at high inoculum density. In contrast, the killing observed with other agents was typically reduced or slowed at high inoculum density. These data demonstrate the rapid bactericidal activity of oritavancin and daptomycin against S. aureus relative to other MRSA agents regardless of bacterial burden.
Rello, J; Torres, A; Ricart, M; Valles, J; Gonzalez, J; Artigas, A; Rodriguez-Roisin, R
All episodes of ventilator-associated pneumonia (VAP) caused by Staphylococcus aureus were prospectively analyzed for a 30-mo period. Methicillin-sensitive S. aureus (MSSA) was isolated in 38 episodes and methicillin-resistant S. aureus (MRSA) in 11 others. The two groups were similar regarding sex, severity of underlying diseases, prior surgery, and presence of renal failure, diabetes, cardiopathy, and coma. MRSA-infected persons were more likely to have received steroids before developing infection (relative risk [RR] = 3.45, 95% confidence interval [CI] = 1.38-8.59), to have been ventilated > 6 d (RR = 2.03, 95% CI = 1.36-3.03), to have been older than 25 yr (RR = 1.50, 95% CI = 1.09-2.06), and to have had preceding chronic obstructive pulmonary disease (RR = 2.76, 95% CI = 0.89-8.56) than MSSA-infected patients. MSSA-infected persons were more likely than MRSA-infected patients to have cranioencephalic trauma (RR = 1.94, 95% CI = 1.22-3.09). All patients with MRSA VAP had previously received antibiotics, compared with only 21.1% of those with MSSA infection (p < 0.000001). The incidence of empyema was similar in both groups; nevertheless, the presence of bacteremia and septic shock was more frequent in the MRSA group. Finally, mortality directly related to pneumonia was significantly higher among patients with MRSA episodes (RR = 20.72, 95% CI = 2.78-154.35). This analysis was repeated for monomicrobial episodes, and the difference remained statistically significant. We conclude that MRSA and MSSA strains infect patients with different demographic profiles; previous antibiotic therapy is the most important risk factor for developing MRSA infection.(ABSTRACT TRUNCATED AT 250 WORDS)
Fetsch, Alexandra; Roesler, Uwe; Kraushaar, Britta; Friese, Anika
Methicillin-susceptible Staphylococcus (S.) aureus (MSSA) and methicillin-resistant S. aureus (MRSA) are colonizers of skin and mucosa. In humans, MSSA and MRSA compete for colonization space in the anterior nares of pig farmers; however, it was also shown that MSSA/MRSA co-colonization is common and one clone can be found rather than differing types of MSSA and MRSA. We investigated the colonization and clonality of both, MSSA and MRSA in pigs over a longer time. Eighteen sows were nasally sampled three times every ten weeks. Additionally, environmental samples were taken. Samples were investigated for MSSA and MRSA, respectively. The spa type was defined from up to five MRSA and MSSA isolates found per sample and sampling time; selected isolates were further investigated by microarray. Three sows (16.7%) were completely negative for MSSA and MRSA. Twelve pigs (66.7%) were irregularly positive for both, MSSA and MRSA over the time, whereas seven out of them (38.9%) were simultaneously colonized. CC398 (t034, t011) MRSA and CC9 (t337, t1430, and t13816) MSSA associated spa types were exclusively found. In 44.4% (n=8) of sows up to two different types of MSSA were present at the same time and sample. Strains of the same clonal lineage showed a high genetic identity despite their origin. Highly identic clones were present in sows and their environment. As conclusion, MSSA/MRSA may not exclude each other in the anterior nares of pigs. Pigs may also carry different clones at the same time.
Tekin, Alicem; Dal, Tuba; Deveci, Ozcan; Tekin, Recep; Atmaca, Selahattin; Dayan, Saim
In order to detect the methicillin susceptibility and determine the prevalence of constitutive and inducible clindamycin resistance in Staphylococcus aureus strains, a total of 254 S. aureus isolates, including 139 (54.7%) methicillin-sensitive and 115 (45.3%) methicillin-resistant, were tested for clindamycin and erythromycin by Kirby-Bauer disk diffusion method. The disk diffusion induction test or D-test was performed on erythromycin-resistant and clindamycin-sensitive strains using the disk diffusion method. The erythromycin-resistant and clindamycin-sensitive strains with a D-shaped zone around the clindamycin disk were considered positive for inducible clindamycin resistance. If a strain was found to be both erythromycin-resistant and clindamycin-resistant, it was considered to have constitutive clindamycin resistance. Overall, ten (3.9%) strains, including two methicillin-resistant S. aureus and eight methicillin-sensitive S. aureus, were found to be erythromycin-resistant and clindamycin-sensitive according to the disk diffusion method and all of these strains showed inducible clindamycin resistance by the D-test method. Constitutive clindamycin resistance was detected in 100 of all MRSA strains. In conclusion, the presence of inducible clindamycin-resistant strains may lead to clindamycin treatment failure in patients with S. aureus infection. The D-test method should be used for antimicrobial susceptibility testing of clindamycin.
Martins, Marta; Ordway, Diane; Kristiansen, Malthe; Viveiros, Miguel; Leandro, Clara; Molnar, Joseph; Amaral, Leonard
The Carpobrotus edulis methanol extract, inactive against the methicillin-resistant Staphylococcus aureus or the multidrug-resistant Mycobacterium tuberculosis, does inhibit the growth of these two bacteria once they are phagocytosed by monocyte derived human macrophages.
Hu, Baofeng; Carroll, Karen C.; Bilker, Warren B.; Tolomeo, Pam; Cluzet, Valerie C.; Baron, Patrick; Ferguson, Jacqueline M.; Morris, Daniel O.; Rankin, Shelley C.; Lautenbach, Ebbing
Two screening methods to detect staphylococcal colonization in humans were compared. Direct plating to CHROMagar (BD Diagnostics) was compared to a broth preenrichment followed by plating to Baird-Parker agar. The broth-enrichment method was comparable to CHROMagar for methicillin-resistant Staphylococcus aureas (MRSA) detection, but the enrichment method was optimum for recovery of coagulase-positive Staphylococcus spp. PMID:27122377
Davis, Meghan F; Hu, Baofeng; Carroll, Karen C; Bilker, Warren B; Tolomeo, Pam; Cluzet, Valerie C; Baron, Patrick; Ferguson, Jacqueline M; Morris, Daniel O; Rankin, Shelley C; Lautenbach, Ebbing; Nachamkin, Irving
Two screening methods to detect staphylococcal colonization in humans were compared. Direct plating to CHROMagar (BD Diagnostics) was compared to a broth preenrichment followed by plating to Baird-Parker agar. The broth-enrichment method was comparable to CHROMagar for methicillin-resistant Staphylococcus aureas (MRSA) detection, but the enrichment method was optimum for recovery of coagulase-positive Staphylococcus spp.
Johnson, Kristen N; Andreacchio, Kathleen; Edelstein, Paul H
The accurate performance of the Vitek 2 GP67 card for detecting methicillin-resistant coagulase-negative staphylococci (CoNS) is not known. We prospectively determined the ability of the Vitek 2 GP67 card to accurately detect methicillin-resistant CoNS, with mecA PCR results used as the gold standard for a 4-month period in 2012. Included in the study were 240 consecutively collected nonduplicate CoNS isolates. Cefoxitin susceptibility by disk diffusion testing was determined for all isolates. We found that the three tested systems, Vitek 2 oxacillin and cefoxitin testing and cefoxitin disk susceptibility testing, lacked specificity and, in some cases, sensitivity for detecting methicillin resistance. The Vitek 2 oxacillin and cefoxitin tests had very major error rates of 4% and 8%, respectively, and major error rates of 38% and 26%, respectively. Disk cefoxitin testing gave the best performance, with very major and major error rates of 2% and 24%, respectively. The test performances were species dependent, with the greatest errors found for Staphylococcus saprophyticus. While the 2014 CLSI guidelines recommend reporting isolates that test resistant by the oxacillin MIC or cefoxitin disk test as oxacillin resistant, following such guidelines produces erroneous results, depending on the test method and bacterial species tested. Vitek 2 cefoxitin testing is not an adequate substitute for cefoxitin disk testing. For critical-source isolates, mecA PCR, rather than Vitek 2 or cefoxitin disk testing, is required for optimal antimicrobial therapy.
Hao, Haihong; Dai, Menghong; Wang, Yulian; Huang, Lingli; Yuan, Zonghui
Methicillin-resistant Staphylococcus aureus (MRSA), popularly known as a type of superbug, has been a serious challenge for animal and human health. S. aureus has developed methicillin resistance mainly by expression of β-lactamase and PBP2a, which is regulated by the blaZ-blaI-blaR1 and mecA-mecI-mecRI systems. Other genetic elements, including murE and femA, also participate in expression of methicillin resistance, but the mechanism remains unclear. The evolution of the staphylococcal cassette chromosome mec determines the epidemiological risk of MRSA. The plasmid-located gene cfr might contribute to multiresistance and transmission of MRSA. Some virulence factors, including Panton-Valentine leukocidin, phenol-soluble modulin, arginine catabolic mobile element and other toxin elements enhance the pathogenesis and fitness of MRSA. Two-component regulation systems (agr, saeRS and vraRS) are closely associated with pathogenesis and drug resistance of MRSA. The systematic exploration of key genetic elements and regulation systems involved in multidrug resistance/pathogenesis/transmission of MRSA is conclusively integrated into this review, providing fundamental information for the development of new antimicrobial agents and the establishment of reasonable antibiotic stewardship to reduce the risk of this superbug.
Hirschl, A; Stanek, G; Rotter, M
Coagulase-positive staphylococci (41 methicillin-resistant strains, 28 penicillinase-positive and 12 penicillinase-negative strains) were tested against most types of penicillines commercially available on the Austrian market using both broth-dilution test (incubated during 48 hours at 35 degrees C) and agardiffusion tests (incubated during 24 hours at 30, 35 and 37 degrees C) employing Mueller-Hinton-broth and -agar, respectively, in order to find out the most convenient way of detecting methicillin-resistant strains. Consecutively, the conclusions drawn from these experiments were verified for tests on Isosensitest-agar (Oxoid). It was demonstrated that methicillin-resistant strains could be detected easily with discs of methicillin and oxacillin at 30 degrees C (Fig. 1). At 35 degrees C this was nearly as easily possible for methicillin but oxacillin discs had to be used at amounts of 1 microgram instead of 5 microgram. Excepting penicillin G-discs with the other penicillines differing numbers of methicillin-resistant strains would have been missed at 37 degrees C (Fig. 1, 2). Only with discs containing 6 microgram penicillin G methicillin-resistant strains were unequivocally identifiable in the agardiffusion test at all 3 incubation-temperatures (Fig. 3), the largest inhibition zone diameter being 12 mm. Penicillinase-positive but methicillin-sensitive strains always produced larger inhibition zones up to 30 mm. From these strains again penicillinase-negative strains were equally well distinguishable by much larger inhibition zones. So, the conclusion was drawn that on Mueller-Hinton agar one disc loaded with 6 microgram of penicillin G allows proper distinction not only of methicillin-resistant and -sensitive but also of penicillinase-positive and -negative strains of staphylococci. On Isosensitest-agar this is true for an incubation-temperature of 35 but not 37 degrees C (Tab. 1).
Bassinger, V. J.; Fontenot, S. L.; Castro, V. A.; Ott, C.; Healy, M.; Pierson, D. L.
Background: Within the past few years, methicillin-resistant Staphylococcus aureus has emerged in environments with susceptible hosts in close proximity, such as hospitals and nursing homes. As the International Space Station (ISS) represents a semi-closed environment with a high level of crewmember interaction, an evaluation of isolates of clinical and environmental Staphylococcus aureus and coagulase negative Staphylococcus was performed to determine if this trend was also present in astronauts occupying ISS or on surfaces of the space station itself. Methods: Identification of isolates was completed using VITEK (GPI cards, BioMerieux), 16S ribosomal DNA analysis (MicroSeq 500, ABI), and Rep-PCR DNA fingerprinting (Divemilab, Bacterial Barcodes). Susceptibility tests were performed using VITEK (GPS-105 cards, BioMerieux) and resistance characteristics were evaluated by testing for the presence of the mecA gene (PBP2' MRSA test kit, Oxoid). Results: Rep-PCR analysis indicated the transfer of S. aureus between crewmembers and between crewmembers and ISS surfaces. While a variety of S. aureus were identified from both the crewmembers and environment, evaluations of the microbial population indicated minimal methicillin resistance. Results of this study indicated that within the semi-closed ISS environment, transfer of bacteria between crewmembers and their environment has been occurring, although there was no indication of a high concentration of methicillin resistant Staphylococcus species. Conclusions: While this study suggests that the spread of methicillin resistant S. aureus is not currently a concern aboard ISS, the increasing incidence of Earth-based antibiotic resistance indicates a need for continued clinical and environmental monitoring.
Kluytmans, Jan; Van Griethuysen, Arjanne; Willemse, Piet; Van Keulen, Peter
Two new selective media, oxacillin resistance screening agar base (ORSAB) and CHROMagar Staph aureus (CSA), were evaluated for identification of Staphylococcus aureus and for screening of methicillin resistance by addition of antimicrobial agents to these media. A well-defined collection consisting of 1,140 staphylococci was used. A total of 624 were S. aureus, of which 358 were methicillin susceptible and 266 were methicillin resistant, and 516 were coagulase-negative staphylococci. The methicillin-resistant S. aureus (MRSA) strains were selected based on the results of phage typing; 247 different types were included in the analysis. For identification of S. aureus, both media performed better after 24 h than after 48 h. The sensitivities at 24 h were comparable (CSA, 98.6%; ORSAB, 97.1%), but the specificity of CSA was significantly higher (CSA, 97.1%; ORSAB, 92.1%). For screening of methicillin resistance, antibiotic supplements were added to both media. The sensitivity was lower after 24 h (CSA, 58.6%; ORSAB, 84.2%) and increased significantly after 48 h (CSA, 77.5%; ORSAB, 91.4%). At both time intervals ORSAB was significantly more sensitive than CSA. However, the specificities of both media were high after 24 h (CSA, 99.1%; ORSAB, 98.3%) and decreased significantly after 48 h of incubation (CSA, 94.7%; ORSAB, 95.5%). In conclusion, for identification of S. aureus, CSA is more accurate than ORSAB because of a significantly higher specificity. For screening of MRSA, ORSAB performs better than CSA, but the usefulness in clinical practice is limited because a significant number of strains are not detected. PMID:12089266
Kampf, G; Jarosch, R; Rüden, H
In order to determine the efficacy of hand disinfectants based on alcohol against three MRSA strains and 3 methicillin-susceptible S. aureus strains (MSSA), 1-propanol (60%) as well as Sterillium and Spitaderm were investigated in the quantitative suspension test at various dilutions and reactions times (15, 30 and 60s). All undiluted disinfectants revealed reduction factors > 6 against MRSA and MSSA after 30s. Diluted disinfectants (50%) were significantly less effective against MRSA at short reaction times (15 s) (p < 0.05). Sterillium in a dilution of 50% did not reach 5 reduction factors against either MRSA or MSSA after 30 s. The impact of an appropriate use of hand disinfectants in order to break chains of infections with MRSA is obvious.
Foucault, Marie-Laure; Courvalin, Patrice; Grillot-Courvalin, Catherine
We have quantified the biological cost of VanA-type glycopeptide resistance due to the acquisition of the resistance operon by methicillin-resistant Staphylococcus aureus (MRSA) from Enterococcus sp. Exponential growths of recipient strain HIP11713, its transconjugant VRSA-1, VRSA-5, and VRSA-6 were compared in the absence or, except for HIP11713, in the presence of vancomycin. Induction of resistance was performed by adding vancomycin in both the preculture and the culture or the culture at only 1/50 the MIC. In the absence of vancomycin, the growth rates of the vancomycin-resistant S. aureus (VRSA) strains were similar to that of susceptible MRSA strain HIP11713. When resistance was induced, and under both conditions, there was a significant reduction of the growth rate of the VRSA strains relative to that of HIP11713 and to those of their noninduced counterparts, corresponding to a ca. 20% to 38% reduction in fitness. Competition experiments between isogenic VRSA-1 and HIP11713 mixed at a 1:1, 1:100, or 100:1 ratio revealed a competitive disadvantage of 0.4% to 3% per 10 generations of the transconjugant versus the recipient. This slight fitness burden can be attributed to the basal level of expression of the van genes in the absence of induction combined with a gene dosage effect due to the presence of the van operon on multicopy plasmids. These data indicate that VanA-type resistance, when induced, is highly costly for the MRSA host, whereas in the absence of induction, its biological cost is minimal. Thus, the potential for the dissemination of VRSA clinical isolates should not be underestimated.
Worldwide, reports document the increasing frequency of methicillin resistant Staphylococcus aureus (MRSA) infections. Other human pathogens are recognized as unresponsive to antibiotics of last resort. These previously treatable infections now account for increased numbers of human disease and de...
Worldwide, reports document the increasing frequency of methicillin resistant Staphylococcus aureus (MRSA) infections. Other human pathogens are recognized as unresponsive to antibiotics of last resort. These previously treatable infections now account for increased numbers of human disease and de...
Falcão, M H; Texeira, L A; Ferreira-Carvalho, B T; Borges-Neto, A A; Figueiredo, A M
Many methods have been described for the detection of methicillin-resistant Staphylococcus aureus (MRSA), but the homogeneous or heterogeneous expression of methicillin resistance affects the reliability of those methods. This study demonstrates that close association between methicillin-susceptible S. aureus (MSSA) and MRSA strains in the host colonisation site can present additional problems for the detection of MRSA in clinical laboratories, which may contribute to failure in the control of MRSA infection in hospital. Worse, this association may also account for the emergence of MRSA during antibiotic therapy.
Twenty-five year epidemiology of invasive methicillin- resistant Staphylococcus aureus (MRSA) isolates recovered at a burn center§ Clinton K. Murray...history: Accepted 12 February 2009 Keywords: Burn center Epidemiology Methicillin-resistant Staphylococcus aureus Antimicrobial susceptibility a b s t r...invasive MRSA isolates over 25 years at a single burn unit. Isolates were tested by pulsed-field gel electrophoresis (PFGE), broth microdilution
Sallam, Khalid Ibrahim; Abd-Elghany, Samir Mohammed; Elhadidy, Mohamed; Tamura, Tomohiro
The emergence of livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) in food-producing animals is of increasing interest, raising questions about the presence of MRSA in food of animal origin and potential sources of transmission to humans via the food chain. In this study, the prevalence, molecular characterization, virulence factors, and antimicrobial susceptibility patterns of MRSA isolates from 200 retail raw chicken samples in Egypt were determined. MRSA was detected by positive amplification of the mecA gene in 38% (76 of 200) of chicken samples analyzed. This represents a potential public health threat in Egypt, as this contamination rate seems to be the highest among other studies reported worldwide. Furthermore, genes encoding α-hemolysin (hla) and staphylococcal enterotoxins (sea, seb, and sec) were detected in all of the 288 MRSA isolates. Nonetheless, none of the strains tested carried tst, the gene encoding toxic shock syndrome toxin 1. Antimicrobial resistance of MRSA isolates was most frequently detected against penicillin (93.4%), ampicillin (88.9%), and cloxacillin (83.3%). These results suggest that retail chicken might be a significant potential source for transmission of multidrug-resistant and toxigenic S. aureus in Egypt. This underlines the need for stricter hygienic measures in chicken production in Egypt to minimize the risk of transmission of these strains to consumers. To the best of our knowledge, this is the first study that reports the isolation and molecular characterization of MRSA in retail chicken samples in Egypt.
Traversa, A; Gariano, G R; Gallina, S; Bianchi, D M; Orusa, R; Domenis, L; Cavallerio, P; Fossati, L; Serra, R; Decastelli, L
Following the detection of methicillin-resistant Staphylococcus aureus (MRSA) ST398 in food-producing animals, both livestock and wildlife, and derived products, are considered potential sources of MRSA in humans. There is a paucity of data on MRSA in foods in Italy, and the data regarding wild animals are particularly scarce. A total of 2162 food samples collected during official monitoring activities in 2008 were analyzed for the detection of S. aureus. Also, samples from 1365 wild animals collected by the National Reference Center for Wild Animal Diseases in 2003-2009 were subjected to anatomopathological examination. S. aureus isolates were processed for phenotypic and molecular methicillin resistance determinations. S. aureus was found in 2.0% of wild animal carcasses and in 3.2% of wild boar lymph nodes: none showed methicillin resistance. The prevalence of S. aureus in food was 17.1%. Two MRSA strains, both from bulk tank milk (prevalence 0.77%) were isolated: the strains were resistant to tetracycline, had spa-type t899, and were negative for the Panton-Valentine leukocidin gene. The low prevalence of MRSA suggests that the risk of transmission to humans via food is limited. However, attention should be paid to the cattle food chain, which may be a potential route of transmission of LA-MRSA.
Talebi, Malihe; Shafiee, Mohammad; Sadeghi, Javad; Moghadam, Nasrin Asghari; Saifi, Mahnaz; Pourshafie, Mohammad R
We investigated the prevalence of methicillin-resistant coagulase-negative staphylococci (MRCoNS) isolated from hospitalized patients and outpatients (OP). Out of 350 staphylococcal isolates collected from three hospitals, 190 were coagulase-negative staphylococci (CoNS). These isolates were subjected to antimicrobial susceptibility tests, detection of mecA, and pulsed-field gel electrophoresis (PFGE) typing. Among the 190 isolated CoNS, Staphylococcus epidermidis (47.3%) and Staphylococcus haemolyticus (44.2%) were the most prevalent species. Other CoNS species that were isolated were Staphylococcus saprophyticus (2.1%), Staphylococcus warneri (2.1%), Staphylococcus simulans (1.6%), Staphylococcus capitis (1.1%), Staphylococcus schleiferi (1.1%), and Staphylococcus hominis (0.5%). The rate of resistance to methicillin was 60% with 58 (50%) S. epidermidis and 55 (49%) S. haemolyticus. The rate of resistance to 13 antibiotics tested with the lowest and highest to chloramphenicol and penicillin, respectively. High clonal diversity with different PFGE patterns was obtained for methicillin-resistant S. epidermidis and S. haemolyticus by 32 and 31 types, respectively. Our results indicated that the dissemination of MRCoNS is widespread in Tehran. The majority of these isolates showed distinct genotyping patterns. At the same time, the common patterns were found among the MRCoNS obtained from outpatient and inpatient isolates, suggestive of an epidemiological link.
Ott, C. M.; Bassinger, V. J.; Fontenot, S. L.; Castro, V. A.; Pierson, D. L.
The International Space Station (ISS) represents a semi-closed environment with a high level of crewmember interaction. As community-acquired methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a health concern in environments with susceptible hosts in close proximity, an evaluation of isolates of clinical and environmental Staphylococcus aureus and coagulase negative Staphylococcus was performed to determine if this trend was also present in astronauts aboard ISS or the space station itself. Rep-PCR fingerprinting analysis of archived ISS isolates confirmed our earlier studies indicating a transfer of S. aureus between crewmembers. In addition, this fingerprinting also indicated a transfer between crewmembers and their environment. While a variety of S. aureus were identified from both the crewmembers and the environment, phenotypic evaluations indicated minimal methicillin resistance. However, positive results for the Penicillin Binding Protein, indicative of the presence of the mecA gene, were detected in multiple isolates of archived Staphylococcus epidermidis and Staphylococcus haemolyticus. Phenotypic analysis of these isolates confirmed their resistance to methicillin. While MRSA has not been isolated aboard ISS, the potential exists for the transfer of the gene, mecA, from coagulase negative environmental Staphylococcus to S. aureus creating MRSA strains. This study suggests the need to expand environmental monitoring aboard long duration exploration spacecraft to include antibiotic resistance profiling.
Kali, Arunava; Stephen, Selvaraj; Umadevi, Sivaraman
Although conventional antibiotic susceptibility tests are most commonly performed for methicillin-resistant Staphylococcus aureus (MRSA), the results of these phenotypic tests are dependent on the standardization of the culture conditions. The aim of the study was to evaluate the conventional phenotypic screening tests in comparison to the mecA gene polymerase chain reaction (PCR). One hundred and two clinical isolates of MRSA identified by the oxacillin disk diffusion were subjected to PCR for the mecA gene and by the cefoxitin disk diffusion test and culture on oxacillin screen agar, mannitol salt agar, and methicillin-resistant Staphylococcus aureus Agar (MeReSA) selective medium, for MRSA. Although all 102 isolates were resistant in oxacillin and cefoxitin disk diffusion, 92 (90.1%) isolates were positive for the mecA gene. The sensitivities of the mannitol salt agar, MeReSA agar, and oxacillin screen agar were 89.13, 97.82, and 98.91%, respectively. The oxacillin screen agar may be recommended for confirming methicillin resistance in the disk diffusion test in resource-poor settings, where molecular methods are not available.
Robinson, Gwen L; Harris, Anthony D; Morgan, Daniel J; Pineles, Lisa; Belton, Beverly M; Johnson, J Kristie
This study determined the survivability of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) for extended periods of time and temperatures using a standard swab for assessment. Our study showed that transportation in Liquid Amies medium could be performed at room temperature or 4°C for up to 14 days without a decrease in recovery of MRSA or VRE.
Robinson, Gwen L.; Harris, Anthony D.; Morgan, Daniel J.; Pineles, Lisa; Belton, Beverly M.
This study determined the survivability of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) for extended periods of time and temperatures using a standard swab for assessment. Our study showed that transportation in Liquid Amies medium could be performed at room temperature or 4°C for up to 14 days without a decrease in recovery of MRSA or VRE. PMID:22535993
Garbacz, Katarzyna; Żarnowska, Sabina; Piechowicz, Lidia; Haras, Krystyna
The aim of this study was to compare the spread of multidrug-resistant (MDR) and methicillin-resistant (MR) staphylococci in healthy dogs and in dogs with evident symptoms of infection. The samples from 172 healthy and 197 infected dogs were examined. The staphylococci were identified with conventional methods and by means of the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method (MboI). Susceptibility to 15 antibiotics from 10 different antimicrobial classes was tested. Resistance to methicillin was confirmed by the presence of Staphylococcus aureus mecA and S. sciuri mecA genes. Multidrug resistance was defined as resistance to three or more antimicrobial classes. The oral mucosa to be the most frequent site of staphylococcal colonization (55.8 %), followed by nasal cavity (44.2 %), and anus (32.6 %). The prevalence of MDR staphylococci in infected dogs was significantly higher than in the healthy animals (74/137 vs. 34/95, P = 0.006). The MR strains of S. pseudintermedius (2.9 %) originated solely from infected dogs. In contrast, the MR coagulase-negative strains (7.4 %) were isolated solely from healthy dogs. S. aureus strains originated from nasal swabs, MRSA strains were not isolated. MDR staphylococci and MR S. pseudintermedius are more common among infected dogs, but coagulase-negative staphylococci (mostly S. sciuri) seem to be a reservoir of methicillin resistance in healthy dogs.
Snyder, Ashley Hall; Srivastava, Ruma; Rybak, Michael J.; McGrath, Eric
Ceftaroline, an advanced generation cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA), may present a new therapeutic alternative for treating lung infections among patients with cystic fibrosis. We report a case of ceftaroline therapy in a pediatric patient with cystic fibrosis, whose dose was increased from 9.7 mg/kg/dose every 12 hours to 10.8 mg/kg/dose every 8 hours by using pharmacokinetic analyses. PMID:25024675
Swe, Thein; Naing, Akari Thein; Baqui, AAMA; Khillan, Ratesh
To our knowledge and literature search, Staphylococcus schleiferi subspecies coagulans infection in human beings has rarely been described in the medical literature. Furthermore, we believe that this is a first detailed case report of methicillin-resistant Staphylococcus schleiferi subspecies coagulans infection in a patient with hepatocellular carcinoma. Because of the possible association of Staphylococcus schleiferi infection and immunosuppression, any isolates of this bacterium in human beings should be presumed to be pathogenic, unless proven otherwise. PMID:27734018
Abreu, Diego; Arroyo, Carlos; Suarez, Ruben; Campolo, Horacio; Izaguirre, Juan; Decía, Ricardo; Machado, Miguel; Carvalhal, Gustavo Franco; Clavijo, Jorge
Prostatic abscess is rare. Its potentially serious course requires a high level of clinical suspicion and prompt and effective treatment. The causative germs are usually either enterobacteria or Enterococcus. The authors highlight the importance of considering epidemiological and clinical aspects in the early diagnosis and treatment. Prostatic abscess due to community-acquired methicillin resistant Staphylococcus has three typical characteristics: skin entry point, periprostatic compromise, and anaemia and low prothrombin. PMID:22696740
Giannini, Mary Anne; Nance, Donna; McCullers, Jonathan A
We studied the bacterial burden on toilet seats in a children's cancer hospital to validate a policy requesting that immunocompromised children use alcohol wipes on the seats prior to use of the toilets. Methicillin-resistant Staphylococcus aureus (MRSA) was recovered from 3.3% of hospital toilets when wipes were not in use. Use of wipes resulted in a 50-fold reduction in mean daily bacterial counts and eliminated MRSA.
Krishnakumar, R.; Renjitkumar, J.
Indwelling epidural catheters are frequently used to manage postoperative pain. This report describes a patient who developed methicillin-resistant Staphylococcus aureus (MRSA) vertebral osteomyelitis of the lumbar spine following epidural catheterization. To the best of our knowledge, this is the first reported case of MRSA vertebral osteomyelitis secondary to epidural catheter use in the English orthopedic literature. The patient and his family consented to publishing the data. PMID:24353973
Yu, Ryan; Dale, Suzanne E; Yamamura, Deborah; Stankus, Vida; Lee, Christine
Due to the emergence of Staphylococcus aureus with reduced vancomycin susceptibility, newer antibiotics, including daptomycin, have been used to treat methicillin-resistant S aureus infections. Daptomycin is a cyclic lipopeptide that is approved to treat S aureus bacteremia and right-sided endocarditis, and reports of S aureus with reduced susceptibility to daptomycin are infrequent. To our knowledge, the present report describes the first Canadian case of daptomycin-nonsusceptible, vancomycin-intermediate S aureus infection. PMID:23730321
dos Santos, Danielle Caldeira Martins; da Costa, Thaina Miranda; Rabello, Renata Fernandes; Alves, Fábio Aguiar; de Mondino, Silvia Susana Bona
The isolation of mannitol-negative methicillin-resistant Staphylococcus aureus from nasal swabs is reported. Among the 59 isolates, 9 (15%) isolates were mannitol-negative; all of these isolates were categorized as staphylococcal cassette chromosome mec (SCCmec) type IVa. This report emphasizes that mannitol fermentation on mannitol salt agar should not be used as the sole criterion when screening nasal swab specimens for S. aureus. PMID:26273270
Pedro, Ana Luisa; Sousa-Uva, António; Pina, Elaine
Dissemination of methicillin-resistant Staphylococcus aureus (MRSA) remains one of the most difficult challenges for prevention, control, and treatment of health care-associated infections. A survey and interviews were conducted on nurses from a hospital center. We found that most nurses' perceived risk of acquiring MRSA related to themselves (72%), other nurses (88.5%), and patients (97.8%). This perception influences attitudes, leading to compliance with the existing recommendations.
El-Sayed, Mardia Telep; Zoraghi, Roya; Reiner, Neil; Suzen, Sibel; Ohlsen, Knut; Lalk, Michael; Altanlar, Nurten; Hilgeroth, Andreas
Novel bisindolyl-cycloalkane indoles resulted from the reaction of aliphatic dialdehydes and indole. As bisindolyl-natural alkaloid compounds have recently been reported as inhibitors of the methicillin-resistant Staphylococcus aureus (MRSA)-pyruvate kinase (PK), we tested our novel compounds as MRSA PK inhibitors and now report first inhibiting activities. We discuss structure-activity relationships of structurally varied compounds. Activity influencing substituents have been characterized and relations to antibacterial activities of the most active compounds have been proved.
Caelli, M; Porteous, J; Carson, C F; Heller, R; Riley, T V
The combination of a 4% tea tree oil nasal ointment and 5% tea tree oil body wash was compared with a standard 2% mupirocin nasal ointment and triclosan body wash for the eradication of methicillin-resistant Staphylococcus aureus carriage. The tea tree oil combination appeared to perform better than the standard combination, although the difference was not statistically significant due to the small number of patients.
Infections caused by Staphylococcus aureus strains with Methicillin resistance are associated with increased mortality and morbidity, aggressive course, multiple drug resistance and hospital outbreaks. Several first and second line antibiotics are rapidly becoming ineffective for treatment due to emergence of resistance. Extracts of medicinal plants are rich source of unique phytochemicals. Plants used in traditional medicine have been reported to have significant anti-MRSA activity. The objective of this review is to provide a brief overview of antibiotics as well as anti-MRSA natural products and their future prospect.
Ruzauskas, M; Couto, N; Belas, A; Klimiene, I; Siugzdiniene, R; Pomba, C
During 2011, 160 nasal samples were taken from pigs on 8 different farms in Lithuania. Four methicillin-resistant Staphylococcus aureus (MRSA) isolates were obtained. The isolates were ST398, spa type t011 and SCCmec V and none carried the lukF/lukS genes. Strains were resistant to tetracycline, attributed to tetK and tetM genes, and to erythromycin owing to the ermB gene. One MRSA strain was resistant to trimethoprim/sulfamethoxazole and carried the dfrK gene. This is the first report on the presence and characteristics of livestock-associated MRSA isolated from pigs in Lithuania.
Park, Young Kyung; Paik, Young Hwan; Yoon, Jang Won; Fox, Lawrence K.
The sequences of the ccrAB genes from bovine-, canine- and chicken-originating methicillin-resistant Staphylococcus (S.) epidermidis (MRSE) and bovine methicillin-resistant Staphylococcus (S.) aureus (MRSA) were compared to investigate the frequency of intra-species horizontal transfer of the staphylococcal cassette chromosome mec (SCCmec) complex. Nineteen MRSE strains were isolated from bovine milk, chickens, and dogs, and their genetic characteristics were investigated by multilocus sequence typing and SCCmec typing. Among the animal MRSE strains, the most frequent SCCmec type was type IV, which consisted of the type B mec complex and ccrAB type 2. The ccrA2 and ccrB2 genes were sequenced from the bovine, chicken and canine MRSE strains and compared with those of the bovine MRSA strains. The sequences generally clustered as MRSA and MRSE groups, regardless of the animal source. Additionally, no bovine MRSE sequence was associated with the bovine MRSA groups. Although most of the bovine MRSE and MRSA isolates possessed SCCmec type IV sequences, our results suggest that the intra-species gene transfer of the SCCmec complex between bovine S. aureus and bovine S. epidermidis strains is not a frequent event. PMID:23820199
Park, Young Kyung; Paik, Young Hwan; Yoon, Jang Won; Fox, Lawrence K; Hwang, Sun Young; Park, Yong Ho
The sequences of the ccrAB genes from bovine-, canine- and chicken-originating methicillin-resistant Staphylococcus (S.) epidermidis (MRSE) and bovine methicillin-resistant Staphylococcus (S.) aureus (MRSA) were compared to investigate the frequency of intra-species horizontal transfer of the staphylococcal cassette chromosome mec (SCCmec) complex. Nineteen MRSE strains were isolated from bovine milk, chickens, and dogs, and their genetic characteristics were investigated by multilocus sequence typing and SCCmec typing. Among the animal MRSE strains, the most frequent SCCmec type was type IV, which consisted of the type B mec complex and ccrAB type 2. The ccrA2 and ccrB2 genes were sequenced from the bovine, chicken and canine MRSE strains and compared with those of the bovine MRSA strains. The sequences generally clustered as MRSA and MRSE groups, regardless of the animal source. Additionally, no bovine MRSE sequence was associated with the bovine MRSA groups. Although most of the bovine MRSE and MRSA isolates possessed SCCmec type IV sequences, our results suggest that the intra-species gene transfer of the SCCmec complex between bovine S. aureus and bovine S. epidermidis strains is not a frequent event.
Tashiro, Masato; Izumikawa, Koichi; Ashizawa, Nobuyuki; Narukawa, Munetoshi; Yamamoto, Yoshihiro
Distinguishing true coagulase-negative staphylococci bacteremia from contamination remains a challenge. We conducted a retrospective analysis of 183 patients with methicillin-resistant coagulase-negative staphylococci (MR-CoNS)-positive and methicillin-resistant Staphylococcus aureus-positive cultures obtained from sterile sites such as blood, synovial fluid, ascitic fluid, and cerebrospinal fluid. Of the 209 MR-CoNS isolates, 83 (39.7%) were considered infection associated, and 126 (60.3%) were considered contamination. MR-CoNS isolates cultured from synovial fluid were more likely to be infection associated (P = 0.009). The median interval from insertion of a central venous catheter to onset of infection tended to be longer in MR-CoNS infection cases than in methicillin-resistant S. aureus infection cases (41 days versus 14 days, P = 0.055). In conclusion, our results suggest that the proportion of cases of true MR-CoNS infection may be higher than previously reported.
Dağı, Hatice Türk; Fındık, Duygu; Demirel, Gamze; Arslan, Uğur
Background: Staphylococus aureus can be found as a commensal on skin and nasal flora or it may cause local and invasive infections. S. aureus has a large number of virulence factors. Aims: To investigate the methicillin resistance and frequency of various virulence factors in S. aureus nasal isolates. Study Design: Descriptive study. Methods: Nasal samples collected from university students were cultured in media. S. aureus was identified by conventional methods and the Staphyloslide latex test (Becton Dickinson, Sparks, USA). Antibiotic susceptibility tests were conducted, and the methicillin resistance was determined. The mecA, nuc, pvl and staphylococcal toxin genes were examined by polymerase chain reaction (PCR). Results: S. aureus was isolated in 104 of 600 (17.3%) nasal samples. In total, 101 (97.1%) S. aureus isolates were methicillin-sensitive and the remaining 3 (2.9%) were methicillin-resistant. Furthermore, all but five isolates carried at least one staphylococcal enterotoxin gene, with seg being predominant. The tst and eta genes were determined in 29 (27.9%), and 3 (2.9%) isolates, respectively. None of the S. aureus isolates harbored see, etb, and pvl genes. Conclusion: A moderate rate of S. aureus carriage and low frequency of MRSA were detected in healthy students. S. aureus isolates had a high prevalence of staphylococcal enterotoxin genes and the tst gene. In this study, a large number of virulence factors were examined in S. aureus nasal isolates, and the data obtained from this study can be used for monitoring the prevalence of virulence genes in S. aureus strains isolated from nasal carriers. PMID:26167341
Li, Longping; Zhou, Luoxiong; Wang, Lihong; Xue, Huping; Zhao, Xin
Emergence of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS) in bovine milk is a major public health concern. The primary purpose of this research was to determine molecular genetic characteristics and antibiotic resistance of staphylococcal isolates recovered from milk of mastitic cows in the Shaanxi Province in Northwestern China. One hundred and thirteen methicillin-susceptible Staphylococcus aureus (MSSA), one mecA-positive and phenotype-positive MRSA, seven mecA- and mecC- negative but phenotype-positive MRSA and two MR-CoNS including one oxacillin-susceptible mecA-positive Staphylococcus haemolyticus (OS-MRSH) and one mecA-positive and methicillin-resistant Staphylococcus epidermidis (MRSE) isolates were recovered from 214 quarter milk samples on 4 dairy farms. All above 123 isolates were subjected to antibiotic resistance profiling. S. aureus isolates were also genotyped using the spa typing and the multilocus sequence typing (MLST). Eight MRSA and 2 MR-CoNS isolates were additionally tested for SCCmec types. Resistance was common among isolates against ampicillin or penicillin (80.5%), kanamycin (68.3%), gentamicin (67.5%), tetracycline (43.9%) and chloramphenicol (30.1%). However, no isolate was resistant to vancomycin or teicoplanin. Twenty, 29 and 58 isolates showed resistance to 1, 2 or more than 2 antibiotics, respectively. The predominant multidrug resistance profile was penicillin/ampicillin/kanamycin/gentamicin/tetracycline (46 isolates). Most S. aureus isolates belonged to spa types t524 (n = 63), t11772 (a new type, n = 31) and t4207 (n = 15). At the same time, MLST types ST71 (n = 67) and ST2738 (a new type, n = 45) were identified as dominant sequence types. The mecA-positive and phenotype-positive MRSA isolate had a composite genotype t524-ST71-SCCmecIVa, while 7 mecA-negative but phenotype-positive MRSA isolates were all t524-ST71. The OS-MRSH isolate contained a type V
Cercenado, Emilia; Morosini, María Isabel
Because of the increase in bacterial resistance, there is a need for new antimicrobial agents. In particular, Staphylococcus aureus is a frequent cause of severe infections and has an extraordinary capacity to develop antibiotic multiresistance, including resistance to glycopeptides, linezolid, and daptomycin. Although the incidence of methicillin-resistant S. aureus (MRSA) seems to have stabilized in the last few years, its wide dissemination in healthcare settings and in the community is a cause of concern. Ceftaroline is a new broad-spectrum cephalosporin with bactericidal activity against Gram-positive bacteria, including MRSA and multidrug-resistant Streptococcus pneumoniae. In addition, this drug is active against staphylococci showing resistance to glycopeptides, linezolid, and daptomycin. The ceftaroline MIC90 against MRSA ranges from 0.5-2mg/L and that against methicillin-resistant coagulase-negative staphylococci is 0.5mg/L. Ceftaroline has also good activity against respiratory pathogens including Haemophilus influenzae and Moraxella catarrhalis. Although this drug is active against Enterobacteriaceae, it does not retain activity when these isolates produce extended-spectrum beta-lactamases, carbapenemases or hyperproduce AmpC. Ceftaroline is not active against nonfermentative Gram-negative bacilli. Ceftaroline is an interesting addition to the therapeutic armamentarium against MRSA and constitutes an important option for the treatment of polymicrobial infections caused by multidrug-resistant Gram-positive microorganisms.
Paul, N C; Moodley, A; Ghibaudo, G; Guardabassi, L
Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is increasingly reported in small animals and cases of human infections have already been described despite its recent emergence in veterinary practice. We investigated the prevalence of MRSP and methicillin-resistant Staphylococcus aureus (MRSA) among small animal dermatologists attending a national veterinary conference in Italy. Nasal swabs were obtained from 128 veterinarians, seven of which harboured MRSP (n = 5; 3.9%) or MRSA (n = 2; 1.6%). A follow-up study of two carriers revealed that MRSP persisted for at least 1 month in the nasal cavity. Methicillin-susceptible S. aureus (MSSA) was isolated from 32 (25%) conference participants, whereas methicillin-susceptible S. pseudintermedius (MSSP) was not detected, suggesting that MRSP may have a particular ability to colonize humans compared to MSSP. All isolates were characterized by spa typing. Methicillin-resistant isolates were further typed by antimicrobial susceptibility testing, SCCmec and multi-locus sequence typing. Two lineages previously associated with pets were identified among the five MRSP isolates; the European epidemic clone ST71-SCCmec II-III and ST106-SCCmec IV. One of the two MRSA isolates displayed a genotype (ST22- SCCmecIV) frequently reported in dogs and cats. MRSP isolates were resistant to more antimicrobial agents compared with MRSA isolates and displayed the typical multidrug resistance patterns of MRSP in pets. The 32 MSSA isolates belonged to 20 spa types and the most frequent types (t12, t15 and t166) were associated with common S. aureus lineages in humans (CC30 and CC45). Although low, the 3.9% MRSP carriage rate found among small animal dermatologists was surprising in consideration of the rare occurrence of S. pseudintermedius in humans, the lack of MSSP detection and the recent appearance of MRSP in Europe. As cases of human MRSP infection have been linked with pets, veterinarians should be aware of this zoonotic risk
Shan, Weiguang; Li, Jiaping; Fang, Ying; Wang, Xuan; Gu, Danxia; Zhang, Rong
A rapid, sensitive, and accurate Vitek MS assay was developed to distinguish clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) from clinical isolates of methicillin-sensitive Staphylococcus aureus (MSSA) by developing an in-house knowledgebase of SuperSpectra. Three unique peaks, including peaks at 2305.6 and 3007.3 Da specific to MRSA, and 6816.7 Da specific to MSSA, were selected for differentiating MRSA and MSSA. This assay accurately identified 84 and 91% of clinical MRSA and MSSA strains out of the total 142 clinically acquired S. aureus strains that were tested. This method will greatly improve the efficiency of single clinical sample identification of MRSA, thereby facilitating a reduction in the transmission of MRSA in clinical settings.
Rahman, Muhammad M.; Hunter, Howard N.; Prova, Shamina; Verma, Vidhu; Qamar, Aneela
ABSTRACT The methicillin resistance factor encoded by fmtA is a core member of the Staphylococcus aureus cell wall stimulon, but its function has remained elusive for the past two decades. First identified as a factor that affects methicillin resistance in S. aureus strains, FmtA was later shown to interact with teichoic acids and to localize to the cell division septum. We have made a breakthrough in understanding FmtA function. We show that FmtA hydrolyzes the ester bond between d-Ala and the backbone of teichoic acids, which are polyglycerol-phosphate or polyribitol-phosphate polymers found in the S. aureus cell envelope. FmtA contains two conserved motifs found in serine active-site penicillin-binding proteins (PBPs) and β-lactamases. The conserved SXXK motif was found to be important for the d-amino esterase activity of FmtA. Moreover, we show that deletion of fmtA (ΔfmtA) led to higher levels of d-Ala in teichoic acids, and this effect was reversed by complementation of ΔfmtA with fmtA. The positive charge on d-Ala partially masks the negative charge of the polyol-phosphate backbone of teichoic acids; hence, a change in the d-Ala content will result in modulation of their charge. Cell division, biofilm formation, autolysis, and colonization are among the many processes in S. aureus affected by the d-Ala content and overall charge of the cell surface teichoic acids. The esterase activity of FmtA and the regulation of fmtA suggest that FmtA functions as a modulator of teichoic acid charge, thus FmtA may be involved in S. aureus cell division, biofilm formation, autolysis, and colonization. PMID:26861022
Sianglum, Wipawadee; Srimanote, Potjanee; Taylor, Peter W; Rosado, Helena; Voravuthikunchai, Supayang P
Rhodomyrtone, purified from Rhodomyrtus tomentosa (Aiton) Hassk, exhibits a high degree of potency against methicillin-resistant Staphylococcus aureus (MRSA). We recently demonstrated that exposure of MRSA to a subinhibitory concentration (0.174 µg/ml) of rhodomyrtone resulted in the alteration of expression of several functional classes of bacterial proteins. To provide further insight into the antibacterial mode of action of this compound, we determined the impact of exposure to rhodomyrtone on the gene transcriptional profile of MRSA using microarray analysis. Exposure of MRSA to subinhibitory concentrations (0.5MIC; 0.5 µg/ml) of rhodomyrtone revealed significant modulation of gene expression, with induction of 64 genes and repression of 35 genes. Prominent changes in response to exposure to rhodomyrtone involved genes encoding proteins essential to metabolic pathways and processes such as amino acid metabolism, membrane function, ATP-binding cassette (ABC) transportation and lipoprotein and nucleotide metabolism. Genes involved in the synthesis of the aspartate family of amino acids, in particular proteins encoded by the dap operon were prominent. The diaminopimelate (DAP) biosynthetic pathway is the precursor of lysine synthesis and is essential for peptidoglycan biosynthesis. However, phenotypic analysis of the peptidoglycan amino acid content of rhodomyrtone-treated MRSA did not differ significantly from that extracted from control cells. Genes involved in the biosynthesis of amino acids and peptidoglycan, and a high affinity ATP-driven K ((+)) transport system, were investigated by quantitative reverse transcription-PCR (qRT-PCR) using EMRSA-16 1, 4, or 18 h after exposure to rhodomyrtone and in general the data concurred with that obtained by microarray, highlighting the relevance of the DAP biosynthetic pathway to the mode of action of rhodomyrtone.
Sianglum, Wipawadee; Srimanote, Potjanee; Taylor, Peter W.; Rosado, Helena; Voravuthikunchai, Supayang P.
Rhodomyrtone, purified from Rhodomyrtus tomentosa (Aiton) Hassk, exhibits a high degree of potency against methicillin-resistant Staphylococcus aureus (MRSA). We recently demonstrated that exposure of MRSA to a subinhibitory concentration (0.174 µg/ml) of rhodomyrtone resulted in the alteration of expression of several functional classes of bacterial proteins. To provide further insight into the antibacterial mode of action of this compound, we determined the impact of exposure to rhodomyrtone on the gene transcriptional profile of MRSA using microarray analysis. Exposure of MRSA to subinhibitory concentrations (0.5MIC; 0.5 µg/ml) of rhodomyrtone revealed significant modulation of gene expression, with induction of 64 genes and repression of 35 genes. Prominent changes in response to exposure to rhodomyrtone involved genes encoding proteins essential to metabolic pathways and processes such as amino acid metabolism, membrane function, ATP-binding cassette (ABC) transportation and lipoprotein and nucleotide metabolism. Genes involved in the synthesis of the aspartate family of amino acids, in particular proteins encoded by the dap operon were prominent. The diaminopimelate (DAP) biosynthetic pathway is the precursor of lysine synthesis and is essential for peptidoglycan biosynthesis. However, phenotypic analysis of the peptidoglycan amino acid content of rhodomyrtone-treated MRSA did not differ significantly from that extracted from control cells. Genes involved in the biosynthesis of amino acids and peptidoglycan, and a high affinity ATP-driven K (+) transport system, were investigated by quantitative reverse transcription-PCR (qRT-PCR) using EMRSA-16 1, 4, or 18 h after exposure to rhodomyrtone and in general the data concurred with that obtained by microarray, highlighting the relevance of the DAP biosynthetic pathway to the mode of action of rhodomyrtone. PMID:23029218
Frey, K.; Lombardo, M; Wright, D; Anderson, A
Resistance to therapeutics such as trimethoprim-sulfamethoxazole has become an increasing problem in strains of methicillin-resistant Staphylococcus aureus (MRSA). Clinically isolated trimethoprim-resistant strains reveal a double mutation, H30N/F98Y, in dihydrofolate reductase (DHFR). In order to develop novel and effective therapeutics against these resistant strains, we evaluated a series of propargyl-linked antifolate lead compounds for inhibition of the mutant enzyme. For the propargyl-linked antifolates, the F98Y mutation generates minimal (between 1.2- and 6-fold) losses of affinity and the H30N mutation generates greater losses (between 2.4- and 48-fold). Conversely, trimethoprim affinity is largely diminished by the F98Y mutation (36-fold) and is not affected by the H30N mutation. In order to elucidate a mechanism of resistance, we determined a crystal structure of a complex of this double mutant with a lead propargyl-linked antifolate. This structure suggests a resistance mechanism consistent both for the propargyl-linked class of antifolates and for trimethoprim that is based on the loss of a conserved water-mediated hydrogen bond.
Arêde, Pedro; Botelho, Tiago; Guevara, Tibisay; Usón, Isabel; Oliveira, Duarte C; Gomis-Rüth, F Xavier
Methicillin resistance in Staphylococcus aureus is elicited by the MecI-MecR1-MecA axis encoded by the mec locus. Recently, MecR2 was also identified as a regulator of mec through binding of the methicillin repressor, MecI. Here we show that plasmid-encoded full-length MecR2 restores resistance in a sensitive S. aureus mecR2 deletion mutant of the resistant strain N315. The crystal structure of MecR2 reveals an N-terminal DNA-binding domain, an intermediate scaffold domain, and a C-terminal dimerization domain that contributes to oligomerization. The protein shows structural similarity to ROK (repressors, open reading frames, and kinases) family proteins, which bind DNA and/or sugar molecules. We found that functional cell-based assays of three point mutants affecting residues participating in sugar binding in ROK proteins had no effect on the resistance phenotype. By contrast, MecR2 bound short double-stranded DNA oligonucleotides nonspecifically, and a deletion mutant affecting the N-terminal DNA-binding domain showed a certain effect on activity, thus contributing to resistance less than the wild-type protein. Similarly, a deletion mutant, in which a flexible segment of intermediate scaffold domain had been replaced by four glycines, significantly reduced MecR2 function, thus indicating that this domain may likewise be required for activity. Taken together, these results provide the structural basis for the activity of a methicillin antirepressor, MecR2, which would sequester MecI away from its cognate promoter region and facilitate its degradation.
Seidl, Kati; Leemann, Michèle; Palheiros Marques, Miguel; Rachmühl, Carole; Leimer, Nadja; Andreoni, Federica; Achermann, Yvonne; Zinkernagel, Annelies S
There has been controversy about the intrinsic virulence of methicillin-resistant Staphylococcus aureus (MRSA) as compared to methicillin-susceptible S. aureus (MSSA). To address this discrepancy, the intrinsic virulence of 42 MRSA and 40 MSSA clinical isolates was assessed by testing endothelial cell (EC) damage, a surrogate marker for virulence in blood stream infections. Since these clinical isolates represent a heterogeneous group, well characterized S. aureus laboratory strains with SCCmec loss- and gain-of-function mutations were used in addition. The clinical MRSA isolates carrying typical hospital acquired SCCmec types (I, II or III) induced significantly less damage (47.8%) as compared to isolates with other SCCmec types (62.3%, p=0.03) and MSSA isolates (64.2%, p<0.01). There was a strong inverse correlation between high-level oxacillin resistance and low EC damage induction (R(2)=0.4464, p<0.001). High-level oxacillin resistant strains (MIC >32μ/ml) grew significantly slower as compared to isolates with low-level resistance (p=0.047). The level of EC damage positively correlated with α- and δ-toxin production (p<0.0001 and p<0.05, respectively) but not with β-toxin production. Invasive MRSA isolates (n=21, 56.3%) were significantly less cytotoxic as compared to invasive MSSA isolates (n=20, 68.0%, p<0.05). There was no difference between EC damage induced by superficial versus invasive isolates in either MRSA or MSSA strains. Our data suggest that the intrinsic virulence of MRSA is similar or even reduced as compared to MSSA strains but is linked to the level of methicillin resistance.
Lovering, Andrew L.; Gretes, Michael C.; Safadi, Susan S.; Danel, Franck; de Castro, Liza; Page, Malcolm G. P.; Strynadka, Natalie C. J.
Methicillin-resistant Staphylococcus aureus (MRSA) is an antibiotic-resistant strain of S. aureus afflicting hospitals and communities worldwide. Of greatest concern is its development of resistance to current last-line-of-defense antibiotics; new therapeutics are urgently needed to combat this pathogen. Ceftobiprole is a recently developed, latest generation cephalosporin and has been the first to show activity against MRSA by inhibiting essential peptidoglycan transpeptidases, including the β-lactam resistance determinant PBP2a, from MRSA. Here we present the structure of the complex of ceftobiprole bound to PBP2a. This structure provides the first look at the molecular details of an effective β-lactam-resistant PBP interaction, leading to new insights into the mechanism of ceftobiprole efficacy against MRSA. PMID:22815485
Rudkin, Justine K.; Schaeffer, Carolyn R.; Lohan, Amanda J.; Tong, Pin; Loftus, Brendan J.; Pier, Gerald B.; Fey, Paul D.; Massey, Ruth C.; O'Gara, James P.
Clinical isolates of Staphylococcus aureus can express biofilm phenotypes promoted by the major cell wall autolysin and the fibronectin-binding proteins or the icaADBC-encoded polysaccharide intercellular adhesin/poly-N-acetylglucosamine (PIA/PNAG). Biofilm production in methicillin-susceptible S. aureus (MSSA) strains is typically dependent on PIA/PNAG whereas methicillin-resistant isolates express an Atl/FnBP-mediated biofilm phenotype suggesting a relationship between susceptibility to β-lactam antibiotics and biofilm. By introducing the methicillin resistance gene mecA into the PNAG-producing laboratory strain 8325-4 we generated a heterogeneously resistant (HeR) strain, from which a homogeneous, high-level resistant (HoR) derivative was isolated following exposure to oxacillin. The HoR phenotype was associated with a R602H substitution in the DHHA1 domain of GdpP, a recently identified c-di-AMP phosphodiesterase with roles in resistance/tolerance to β-lactam antibiotics and cell envelope stress. Transcription of icaADBC and PNAG production were impaired in the 8325-4 HoR derivative, which instead produced a proteinaceous biofilm that was significantly inhibited by antibodies against the mecA-encoded penicillin binding protein 2a (PBP2a). Conversely excision of the SCCmec element in the MRSA strain BH1CC resulted in oxacillin susceptibility and reduced biofilm production, both of which were complemented by mecA alone. Transcriptional activity of the accessory gene regulator locus was also repressed in the 8325-4 HoR strain, which in turn was accompanied by reduced protease production and significantly reduced virulence in a mouse model of device infection. Thus, homogeneous methicillin resistance has the potential to affect agr- and icaADBC-mediated phenotypes, including altered biofilm expression and virulence, which together are consistent with the adaptation of healthcare-associated MRSA strains to the antibiotic-rich hospital environment in which they are
Bemis, David A; Jones, Rebekah D; Videla, Ricardo; Kania, Stephen A
Cefoxitin disk diffusion susceptibility testing is a recommended screening method for the detection of methicillin resistance in human isolates of Staphylococcus aureus and coagulase-negative staphylococci. A retrospective analysis of 1,146 clinical isolates of Staphylococcus pseudintermedius from dogs was conducted to determine if screening by the cefoxitin disk method can be similarly useful with S. pseudintermedius. The distribution of cefoxitin growth inhibition zone diameters within this collection was bimodal and correlated well with the results of methicillin resistance gene (mecA) detection by polymerase chain reaction. Of the isolates, 5% had discordant results and, when retested, 84% of these were in agreement. While a greater diversity of isolates and interlaboratory comparisons must be tested, the current study suggests that an epidemiological breakpoint (of approximately ≤ 30 mm = resistant; ≥ 31 = susceptible) can be established to predict methicillin resistance in S. pseudintermedius.
Drew, W L; Barry, A L; O'Toole, R; Sherris, J C
The resistance of Staphylococcus aureus to methicillin and related drugs can be reliably determined by using the Kirby-Bauer method of susceptibility testing if the incubation temperature is 35 C or below, but resistance may be missed at 37 C. The 1-mug discs of oxacillin and nafcillin or the 5-mug discs of methicillin may be used for this purpose but not the 1-mug discs of cloxacillin. The latter fail to discriminate between sensitive and resistant staphylococci by zone measurement; some resistant strains of staphylococci may show larger zones of inhibition than sensitive strains. Stability of these antibiotic-containing discs was studied under conditions of temperature and humidity variation that might be encountered in a clinical laboratory refrigerator. Oxacillin discs were the most stable and are to be preferred for susceptibility testing. Nafcillin discs were less stable, and methicillin discs lose their potency rapidly unless carefully stored in a refrigerator with a desiccant.
Ishihara, Kanako; Shimokubo, Natsumi; Sakagami, Akie; Ueno, Hiroshi; Muramatsu, Yasukazu; Kadosawa, Tsuyoshi; Yanagisawa, Chie; Hanaki, Hideaki; Nakajima, Chie; Suzuki, Yasuhiko; Tamura, Yutaka
Recently, methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP) have been increasingly isolated from veterinarians and companion animals. With a view to preventing the spread of MRSA and MRSP, we evaluated the occurrence and molecular characteristics of each in a veterinary college. MRSA and MRSP were isolated from nasal samples from veterinarians, staff members, and veterinary students affiliated with a veterinary hospital. Using stepwise logistic regression, we identified two factors associated with MRSA carriage: (i) contact with an identified animal MRSA case (odds ratio [OR], 6.9; 95% confidence interval [95% CI], 2.2 to 21.6) and (ii) being an employee (OR, 6.2; 95% CI, 2.0 to 19.4). The majority of MRSA isolates obtained from individuals affiliated with the veterinary hospital and dog patients harbored spa type t002 and a type II staphylococcal cassette chromosome mec (SCCmec), similar to the hospital-acquired MRSA isolates in Japan. MRSA isolates harboring spa type t008 and a type IV SCCmec were obtained from one veterinarian on three different sampling occasions and also from dog patients. MRSA carriers can also be a source of MRSA infection in animals. The majority of MRSP isolates (85.2%) carried hybrid SCCmec type II-III, and almost all the remaining MRSP isolates (11.1%) carried SCCmec type V. MRSA and MRSP were also isolated from environmental samples collected from the veterinary hospital (5.1% and 6.4%, respectively). The application of certain disinfection procedures is important for the prevention of nosocomial infection, and MRSA and MRSP infection control strategies should be adopted in veterinary medical practice.
Samanta, Dhritiman; Batte, Justin L; Brown, Stephanie N; Crosby, Angela G; Marcos, Luis A; Elasri, Mohamed O
Staphylococcus aureus is the predominant cause of bacteremia worldwide. We assessed the molecular epidemiology and antibiotic resistance of methicillin-resistant S aureus isolates causing bacteremia in southern Mississippi. Diverse genetic backgrounds in terms of staphylococcal cassette chromosome mec, pulsed-field gel electrophoresis, and multilocus sequence typing types of methicillin-resistant S aureus were identified as causing bacteremia in Mississippi. A strong association of Panton-Valentine leukocidin genes with elevated vancomycin minimum inhibitory concentration is one of the important findings of our study.
Thiébaut, Anne C. M.; Lawrence, Christine; Bouchiat, Coralie; Herrmann, Jean-Louis; Salomon, Jérôme
Staphylococcus aureus nasal carriage is a risk factor for subsequent infection. Estimates of colonization duration vary widely among studies, and factors influencing the time to loss of colonization, especially the impact of antibiotics, remain unclear. We conducted a prospective study on patients naive for S. aureus colonization in 4 French long-term-care facilities. Data on nasal colonization status and potential factors for loss of colonization were collected weekly. We estimated methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) colonization durations using the Kaplan-Meier method and investigated factors for loss of colonization using shared-frailty Cox proportional hazards models. A total of 285 S. aureus colonization episodes were identified in 149 patients. The median time to loss of MRSA or MSSA colonization was 3 weeks (95% confidence interval, 2 to 8 weeks) or 2 weeks (95% confidence interval, 2 to 3 weeks), respectively. In multivariable analyses, the methicillin resistance phenotype was not associated with S. aureus colonization duration (P = 0.21); the use of fluoroquinolones (hazard ratio, 3.37; 95% confidence interval, 1.31 to 8.71) and having a wound positive for a nonnasal strain (hazard ratio, 2.17; 95% confidence interval, 1.15 to 4.07) were associated with earlier loss of MSSA colonization, while no factor was associated with loss of MRSA colonization. These results suggest that the methicillin resistance phenotype does not influence the S. aureus colonization duration and that fluoroquinolones are associated with loss of MSSA colonization but not with loss of MRSA colonization. PMID:26416866
Nienhoff, Ulrike; Kadlec, Kristina; Chaberny, Iris F; Verspohl, Jutta; Gerlach, Gerald-F; Kreienbrock, Lothar; Schwarz, Stefan; Simon, Daniela; Nolte, Ingo
The aim of this study was to determine the frequency of carriage of methicillin-resistant Staphylococcus pseudintermedius (MRSP) among dogs admitted to a small animal hospital during a 17-month period, to characterize these isolates and to initially screen for possible factors associated with MRSP carriage. Swabs were taken from the nose/pharynx and the perineum as well as from wounds and skin infections (if present) of 814 dogs before entering the small animal hospital. A questionnaire for background information was completed. The staphylococcal species and methicillin resistance were confirmed pheno- and genotypically. The identified MRSP isolates were characterized by SCCmec typing, testing for susceptibility to 25 antimicrobial agents and SmaI-directed pulsed-field gel electrophoresis. A first screening for possible risk factors for MRSP carriage was performed by means of unifactorial contingency tables and CART analysis. Sixty (7.4%) dogs were positive for MRSP. All MRSP isolates harboured a type II-III SCCmec cassette and showed extended resistance to antimicrobial agents. Fifteen different SmaI patterns were observed. The major factors that clustered with MRSP carriage were former hospitalization and antibiotic treatment within the last six months before sampling. This study showed that only a minor part of the sampled dogs carried multi-resistant MRSP isolates. The facts that prior hospitalization and/or antibiotic therapy are potential associated factors for MRSP carriage underline the necessity of a judicious use of antibiotics in small animal medicine.
Couto, Natacha; Belas, Adriana; Tilley, Paula; Couto, Isabel; Gama, Luís T; Kadlec, Kristina; Schwarz, Stefan; Pomba, Constança
The aim of this study was to evaluate the biocide and antimicrobial susceptibility of methicillin-resistant staphylococcal isolates from horses. Fourteen methicillin-resistant staphylococci (MRS) were subjected to an extensive genotype characterization, including SCCmec, dru, spa, PFGE and MLST typing. Antimicrobial susceptibility testing was performed and resistance genes were detected by PCR. Minimum bactericidal concentrations (MBCs) of four biocides [chlorhexidine acetate (CHA), benzalkonium chloride (BAC), triclosan (TCL) and glutaraldehyde (GLA)] were determined following the recommendations of document NF EN 1040. The presence of qac and sh-fabI genes was investigated by PCR. Several antimicrobial resistance patterns and genes were detected. When MRS strains were exposed for a longer period of time, a lower concentration of the biocide was needed to achieve lethality. TCL had the lowest MBC values. All MBC values were lower than the recommended in-use concentrations for veterinary medicine. S. haemolyticus and S. cohnii subsp. cohnii carried plasmid-borne qacA and sh-fabI or qacB and a qacH-like genes, respectively. Biocides appear to be a reliable antiseptic option against MRS, since even in the presence of bacterial efflux mechanisms, the recommended concentration is much higher than the in vitro MBC.
Strohsahl, Christopher M.; Miller, Benjamin L.; Krauss, Todd D.
Staphylococcus aureus is a leading cause of human illness, and has developed the remarkable ability to resist the bactericidal capabilities of many of the world's leading antibiotics (i.e. MRSA). In an effort to enable rapid detection and treatment of MRSA infections, we have developed a DNA detection technology termed the NanoLantern(TM). The NanoLantern(TM) biosensor technology is based on the simple immobilization of a fluorophore-terminated DNA hairpin onto a gold chip. This produces a label-free sensor that allows for a positive response to be obtained without extensive processing of the sample, saving cost and increasing accuracy. We will also discuss a newly developed method of partial gene analysis, used to develop a DNA hairpin probe that is capable of detecting the presence of the mecR gene, a gene necessary for methicillin resistance to be present in S. aureus, with 100% sequence specificity. The successful incorporation of this probe into the NanoLantern(TM) platform, along with the concomitant development of the paired PCR assay has allowed for the successful detection of methicillin-resistance directly from a culture of S. aureus. These results represent an important step forward in terms of developing the ability to rapidly and effectively detect the presence of antibiotic resistance in bacterial infections.
Schissler, Jennifer R; Hillier, Andrew; Daniels, Joshua B; Cole, Lynette K; Gebreyes, Wondwossen A
The Clinical and Laboratory Standards Institute published in 2008 new interpretive criteria for the identification of methicillin resistance in staphylococci isolated from animals. The sensitivity of the 2008 interpretive criteria for mecA gene-positive Staphylococcus pseudintermedius, compared with the previous criteria of 2004, was investigated. Thirty clinical isolates of methicillin-resistant S. pseudintermedius from dogs were used. The presence of the mecA gene was determined by polymerase chain reaction. The minimum inhibitory concentration for oxacillin was determined by broth microdilution. The 2008 breakpoint of >or=4 microg/ml for methicillin resistance resulted in a diagnostic sensitivity of 73.3% (22/30). The 2004 breakpoint guideline of >or=0.5 microg/ml resulted in a diagnostic sensitivity of 97% (29/30). For oxacillin disk diffusion, the 2008 interpretive criterion of
Ammar, A M; Attia, A M; Abd El-Hamid, M I; El-Shorbagy, I M; Abd El-Kader, S A
Antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem for clinicians worldwide. The present study attempted to evaluate the susceptibility patterns of MRSA to various antimicrobials and the prevalence of inducible clindamycin resistance as well as the relevant antibiotic and antiseptic resistance genes among these isolates. Totally, 40 MRSA isolates were recovered from examined milk and meat product samples (18.60%). Multi-drug resistance (MDR) was remarkably observed among 85% of these isolates. There was a good correlation between phenotypic determination of methicillin, amoxicillin/clavulinic acid and tetracycline resistances and PCR detections of mecA, blaZ and tet(K) genes, respectively, but norA gene was not detected in the four ciprofloxacin resistant isolates. Although, 55% of MRSA expressed resistance to benzalkonium chloride (BC), neither qacA/B nor smr gene was detected. Of 20 isolates exhibiting erythromycin- clindamycin discordant resistance pattern, 8 displayed positive double disk diffusion (D-zone) test denoting inducible macrolide-lincosamide-streptogramin B (MLSB) resistance phenotype with the inducibly expressed erm(A) and erm(C) genes in 87.5% of these isolates. Besides, the remaining 12 isolates showed MS phenotype (resistant to macrolides and type B streptogramins only) with a variety of erm(A), mph(C), msr(A) or a combination of these genes including erm(C). Finally, the constitutive MLSB phenotype with the constitutive expression of erm(A), erm(B) and erm(C) genes was comprised in 2 isolates with higher minimum inhibitory concentration (MIC) values for erythromycin (512 and 1024 µg/ml) and clindamycin (16 and 32 µg/ml). These findings suggested the importance of monitoring the evolution of MRSA resistance.
Ugwu, Clifford C.; Gomez-Sanz, Elena; Agbo, Ifeoma C.; Torres, Carmen; Chah, Kennedy F.
This study was conducted to determine the species distribution, antimicrobial resistance pheno- and genotypes and virulence traits of mannitol-positive methicillin-resistant staphylococci (MRS) isolated from pigs in Nsukka agricultural zone, Nigeria. Twenty mannitol-positive methicillin-resistant coagulase-negative staphylococcal (MRCoNS) strains harboring the mecA gene were detected among the 64 Staphylococcus isolates from 291 pigs. A total of 4 species were identified among the MRCoNS isolates, namely, Staphylococcus sciuri (10 strains), Staphylococcus lentus (6 strains), Staphylococcus cohnii (3 strains) and Staphylococcus haemolyticus (one strain). All MRCoNS isolates were multidrug-resistant. In addition to β-lactams, the strains were resistant to fusidic acid (85%), tetracycline (75%), streptomycin (65%), ciprofloxacin (65%), and trimethoprim/sulphamethoxazole (60%). In addition to the mecA and blaZ genes, other antimicrobial resistance genes detected were tet(K), tet(M), tet(L), erm(B), erm(C), aacA-aphD, aphA3, str, dfrK, dfrG, cat pC221, and cat pC223. Thirteen isolates were found to be ciprofloxacin-resistant, and all harbored a Ser84Leu mutation within the QRDR of the GyrA protein, with 3 isolates showing 2 extra substitutions, Ser98Ile and Arg100Lys (one strain) and Glu88Asp and Asp96Thr (2 strains). A phylogenetic tree of the QRDR nucleotide sequences in the gyrA gene revealed a high nucleotide diversity, with several major clusters not associated with the bacterial species. Our study highlights the possibility of transfer of mecA and other antimicrobial resistance genes from MRCoNS to pathogenic bacteria, which is a serious public health and veterinary concern. PMID:26413075
Saphyakhajon, Phisit; Joshi, Avni Y; Huskins, W Charles; Henry, Nancy K; Boyce, Thomas G
The bacterial agents causing bone and joint infections have been changing. Currently, methicillin-resistant Staphylococcus aureus (MRSA) and Kingella kingae are emerging pathogens. For treatment of MRSA infections, clindamycin, vancomycin, and linezolid are commonly prescribed antibiotics. Kingella are sensitive to most penicillins and cephalosporins. Because MRSA osteoarticular infections tend to be severe, longer periods of antibiotic treatment with more frequent monitoring of inflammatory markers are sometimes required to obtain a complete cure with no residual complications. To assist management, we have included a clinical decision tree with antibiotic treatment protocols.
Bettenworth, Dominik; Nowacki, Tobias M; Friedrich, Alexander; Becker, Karsten; Wessling, Johannes; Heidemann, Jan
We report on a 24-year-old male patient with history of bloody diarrhea, abdominal pain and vomiting. Endoscopy revealed massive ulcerative discontinuous proctosigmoiditis with deep, sharply demarcated epithelial denudations and enterotoxigenic methicillin-resistant Staphylococcus aureus (MRSA) was detected in mucosal biopsies. After treatment with linezolide and steroids, a significant amelioration of colitis was detected and testing for MRSA became negative. In face of the case presented here, we suggest that in patients with refractory inflammatory bowel disease (IBD), microbiological assessment should be performed to detect a possible Staphylococcus aureus infection in order to initiate an antimicrobial treatment in addition to IBD-specific treatment.
Anusionwu, Obiora F; Smith, Cheri; Cheng, Alan
Methicillin resistant Staphylococcus aureus (MRSA) septicemia is associated with high morbidity and mortality especially in patients with immunosuppression, diabetes, renal disease and endocarditis. There has been an increase in implantation of cardiac implantable electronic devices (CIED) with more cases of device-lead associated endocarditis been seen. A high index of suspicion is required to ensure patient outcomes are optimized. The excimer laser has been very efficient in helping to ensure successful lead extractions in patients with CIED infections. We present an unusual case report and literature review of MRSA septicemia from device-lead endocarditis and the importance of early recognition and prompt treatment. PMID:22905295
Furukawa, Kenei; Tsutsumi, Jun; Takayama, Sumio; Mashiko, Hiroshi; Shiba, Hiroaki; Yanaga, Katsuhiko
A 68-year-old man underwent total gastrectomy and splenectomy for adenocarcinoma and low anterior resection for carcinoma in tubulo-villous adenoma of the rectum. Broad-spectrum antibiotics were administered for postoperative pancreatic fistula. Methicillin-resistant Staphylococcus aureus enteritis occurred on the 50th postoperative day and cecal perforation followed. The patient underwent construction of cecostomy with peritoneal drainage, and vancomycin was administered orally and per cecostomy for 2 weeks. The patient recovered well and was discharged at 35 days after re-operation in good general condition. PMID:25785319
Daum, Luke T; Bumah, Violet V; Masson-Meyers, Daniela S; Khubbar, Manjeet; Rodriguez, John D; Fischer, Gerald W; Enwemeka, Chukuka S; Gradus, Steve; Bhattacharyya, Sanjib
We report here the whole-genome sequence of the USA300 strain of methicillin-resistant Staphylococcus aureus (MRSA), designated ATCC BAA-1680, and commonly referred to as community-associated MRSA (CA-MRSA). This clinical MRSA isolate is commercially available from the American Type Culture Collection (ATCC) and is widely utilized as a control strain for research applications and clinical diagnosis. The isolate was propagated in ATCC medium 18, tryptic soy agar, and has been utilized as a model S. aureus strain in several studies, including MRSA genetic analysis after irradiation with 470-nm blue light.
Baillargeon, Jacques; Kelley, Michael F; Leach, Charles T; Baillargeon, Gwen; Pollock, Brad H
Recent reports indicate that correctional facility inmates may be at elevated risk for contracting methicillin-resistant Staphylococcus aureus (MRSA) infection because of overcrowding, poor hygiene, and high rates of diseases causing immunosuppression. The present study of 299,179 Texas inmates who were incarcerated between 1999-2001 indicated an incidence of 12 MRSA infections/1000 person-years. Inmates with circulatory disease, cardiovascular disease, diabetes, end-stage liver disease, end-stage renal disease, human immunodeficiency virus infection or acquired immunodeficiency syndrome, and skin diseases all exhibited elevated rates of MRSA infection.
Venniyil, Prasanth V.; Ganguly, Satyaki; Kuruvila, Sheela; Devi, Sheela
Background: Health care–associated methicillin-resistant Staphylococcus aureus(HA-MRSA) are resistant to multiple antibiotics, therefore infections caused by them are difficult to treat resulting in high morbidity and mortality. While most of the research activities and public health initiatives are focused on HA-MRSA, the newly emerging pathogen, community-associated methicillin-resistant Staphylococcus aureus(CA-MRSA) is gaining in significance in respect to patient morbidity. There is a significant paucity of data regarding CA-MRSA in the developing parts of the world. Aim: To study the proportions of HA-MRSA and CA-MRSA infections among patients with culture-proven S. aureus infection and to find out how many of these patients showed presence of MRSA in nasal cultures of healthy contacts. Materials and Methods: Clinical details of 227 patients were recorded in the study, such as the duration and recurrence of the infection, history of antibiotic intake, and the presence of other medical illnesses. A pus swab was taken from each lesion and sent for culture and sensitivity. If the culture grew S. aureus, they were screened for methicillin resistance. A swab from the anterior nares of the healthy contact of each patient, whenever available, was collected and it was screened for MRSA. Results: Furunculosis was most common among the primary pyodermas (53/134; 39. 5%). Out of 239 pus culture samples obtained from 227 patients, 192 (84.58%) grew S. aureus; of these 150 (78.12%) were methicillin-sensitive S. aureus (MSSA), whereas 42 (21.98%) were MRSA. Out of the 42 MRSA isolated, 33 turned out to be CA-MRSA (78%) and 9 (22%) were HA-MRSA. Nasal swabs of healthy contacts of 34 MRSA patients were cultured. Out of them, two grew MRSA in the culture. Conclusion: The isolation rate of S. aureus was high in our study. Furthermore, our study, although hospital based, clearly indicated the substantial magnitude of the CA-MRSA problem in the local population. PMID:27294048
Jana, Tanima; Machicado, Jorge D.; Davogustto, Giovanni E.; Pan, Jen-Jung
Prostatic abscesses are usually related to gram-negative bacilli. However, methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a substantial cause of prostatic abscesses in recent years. Herein, we report the case of a 31-year-old man with a history of orthotopic liver transplantation 10 years ago who presented with acute onset dysuria and abdominal pain and was diagnosed with a MRSA prostatic abscess. To our knowledge, this is the first case describing a prostatic abscess in a liver transplant recipient and the first reporting MRSA as the causative organism of a prostatic abscess in a solid organ transplant recipient. PMID:25389507
Yamada, Y; Sugai, M; Woo, M; Nishida, N; Sugimoto, T
Local infection of the trachea in intubated neonates is one of the main risk factors for development of acquired subglottic stenosis, although its role in the pathogenesis is unclear. Methicillin resistant Staphylococcus aureus (MRSA) is often the cause of critical illness in neonatal patients. Two cases are reported of acquired subglottic stenosis following bacterial infection of the trachea, suggesting an association with the staphylococcal exotoxin, epidermal cell differentiation inhibitor (EDIN). EDIN-producing MRSA were isolated from purulent tracheal secretions from both infants. Acquired subglottic stenosis in both cases was probably caused by delayed wound healing as the result of EDIN inhibition of epithelial cell migration. PMID:11124922
Kim, Tae-Hee; Kim, Soo Ah; Heo, Gyeong-Eun
Infections of the vulva can present a complex differential to the gynecologist, ranging from superficial skin infections to lifethreatening necrotizing fasciitis. Recognition and timely treatment remain universal to skin and soft-tissue infections as the subcutaneous anatomy of the vulva can facilitate rapid spread to other tissues with significant morbidity and mortality. Employing a multidisciplinary team approach to care for vulvar cellulitis and abscess can guide treatment from antibiotic therapies to more aggressive surgical debridement. In this report, we describe a case of vulvar abscess caused by Methicillin-resistant staphylococcus aureus (MRSA) in a postmenopausal woman with underlying diseases of bronchiectasis and atelectasis. PMID:27617247
Furukawa, Kenei; Tsutsumi, Jun; Takayama, Sumio; Mashiko, Hiroshi; Shiba, Hiroaki; Yanaga, Katsuhiko
A 68-year-old man underwent total gastrectomy and splenectomy for adenocarcinoma and low anterior resection for carcinoma in tubulo-villous adenoma of the rectum. Broad-spectrum antibiotics were administered for postoperative pancreatic fistula. Methicillin-resistant Staphylococcus aureus enteritis occurred on the 50th postoperative day and cecal perforation followed. The patient underwent construction of cecostomy with peritoneal drainage, and vancomycin was administered orally and per cecostomy for 2 weeks. The patient recovered well and was discharged at 35 days after re-operation in good general condition.
Skov, Robert; Gudlaugsson, Olafur; Hardardottir, Hjordis; Harthug, Stig; Jakobsen, Trond; Kolmos, Hans Jørn; Olsson-Liljequist, Barbro; Peltonen, Reijo; Tveten, Yngvar; Vuopio-Varkila, Jaana; Ahrén, Christina
The recent increase in the incidence of methicillin-resistant Staphylococcus aureus in all the Nordic countries prompted the Scandinavian Society for Antimicrobial Chemotherapy (SSAC) to create the 'SSAC Working Party on MRSA' with the objective to identify methods to keep the invasive MRSA infections in the Nordic countries below 1%. The lack of common definitions was recognized as a major obstacle for a joint Nordic effort to combat MRSA. The aim of this publication is to present proposals for epidemiological definitions of individual cases, for how to report MRSA frequency per country, and for communication of MRSA strain characteristics between the countries.
Vandenbroucke-Grauls, C M J E; Beaujean, D J M A
It was recently observed that pig breeders in The Netherlands often carry methicillin-resistant Staphylococus aureus (MRSA). These MRSA strains are related to MRSA strains found in pigs. A case-control study showed that not only pig breeders but also cattle breeders are at risk of carrying MRSA. It is advised to keep pig breeders, if they are admitted to a hospital, in isolation until surveillance cultures are proven negative. This also applies to veterinarians and slaughterhouse personnel. For cattle breeders screening without isolation on admission to a hospital is sufficient.
Ito, Teruyo; Tsubakishita, Sae; Sasaki, Takashi; Takeuchi, Fumihiko; Morimoto, Yuh; Katayama, Yuki; Matsuo, Miki; Kuwahara-Arai, Kyoko; Hishinuma, Tomomi; Baba, Tadashi
Since the discovery of the first strain in 1961 in England, MRSA, the most notorious multidrug-resistant hospital pathogen, has spread all over the world. MRSA repeatedly turned down the challenges by number of chemotherapeutics, the fruits of modern organic chemistry. Now, we are in short of effective therapeutic agents against MRSA prevailing among immuno-compromised patients in the hospital. On top of this, we recently became aware of the rise of diverse clones of MRSA, some of which have increased pathogenic potential compared to the classical hospital-associated MRSA, and the others from veterinary sources. They increased rapidly in the community, and started menacing otherwise healthy individuals by causing unexpected acute infection. This review is intended to provide a whole picture of MRSA based on its genetic makeup as a versatile pathogen and our tenacious colonizer. PMID:24265961
Guntupalli, Rajesh; Sorokulova, Iryna; Olsen, Eric; Globa, Ludmila; Pustovyy, Oleg; Moore, Timothy; Chin, Bryan; Barbaree, James; Vodyanoy, Vitaly
Discrimination of methicillin resistant (MRSA) and sensitive (MSSA) strains of Staphylococcus aureus, was achieved by the specially selected lytic bacteriophage with a wide host range of S. aureus strains and a penicillin-binding protein (PBP 2a) specific antibody. A quartz crystal microbalance with dissipation monitoring (QCM-D) was employed to analyze bacteria-phage interactions. The lytic phages were transformed into phage spheroids by exposure to water-chloroform interface. Phage spheroid monolayers were transferred onto QCM-D sensors by Langmuir-Blodgett (LB) technique. Biosensors were tested in the flow mode with bacterial water suspensions, while collecting frequency and energy dissipation changes. Bacteria-spheroid interactions resulted in decreased resonance frequency and an increase in dissipation energy for both MRSA and MSSA strains. Following the bacterial binding, these sensors were further exposed to a flow of the penicillin-binding protein (PBP 2a) specific antibody conjugated latex beads. Sensors tested with MRSA responded to PBP 2a antibody beads; while sensors examined with MSSA gave no response. This experimental difference establishes an unambiguous discrimination between methicillin resistant and sensitive S. aureus strains. Both free and immobilized bacteriophages strongly inhibit bacterial growth on solid/air interfaces and in water suspensions. After lytic phages are transformed into spheroids, they retain their strong lytic activity and demonstrate high bacterial capture efficiency. The phage and phage spheroids can be used for screening and disinfection of antibiotic resistant bacteria. Other applications may include use on biosensors, bacteriophage therapy, and antimicrobial surfaces.
Gómez-Sanz, Elena; Torres, Carmen; Lozano, Carmen; Sáenz, Yolanda; Zarazaga, Myriam
The objective was to identify the methicillin-resistant coagulase-positive staphylococci (MRCoPS) nasal carriage rate of healthy dogs in La Rioja (Spain) and to characterize the recovered isolates by different molecular techniques. Nasal samples from 196 dogs were obtained (98 household-dogs, 98 pound-dogs). Isolates were identified and characterized by spa-, SCCmec- and MLST-typing, SmaI-PFGE, antimicrobial susceptibility, determination of antimicrobial resistance and toxin genes profiling. S. pseudintermedius was the only species recovered. Nine methicillin-resistant S. pseudintermedius (MRSP) were obtained from 9 of 196 sampled dogs (8% pound-dogs, 1% household-dogs). MRSP isolates were typed (MLST/PFGE/spa/SCCmec) as: ST71/A/t02/II-III (7 isolates), ST92/C/t06/V (1 isolate), and ST26/B/non-typable/non-typable (1 isolate). All MRSP were resistant to [resistance gene/number isolates]: β-lactams [mecA+blaZ/9], tetracycline [tet(K)/7, tet(M)/2], macrolides and lincosamides [erm(B)/9], aminoglycosides [aacA-aphD+aadE+aphA-3/9], and co-trimoxazol [dfr(G)/9]. Eight MRSP isolates showed also resistance to fluoroquinolones and amino acid changes in GyrA [Ser84Leu+Glu714Lys, 7 isolates; Ser84Leu, 1 isolate] and GrlA [Ser80Ile, 8 isolates] proteins were detected. The remaining isolate was chloramphenicol resistant and harboured cat(pC221) gene. All MRSP isolates harboured the aadE-sat4-aphA-3 multiresistance-gene-cluster linked to erm(B) gene as well as the siet, si-ent and lukS/F-I toxin genes. MRSP is a moderately common (4.6%) colonizer of healthy dogs in Spain. A major MRSP lineage (ST71) was detected and its future evolution should be tracked.
Glaeser, Stefanie P; Sowinsky, Olivia; Brunner, Jana S; Dott, Wolfgang; Kämpfer, Peter
Vancomycin-resistant enterococci (VRE) and methicillin-resistant staphylococci (MRS) were detected in two mesophilic German biogas plants (BGPs) using selective pre-enrichment methods combined with cultivation on CHROMagar media and antibiotic resistance gene screening. Genetic fingerprinting and 16S rRNA gene sequencing showed the presence of enterococci isolated by the VRE selective cultivation (67 isolates) in input and output samples of BGPs. In contrast, MRS (44 isolates) were detected in input, but in none of the output samples. Enterococcus isolates showed highest 16S rRNA gene sequence similarity (>99.8%) to E. lemanii, E. casseliflavus/E. gallinarium or E. devriesei/E. pseudoavium/E. viikkiensis and carried vanA, vanB and/or vanC1 genes. Enterococcus faecium and E. faecalis VRE were not detected, but isolates closely related to those species (>99.9% 16S rRNA gene sequence similarity) were detected by the MRS selective cultivation methods. Staphylococcus isolates shared highest 16S rRNA gene sequence similarity (>99.9%) with S. haemolyticus, S. lentus and S. sciuri and carried mecA genes. Methicillin-resistant S. aureus (MRSA) were not detected. In summary, manure from livestock husbandry contained both, VRE and MRS. VRE were also detected in output samples, indicating that enterococci with vancomycin resistance genes could be release into the environment by the application of BGP output material as biofertilizers.
Lim, Daniel; Strynadka, Natalie C J
The multiple antibiotic resistance of methicillin-resistant strains of Staphylococcus aureus (MRSA) has become a major clinical problem worldwide. The key determinant of the broad-spectrum beta-lactam resistance in MRSA strains is the penicillin-binding protein 2a (PBP2a). Because of its low affinity for beta-lactams, PBP2a provides transpeptidase activity to allow cell wall synthesis at beta-lactam concentrations that inhibit the beta-lactam-sensitive PBPs normally produced by S. aureus. The crystal structure of a soluble derivative of PBP2a has been determined to 1.8 A resolution and provides the highest resolution structure for a high molecular mass PBP. Additionally, structures of the acyl-PBP complexes of PBP2a with nitrocefin, penicillin G and methicillin allow, for the first time, a comparison of an apo and acylated resistant PBP. An analysis of the PBP2a active site in these forms reveals the structural basis of its resistance and identifies features in newly developed beta-lactams that are likely important for high affinity binding.
Molinari, John A
The ability of MRSA and other staphylococci to colonize, persist, and adapt to multiple environmental and tissue conditions has allowed for these bacteria to be virtually ubiquitous in their distribution. The effectiveness of commonly used antibiotics, such as penicillin, has continued to decline against infections caused by MRSA and increasingly resistant strains. The challenge for both dental and medical health professionals is to routinely apply proven, evidence-based infection control precautions. As mentioned earlier in this discussion, when compliance with effective aseptic technique practices improves, the patients and environments. Ensure that reusable equipment is not used for the care of another patient until it has been appropriately cleaned and reprocessed and that single-use items are properly discarded. Clean and disinfect clinical contact surfaces that are not barrier-protected by using an environmental protection agency-registered hospital disinfectant with a low- (ie, HIV and HBV label claims) to intermediate-level (ie, tuberculocidal claim) activity after each patient. When the surface is visibly contaminated with blood, an intermediate-incidence of detectable infections can be reduced. Microorganisms will continue to evolve and adapt in order to survive and thrive, sometimes at the expense of susceptible human hosts. The struggle is to constantly remain aware of impending infectious threats which may challenge current precautions, and maintain and improve the quality of infection control to minimize the potential for disease.
Peng, Qing; Huang, Yuanchun; Hou, Bing; Hua, Dexing; Yao, Fen; Qian, Yuanshu
Tea (Camellia sinensis) has been known for its modulation of resistance of methicillin-resistant Staphylococcus aureus (MRSA) to beta-lactam antibiotics in vitro. This study aimed to confirm the in vitro effect of green tea extracts with beta-lactams and to determine whether green tea extracts can reduce the minimum inhibitory concentrations (MICs) of amoxicillin in MRSA-infected mice. The catechins in the test tea that account for the reduced resistance to beta-lactams were quantitatively determined by high-performance liquid chromatography. The MICs of the ampicillin, cefazolin, amoxicillin, oxacillin, tea extract alone and tea extract in combination with beta-lactams were determined. Proportions of tea extracts and amoxicillin-tea extract combinations were administered to groups of mice enterally. The in vitro experiment showed that the MICs of four beta-lactams were greatly decreased in the presence of 0.25% tea extract. However, in an in vivo experiment, amoxicillin in combination with 5% tea extract conferred a higher ED(50) than that of antibiotic alone. Green tea extract, alone or in combination with amoxicillin, does not have protective benefits in MRSA-infected mice. This study concluded that tea extract weakened the antibacterial effect of amoxicillin in MRSA infected mice. Tea drinking is not recommended in combination with amoxicillin treatment.
Chow, Kin-Hung; Lai, Eileen L.; Law, Pierra Y. T.; Chan, Pui-Ying; Ho, Alex Y. M.; Ng, Tak-Keung; Yam, Wing-Cheong
Out of 3,081 animals studied, 24.9% of pigs, 4.7% of chickens, 6.3% of dogs, 10.5% of cats, and 7.1% of rodents were Staphylococcus aureus positive. Prevalence of methicillin-resistant S. aureus (MRSA) was high in pigs (animals, 21.3%; batches, 46.5%), with all MRSA isolates and most methicillin-sensitive S. aureus isolates belonging to clonal complex 9 (CC9) and being multidrug resistant. The predominant S. aureus CCs among dog and cat isolates were similar. Among rodent isolates, CC398 predominated, with spa t034 the most frequent spa type detected. PMID:22895044
Ho, Pak-Leung; Chow, Kin-Hung; Lai, Eileen L; Law, Pierra Y T; Chan, Pui-Ying; Ho, Alex Y M; Ng, Tak-Keung; Yam, Wing-Cheong
Out of 3,081 animals studied, 24.9% of pigs, 4.7% of chickens, 6.3% of dogs, 10.5% of cats, and 7.1% of rodents were Staphylococcus aureus positive. Prevalence of methicillin-resistant S. aureus (MRSA) was high in pigs (animals, 21.3%; batches, 46.5%), with all MRSA isolates and most methicillin-sensitive S. aureus isolates belonging to clonal complex 9 (CC9) and being multidrug resistant. The predominant S. aureus CCs among dog and cat isolates were similar. Among rodent isolates, CC398 predominated, with spa t034 the most frequent spa type detected.
Maluping, R P; Paul, N C; Moodley, A
Staphylococcus pseudintermedius is a leading aetiologic agent of pyoderma and other body tissue infections in dogs and cats. In recent years, an increased prevalence of methicillin-resistant S. pseudintermedius (MRSP) has been reported. Isolation of MRSP in serious infections poses a major therapeutic challenge as strains are often resistant to all forms of systemic antibiotic used to treat S. pseudintermedius -related infections. This study investigates the occurrence of MRSP from a total of 7183 clinical samples submitted to the authors' laboratories over a 15-month period. Identification was based on standard microbiological identification methods, and by S. pseudintermedius-specific nuc polymerase chain reaction (PCR). Methicillin resistance was confirmed by PBP2a latex agglutination and mecA PCR. Susceptibility against non-beta-lactam antibiotics was carried out using a disc-diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines. In addition, susceptibility to pradofloxacin--a new veterinary fluoroquinolone--was also investigated. SCCmec types were determined by multiplex PCR. Staphylococcus pseudintermedius was isolated from 391 (5%) samples and 20 were confirmed as MRSP from cases of pyoderma, otitis, wound infections, urinary tract infection and mastitis in dogs only. All 20 isolates were resistant to clindamycin and sulphamethoxazole/trimethoprim. Nineteen were resistant to chloramphenicol, enrofloxacin, gentamicin, marbofloxacin and pradofloxacin; additionally, seven isolates were resistant to tetracycline. Fifteen isolates carried SCCmec type II-III, four isolates had type V and one harboured type IV. To date, only a few scientific papers on clinical MRSP strains isolated from the UK have been published, thus the results from this study would provide additional baseline data for further investigations.
Bonesso, Mariana Fávero; Marques, Silvio Alencar; Camargo, Carlos Henrique; Fortaleza, Carlos Magno Castelo Branco; da Cunha, Maria de Lourdes Ribeiro de Souza
The aim of this study was to determine the prevalence of Staphylococcus aureus and risk factors for the acquisition of MRSA (Methicillin Resistant Staphylococcus aureus) as the main cause of skin and soft tissue infections. S. aureus were characterized for the presence of PVL, TSST-1 and mecA genes. SCCmec typing was carried out in mecA positive strains and PFGE was performed only in these strains. During the study period, 127 outpatients attending a dermatology clinical the Botucatu Medical School, a regional tertiary hospital in Botucatu, Sao Paulo, Brazil, were diagnosed with active skin infections. A total 66 (56.9%) S. aureus strains were isolated. The methicillin resistance gene mecA was detected in seven (10.6%) S. aureus strains. The SCCmec types detected in the seven mecA-positive S. aureus strains were type Ia in one, type II in three, and type IV in three. The PVL gene was detected in 10 (15.1%) in sensitive strains. Pulsed field gel electrophoresis revealed non-clonal diversity among the isolates. The risk factors associated with MRSA acquisition in this study were previous ciprofloxacin use and working in a healthcare environment. The risk factors indicate plausible routes of CA-MRSA transmission among the subjects studied.
Context: Community-associated methicillin-resistant Staphlococcus aureus (CA-MRSA) has become of increasing concern in the athletic setting. Appropriate recognition, treatment, and prevention measures are all paramount to protect individual athletes and teamwide outbreaks. Evidence Acquisition: Relevant electronic databases (Medline or PubMed) through 2008 were searched. Articles and studies relevant to this topic were reviewed for pertinent clinical information. Study Type: Clinical review. Results: CA-MRSA is an increasing problem both in the community at large and in the athletic population. Conclusion: Early infections based on methicillin-resistant Staphlococcus aureus are often misidentified, leading to delay in appropriate treatment. A high level of suspicion, prompt recognition, and appropriate treatment can minimize morbidity associated with CA-MRSA. Careful selection of antibiotics in suspected cases is important, with more severe infections requiring hospitalization and intravenous antibiotics. Eradication of bacteria in colonized patients has not yet proven to be effective. Prevention of infections is multifaceted, and it includes education, proper personal hygiene, routine cleaning of equipment, and proper wound care. PMID:23015900
Fogarty, Lisa R; Haack, Sheridan K; Johnson, Heather E; Brennan, Angela K; Isaacs, Natasha M; Spencer, Chelsea
Methicillin-resistant Staphylococcus aureus (MRSA) are a threat to human health worldwide, and although detected at marine beaches, they have been largely unstudied at freshwater beaches. Genes indicating S. aureus (SA; femA) and methicillin resistance (mecA) were detected at 11 and 12 of 13 US Great Lakes beaches and in 18% or 27% of 287 recreational water samples, respectively. Eight beaches had mecA+femA (potential MRSA) detections. During an intensive study, higher bather numbers, staphylococci concentrations, and femA detections were found in samples collected after noon than before noon. Local population density, beach cloud cover, and beach wave height were significantly correlated with SA or MRSA detection frequency. The Panton-Valentine leukocidin gene, associated with community-acquired MRSA, was detected in 12 out of 27 potential MRSA samples. The femA gene was detected less frequently at beaches that met US enterococci criteria or EU enterococci 'excellent' recreational water quality, but was not related to Escherichia coli-defined criteria. Escherichia coli is often the only indicator used to determine water quality at US beaches, given the economic and healthcare burden that can be associated with infections caused by SA and MRSA, monitoring of recreational waters for non-fecal bacteria such as staphylococci and/or SA may be warranted.
Romero-Urbina, Dulce G; Lara, Humberto H; Velázquez-Salazar, J Jesús; Arellano-Jiménez, M Josefina; Larios, Eduardo; Srinivasan, Anand; Lopez-Ribot, Jose L
Summary Silver nanoparticles offer a possible means of fighting antibacterial resistance. Most of their antibacterial properties are attributed to their silver ions. In the present work, we study the actions of positively charged silver nanoparticles against both methicillin-sensitive Staphylococcus aureus and methicillin-resistant Staphylococcus aureus. We use aberration-corrected transmission electron microscopy to examine the bactericidal effects of silver nanoparticles and the ultrastructural changes in bacteria that are induced by silver nanoparticles. The study revealed that our 1 nm average size silver nanoparticles induced thinning and permeabilization of the cell wall, destabilization of the peptidoglycan layer, and subsequent leakage of intracellular content, causing bacterial cell lysis. We hypothesize that positively charged silver nanoparticles bind to the negatively charged polyanionic backbones of teichoic acids and the related cell wall glycopolymers of bacteria as a first target, consequently stressing the structure and permeability of the cell wall. This hypothesis provides a major mechanism to explain the antibacterial effects of silver nanoparticles on Staphylococcus aureus. Future research should focus on defining the related molecular mechanisms and their importance to the antimicrobial activity of silver nanoparticles. PMID:26734530
Weaver, Alan J.; Shepard, Joyce B.; Wilkinson, Royce A.; Watkins, Robert L.; Walton, Sarah K.; Radke, Amanda R.; Wright, Thomas J.; Awel, Milat B.; Cooper, Catherine; Erikson, Elizabeth; Labib, Mohamed E.; Voyich, Jovanka M.; Teintze, Martin
This study investigated the potential antibacterial activity of three series of compounds synthesized from 12 linear and branched polyamines with 2–8 amino groups, which were substituted to produce the corresponding guanides, biguanides, or phenylguanides, against Acinetobacter baumannii, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Antibacterial activity was measured for each compound by determining the minimum inhibitory concentration against the bacteria, and the toxicity towards mammalian cells was determined. The most effective compound, THAM trisphenylguanide, was studied in time-to-kill and cytoplasmic leakage assays against methicillin-resistant Staphylococcus aureus (MRSA, USA300) in comparison to chlorhexidine. Preliminary toxicity and MRSA challenge studies in mice were also conducted on this compound. THAM trisphenylguanide showed significant antibacterial activity (MIC ∼1 mg/L) and selectivity against MRSA relative to all the other bacteria examined. In time-to-kill assays it showed increased antimicrobial activity against MRSA versus chlorhexidine. It induced leakage of cytoplasmic content at concentrations that did not reduce cell viability, suggesting the mechanism of action may involve membrane disruption. Using an intraperitoneal mouse model of invasive MRSA disease, THAM trisphenylguanide reduced bacterial burden locally and in deeper tissues. This study has identified a novel guanide compound with selective microbicidal activity against Staphylococcus aureus, including a methicillin-resistant (MRSA) strain. PMID:24840307
Romero-Urbina, Dulce; Velazquez-Salazar, J. Jesus; Lara, Humberto H.; Arellano-Jimenez, Josefina; Larios, Eduardo; Yuan, Tony T.; Hwang, Yoon; Desilva, Mauris N.; Jose-Yacaman, Miguel
The increased number of infections due to antibiotic-resistant bacteria is a major concern to society. The objective of this work is to determine the effect of positively charged AgNPs on methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus(MRSA) cell wall using advanced electron microscopy techniques. Positively charged AgNPs suspensions were synthesized via a microwave heating technique. The suspensions were then characterized by Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM) showing AgNPs size range from 5 to 30 nm. MSSA and MRSA were treated with positively charged AgNPs concentrations ranging from 0.06 mM to 31 mM. The MIC50 studies showed that viability of MSSA and MRSA could be reduced by 50% at a positively charged AgNPs concentration of 0.12 mM supported by Scanning-TEM (STEM) images demonstrating bacteria cell wall disruption leading to lysis after treatment with AgNPs. The results provide insights into one mechanism in which positively charged AgNPs are able to reduce the viability of MSSA and MRSA. This research is supported by National Institute on Minority Health and Health Disparities (G12MD007591) from NIH, NSF-PREM Grant No. DMR-0934218, The Welch Foundation and NAMRU-SA work number G1009.
Abd El-Hamid, Marwa I; Bendary, M M
The present study was designed to elucidate the phenotypic and genotypic characterization of S. aureus isolates in Egypt. The antibiotic susceptibility pattern of 133 identified S. aureus isolates revealed that over 70% of the isolates were multi-drug resistant (MDR). Fifty MDR isolates were characterized using antibiotyping and different molecular typing methods. Amplification of mecA gene confirmed 30 strains as methicillin-resistant S. aureus (MRSA) and 20 as methicillin-sensitive S. aureus (MSSA). Generally, 22 MRSA (73.33%) and ten MSSA (50%) harbored Panton-Valentine leukocidin (pvl) gene with no statistically significant difference (p=0.093). Staphylococcal cassette chromosome (SCC) mec typing revealed that 48% of the typeable MRSA isolates possessed SCCmec type IV. SCCmec and antimicrobial susceptibility typing signified the presence of community-acquired (CA)-MRSA in Egypt. Surprisingly, all isolates of SCCmec types I, II and III and 50% of both MRSA isolates of SCCmec types IV and V possessed pvl gene. It was clear that staphylococcal protein A (spa) and coagulase (coa) typing discriminated the isolates into eight different groups, whilst polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of coagulase gene yielded ten distinct RFLP banding patterns. This study presented the first baseline information on the characterization of MRSA and MSSA in Egypt.
Fogarty, Lisa R.; Haack, Sheridan K.; Johnson, Heather E.; Brennan, Angela K.; Isaacs, Natasha M.; Spencer, Chelsea
Methicillin-resistant Staphylococcus aureus (MRSA) are a threat to human health worldwide, and although detected at marine beaches, they have been largely unstudied at freshwater beaches. Genes indicating S. aureus (SA; femA) and methicillin resistance (mecA) were detected at 11 and 12 of 13 US Great Lakes beaches and in 18% or 27% of 287 recreational water samples, respectively. Eight beaches had mecA + femA (potential MRSA) detections. During an intensive study, higher bather numbers, staphylococci concentrations, and femA detections were found in samples collected after noon than before noon. Local population density, beach cloud cover, and beach wave height were significantly correlated with SA or MRSA detection frequency. The Panton-Valentine leukocidin gene, associated with community-acquired MRSA, was detected in 12 out of 27 potential MRSA samples. The femA gene was detected less frequently at beaches that met US enterococci criteria or EU enterococci ‘excellent’ recreational water quality, but was not related to Escherichia coli-defined criteria. Escherichia coli is often the only indicator used to determine water quality at US beaches, given the economic and healthcare burden that can be associated with infections caused by SA and MRSA, monitoring of recreational waters for non-fecal bacteria such as staphylococci and/or SA may be warranted.
Romero-Urbina, Dulce G; Lara, Humberto H; Velázquez-Salazar, J Jesús; Arellano-Jiménez, M Josefina; Larios, Eduardo; Srinivasan, Anand; Lopez-Ribot, Jose L; Yacamán, Miguel José
Silver nanoparticles offer a possible means of fighting antibacterial resistance. Most of their antibacterial properties are attributed to their silver ions. In the present work, we study the actions of positively charged silver nanoparticles against both methicillin-sensitive Staphylococcus aureus and methicillin-resistant Staphylococcus aureus. We use aberration-corrected transmission electron microscopy to examine the bactericidal effects of silver nanoparticles and the ultrastructural changes in bacteria that are induced by silver nanoparticles. The study revealed that our 1 nm average size silver nanoparticles induced thinning and permeabilization of the cell wall, destabilization of the peptidoglycan layer, and subsequent leakage of intracellular content, causing bacterial cell lysis. We hypothesize that positively charged silver nanoparticles bind to the negatively charged polyanionic backbones of teichoic acids and the related cell wall glycopolymers of bacteria as a first target, consequently stressing the structure and permeability of the cell wall. This hypothesis provides a major mechanism to explain the antibacterial effects of silver nanoparticles on Staphylococcus aureus. Future research should focus on defining the related molecular mechanisms and their importance to the antimicrobial activity of silver nanoparticles.
Hatcher, S M; Myers, K W; Heaney, C D; Larsen, J; Hall, D; Miller, M B; Stewart, J R
Industrial hog operations (IHOs) have been identified as a source of antibiotic-resistant Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA). However, few studies have investigated the presence of antibiotic-resistant S. aureus in the environment near IHOs, specifically surface waters proximal to spray fields where IHO liquid lagoon waste is sprayed. Surface water samples (n=179) were collected over the course of approximately one year from nine locations in southeastern North Carolina and analyzed for the presence of presumptive MRSA using CHROMagar MRSA media. Culture-based, biochemical, and molecular tests, as well as matrix-assisted laser desorption/ionization-time of flight mass spectrometry were used to confirm that isolates that grew on CHROMagar MRSA media were S. aureus. Confirmed S. aureus isolates were then tested for susceptibility to 16 antibiotics and screened for molecular markers of MRSA (mecA, mecC) and livestock adaptation (absence of scn). A total of 12 confirmed MRSA were detected in 9 distinct water samples. Nine of 12 MRSA isolates were also multidrug-resistant (MDRSA [i.e., resistant to ≥3 antibiotic classes]). All MRSA were scn-positive and most (11/12) belonged to a staphylococcal protein A (spa) type t008, which is commonly associated with humans. Additionally, 12 confirmed S. aureus that were methicillin-susceptible (MSSA) were recovered, 7 of which belonged to spa type t021 and were scn-negative (a marker of livestock-adaptation). This study demonstrated the presence of MSSA, MRSA, and MDRSA in surface waters adjacent to IHO lagoon waste spray fields in southeastern North Carolina. To our knowledge, this is the first report of waterborne S. aureus from surface waters proximal to IHOs.
Chung, Pooi Yin; Chung, Lip Yong; Navaratnam, Parasakthi
The evolution of antibiotic resistance in Staphylococcus aureus showed that there is no long-lasting remedy against this pathogen. The limited number of antibacterial classes and the common occurrence of cross-resistance within and between classes reinforce the urgent need to discover new compounds targeting novel cellular functions not yet targeted by currently used drugs. One of the experimental approaches used to discover novel antibacterials and their in vitro targets is natural product screening. Three known pentacyclic triterpenoids were isolated for the first time from the bark of Callicarpa farinosa Roxb. (Verbenaceae) and identified as α-amyrin [3β-hydroxy-urs-12-en-3-ol], betulinic acid [3β-hydroxy-20(29)-lupaene-28-oic acid], and betulinaldehyde [3β-hydroxy-20(29)-lupen-28-al]. These compounds exhibited antimicrobial activities against reference and clinical strains of methicillin-resistant (MRSA) and methicillin-sensitive S. aureus (MSSA), with minimum inhibitory concentration (MIC) ranging from 2 to 512 μg/mL. From the genome-wide transcriptomic analysis to elucidate the antimicrobial effects of these compounds, multiple novel cellular targets in cell division, two-component system, ABC transporters, fatty acid biosynthesis, peptidoglycan biosynthesis, aminoacyl-tRNA synthetases, ribosomes and β-lactam resistance pathways are affected, resulting in destabilization of the bacterial cell membrane, halt in protein synthesis, and inhibition of cell growth that eventually lead to cell death. The novel targets in these essential pathways could be further explored in the development of therapeutic compounds for the treatment of S. aureus infections and help mitigate resistance development due to target alterations.
Iyamba, Jean-Marie Liesse; Wambale, José Mulwahali; Lukukula, Cyprien Mbundu; Takaisi-Kikuni, Ntondo za Balega
Introduction Surgical site infections (SSIs) after surgery are usually caused by Staphylococcus aureus and coagulase-negative staphylococci (CNS). In low income countries, methicillin resistant Staphylococcus aureus (MRSA) and methicillin resistant coagulase-negative staphylococci (MR-CNS) surgical site infections are particularly associated with high treatment cost and remain a source of mortality and morbidity. This study aimed to determine the prevalence and the sensitivity to antibiotics of MRSA and MR-CNS isolated from SSIs. Methods Wound swabs were collected from 130 hospitalized surgical patients in two major hospitals of Kinshasa. S. aureus and CNS strains were identified by standard microbiological methods and latex agglutination test (Pastorex Staph-Plus). The antibiotic susceptibility of all staphylococcal strains was carried out using disk-diffusion method. Results Eighty nine staphylococcal strains were isolated. Out of 74 S. aureus and 15 CNS isolated, 47 (63.5%) and 9 (60%) were identified as MRSA and MR-CNS respectively. Among the MRSA strains, 47 strains (100%) were sensitive to imipenem, 39 strains (89%) to amoxycillin-clavulanic acid and 38 strains (81%) to vancomycin. All MR-CNS were sensitive to imipenem, amoxycillin-clavulanic acid and vancomycin. The isolated MRSA and MR-CNS strains showed multidrug resistance. They were both resistant to ampicillin, cotrimoxazole, erythromycin, clindamycin, ciprofloxacin, cefotaxime and ceftazidime. Conclusion The results of the present study showed a high prevalence of MRSA and MR-CNS. Imipenem, amoxycillin-clavulanic acid and vancomycin were the most active antibiotics. This study suggests that antibiotic surveillance policy should become national priority as MRSA and MR-CNS were found to be multidrug resistant. PMID:25478043
Rajkhowa, S; Sarma, D K; Pegu, S R
Staphylococcus aureus is one of the most important pathogens of both humans and animal. Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen that causes serious infections both in hospitals and communities due to its multidrug resistance tendency. This study was undertaken to characterize the MRSA isolates from pigs and to determine the antimicrobial resistance of these isolates. Forty nine MRSA strains (one strain per positive pig) isolated from pigs of Northeast India were characterized by SCCmec typing and antimicrobial resistance. The overall prevalence of MRSA was 7.02 % with the highest prevalence recorded in pigs aged 1-3 months (P = 0.001) and in nasal samples (P = 0.005). Two SCC mec types (type III and V) were found in Indian pigs with predominance of type V. All isolates were resistant to penicillin. Seventeen resistance groups were observed where 87.75 % isolates showed multidrug resistance (showed resistance to three or more classes of antimicrobials). The most predominant resistance pattern observed was Oxytetracycline + Penicillin + Sulfadiazine + Tetracycline accounting 12.24 % of the isolates. The present study contributes to the understanding of characteristics and antimicrobial resistance of porcine MRSA isolates which in turn will help in devising strategy for the control of this pathogen. Findings of the study also throw light on multidrug resistance MRSA and emphasize the need for judicious use of antimicrobials in animal practice.
Cui, Haiying; Li, Wei; Li, Changzhu; Vittayapadung, Saritporn; Lin, Lin
The global burden of bacterial disease remains high and is set against a backdrop of increasing antimicrobial resistance. There is a pressing need for highly effective and natural antibacterial agents. In this work, the anti-biofilm effect of cinnamon oil on methicillin-resistant Staphylococcus aureus was evaluated. Then, cinnamon oil was encapsulated in liposomes to enhance its chemical stability. The anti-biofilm activities of the liposome-encapsulated cinnamon oil against MRSA biofilms on stainless steel, gauze, nylon membrane and non-woven fabrics were evaluated by colony forming unit determination. Scanning electron microscopy and laser scanning confocal microscopy analyses were employed to observe the morphological changes in MRSA biofilms treated with the encapsulated cinnamon oil. As a natural and safe spice, the cinnamon oil exhibited a satisfactory antibacterial performance on MRSA and its biofilms. The application of liposomes further improves the stability of antimicrobial agents and extends the action time.
Miao, Jian; Chen, Lequn; Wang, Jingwen; Wang, Wenxin; Chen, Dingqiang; Li, Lin; Li, Bing; Deng, Yang; Xu, Zhenbo
Methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen in hospitals and the community. As the rapid spread and wide distribution of antimicrobial resistance (such as MRSA), treatment for infectious diseases caused by microorganisms has become a vital threat. Thus, early identification and genotyping are essential for further therapeutic treatment and the control of rapid expansion of MRSA. In combination with applications and data feedbacks, this review focused on the currently available molecular-based assays on their utility and performance for rapid typing of MRSA, especially on effective molecular-based methods. Besides, a common mobile element SCCmec and prevalence of HA-MRSA, LA-MRSA and CA-MRSA were introduced in this review in order to provide a more complete profile of MRSA.
Feng, Yanyan; Tian, Wei; Lin, Dachuan; Luo, Qianyi; Zhou, Yingze; Yang, Tong; Deng, Yuting; Liu, Ya-Hong; Liu, Jian-Hua
The aim of this study was to determine the presence of and characterize methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolated from pets in South China. From 2007 to 2009, 898 samples were collected from 785 pets in Guangdong Province. The identity of staphylococcal species and the presence of methicillin resistance were confirmed by phenotypic and genotypic assays. The genetic relationships of MRSP isolates were determined by multilocus sequence typing (MLST), PFGE and spa typing. SCCmec elements and antimicrobial resistance genes profiling were characterized by PCR amplification. A total of 144 S. pseudintermedius isolates were recovered from the dogs and cats tested, and 69 (47.9%) of these isolates were identified as MRSP. Most of the MRSP isolates exhibited simultaneous resistance to four or more different antimicrobial agents. However, valnemulin showed robust activity against MRSP (MIC(90)=1 μg/ml). Integron 1, 2 and 3 were not detected in MRSP isolates. Twenty-four different multilocus sequence types were found among the MRSP isolates, with ST4 (n=9), ST5 (n=8), and ST95 (n=7) being dominant sequence types. In addition, 8 new sequence types (ST134, 135, 136, 137, 138, 139, 140 and 148) were identified. Of the 69 MRSP isolates, SCCmecV was the most prevalent type (n=33), followed by SCCmecVII (n=13), SCCmecII-III (n=7), and SCCmecIII (n=4). This study demonstrates for the first time that the occurrence of MRSP in healthy pets in China and shows that MRSP in South China has high genetic diversity.
Mohammad, Haroon; Reddy, P V Narasimha; Monteleone, Dennis; Mayhoub, Abdelrahman S; Cushman, Mark; Hammac, G Kenitra; Seleem, Mohamed N
Staphylococcus pseudintermedius is a commensal organism of companion animals that is a significant source of opportunistic infections in dogs. With the emergence of clinical isolates of S. pseudintermedius (chiefly methicillin-resistant S. pseudintermedius (MRSP)) exhibiting increased resistance to nearly all antibiotic classes, new antimicrobials and therapeutic strategies are urgently needed. Thiazole compounds have been previously shown to possess potent antibacterial activity against multidrug-resistant strains of Staphylococcus aureus of human and animal concern. Given the genetic similarity between S. aureus and S. pseudintermedius, this study explores the potential use of thiazole compounds as novel antibacterial agents against methicillin-sensitive S. pseudintermedius (MSSP) and MRSP. A broth microdilution assay confirmed these compounds exhibit potent bactericidal activity (at sub-microgram/mL concentrations) against both MSSA and MRSP clinical isolates while the MTS assay confirmed three compounds (at 10 μg/mL) were not toxic to mammalian cells. A time-kill assay revealed two derivatives rapidly kill MRSP within two hours. However, this rapid bactericidal activity was not due to disruption of the bacterial cell membrane indicating an alternative mechanism of action for these compounds against MRSP. A multi-step resistance selection analysis revealed compounds 4 and 5 exhibited a modest (two-fold) shift in activity over ten passages. Furthermore, all six compounds (at a subinihibitory concentration) demonstrated the ability to re-sensitize MRSP to oxacillin, indicating these compounds have potential use for extending the therapeutic utility of β-lactam antibiotics against MRSP. Metabolic stability analysis with dog liver microsomes revealed compound 3 exhibited an improved physicochemical profile compared to the lead compound. In addition to this, all six thiazole compounds possessed a long post-antibiotic effect (at least 8 hours) against MRSP
Mohammad, Haroon; Reddy, P. V. Narasimha; Monteleone, Dennis; Mayhoub, Abdelrahman S.; Cushman, Mark; Hammac, G. Kenitra; Seleem, Mohamed N.
Staphylococcus pseudintermedius is a commensal organism of companion animals that is a significant source of opportunistic infections in dogs. With the emergence of clinical isolates of S. pseudintermedius (chiefly methicillin-resistant S. pseudintermedius (MRSP)) exhibiting increased resistance to nearly all antibiotic classes, new antimicrobials and therapeutic strategies are urgently needed. Thiazole compounds have been previously shown to possess potent antibacterial activity against multidrug-resistant strains of Staphylococcus aureus of human and animal concern. Given the genetic similarity between S. aureus and S. pseudintermedius, this study explores the potential use of thiazole compounds as novel antibacterial agents against methicillin-sensitive S. pseudintermedius (MSSP) and MRSP. A broth microdilution assay confirmed these compounds exhibit potent bactericidal activity (at sub-microgram/mL concentrations) against both MSSA and MRSP clinical isolates while the MTS assay confirmed three compounds (at 10 μg/mL) were not toxic to mammalian cells. A time-kill assay revealed two derivatives rapidly kill MRSP within two hours. However, this rapid bactericidal activity was not due to disruption of the bacterial cell membrane indicating an alternative mechanism of action for these compounds against MRSP. A multi-step resistance selection analysis revealed compounds 4 and 5 exhibited a modest (two-fold) shift in activity over ten passages. Furthermore, all six compounds (at a subinihibitory concentration) demonstrated the ability to re-sensitize MRSP to oxacillin, indicating these compounds have potential use for extending the therapeutic utility of β-lactam antibiotics against MRSP. Metabolic stability analysis with dog liver microsomes revealed compound 3 exhibited an improved physicochemical profile compared to the lead compound. In addition to this, all six thiazole compounds possessed a long post-antibiotic effect (at least 8 hours) against MRSP
Rashid, Zetti Zainol; Bahari, Norazlah; Othman, Amizah; Jaafar, Roslinda; Mohamed, Nurul Azmawati; Jabbari, Idimaz; Sulong, Anita; Hashim, Rohaidah; Ahmad, Norazah
Abstract. Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is a pathogen recognized to be distinct in both phenotype and genotype from hospital-acquired MRSA. We have identified CA-MRSA cases in Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia, including their antibiotic susceptibility patterns and genotypic characteristics. Cases were identified during January to December 2009 from routine clinical specimens, where culture and antibiotic susceptibility results yielded pauci-resistant MRSA isolates suspected as being CA-MRSA. The patients' clinical data were collected and their specimens were sent for molecular confirmation and analysis. Five cases of CA-MRSA were identified, which had a multi-sensitive pattern on antibiotic susceptibility tests and were resistant to only penicillin and oxacillin. All cases were skin and soft-tissue infections, including diabetic foot with gangrene, infected scalp hematoma, philtrum abscess in a healthcare worker, thrombophlebitis complicated with abscess and infected bedsore. All five cases were confirmed MRSA by detection of mecA. SCCmec typing (ccr and mec complex) revealed SCCmec type IV for all cases except the infected bedsore case. Panton-Valentine leukocidin gene was positive in all isolates. As clinical features among methicillin-sensitive Staphylococcus aureus, CA-MRSA and "nosocomial CA-MRSA" are indistinct, early recognition is necessary in order to initiate appropriate antibiotics and infection control measures. Continual surveillance of pauci-resistant MRSA and molecular analysis are necessary in order to identify emerging strains as well as their epidemiology and transmission, both in the community and in healthcare setting.
Boa, Tyler T; Rahube, Teddie O; Fremaux, Bastien; Levett, Paul N; Yost, Christopher K
Methicillin-resistant Staphylococcus aureus (MRSA) is a public health threat within the general community, thereby warranting identification of MRSA reservoirs within the community. Computer terminals in schools were sampled for S. aureus and methicillin-resistant staphylococci. The overall prevalence of MRSA on computer keyboards was low: 0.68% for a postsecondary institution and 2% and 0% for two secondary institutes. The MRSA isolate from the postsecondary institution did not correspond to the Canadian epidemic clusters, but is related to the USA 700 cluster, which contains strains implicated in outbreaks within the U.S. The isolate from the secondary institute's keyboard was typed as CMRSA7 (USA 400), a strain that has been implicated in both Canadian and U.S. epidemics. Methicillin-resistant S. haemolyticus and S. epidermidis were also isolated from keyboards, indicating that a mixed community of methicillin-resistant staphylococci can be present on keyboards. Although the prevalence was low, the presence of MRSA combined with the high volume of traffic on these student computer terminals demonstrates the potential for public-access computer terminals and computer rooms at educational institutes to act as reservoirs.
Campos-Peña, E; Martín-Nuñez, E; Pulido-Reyes, G; Martín-Padrón, J; Caro-Carrillo, E; Donate-Correa, J; Lorenzo-Castrillejo, I; Alcoba-Flórez, J; Machín, F; Méndez-Alvarez, S
We describe a new, efficient, sensitive, and fast single-tube multiple-PCR protocol for the identification of the most clinically significant Staphylococcus spp. and the simultaneous detection of the methicillin and mupirocin resistance loci. The protocol identifies at the species level isolates belonging to S. aureus, S. epidermidis, S. haemolyticus, S. hominis, S. lugdunensis, and S. saprophyticus.
MRS926 is a livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) strain of sequence type (ST) 398. In order to facilitate in vitro and in vivo studies of this strain, we sought to tag it with a fluorescent marker. We cloned a codon-optimized gene for TurboGFP into a shuttle vector...
Hardy, Katherine J; Oppenheim, Beryl A; Gossain, Savita; Gao, Fang; Hawkey, Peter M
Staphylococcal interspersed repeat unit typing has previously been shown to have the ability to discriminate between epidemic methicillin-resistant Staphylococcus aureus strains in the United Kingdom. The current study illustrates its ability to distinguish between strains within an endemic setting thereby providing a rapid transportable typing method for the identification of transmission events.
Bramble, Manuel; Morris, Daniel; Tolomeo, Pam; Lautenbach, Ebbing
In this narrative review, we found numerous reports suggesting that dogs and cats may play a role in household methicillin-resistant Staphylococcus aureus (MRSA) transmission and recurrent MRSA infection in human contacts. Future work should emphasize elucidating more clearly the prevalence of MRSA in household pets and characterize transmission dynamics of MRSA humans and pet animals.
Sunkesula, Venkata; Kundrapu, Sirisha; Macinga, David R; Donskey, Curtis J
Of 82 patients with methicillin-resistant Staphylococcus aureus (MRSA) colonization, 67 (82%) had positive hand cultures for MRSA. A single application of alcohol gel (2 mL) consistently reduced the burden of MRSA on hands. However, incomplete removal of MRSA was common, particularly in those with a high baseline level of recovery.
There is increasing interest in the presence of Staphylococcus aureus, specifically methicillin-resistant S. aureus (MRSA), on retail meat products. In this study, staphylococci were isolated from retail pork and retail beef in Georgia and MRSA from the products were compared to human MRSA from the...
Patel, Mukesh; Kumar, Ritu A; Stamm, Alan M; Hoesley, Craig J; Moser, Stephen A; Waites, Ken B
Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) strains are increasingly recovered from nosocomial settings. We conducted a retrospective study of surgical site infections (SSI) during 2004 and 2005 to determine the prevalence of CA-MRSA; 57% of MRSA strains tested belonged to the USA300 genotype. CA-MRSA has become a prominent cause of SSI at our institution.
Anderson, Deverick J.; Harris, Simon R.; Godofsky, Eliot; Toriscelli, Todd; Rude, Thomas H.; Elder, Kevin; Sexton, Daniel J.; Pellman, Elliot J.; Mayer, Thom; Fowler, Vance G.; Peacock, Sharon J.
Two American football players on the same team were diagnosed with methicillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections on the same day. Our investigation, including whole genome sequencing, confirmed that players did not transmit MRSA to one another nor did they acquire the MRSA from a single source within the training facility. PMID:25734164
Moure, Raquel; Cañizares, Ángeles; Muíño, María; Lobato, Margarita; Fernández, Ana; Rodríguez, María; Gude, Maria José; Tomás, Maria; Bou, Germán
The new cobas® Cdiff and cobas® MRSA/SA tests were compared with conventional methods for the rapid detection of toxigenic Clostridium difficile and methicillin-resistant Staphylococcus aureus. The final concordance between cobas Cdiff Test and GDH/toxin gene screening was 97.62% and between cobas MRSA/SA Test and chromogenic culture, 91.30%, respectively.
Methicillin-resistant Staphylococcus aureus (MRSA) colonization of livestock animals is common and prevalence rates for pigs have been reported to be as high as 49%. One hypothesis to explain the high prevalence of MRSA in swine herds is the ability of these organisms to exist as biofilms. To invest...
Morcillo, Ana; Castro, Beatriz; Rodríguez-Álvarez, Cristobalina; Abreu, Rossana; Aguirre-Jaime, Armando; Arias, Angeles
Background: Livestock-associated methicillin-resistant Staphylococcus aureus (MRSA) such as the MRSA ST398 strain has spread all over the World and the most worrying aspect of this fact appears to be its capacity to easily spread to humans. The excessive use of antibiotics has made swine a reservoir of MRSA. The aim of the present study was to determine the antibiotic resistance profile of MRSA samples isolated from healthy swine of the island of Tenerife (Spain). Methods: A total of 256 MRSA isolates from swine samples and five MRSA isolates from pig worker samples were investigated for MRSA antibiotic resistant patterns. Results: Analysis of the susceptibility status of MRSA pig isolates revealed that 39 isolates were resistant to one antibiotic, 71 isolates were resistant to two antibiotics and 96 isolates were resistant to three or more antibiotics. SCCmec typing revealed the presence of types IV and V. Isolates having SCCmec IV had an increased resistance to the antimicrobial agents tested than those having SCCmec V. We observed significant differences when comparing the most common resistance patterns and SCCmec type. Conclusions: MRSA isolated from humans showed similar resistance to those isolated from pigs, excepting erythromycin, since all the workers’ isolates were sensitive to this antibiotic. The evolution of new MRSA clones has emphasized the need for infection control practices in animals and humans in close contact. PMID:25588155
Shariq, Ali; Tanvir, Syed Bilal; Zaman, Atif; Khan, Salman; Anis, Armeena; Khan, Misha Aftab; Ahmed, Sumaira
Objective: To determine the resistance and sensitivity pattern of methicillin-resistant Staphylococcus aureus (MRSA) isolates to linezolid (LZD) along with its prevalence in a tertiary care hospital of Karachi, Pakistan. Materials and Methods: A cross-sectional study was carried out. This study lasted for about 1 year. Prevalence and sensitivity of LZD, vancomycin, and oxacillin was tested against isolates of MRSA. Results: Out of total 369 specimens 165 were found to be MRSA making the prevalence in our study 44.7%. All of the isolates which were tested positive for MRSA were susceptible to LZD and no resistance was noted when compared with previous studies performed in Europe and USA. Conclusion: Stringent implementation of infection control measures along with screening for resistance in patients on prolonged LZD therapy or who previously went under LZD therapy should be performed, coupled with judicious usage of the aforementioned antibiotic should be undertaken, as sufficient data is not available at this point for the clinical spectrum of LZD resistant S. aureus, antimicrobial resistance. PMID:28293153
Bendary, M M; Solyman, S M; Azab, M M; Mahmoud, N F; Hanora, A M
Staphylococcus aureus (S. aureus) has been one of the most problematic pathogens. Methicillin Resistant S. aureus (MRSA) has emerged as a major concern for both human and animal. Antibiotic resistance genes dissemination might be possible between human and animal bacteria. The aim of this study is to show phenotypic and genotypic diversity of human and animal MRSA isolates. Antibiogram typing and biofilm production were used as a primary phenotypic typing tool for the characterization of (40) animal and (38) human MRSA isolates. Genetic typing based on sequencing of 16S rRNA gene and virulence gene profiles were done. Antimicrobial resistance profiles of the animal isolates showed little evidence of widespread of resistance, although this was seen in many human isolates. The biofilm production was detected in higher percentage among animal isolates. Based on the genetic typing and multiple antibiotic resistance (MAR) index, the majority of animal isolates clustered into lineages that were not found in human isolates. Animal and human MRSA isolates showed diversity in antibiotic resistance and virulence gene profiles may be due to host adaptation or chances for contamination between the two hosts were not present in our study.
Sasatsu, M; Shimizu, K; Noguchi, N; Kono, M
The relationship between an effective concentration and the duration of exposure to antiseptics was evaluated in strains of Staphylococcus aureus with a known genetic background, which include methicillin-resistant strains, using a modified version of the phenol coefficient method as part of an effort to investigate the antiseptic resistance of S. aureus. Chlorhexidine digluconate killed an antiseptic-sensitive strain within 1.5 min at 22 degrees C at a standard concentration (0.1%), whereas resistant strains still survived after 1.5 min. Povidone-iodine killed the sensitive strain within 1.5 min at a concentration of 0.1%, whereas it took this agent 3.0 and 4.5 min to kill low- and high-level resistant strains, respectively, at a concentration of 0.8%. These results indicate that the modified phenol coefficient method used is suitable for the evaluation of the sensitivity of microorganisms to antiseptics. An antiseptic-resistant chain that was associated with the ebr gene exhibited cross-resistance to povidone-iodine.
Groves, Mitchell D.; Crouch, Bethany; Coombs, Geoffrey W.; Jordan, David; Pang, Stanley; Barton, Mary D.; Giffard, Phil
This work investigated the molecular epidemiology and antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) isolated from veterinarians in Australia in 2009. The collection (n = 44) was subjected to extensive molecular typing (MLST, spa, SCCmec, dru, PFGE, virulence and antimicrobial resistance genotyping) and antimicrobial resistance phenotyping by disk diffusion. MRSA was isolated from Australian veterinarians representing various occupational emphases. The isolate collection was dominated by MRSA strains belonging to clonal complex (CC) 8 and multilocus sequence type (ST) 22. CC8 MRSA (ST8-IV [2B], spa t064; and ST612-IV [2B], spa variable,) were strongly associated with equine practice veterinarians (OR = 17.5, 95% CI = 3.3–92.5, P < 0.001) and were often resistant to gentamicin and rifampicin. ST22-IV [2B], spa variable, were strongly associated with companion animal practice veterinarians (OR = 52.5, 95% CI = 5.2–532.7, P < 0.001) and were resistant to ciprofloxacin. A single pig practice veterinarian carried ST398-V [5C2], spa t1451. Equine practice and companion animal practice veterinarians frequently carried multiresistant-CC8 and ST22 MRSA, respectively, whereas only a single swine specialist carried MRSA ST398. The presence of these strains in veterinarians may be associated with specific antimicrobial administration practices in each animal species. PMID:26735694
Zubeir, I E M El; Kanbar, T; Alber, J; Lämmler, C; Akineden, O; Weiss, R; Zschöck, M
Methicillin/oxacillin resistance of 10 S. intermedius strains was investigated by conventional and molecular methods. The strains tested had been isolated in Germany during routine veterinary microbiological examinations of specimens from a small animal clinic between May and September 2005. Epidemiological relationships of the strains were studied by macrorestriction analysis of their chromosomal DNA using pulsed field gel electrophoresis (PFGE). Species identity of the 10 S. intermedius strains was confirmed by conventional methods and by PCR mediated amplification of S. intermedius specific segments of thermonuclease encoding gene nuc. As controls, four methicillin/oxacillin resistant S. intermedius (MRSI) strains obtained from specimens sent by four veterinarians and three selected methicillin/oxacillin sensitive S. intermedius (MSSI), also obtained from the small animal clinic, were tested. The 10 strains, representing approximately 6% of all S. intermedius isolated from the clinic throughout the time period mentioned above, and the four MRSI obtained from veterinarians, were methicillin/oxacillin and penicillin resistant using disk diffusion tests and could be cultivated on oxacillin resistant screening agar base (ORSAB). Both resistances could be confirmed by multiplex PCR detecting the resistance genes mecA and blaZ. The three MSSI were methicillin/oxacillin sensitive in all tests. Epidemiological investigation by macrorestriction analysis of the chromosomal DNA of the strains by pulsed field gel electrophoresis revealed that all 10 MRSI strains obtained from the clinic and the four MRSI strains obtained from veterinarians, in contrast to the three MSSI strains, represent identical or closely related bacterial clones possibly indicating a cross-infection of the animals in the clinic and the distribution of a single MRSI clone in the pet population.
Bakthavatchalam, Yamuna Devi; Nabarro, Laura E B; Veeraraghavan, Balaji
The dissemination of methicillin-resistant (MR) Staphylococcus aureus (SA) in community and health-care settings is of great concern and associated with high mortality and morbidity. Rapid detection of MRSA with short turnaround time can minimize the time to initiate appropriate therapy and further promote infection control. Early detection of MRSA directly from clinical samples is complicated by the frequent association of MRSA with methicillin-susceptible SA (MSSA) and coagulase-negative Staphylococcus (CoNS) species. Infection associated with true MRSA or MSSA is differentiated from CoNS, requires target specific primers for the presence of SA and mec A or nuc or fem A gene for confirmation of MR. Recently, livestock-associated MRSA carrying mec C variant complicates the epidemiology of MRSA further. Several commercial rapid molecular kits are available with a different combination of these targets for the detection of MRSA or MSSA. The claimed sensitivity and specificity of the currently available commercial kits is varying, because of the different target combination used for detection of SA and MR. PMID:28250621
Cheng, Nai-Chen; Wang, Jann-Tay; Chang, Shan-Chwen; Tai, Hao-Chih; Tang, Yueh-Bih
Staphylococcus aureus is an uncommon causative agent of monomicrobial necrotizing fasciitis, but we have noted several cases over the years. The patients treated for necrotizing fasciitis between January 1998 and December 2008 in our institution were identified, and their medical records were reviewed. Of 105 necrotizing fasciitis cases during the study period, 18 were caused by monomicrobial S. aureus infection (17%). The median age was 62 years (range, 12-81 years). Among this cohort, 10 patients had coexisting medical conditions or risk factors, including diabetes and hypertension. Lower limbs and upper limbs are the most commonly involved sites. Among the bacterial isolates from these cases, 8 were methicillin-sensitive S. aureus (MSSA) and 10 were methicillin-resistant S. aureus (MRSA). One patient died in the MSSA group, and 5 patients died in the MRSA group. The mortality rate and other clinical characteristics were not significantly different between the 2 groups. However, all MRSA necrotizing fasciitis developed after the year 2000, and it was significantly different from MSSA necrotizing fasciitis that predominantly took place before the year 2000. In conclusion, S. aureus is an important pathogen of monomicrobial necrotizing fasciitis, and MRSA has emerged as the predominant causative agent in recent years. Therefore, MRSA-directed antibiotic therapy should be considered when treating patients suspected with necrotizing fasciitis in endemic areas.
Wirth, Thierry; Andersen, Paal S.; Skov, Robert L.; De Grassi, Anna; Simões, Patricia Martins; Tristan, Anne; Petersen, Andreas; Aziz, Maliha; Kiil, Kristoffer; Cirković, Ivana; Udo, Edet E.; del Campo, Rosa; Vuopio-Varkila, Jaana; Ahmad, Norazah; Tokajian, Sima; Peters, Georg; Schaumburg, Frieder; Olsson-Liljequist, Barbro; Givskov, Michael; Driebe, Elizabeth E.; Vigh, Henrik E.; Shittu, Adebayo; Ramdani-Bougessa, Nadjia; Rasigade, Jean-Philippe; Price, Lance B.; Vandenesch, Francois; Larsen, Anders R.; Laurent, Frederic
ABSTRACT Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) was recognized in Europe and worldwide in the late 1990s. Within a decade, several genetically and geographically distinct CA-MRSA lineages carrying the small SCCmec type IV and V genetic elements and the Panton-Valentine leukocidin (PVL) emerged around the world. In Europe, the predominant CA-MRSA strain belongs to clonal complex 80 (CC80) and is resistant to kanamycin/amikacin and fusidic acid. CC80 was first reported in 1993 but was relatively rare until the late 1990s. It has since been identified throughout North Africa, the Middle East, and Europe, with recent sporadic reports in sub-Saharan Africa. While strongly associated with skin and soft tissue infections, it is rarely found among asymptomatic carriers. Methicillin-sensitive S. aureus (MSSA) CC80 strains are extremely rare except in sub-Saharan Africa. In the current study, we applied whole-genome sequencing to a global collection of both MSSA and MRSA CC80 isolates. Phylogenetic analyses strongly suggest that the European epidemic CA-MRSA lineage is derived from a PVL-positive MSSA ancestor from sub-Saharan Africa. Moreover, the tree topology suggests a single acquisition of both the SCCmec element and a plasmid encoding the fusidic acid resistance determinant. Four canonical SNPs distinguish the derived CA-MRSA lineage and include a nonsynonymous mutation in accessory gene regulator C (agrC). These changes were associated with a star-like expansion into Europe, the Middle East, and North Africa in the early 1990s, including multiple cases of cross-continent imports likely driven by human migrations. PMID:25161186
Kang, Kyoung-Mi; Mishra, Nagendra N; Park, Kun Taek; Lee, Gi-Yong; Park, Yong Ho; Bayer, Arnold S; Yang, Soo-Jin
Daptomycin (DAP) has potent activity in vitro and in vivo against both methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) strains. DAP-resistance (DAP-R) in S. aureus has been mainly observed in MRSA strains, and has been linked to single nucleotide polymorphisms (SNPs) within the mprF gene leading to altered cell membrane (CM) phospholipid (PL) profiles, enhanced positive surface charge, and changes in CM fluidity. The current study was designed to delineate whether these same genotypic and phenotypic perturbations are demonstrated in clinically-derived DAP-R MSSA strains. We used three isogenic DAP-susceptible (DAP-S)/DAP-R strainpairs and compared: (i) presence of mprF SNPs, (ii) temporal expression profiles of the two key determinants (mprF and dltABCD) of net positive surface charge, (iii) increased production of mprF-dependent lysinylated-phosphatidylglycerol (L-PG), (iv) positive surface charge assays, and (v) susceptibility to cationic host defense peptides (HDPs) of neutrophil and platelet origins. Similar to prior data in MRSA, DAP-R (vs DAP-S) MSSA strains exhibited hallmark hot-spot SNPs in mprF, enhanced and dysregulated expression of both mprF and dltA, L-PG overproduction, HDP resistance and enhanced positive surface charge profiles. However, in contrast to most DAP-R MRSA strains, there were no changes in CM fluidity seen. Thus, charge repulsion via mprF-and dlt-mediated enhancement of positive surface charge may be the main mechanism to explain DAP-R in MSSA strains.
Bullous impetigo in children infected with methicillin-resistant Staphylococcus aureus alone or in combination with methicillin-susceptible S. aureus: analysis of genetic characteristics, including assessment of exfoliative toxin gene carriage.
Shi, Da; Higuchi, Wataru; Takano, Tomomi; Saito, Kohei; Ozaki, Kyoko; Takano, Misao; Nitahara, Yoshiyuki; Yamamoto, Tatsuo
Among bullous impetigo isolates, exfoliative toxin (ET) gene carriage was found in 61.5% of methicillin-resistant Staphylococcus aureus (MRSA) isolates versus 90.6% of methicillin-susceptible S. aureus (MSSA) isolates. MRSA-only cases were ETB or ETA positive, while MRSA/MSSA coinfection cases were ET negative for MRSA but ETA positive for MSSA. Collagen adhesin may facilitate some MRSA infections.
Tuchina, Elena S.; Petrov, Pavel O.; Ratto, Fulvio; Centi, Sonia; Pini, Roberto; Tuchin, Valery V.
The effect of NIR laser radiation (808 nm) on methicillin-sensitive and methicillin resistant strains of Staphylococcus aureus incubated with gold nanorods is studied. Nanorods having length of 44 (± 4) nm and diameter of 10 (± 3) nm with the absorption maximum in the NIR (800 nm), functionalized with human immunoglobulins IgA and IgG, were synthesized and used in the studies. The killing ability up to 97% of the microorganism populations by using this nanotechnology was shown.
Long, S. Wesley; Olsen, Randall J.; Mehta, Shrenik C.; Palzkill, Timothy; Cernoch, Patricia L.; Perez, Katherine K.; Musick, William L.; Rosato, Adriana E.
Ceftaroline is the first member of a novel class of cephalosporins approved for use in the United States. Although prior studies have identified eight ceftaroline-resistant methicillin-resistant Staphylococcus aureus (MRSA) isolates in Europe and Asia with MICs ranging from 4 to 8 mg/liter, high-level resistance to ceftaroline (>32 mg/liter) has not been described in MRSA strains isolated in the United States. We isolated a ceftaroline-resistant (MIC > 32 mg/liter) MRSA strain from the blood of a cystic fibrosis patient and five MRSA strains from the respiratory tract of this patient. Whole-genome sequencing identified two amino acid-altering mutations uniquely present in the ceftaroline-binding pocket of the transpeptidase region of penicillin-binding protein 2a (PBP2a) in ceftaroline-resistant isolates. Biochemical analyses and the study of isogenic mutant strains confirmed that these changes caused ceftaroline resistance. Thus, we identified the molecular mechanism of ceftaroline resistance in the first MRSA strain with high-level ceftaroline resistance isolated in the United States. PMID:25155594
Argudín, M Angeles; Vanderhaeghen, Wannes; Butaye, Patrick
In this study we determined whether methicillin-resistant non-Staphylococcus aureus (MRNAS) from veal calves may be a potential reservoir of antimicrobial-resistance and virulence genes. Fifty-eight MRNAS were studied by means of DNA-microarray and PCR for detection of antimicrobial resistance and virulence genes. The isolates carried a variety of antimicrobial-resistance genes [aacA-aphD, aadD, aph3, aadE, sat, spc, ampA, erm(A), erm(B), erm(C), erm(F), erm(T), lnu(A), msr(A)-msr(B), vga(A), mph(C), tet(K), tet(M), tet(L), cat, fexA, dfrA, dfrD, dfrG, dfrK, cfr, fusB, fosB, qacA, qacC, merA-merB]. Some isolates carried resistance genes without showing the corresponding resistance phenotype. Most MRNAS carried typical S. aureus virulence factors like proteases (sspP) and enterotoxins (seg) genes. Most Staphylococcus epidermidis isolates carried the arginine catabolic element, and nearly 40% of the Staphylococcus sciuri isolates carried leukocidins, and/or fibronectin-binding protein genes. MRNAS were highly multi-resistant and represent an important reservoir of antimicrobial resistance and virulence genes.
Nomura, S; Hanaki, H; Unemi, N
The in vitro activity of TOC-50, a new parenteral cephalosporin, was assessed against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis (MRSE). TOC-50 showed excellent activity, which was stronger than that of methicillin, cloxacillin, the cephalosporins tested, imipenem, gentamycin, minocycline, ofloxacin and ciprofloxacin against MRSA and had a minimum inhibitory concentration (MIC) comparable to that of vancomycin (the MICs of TOC-50 and vancomycin for growth inhibition of 90% of the strains tested were 3.13 and 1.56 micrograms/ml, respectively). Against MRSE, TOC-50 exhibited excellent activity, which was stronger than that of methicillin, ampicillin, the cephalosporins tested and imipenem, and was twice as active as vancomycin. In terms of the bactericidal effect against MRSA, TOC-50 was superior to vancomycin.
Kaiser, Michael; Gil, Joel; Treu, Ryan; Valdes, Jose; Davis, Stephen
Infections of acute and chronic wounds have a substantial negative impact on patient outcomes. Because bacterial resistance to traditional antimicrobials continues to increase, an in vitro study was conducted to examine current sensitivities of various methicillin-resistant and methicillin-sensitive strains of Staphylococcus aureus (MRSA and MSSA) to commonly used topical antimicrobial agents. Using fresh cultures of eight strains of MRSA and MSSA, the area of the zone of inhibition produced by various topical antimicrobials, including an aminoglycoside antibiotic, monocarboxylic acid antibiotic, pleuromutilin antibiotic, triple antibiotic ointment, and petrolatum ointment, was examined. Six culture plates per antimicrobial were prepared using the Kirby Bauer method; soy blood culture plates were inoculated with the bacteria, incubated for 24 hours at 37˚ C, and their zones of inhibition measured and calculated. Data were analyzed using ANOVA testing. Mupirocin treatment was the most effective antimicrobial, with areas of inhibition ranging from 30.34 cm2 to 61.70 cm2 (P <0.05), as compared to the next most effective, retapamulin, with areas of inhibition ranging from 11.97 cm2 to 23.54 cm2. This study provides current scientific data to help the development of a thoughtful rationale for the use of topical antimicrobials in wounds. Additional in vivo studies to substantiate these findings are needed.
Kadlec, Kristina; Weiß, Sonja; Wendlandt, Sarah; Schwarz, Stefan; Tonpitak, Walaiporn
Methicillin-resistant Staphylococcus pseudintermedius (MRSP) in small animal practice are very difficult to treat due to multi-resistance. In contrast to other countries, little is known about MRSP from Thailand. In particular, information on feline MRSP isolates in general is rare. In total, 39 MRSP isolates from dogs (n=28) and cats (n=11) from Thailand collected from independent clinical cases were used. Oxacillin resistance and presence of the mecA gene was confirmed. Susceptibility to additional 29 antimicrobial agents was tested according to CLSI recommendations. Antimicrobial resistance genes were detected by PCR assays. Molecular typing comprised spa typing, dru typing and macrorestriction analysis with subsequent pulsed-field gel electrophoresis (PFGE). For selected isolates, multi-locus sequence typing (MLST) was performed. All isolates were multi-resistant with resistance to at least six classes of antimicrobial agents. In all cases corresponding resistance genes were detected. In addition to mecA, the genes blaZ, catpC221, aacA/aphD, erm(B), dfrG, tet(M) and tet(K) were identified. Six spa types (t02, t05, t09, t10, t23, t72), eleven dru types (dt8ak, dt10ao, dt10cp, dt10cq, dt11a, dt11bo, dt11cb, dt11cj, dt11v, dt11y, dt11z) and 27 PFGE types (designated as A1-A10, B1-B8, C1-C2, D, E, F, G, H, I, J) were identified. MLST for one isolate of each main PFGE pattern A-J revealed seven types [ST45 (n=3), ST112, ST155, ST282 and the novel types ST432, ST433 (n=2) and ST434]. This study showed that MRSP isolates from clinical cases in individual dogs and cats in Thailand are multi-resistant with similar resistance genes and characteristics as isolates from Europe and North America.
Zoraghi, Roya; See, Raymond H; Gong, Huansheng; Lian, Tian; Swayze, Rick; Finlay, B Brett; Brunham, Robert C; McMaster, William R; Reiner, Neil E
Novel antimicrobial targets are urgently needed to overcome rising antibiotic resistance of important human pathogens including methicillin-resistant Staphylococcus aureus (MRSA). Here we report the essentiality and kinetic properties of MRSA pyruvate kinase (PK). Targetron-mediated gene disruption demonstrated PK is essential for S. aureus growth and survival, suggesting that this protein may be a potential drug target. The presence of the pfk (6-phosphofructokinase)-pyk operon in MRSA252, and the nonessential nature of PFK shown by targetron, further emphasized the essential role of PK in cell viability. The importance of PK in bacterial growth was confirmed by showing that its enzymatic activity peaked during the logarithmic phase of S. aureus growth. PK from Staphylococcus and several other species of bacteria have an extra C-terminal domain (CT) containing a phosphoenolpyruvate (PEP) binding motif. To elucidate the possible structure and function of this sequence, the quaternary structures and kinetic properties of the full-length MRSA PK and truncated MRSA PK lacking the CT domain were characterized. Our results showed that (1) MRSA PK is an allosteric enzyme with homotetramer architecture activated by AMP or ribose 5-phosphate (R5P), but not by fructose 1,6-bisphosphate (FBP), which suggests a different mode of allosteric regulation when compared with human isozymes, (2) the CT domain is not required for the tetramerization of the enzyme; homotetramerization occurred in a truncated PK lacking the domain, (3) truncated enzyme exhibited high affinity toward both PEP and ADP and exhibited hyperbolic kinetics toward PEP in the presence of activators (AMP and R5P) consistent with kinetic properties of full-length enzyme, indicating that the CT domain is not required for substrate binding or allosteric regulation observed in the holoenzyme, (4) the kinetic efficiency (k(cat)/S(0.5)) of truncated enzyme was decreased by 24- and 16-fold, in ligand-free state, toward
Taneike, Ikue; Otsuka, Taketo; Dohmae, Soshi; Saito, Kohei; Ozaki, Kyoko; Takano, Misao; Higuchi, Wataru; Takano, Tomomi; Yamamoto, Tatsuo
Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) with Panton-Valentine leukocidin (PVL) genes is increasing worldwide. Nosocomial outbreak-derived (hospital-acquired) MRSA (HA-MRSA) in Japan in the 1980s was also largely PVL(+). PVL(+) HA-MRSA and CA-MRSA shared the same multi-locus sequence type (ST30) and methicillin resistance cassette (SCCmecIV), but were divergent in oxacillin resistance, spa typing, PFGE analysis or clfA gene analysis. PVL(+) HA-MRSA, which probably originated in PVL(+)S. aureus ST30, was highly adhesive (carrying cna and bbp genes), highly-toxic (carrying luk(PV) and sea genes) and highly drug-resistant. PVL(+) HA-MRSA was once replaced by other PVL(-) HA-MRSA (e.g., ST5), and is re-emerging as CA-MRSA.
Wettstein, K; Descloux, S; Rossano, A; Perreten, V
Methicillin resistance has emerged in clinical isolates of Staphylococcus pseudintermedius from cats in Switzerland. Three cats suffering from urinary tract infections were infected with methicillin-resistant S. pseudintermedius (MRSP). Phenotypic and genotypic characterization of the resistance profile showed that the isolates displayed resistance to all beta-lactams and cephalosporins (blaZ, mecA), fluoroquinolones, tetracyclines [tet(K)], macrolides, lincosamides and streprogramins B [erm(B)], chloramphenicol (catpC221), trimethoprim [dfr(G)] and the aminoglycosides gentamicin [aac(6')-Ie-aph(2')-Ia], kanamycin and neomycin [aph(3')-III] and streptomycin [ant(6)-Ia]. They also harbor the leukocidin gene lukS-I. MRSP represents a new challenge for antibiotic therapy and this zoonotic bacteria may rapidly spread to animals and humans.
Qadir, Muhammad Imran; Abbas, Khizar; Younus, Adnan; Shaikh, Rehan Sadiq
Objective of the present study was to investigate the antibacterial activity of Sea buckthorn (Hippophae rhamnoides L.) berries and leaves against methicillin resistant Staphylococcus aureus (MRSA) by using the standard disc diffusion method. Chloroform, n-hexane and aqueous extract of the plant parts were used. Doses of 2mg/ml, 4 mg/ml and 6mg/ml were tested against the microorganism, and the zone of inhibition was compared against the standard drug vancomycin. Results indicated that n-hexane and chloroform extracts of berries and n-hexane extract leaves showed significant (p<0.05) antibacterial activity comparable with vancomycin. It was concluded from the study that extracts berries and leaves of Hippophae rhamnoides have antibacterial activity against MRSA.
Córdova-Guerrero, Jesús; Hernández-Guevara, Esteban; Ramírez-Zatarain, Sandy; Núñez-Bautista, Marco; Ochoa-Terán, Adrián; Muñiz-Salazar, Raquel; Montes-Ávila, Julio; López-Angulo, Gabriela; Paniagua-Michel, Armando; Nuño Torres, Gustavo A.
Staphylococcus aureus is one of the most common causes of nosocomial infections. The purpose of this study was the synthesis and in vitro evaluation of antimicrobial activity of 10 new 3-oxazolidin-2-one analogues on 12 methicillin resistant S. aureus (MRSA) clinical isolates. S. aureus confirmation was achieved via catalase and coagulase test. Molecular characterization of MRSA was performed by amplification of the mecA gene. Antimicrobial susceptibility was evaluated via the Kirby-Bauer disc diffusion susceptibility test protocol, using commonly applied antibiotics and the oxazolidinone analogues. Only (R)-5-((S)-1-dibenzylaminoethyl)-1,3-oxazolidin-2-one (7a) exhibited antibacterial activity at 6.6 μg. These results, allow us to infer that molecules such as 7a can be potentially used to treat infections caused by MRSA strains. PMID:24675696
Sakai, Kent; Koyama, Nobuhiro; Fukuda, Takashi; Mori, Yukiko; Onaka, Hiroyasu; Tomoda, Hiroshi
Staphyloxanthin, a yellow pigment produced by methicillin-resistant Staphylococcus aureus (MRSA), is a virulent factor escaping from the host immune system. A new screening method for inhibitors of staphyloxanthin production by MRSA was established using paper disks. By this screening method, inhibitors of staphyloxanthin production were selected from the natural product library (ca. 300) and from actinomycete culture broths (ca. 1000). From the natural product library, four known inhibitors of lipid metabolism, cerulenin, dihydrobisvertinol, xanthohumol and zaragozic acid, were found to inhibit staphyloxanthin production; however, typical antibiotics used clinically, including vancomycin, had no effect on staphyloxanthin production. From actinomycete culture broths, two known anthraquinones, 6-deoxy-8-O-methylrabelomycin and tetrangomycin, were found to inhibit staphyloxanthin production by MRSA in the paper disk assay. These results suggested that this screening method is useful and effective to find compounds targeting staphyloxanthin production, leading to a new type of chemotherapeutics against MRSA infection.
Pansegrau, Morgan L.; Mengarelli, Eddie; Dersu, Inci Irak
Summary Neovascular glaucoma is commonly treated surgically with implantation of glaucoma drainage devices. We report the case of a 57-year-old man who underwent an uneventful Ahmed glaucoma valve (AGV) placement for radiation-induced neovascular glaucoma but later developed early postoperative infection with tube exposure. The infection was identified 3 weeks postoperatively and antibiotic treatment was immediately initiated. However, the conjunctival melt progressed, and the AGV had to be removed. Culture of the device revealed methicillin-resistant Staphylococcus aureus (MRSA). There is a potential increased risk of postoperative infection and tube exposure following glaucoma valve implantation in patients with previous radiation therapy. To our knowledge, this is the second case in the literature of MRSA causing early postoperative infection following drainage device placement that required explantation. PMID:27330471
Tawil, Nancy; Sacher, Edward; Mandeville, Rosemonde; Meunier, Michel
Early diagnosis and appropriate treatment of Escherichia coli (E. coli) O157:H7 and methicillin-resistant Staphylococcus aureus (MRSA) are key elements in preventing resultant life-threatening illnesses, such as hemorrhagic colitis, hemolytic uremic syndrome, and septicemia. In this report, we describe the use of surface plasmon resonance (SPR) for the biodetection of pathogenic bacteria, using bacteriophages as the recognition elements. T4 bacteriophages were used to detect E. coli, while a novel, highly specific phage was used to detect MRSA. We found that the system permits label-free, real-time, specific, rapid and cost-effective detection of pathogens, for concentrations of 10(3) colony forming units/milliliter, in less than 20 min. This system promises to become a diagnostic tool for bacteria that cause major public concern for food safety, bioterrorism, and nosocomial infections.
Braich, Puneet S.; Aggarwal, Shruti; Mukhtar, Sabrina; Almeida, David RP.
A 47-year-old African-American woman was admitted to the intensive care unit of our community hospital for respiratory failure secondary to severe decompensated heart failure, requiring intubation. In the ensuing days, she developed a methicillin-resistant Staphylococcus aureus (MRSA) infection of the cornea, despite no growth of MRSA in multiple blood, sputum, and urine cultures. This unexpected corneal infection complicated her hospital stay, and increased morbidity and disease-related cost. Risk factors, warning signs, and preventative measures for MRSA keratitis secondary to lagophthalmos (inability to completely close one's eyelids) are outlined in this case report. Implementing simple precautions such as taping eyelids shut or using artificial lubrication may reduce patient morbidity and disease-related costs. These recommendations are directed to non-ophthalmic clinicians who provide care to patients in settings where MRSA colonization is widespread. PMID:26486112
KURODA, Taisuke; KINOSHITA, Yuta; NIWA, Hidekazu; MIZOBE, Fumiaki; UENO, Takanori; KUWANO, Atsutoshi; HATAZOE, Takashi; HOBO, Seiji
ABSTRACT We report the first case of methicillin-resistant Staphylococcus aureus (MRSA) keratitis in a racehorse. A 5-year-old mare developed punctate keratitis after racing. The corneal ulcer continued to expand despite ophthalmic antimicrobial therapy. On day 6, a conjunctival graft surgery was performed. The mare was euthanized, following colitis and laminitis development on day 10. MRSA was isolated from the corneal swab taken at the time of euthanasia. Immunohistochemical analysis demonstrated gram-positive and anti-S. aureus monoclonal antibody-positive cocci infiltration of the corneal stroma; and a diagnosis of MRSA ulcerative keratitis was made. An ophthalmic antimicrobial against the isolated MRSA did not improve the ocular lesion. The MRSA strain was found to be staphylococcal cassette chromosome mec type II, a strain frequently isolated from humans in Japan. PMID:26435683
Bhavsar, Mrugank M.; Devarajan, T. V.; Nembi, P. Senthur; Ramakrishnan, Nagarajan; Mani, Ashwin K.
A 62-year-old male with the previous history of uncontrolled diabetes and hypertension on irregular treatment presented with a history of fever, dysuria, and urinary retention with progressive painful loss of vision over a period of 2 days. His eye examination showed hypopyon, and he was diagnosed to have rapidly progressive endogenous endophthalmitis. He was started on vancomycin and piperacillin-tazobactam empirically. His blood and urine cultures grew methicillin-resistant Staphylococcus aureus (MRSA). Transesophageal echocardiography ruled out infective endocarditis. Intravitreal injection of vancomycin and ceftazidime was given. Vitreous culture also grew MRSA. A workup for possible source revealed multiple prostatic abscesses on the transrectal ultrasound. Antibiotic was changed to daptomycin in view of high vancomycin minimum inhibitory concentration. His vision was improved at the time of discharge.
Lim, Suk-Kyung; Nam, Hyang-Mi; Park, Hyun-Jung; Lee, Hee-Soo; Choi, Min-Jung; Jung, Suk-Chan; Lee, Ji-Yeon; Kim, Young-Cho; Song, Si-Wook; Wee, Sung-Hwan
A total of 2,858 meat samples collected during 2003-2008 in Korea were investigated, and methicillin-resistant Staphylococcus aureus (MRSA) were isolated from 1.0% (9/890) of beef, 0.3% (4/1,055) of pork, and 0.3% (3/913) of chicken meat samples, respectively. MRSA isolates showed the two sequence types (STs), ST72 from beef and pork and ST692 from chicken meat. MRSA isolates from beef and pork were Panton-Valentine leukocidin -negative, staphylococcal cassette chromosome mec type IVa strain with ST 72, which is the most prevalent type of community acquired-MRSA in Korea. An identical pulse-field gel electrophoresis pattern was detected among 10 of 16 MRSA isolates: 9 strains from beef (n=5) and pork (n=4) in 2008, respectively, and one strain from beef in 2005.
Seetharamappa, Jaldappagari; Oke, Muse; Liu, Huanting; McMahon, Stephen A.; Johnson, Kenneth A.; Carter, Lester; Dorward, Mark; Zawadzki, Michal; Overton, Ian M.; van Niekirk, C. A. Johannes; Graham, Shirley; Botting, Catherine H.; Taylor, Garry L.; White, Malcolm F.; Barton, Geoffrey J.; Coote, Peter J.; Naismith, James H.
Sar2676, a pantothenate synthetase with a molecular weight of 31 419 Da from methicillin-resistant Staphylococcus aureus, has been expressed, purified and crystallized at 293 K. The protein crystallizes in a primitive triclinic lattice, with unit-cell parameters a = 45.3, b = 60.5, c = 117.6 Å, α = 87.2, β = 81.2, γ = 68.4°. A complete data set has been collected to 2.3 Å resolution at the ESRF. Consideration of the likely solvent content suggested the asymmetric unit to contain four molecules. This has been confirmed by molecular-replacement phasing calculations, which give a solution with four monomers using a monomer of pantothenate synthetase from Escherichia coli (PDB code 1iho), which is 41% identical to Sar2676, as a search model. PMID:17554169
Perumal, N; Murugesan, S; Krishnan, P
The objective of this study was to determine the distribution of genes encoding aminoglycoside-modifying enzymes (AMEs) and staphylococcal cassette chromosome mec (SCCmec) elements among clinical isolates of methicillin-resistant staphylococci (MRS). Antibiotic susceptibility test was done using Kirby-Bauer disk diffusion method. The presence of SCCmec types and AME genes, namely, aac (6')-Ie-aph (2''), aph (3')-IIIa and ant (4')-Ia was determined using two different multiplex polymerase chain reaction. The most encountered AME genes were aac (6')-Ie-aph (2'') (55.4%) followed by aph (3')-IIIa (32.3%) and ant (4')-Ia gene (9%). SCCmec type I (34%) was predominant in this study. In conclusion, the aac (6')-Ie-aph (2'') was the most common AME gene and SCCmec type I was most predominant among the MRS isolates.
Mensa, J; Barberán, J; Llinares, P; Picazo, Jj; Bouza, E; Alvarez-Lerma, F; Borges, M; Serrano, R; León, C; Guirao, X; Arias, J; Carreras, E; Sanz, Ma; García-Rodríguez, Ja
Infections due to methicillin-resistant Staphylococcus aureus (MRSA) have undergone important changes in the last five years that have influenced the choice of therapy: i) increase of their frequency in hospital-associated settings and, more recently, in community settings; ii) better knowledge of clinical implications of the pharmacokinetic and pharmacodynamic properties of vancomycin; iii) improvement of current standard methods for rapid detection of MRSA in clinical samples; iv) clear evidence that vancomycin is losing efficacy against MRSA with MIC > 1 microg/mL; and v) appearance of new antibiotics suitable for use in these infections (linezolid, daptomycin, tigecyclin). Under this situation guidelines for the treatment of common infections caused by MRSA appear to be necessary to improve the efficacy and reduce the mortality.
Elwell, L P; Wilson, H R; Knick, V B; Keith, B R
The in vitro susceptibilities of 16 independent, geographically distinct clinical isolates of methicillin-resistant Staphylococcus aureus to trimethoprim (TMP) in combination with sulfamethoxazole (SMX) were evaluated. Although methicillin-resistant S. aureus strains appear to be universally resistant to SMX, the combination TMP-SMX was found to be synergistic in vitro (in combination, the MICs of both drugs decreased 6- to 25-fold) as well as in vivo (5- to 6-fold reduction in TMP at 50% effective doses). PMID:3488022
Kelley, KaShonda; Cosman, Angela; Belgrader, Phillip; Chapman, Brenda
Health care-associated infections with methicillin-resistant Staphylococcus aureus (MRSA) contribute to significant hospitalization costs. We report here a droplet digital PCR (ddPCR) assay, which is a next-generation emulsion-based endpoint PCR assay for high-precision MRSA analysis. Reference cultures of MRSA, methicillin-susceptible S. aureus (MSSA), and confounders were included as controls. Copan swabs were used to sample cultures and collect specimens for analysis from patients at a large teaching hospital. Swab extraction and cell lysis were accomplished using magnetic-driven agitation of silica beads. Quantitative PCR (qPCR) (Roche Light Cycler 480) and ddPCR (Bio-Rad QX100 droplet digital PCR system) assays were used to detect genes for the staphylococcal protein SA0140 (SA) and the methicillin resistance (mecA) gene employing standard TaqMan chemistries. Both qPCR and ddPCR assays correctly identified culture controls for MRSA (76), MSSA (12), and confounder organisms (36) with 100% sensitivity and specificity. Analysis of the clinical samples (211 negative and 186 positive) collected during a study of MRSA nasal carriage allowed direct comparison of the qPCR and ddPCR assays to the Cepheid MRSA GeneXpert assay. A total of 397 clinical samples were examined in this study. Cepheid MRSA GeneXpert values were used to define negative and positive samples. Both the qPCR and ddPCR assays were in good agreement with the reference assay. The sensitivities for the qPCR and ddPCR assays were 96.8% (95% confidence interval [CI], 93.1 to 98.5%) and 96.8% (95% CI, 93.1 to 98.5%), respectively. Both the qPCR and ddPCR assays had specificities of 91.9% (95% CI, 87.5 to 94.9%) for qPCR and 91.0% (95% CI, 86.4 to 94.2%) for ddPCR technology. PMID:23596244
Latronico, Francesca; Moodley, Arshnee; Nielsen, Søren Saxmose; Guardabassi, Luca
The recent worldwide spread of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in dogs is a reason for concern due to the typical multidrug resistance patterns displayed by some MRSP lineages such as sequence type (ST) 71. The objective of this study was to compare the in vitro adherence properties between MRSP and methicillin-susceptible (MSSP) strains. Four MRSP, including a human and a canine strain belonging to ST71 and two canine non-ST71 strains, and three genetically unrelated MSSP were tested on corneocytes collected from five dogs and six humans. All strains were fully characterized with respect to genetic background and cell wall-anchored protein (CWAP) gene content. Seventy-seven strain-corneocyte combinations were tested using both exponential- and stationary-phase cultures. Negative binomial regression analysis of counts of bacterial cells adhering to corneocytes revealed that adherence was significantly influenced by host and strain genotype regardless of bacterial growth phase. The two MRSP ST71 strains showed greater adherence than MRSP non-ST71 (p < 0.0001) and MSSP (p < 0.0001). This phenotypic trait was not associated to any specific CWAP gene. In general, S. pseudintermedius adherence to canine corneocytes was significantly higher compared to human corneocytes (p < 0.0001), but the MRSP ST71 strain of human origin adhered equally well to canine and human corneocytes, suggesting that MRSP ST71 may be able to adapt to human skin. The genetic basis of the enhanced in vitro adherence of ST71 needs to be elucidated as this phenotypic trait may be associated to the epidemiological success and zoonotic potential of this epidemic MRSP clone.
Kawakami, Tetsuji; Shibata, Sanae; Murayama, Nobuo; Nagata, Masahiko; Nishifuji, Koji; Iwasaki, Toshiroh; Fukata, Tsuneo
To understand species distribution, trends of antimicrobial susceptibility and prevalence of methicillin resistance in canine staphylococci in Japan, 190 coagulase-positive staphylococci (CoPS) were isolated from dogs with pyoderma in 2 Japanese veterinary referral hospitals. Using a multiplex polymerase chain reaction (M-PCR) method, two CoPS species were identified: 170 Staphylococcus pseudintermedius (89.5%) and 20 S. schleiferi subsp. coagulans isolates (10.5%). In these isolates, susceptibility to 7 antimicrobial agents was determined. Overall, the levels of susceptibility to cefalexin (CEX), amoxicillin/clavulanic acid (CVA/AMPC), minocycline (MINO), ofloxacin (OFLX), norfloxacin (NFLX), lincomycin (LCM) and clindamycin (CLDM) in S. pseudintermedius isolates were 38.2, 52.4, 34.7, 31.2, 34.1, 1.2 and 11.2%, respectively. In S. schleiferi subsp. coagulans isolates, 55% demonstrated susceptibility to CEX, 80% to CVA/AMPC, 70% to MINO, 45% to OFLX or NFLX and 30% to CLDM. None of S. schleiferi subsp. coagulans isolates was susceptible to LCM. To determine the prevalence of methicillin-resistant strains, we used a PCR method, which enabled detection of the fragment of mecA gene in 66.5% (113 of 170) in S. pseudintermedius and 30.0% (6 of 20) in S. schleiferi subsp. coagulans isolates. The frequencies of susceptibility to CEX, CVA/AMPC, OFLX, NFLX and CLDM were significantly lower in methicillin-resistant CoPS than in methicillin-susceptible CoPS isolates. These data suggest a high level of methicillin resistance in staphylococci isolated from dogs with pyoderma in Japan.
Rebic, Velma; Budimir, Ana; Aljicevic, Mufida; Bektas, Sabaheta; Vranic, Sabina Mahmutovic; Rebic, Damir
Background: Methicillin resistant Staphylococcus aureus (MRSA) is responsible for a wide spectrum of nosocomial and community associated infections worldwide. The aim of this study was to analyze MRSA strains from the general population in Canton Sarajevo, B&H. Methods: Our investigation including either phenotypic and genotypic markers such as antimicrobial resistance, pulsed-field gel electrophoresis (PFGE), SCC typing, and Panton-Valentine leukocidin (PVL) detection. Results: Antimicrobial susceptibility: all MRSA isolates were resistant to the β-lactam antibiotics tested, and all isolates were susceptible trimethoprim sulphamethoxazole, rifampicin, fusidic acid, linezolid and vancomycin. Sixty-eight per cent of the MRSA isolates were resistant to erythromycin, 5% to clindamycin, 5% to gentamicin and 4% to ciprofloxacin. After the PFGE analysis, the isolates were grouped into five similarity groups: A-E. The largest number of isolates belonged to one of two groups: C: 60 (60%) and D: 27 (27%). In both groups C and D, SCCmec type IV was predominant (60% and 88, 8%, respectively). A total of 24% of the isolates had positive expression of PVL genes, while 76% showed a statistically significantly greater negative expression of PVL genes. Conclusion: SCCmec type IV, together with the susceptibility profile and PFGE grouping, is considered to be typical of CA-MRSA PMID:27708486
Rahimi, Fateh; Bouzari, Majid
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is known as a common pathogen in nosocomial and community-acquired infections. Sewage acts as an environmental reservoir and may have a significant role in development and dissemination of antibiotic resistance. Objectives: This study was undertaken to determine the epidemiological relatedness between the MRSA isolated from sewage and human infections. Materials and Methods: Samples were collected from a referral hospital and also a sewage treatment plant in Tehran, Iran, during 2010. All the MRSA isolates were identified at the species level and typed using Phene plate (PhP) system and SCCmec typing. Antibiotic susceptibility tests were also performed. Results: Of the 1142 isolates, 200 MRSA strains from the sewage (n = 100) and the clinic (n = 100) were isolated. Distinct PhP types, consisting of 16 common types and 13 single types, and also 3 different staphylococcal cassette chromosome mec (SCCmec) types (III, IVa and IVc) were found amongst the MRSA isolated from the two different sources. The results of antibiotic susceptibility testing showed an increased resistance to penicillin, ciprofloxacin, erythromycin, clindamycin and tetracycline. In addition, none of the isolates showed resistance to vancomycin, quinupristin -dalfopristin and linezolid. Conclusions: The presence of common PhP types and also SCCmec type III, as an indicator for hospital strains, among the isolates, may indicate an epidemiological link between clinical and sewage MRSA isolates in Tehran. PMID:26421131
Ruscher, Claudia; Lübke-Becker, Antina; Wleklinski, Claus-G; Soba, Alexandra; Wieler, Lothar H; Walther, Birgit
In this study the prevalence of Methicillin-resistant Staphylococcus pseudintermedius (MRSP) in clinical specimens of different animal species was defined by investigating a total of 16,103 clinical samples originating from veterinary facilities of five German federal states in 2007. Of all samples examined, 72 were positive for MRSP, giving an overall prevalence of 0.45%. In clinical specimens originating from small animals the prevalence was 0.58% (n=67; dogs n=61 and cats n=6), while samples from equidaes revealed a prevalence of 0.10% (n=5; horses n=4, donkey n=1). Forty-six representative phenotypically identified MRSP were further differentiated by DNA-based species assignment, PCR detection of mecA, SCCmec-typing and MIC determination. As expected, all 46 isolates were unambiguously proven to be MRSP by sequencing of housekeeping genes pta and cpn60 and being positive for mecA. Furthermore, all isolates harboured the mobile staphylococcal cassette chromosome mec (SCCmec) SCCmecIII. Antibiotic susceptibility testing for 20 different conventional antimicrobial agents disclosed a high rate of multidrug-resistant isolates (45 of 46) displaying an identical or at least similar resistance pattern for non-beta-lactam antimicrobials. The recognized prevalence of MRSP, which have already been shown to be potential zoonotic agents, reflects the recently emerging development of these serious and often multidrug-resistant pathogens in Germany.
Vajs, Jure; Proud, Conor; Brozovic, Anamaria; Gazvoda, Martin; Lloyd, Adrian; Roper, David I; Osmak, Maja; Košmrlj, Janez; Dowson, Christopher G
Diaryltriazene derivatives were synthesized and evaluated for their antimicrobial properties. Initial experiments showed some of these compounds to have activity against both methicillin-resistant strains of Staphylococus aureus (MRSA) and Mycobacterium smegmatis, with MICs of 0.02 and 0.03 μg/mL respectively. Those compounds with potent anti-staphylococcal and anti-mycobacterial activity were not found to act as growth inhibitors of mammalian cell lines or yeast. Furthermore, we demonstrated that one of the most active anti-MRSA diaryltriazene derivatives was subject to very low frequencies of resistance at <10(-9). Whole genome sequencing of resistant isolates identified mutations in the enzyme that lysylates phospholipids. This could result in the modification of phospholipid metabolism and consequently the characteristics of the staphylococcal cell membrane, ultimately modifying the sensitivity of these pathogens to triazene challenge. Our work has therefore extended the potential range of triazenes, which could yield novel antimicrobials with low levels of resistance.
Jesline, A.; John, Neetu P.; Narayanan, P. M.; Vani, C.; Murugan, Sevanan
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major nosocomial pathogens responsible for a wide spectrum of infections and the emergence of bacterial resistance to antibiotics has lead to treatment drawbacks towards large number of drugs. Formation of biofilms is the main contributing factor to antibiotic resistance. The development of reliable processes for the synthesis of zinc oxide nanoparticles is an important aspect of nanotechnology today. Zinc oxide and titanium dioxide nanoparticles comprise well-known inhibitory and bactericidal effects. Emergence of antimicrobial resistance by pathogenic bacteria is a major health problem in recent years. This study was designed to determine the efficacy of zinc and titanium dioxide nanoparticles against biofilm producing methicillin-resistant S. aureus. Biofilm production was detected by tissue culture plate method. Out of 30 MRSA isolates, 22 isolates showed strong biofilm production and 2 showed weak and moderate biofilm formation. Two strong and weak biofilm-producing methicillin-resistant S. aureus isolates were subjected to antimicrobial activity using commercially available zinc and titanium dioxide nanoparticles. Thus, the nanoparticles showed considerably good activity against the isolates, and it can be concluded that they may act as promising, antibacterial agents in the coming years.
Methicillin-Resistant Staphylococcus aureus Colonization and Risk of Subsequent Infection in Critically Ill Children: Importance of Preventing Nosocomial Methicillin-Resistant Staphylococcus aureus Transmission
Goldner, Brian W.; Ross, Tracy; Shepard, John W.; Carroll, Karen C.
Background. Methicillin-resistant Staphylococcus aureus (MRSA) colonization is a predictor of subsequent infection in hospitalized adults. The risk of subsequent MRSA infections in hospitalized children colonized with MRSA is unknown. Methods. Children admitted to an academic medical center’s pediatric intensive care unit between March 2007 and March 2010 were included in the study. Anterior naris swabs were cultured to identify children with MRSA colonization at admission. Laboratory databases were queried and National Healthcare Safety Network definitions applied to identify patients with MRSA infections during their hospitalization or after discharge. Results. The MRSA admission prevalence among 3140 children was 4.9%. Overall, 56 children (1.8%) developed an MRSA infection, including 13 (8.5%) colonized on admission and 43 (1.4%) not colonized on admission (relative risk [RR], 5.9; 95% confidence interval [CI], 3.4–10.1). Of those, 10 children (0.3%) developed an MRSA infection during their hospitalization, including 3 of 153 children (1.9%) colonized on admission and 7 of 2987 children (0.2%) not colonized on admission (RR, 8.4; 95% CI, 2.7–25.8). African-Americans and those with public health insurance were more likely to get a subsequent infection (P < .01 and P = .03, respectively). We found that 15 children acquired MRSA colonization in the pediatric intensive care unit, and 7 (47%) developed a subsequent MRSA infection. Conclusions. MRSA colonization is a risk factor for subsequent MRSA infection in children. Although MRSA colonized children may have lower risks of subsequent infection than adults, children who acquire MRSA in the hospital have similarly high rates of infection. Preventing transmission of MRSA in hospitalized children should remain a priority. PMID:21878424
Verstappen, Koen M.; Broens, Els M.; Laarhoven, Laura M.; van Duijkeren, Engeline; Hordijk, Joost; de Heus, Phebe; Spaninks, Mirlin; Timmerman, Arjen J.; Wagenaar, Jaap A.
Methicillin-resistant Staphylococcus pseudintermedius (MRSP), which is often multidrug resistant (MDR), has recently emerged as a threat to canine health worldwide. Knowledge of the temporal distribution of specific MRSP lineages, their antimicrobial resistance phenotypes, and their association with clinical conditions may help us to understand the emergence and spread of MRSP in dogs. The aim of this study was to determine the yearly proportions of MRSP lineages and their antimicrobial-resistant phenotypes in the Netherlands and to examine possible associations with clinical conditions. MRSP was first isolated from a canine specimen submitted for diagnostics to the Faculty of Veterinary Medicine of Utrecht University in 2004. The annual cumulative incidence of MRSP among S. pseudintermedius increased from 0.9% in 2004 to 7% in 2013. MRSP was significantly associated with pyoderma and, to a lesser extent, with wound infections and otitis externa. Multilocus sequence typing (MLST) of 478 MRSP isolates yielded 39 sequence types (ST) belonging to 4 clonal complexes (CC) and 15 singletons. CC71 was the dominant lineage that emerged since 2004, and CC258, CC45, and several unlinked isolates became more frequent during the following years. All but two strains conferred an MDR phenotype, but strains belonging to CC258 or singletons were less resistant. In conclusion, our study showed that MDR CC71 emerged as the dominant lineage from 2004 and onward and that less-resistant lineages were partly replacing CC71. PMID:26582835
Duim, Birgitta; Verstappen, Koen M; Broens, Els M; Laarhoven, Laura M; van Duijkeren, Engeline; Hordijk, Joost; de Heus, Phebe; Spaninks, Mirlin; Timmerman, Arjen J; Wagenaar, Jaap A
Methicillin-resistant Staphylococcus pseudintermedius (MRSP), which is often multidrug resistant (MDR), has recently emerged as a threat to canine health worldwide. Knowledge of the temporal distribution of specific MRSP lineages, their antimicrobial resistance phenotypes, and their association with clinical conditions may help us to understand the emergence and spread of MRSP in dogs. The aim of this study was to determine the yearly proportions of MRSP lineages and their antimicrobial-resistant phenotypes in the Netherlands and to examine possible associations with clinical conditions. MRSP was first isolated from a canine specimen submitted for diagnostics to the Faculty of Veterinary Medicine of Utrecht University in 2004. The annual cumulative incidence of MRSP among S. pseudintermedius increased from 0.9% in 2004 to 7% in 2013. MRSP was significantly associated with pyoderma and, to a lesser extent, with wound infections and otitis externa. Multilocus sequence typing (MLST) of 478 MRSP isolates yielded 39 sequence types (ST) belonging to 4 clonal complexes (CC) and 15 singletons. CC71 was the dominant lineage that emerged since 2004, and CC258, CC45, and several unlinked isolates became more frequent during the following years. All but two strains conferred an MDR phenotype, but strains belonging to CC258 or singletons were less resistant. In conclusion, our study showed that MDR CC71 emerged as the dominant lineage from 2004 and onward and that less-resistant lineages were partly replacing CC71.
González-Domínguez, María; Seral, Cristina; Potel, Carmen; Sáenz, Yolanda; Álvarez, Maximiliano; Torres, Carmen; Castillo, Francisco Javier
A high proportion of methicillin-resistant Staphylococcus aureus isolates recovered in one year period showed high-level mupirocin-resistance (HLMUPR-MRSA) in our environment (27.2%). HLMUPR-MRSA isolates were mainly collected from skin and soft tissue samples, and diabetes was the main related comorbidity condition. These isolates were more frequently found in vascular surgery. HLMUPR-MRSA was more resistant to aminoglycosides than mupirocin-susceptible MRSA, linked to the presence of bifunctional and/or nucleotidyltransferase enzymes with/without macrolide resistance associated with the msr(A) gene. Most of HLMUPR-MRSA isolates belonged to ST125/t067. Nine IS257-ileS2 amplification patterns (p3 was the most frequent) were observed in HLMUPR-MRSA isolates, suggesting the presence of several mupirocin-resistance-carrying plasmids in our environment and promoting the emergence of mupirocin resistance. The presence of the same IS257-ileS2 amplification pattern p3 in 65% of HLMUPR-MRSA, all of them ST125/t067, suggests a clonal spread in our hospital and community environment which could explain the high prevalence of HLMUPR-MRSA during the study period. An outbreak situation or an increase in mupirocin consumption was not observed.
Sudağidan, Mert; Cavuşoğlu, Cengiz; Bacakoğlu, Feza
Staphylococci are the most important agents of nosocomial infections originating from biomaterials. The aim of this study was to investigate the presence of virulence genes and their phenotypic expressions in 11 methicillin-resistant Staphylococcus aureus strains isolated from the surfaces of clinically used biomaterials of 48 thorasic intensive-care unit patients. By the use of specific primers, the presence of genes encoding the attachment and biofilm production (icaA, icaC, bap), methicillin resistance (mecA), enterotoxins A-E (sea, seb, sec, sed, see), toxic shock syndrome toxin (tst), exfoliative toxins A and B (eta and etb), alpha- and beta-hemolysins (hla and hlb), staphylococcal exotoxin-like protein-1 (set1), proteases (sspA, sspB, aur, serine proteaz gene), lipase (geh) and the regulatory genes (sarA and agrCA) were investigated by polymerase chain reaction (PCR). The phenotypic properties of the isolates such as biofilm formation, antibiotic susceptibility, extracellular protease and lipase production were also evaluated. None of the isolates were found to be biofilm and/or slime producers, however, all strains were found to have icaA gene which is responsible for biofilm formation. Nevertheless the presence of icaC and bap genes that are also responsible for biofilm formation were not detected. All the strains have had mecA gene and were resistant to oxacillin, penicilin G and gentamicin, while 10 were also resistant to erythromycin and nine were also resistant to ofloxacin. The isolates were susceptible to vancomycin, teicoplanin and co-trimoxazole. Screening of toxin and regulatory genes revealed that all the strains harboured sea, set1, hla, hlb and sarA genes. The phenotypic tests for the determination of extracellular protease production revealed that all the strains formed very weak zones on skim milk and milk agar plates, and yielded negative results on casein agar plates. Furthermore, all strains were found to harbour sspA, sspB, aur and serine
Lee, G C; Hall, R G; Boyd, N K; Dallas, S D; DU, L C; Treviño, L B; Retzloff, C; Treviño, S B; Lawson, K A; Wilson, J P; Olsen, R J; Wang, Y; Frei, C R
Skin and soft tissue infections (SSTIs) due to Staphylococcus aureus have become increasingly common in the outpatient setting; however, risk factors for differentiating methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) SSTIs are needed to better inform antibiotic treatment decisions. We performed a case-case-control study within 14 primary-care clinics in South Texas from 2007 to 2015. Overall, 325 patients [S. aureus SSTI cases (case group 1, n = 175); MRSA SSTI cases (case group 2, n = 115); MSSA SSTI cases (case group 3, n = 60); uninfected control group (control, n = 150)] were evaluated. Each case group was compared to the control group, and then qualitatively contrasted to identify unique risk factors associated with S. aureus, MRSA, and MSSA SSTIs. Overall, prior SSTIs [adjusted odds ratio (aOR) 7·60, 95% confidence interval (CI) 3·31-17·45], male gender (aOR 1·74, 95% CI 1·06-2·85), and absence of healthcare occupation status (aOR 0·14, 95% CI 0·03-0·68) were independently associated with S. aureus SSTIs. The only unique risk factor for community-associated (CA)-MRSA SSTIs was a high body weight (⩾110 kg) (aOR 2·03, 95% CI 1·01-4·09).
Pinheiro, Luiza; Brito, Carla Ivo; Pereira, Valéria Cataneli; de Oliveira, Adilson; Camargo, Carlos Henrique; da Cunha, Maria de Lourdes Ribeiro de Souza
This study aimed to correlate the presence of ica genes, biofilm formation and antimicrobial resistance in 107 strains of Staphylococcus epidermidis isolated from blood cultures. The isolates were analysed to determine their methicillin resistance, staphylococcal cassette chromosome mec (SCCmec) type, ica genes and biofilm formation and the vancomycin minimum inhibitory concentration (MIC) was measured for isolates and subpopulations growing on vancomycin screen agar. The mecA gene was detected in 81.3% of the S. epidermidis isolated and 48.2% carried SCCmec type III. The complete icaADBC operon was observed in 38.3% of the isolates; of these, 58.5% produced a biofilm. Furthermore, 47.7% of the isolates grew on vancomycin screen agar, with an increase in the MIC in 75.9% of the isolates. Determination of the MIC of subpopulations revealed that 64.7% had an MIC ≥ 4 μg mL-1, including 15.7% with an MIC of 8 μg mL-1 and 2% with an MIC of 16 μg mL-1. The presence of the icaADBC operon, biofilm production and reduced susceptibility to vancomycin were associated with methicillin resistance. This study reveals a high level of methicillin resistance, biofilm formation and reduced susceptibility to vancomycin in subpopulations of S. epidermidis. These findings may explain the selection of multidrug-resistant isolates in hospital settings and the consequent failure of antimicrobial treatment. PMID:25410990
Bes, Michele; Meugnier, Helene; Lina, Gerard; Bozdogan, Bülent; Courvalin, Patrice; Reverdy, Marie-Elisabeth; Enright, Mark C.; Vandenesch, François; Etienne, Jerome
We determined the agr type, multilocus sequence type, protein A gene type (spa typing), toxin gene profile, and antimicrobial drug resistance profile of 469 isolates of Panton-Valentine leukocidin–positive community-acquired methicillin-resistant Staphylococcus aureus isolates (PVL-positive CA-MRSA). The isolates had been collected from around the world from 1999 through 2005 by the French National Reference Center for Staphylococci. We found that some continent-specific clones described in 2003, such as clone ST8, have now spread all over the world. Likewise, some PVL-positive CA-MRSA have spread to several countries on various continents. New clones have emerged (e.g., ST377) on new genetic backgrounds. PVL-positive CA-MRSA that were usually susceptible to most antistaphylococcal antimicrobial agents have acquired new resistance determinants (e.g., to gentamicin) in certain countries. The major trait shared by all these clones is a short staphylococcal chromosomal cassette mec element of type IV or V. PMID:17553275
Guerin, François; Buu-Hoï, Annie; Mainardi, Jean-Luc; Kac, Guillaume; Colardelle, Nathalie; Vaupré, Sabine; Gutmann, Laurent; Podglajen, Isabelle
Epidemiological relationships were investigated between 40 methicillin-resistant Staphylococcus aureus (MRSA) strains with decreased glycopeptide susceptibility isolated from November 1998 to March 1999 from 39 patients (17 infected and 22 colonized patients) in nine wards of the Broussais Hospital, Paris, France. Reduced glycopeptide susceptibility was readily detected on brain heart infusion (BHI) agar containing 6 μg of teicoplanin per ml and on gradient plates, but not by the standard disk diffusion method. The MICs of vancomycin and teicoplanin, determined on BHI agar, were 4 and 8 to 32 μg/ml, respectively (standard antibiotic dilution), and 4 to 8 and 8 to 32 μg/ml, respectively (E-test). All strains were resistant to macrolides, aminoglycosides, tetracycline, rifampin, sulfonamides, and pefloxacin, showed reduced susceptibility to fusidic acid and fosfomycin, and were susceptible to trimethoprim and chloramphenicol. Pulsed-field gel electrophoresis and lysotyping revealed that a multidrug-resistant MRSA clone with decreased susceptibility to glycopeptides has been discretely endemic since at least 1996 in our institution, where it was responsible for an outbreak in November and December 1998. PMID:10921964
Porrero, M Concepción; Mentaberre, Gregorio; Sánchez, Sergio; Fernández-Llario, Pedro; Gómez-Barrero, Susana; Navarro-Gonzalez, Nora; Serrano, Emmanuel; Casas-Díaz, Encarna; Marco, Ignasi; Fernández-Garayzabal, José-Francisco; Mateos, Ana; Vidal, Dolors; Lavín, Santiago; Domínguez, Lucas
Methicillin-resistant Staphylococcus aureus (MRSA) is a life-threatening pathogen in humans and its presence in animals is a public health concern. The aim of this study was to measure the prevalence of MRSA in free-living wild animals. Samples from red deer (n=273), Iberian ibex (n=212), Eurasian Griffon vulture (n=40) and wild boar (n=817) taken from different areas in Spain between June 2008 and November 2011 were analyzed. Characterization of the isolates was performed by spa typing, multi-locus sequence typing (MLST) and antimicrobial susceptibility testing. A low prevalence of MRSA was found with 13 isolates obtained from 12 animals (0.89%; 95% CI: 0.46-1.56). All MRSA sequence types belonged to ST398 (t011 and t1451) and ST1 (t127). Genotypes and antimicrobial susceptibility patterns (tetracycline resistance in ST398 and clindamycin-erythromycin-tetracycline resistance in ST1) suggest that the MRSA found probably originated in livestock (ST398) or humans (ST1). This is the first report of MRSA carriers in free-living wild animals in Europe. Although our data showed that MRSA prevalence is currently low, free-living wild animals might act as reservoir and represent a potential risk for human health.
Pathare, Nirmal A; Asogan, Harshini; Tejani, Sara; Al Mahruqi, Gaitha; Al Fakhri, Salma; Zafarulla, Roshna; Pathare, Anil V
In Oman, the prevalence of health care associated methicillin resistant Staphylococcus aureus [HA-MRSA] is unknown. Therefore, to estimate the prevalence of HA-MRSA, we collected nasal swabs and swabs from cell phones on sterile polyester swabs and immediately inoculated on the mannitol salt agar containing oxacillin from medical students and hospital health care providers. Antibiotic susceptibility testing of the isolates was then performed using the Kirby Bauer's disc diffusion method. Additionally, a brief survey questionnaire was used to acquire demographic data. Amongst the 311 participants enrolled, nasal colonization with HA-MRSA was found in 47 individuals (15.1%, 95% confidence interval [CI]=11.1%, 19.1%). HA-MRSA was also isolated from the cell phone surfaces in 28 participants (9.0%, 95% CI=8.6%, 9.3%). 5 participants (1.6%) showed positive results both from their nasal swabs and from their cell phones. Antibiotic resistance to erythromycin [48%] and clindamycin [29%] was relatively high. 9.3% HA-MRSA isolates were vancomycin resistant [6.6% nasal carriage]. There was no statistically significant correlation between HA-MRSA isolates and the demographic characteristics or the risk factors namely gender, underlying co-morbidities like diabetes, hypertension, skin/soft tissue infections, skin ulcers/wounds, recent exposure to antibiotics, or hospital visits (p>0.05, Chi-square test).
Dinjaski, Nina; Fernández-Gutiérrez, Mar; Selvam, Shivaram; Parra-Ruiz, Francisco J.; Lehman, Susan M.; Román, Julio San; García, Ernesto; García, José L.; García, Andrés J.; Prieto, María Auxiliadora
Biomaterial-associated infections represent a significant clinical problem, and treatment of these microbial infections is becoming troublesome due to the increasing number of antibiotic-resistant strains. Here, we report a naturally functionalized bacterial polyhydroxyalkanoate (PHACOS) with antibacterial properties. We demonstrate that PHACOS selectively and efficiently inhibits the growth of methicillin-resistant Staphylococcus aureus (MRSA) both in vitro and in vivo. This ability has been ascribed to the functionalized side chains containing thioester groups. Significantly less (3.2-fold) biofilm formation of S. aureus was detected on PHACOS compared to biofilms formed on control poly(3-hydroxyoctanoate-co-hydroxyhexanoate) and poly(ethylene terephthalate), but no differences were observed in bacterial adhesion among these polymers. PHACOS elicited minimal cytotoxic and inflammatory effects on murine macrophages and supported normal fibroblast adhesion. In vivo fluorescence imaging demonstrated minimal inflammation and excellent antibacterial activity for PHACOS compared to controls in an in vivo model of implant-associated infection. Additionally, reductions in neutrophils and macrophages in the vicinity of sterile PHACOS compared to sterile PHO implant were observed by immunohistochemistry. Moreover, a similar percentage of inflammatory cells was found in the tissue surrounding sterile PHACOS and S. aureus pre-colonized PHACOS implants, and these levels were significantly lower than S. aureus pre-colonized control polymers. These findings support a contact active surface mode of antibacterial action for PHACOS and establish this functionalized polyhydroxyalkanoate as an infection-resistant biomaterial. PMID:24094939
Liu, Junyan; Chen, Dingqiang; Peters, Brian M; Li, Lin; Li, Bing; Xu, Zhenbo; Shirliff, Mark E
Considered to be a potential "superbug", methicillin-resistant Staphylococcus aureus (MRSA) has been one of the major recent infectious pathogens and thus poses a challenge to hospital infection control. The mobile genetic element staphylococcal chromosomal cassette mec (SCCmec) carries both the mecA or mecC gene, encoding for a novel speciﬁc penicillin-binding protein (PBP2a), and site-speciﬁc recombinase genes ccrAB or/and ccrC. In MRSA, the acquisition of SCCmec leads to the resistance to the β-lactam antibiotics. As SCCmec plays a core role in the antimicrobial resistance characteristics, molecular epidemiology and evolution of MRSA, a thorough summary and comprehensive understanding of the prevalence and structural characteristics of SCCmec may aid in global surveillance, implementation and investigation on MRSA isolates, as well as further development of preventive and therapeutic approaches. Consequently, this review is aimed at describing the history, prevalence, types and subtypes, and current typing methods of SCCmec, with the focus on the typical structures of the SCCmec cassette.
Wang, Lei; Ruan, Shigui
In this work, we investigate the role of environmental contamination on the clinical epidemiology of antibiotic-resistant bacteria in hospitals. Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium that causes infections in different parts of the body. It is tougher to treat than most strains of Staphylococcus aureus or staph, because it is resistant to some commonly used antibiotics. Both deterministic and stochastic models are constructed to describe the transmission characteristics of MRSA in hospital setting. The deterministic epidemic model includes five compartments: colonized and uncolonized patients, contaminated and uncontaminated health care workers (HCWs), and bacterial load in environment. The basic reproduction number R 0 is calculated, and its numerical and sensitivity analysis has been performed to study the asymptotic behavior of the model, and to help identify factors responsible for observed patterns of infections. A stochastic epidemic model with stochastic simulations is also presented to supply a comprehensive analysis of its behavior. Data collected from Beijing Tongren Hospital will be used in the numerical simulations of our model. The results can be used to provide theoretical guidance for designing efficient control measures, such as increasing the hand hygiene compliance of HCWs and disinfection rate of environment, and decreasing the transmission rate between environment and patients and HCWs.
Abstract We retrospectively reviewed the cases of 15 foals and adult horses in which vancomycin was used, alone or in combination with an aminoglycoside, to treat methicillin-resistant staphylococcal and enterococcal infections. Signalment, presenting complaint, history (including history of treatment for the current complaint), results of bacterial culture and antimicrobial susceptibility testing, treatment, and outcome were reviewed. The average vancomycin dosage was 7.5 mg/kg q8h, administered by intravenous infusion over 30 min. The infection resolved in all 7 horses with soft tissue infections and in 6 of the 8 horses with infections involving a bone or a joint, or both. No adverse effects of vancomycin therapy were noted. Although the number of cases is small, our findings suggest that vancomycin, alone or in combination with an aminoglycoside, is safe and effective for the treatment of resistant staphylococcal and enterococcal infections in horses and foals. However, owing to the importance of staphylococci and enterococci in human medicine and the problems with emerging resistance, we recommend that the use of vancomycin in horses be limited to cases in which culture and susceptibility results clearly indicate that this agent is likely to be effective and in which there is no reasonable alternative. PMID:16479726
Haste, Nina M; Thienphrapa, Wdee; Tran, Dan N; Loesgen, Sandra; Sun, Peng; Nam, Sang-Jip; Jensen, Paul R; Fenical, William; Sakoulas, George; Nizet, Victor; Hensler, Mary E
The rapid rise in antimicrobial resistance in bacteria has generated an increased demand for the development of novel therapies to treat contemporary infections, especially those caused by methicillin-resistant Staphylococcus aureus (MRSA). However, antimicrobial development has been largely abandoned by the pharmaceutical industry. We recently isolated the previously described thiopeptide antibiotic nosiheptide from a marine actinomycete strain and evaluated its activity against contemporary clinically relevant bacterial pathogens. Nosiheptide exhibited extremely potent activity against all contemporary MRSA strains tested including multiple drug-resistant clinical isolates, with MIC values 0.25 mg l(-1). Nosiheptide was also highly active against Enterococcus spp. and the contemporary hypervirulent BI/NAP1/027 strain of Clostridium difficile but was inactive against most Gram-negative strains tested. Time-kill analysis revealed nosiheptide to be rapidly bactericidal against MRSA in a concentration- and time-dependent manner, with a nearly 2-log kill noted at 6 h at 10 × MIC. Furthermore, nosiheptide was found to be non-cytotoxic against mammalian cells at >100 × MIC, and its anti-MRSA activity was not inhibited by 20% human serum. Notably, nosiheptide exhibited a significantly prolonged post-antibiotic effect against both healthcare- and community-associated MRSA compared with vancomycin. Nosiheptide also demonstrated in vivo activity in a murine model of MRSA infection, and therefore represents a promising antibiotic for the treatment of serious infections caused by contemporary strains of MRSA.
Warrack, S; Panjikar, P; Duster, M; Safdar, N
Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen of major public health importance. Colonisation precedes infection; thus reducing MRSA carriage may be of benefit for reducing infection. Probiotics represent a novel approach to reducing MRSA carriage. We undertook a pilot feasibility randomised controlled trial of the tolerability and acceptability of probiotics for reducing nasal and intestinal carriage of MRSA. In addition, subjects were screened for vancomycin-resistant enterocococci (VRE). Subjects with a history of MRSA were recruited from a large, academic medical center and randomised to take either a placebo or probiotic (Lactobacillus rhamnosus HN001). Subjects returned to the clinic after four weeks for further testing to determine adherence to the probiotic regimen and colonisation of MRSA. 48 subjects were enrolled and randomised. Nearly 25% were transplant recipients and 30% had diabetes. The probiotic was well tolerated in the study population though minor side effects, such as nausea and bloating, were observed. A majority of the subjects randomised to HN001 had good adherence to the regimen. At the four week time point among subjects randomised to the probiotic, MRSA was detected in 67 and 50% of subjects colonised in the nares and the gastrointestinal tract, respectively. Three subjects who initially tested positive for VRE were negative after four weeks of probiotic exposure. Probiotics were well tolerated in our study population of largely immunocompromised subjects with multiple comorbidities. Adherence to the intervention was good. Probiotics should be studied further for their potential to reduce colonisation by multidrug resistant bacteria.
Lee, Kyung-Min; Lee, Hyun-Soo; Kim, Man-Soo
Considering the popular use of antibiotic-containing eyedrops in Korea, it is important to know the emerging antibiotic-resistant strains of bacteria before treating infectious eye diseases. This is especially important in high-risk groups because of the high incidence of resistant infections and the subsequent treatment requirements. We report two cases of methicillin-resistant Staphylococcus aureus (MRSA) corneal ulcers in high-risk groups. The first case involved a patient who had keratitis after using antibiotic- and steroid-containing eyedrops to treat a corneal opacity that developed after repeated penetrating keratoplasty. The second case involved a patient who used antibiotic-containing eyedrops and a topical lubricant on a regular basis for >1 month to treat exposure keratitis due to lagophthalmos. The second patient's problems, which included a persistent superficial infiltration, developed after brain tumor surgery. Both cases showed MRSA on corneal culture, and the corneal ulcers improved in both patients after the application of vancomycin-containing eyedrops. In conclusion, MRSA infection should be considered in corneal ulcers that have a round shape, mild superficial infiltration, and slow progression, especially in high-risk groups. This report includes descriptions of the characteristic features, antibiotic sensitivities, prevention, and successful treatment with vancomycin-containing eyedrops for MRSA corneal ulcers.
Spread of methicillin-resistant Staphylococcus aureus belonging to the Brazilian epidemic clone in a general hospital and emergence of heterogenous resistance to glycopeptide antibiotics among these isolates.
dos Santos Soares, M J; da Silva-Carvalho, M C; Ferreira-Carvalho, B T; Figueiredo, A M
Methicillin-resistant Staphylococcus aureus (MRSA) infections have been increasing at an alarming rate world-wide. MRSA epidemics due to the clonal spread of multi-resistant isolates have been described. In this paper we show the absolute predominance of MRSA strains from the Brazilian epidemic clone in a hospital in the Northeast region of Brazil and the emergence of a vancomycin and teicoplanin heterogeneous resistant subpopulation among these isolates.
Friendship, Robert M.; Weese, J. Scott
Methicillin-resistant Staphylococcus aureus (MRSA) originating from swine is concerning for public health, but an understanding of the emergence and persistence of MRSA in nursery herds is lacking. The aim of this study was to determine whether MRSA in nursery pigs is associated with particular herd-level parameters, including the use of antimicrobials, disinfectants, and heavy metals, which may be driving the selection and persistence of antimicrobial resistance. Nasal cultures for MRSA were completed for 390 pigs from 26 farms at the end of the suckling phase and again at 3 weeks postweaning. Herd-level information was collected, and a random subset of MRSA isolates was screened for resistance to zinc and quaternary ammonium compounds (QACs). Multivariate analysis revealed that in-feed concentrations of zinc (P < 0.001) and frequent disinfection of nursery pens (P < 0.001) are associated with MRSA shedding in nursery pigs. Furthermore, 62.5% (25/40) of MRSA isolates carried the zinc resistance gene czrC and demonstrated decreased susceptibility to zinc. All MRSA isolates carried at least 1 QAC resistance gene. The most common genotype was qacG qacH smr, which occurred in 32.5% (13/40) of isolates. Seven isolates (17.5%) demonstrated a significant tolerance to benzalkonium chloride, indicating a potential to survive commercial QAC exposure in the presence of organic matter. Overall, these findings indicate that high levels of in-feed zinc and QAC-based disinfectants are important drivers in the selection and persistence of MRSA in commercial swine herds, and these agents may be coselecting for other antimicrobial resistance genes. PMID:25662976
Edwards-Jones, V; Buck, R; Shawcross, S G; Dawson, M M; Dunn, K
Patchouli, tea tree, geranium, lavender essential oils and Citricidal (grapefruit seed extract) were used singly and in combination to assess their anti-bacterial activity against three strains of Staphylococcus aureus: Oxford S. aureus NCTC 6571 (Oxford strain), Epidemic methicillin-resistant S. aureus (EMRSA 15) and MRSA (untypable). The individual essential oils, extracts and combinations were impregnated into filter paper discs and placed on the surface of agar plates, pre-seeded with the appropriate strain of Staphylococcus. The effects of the vapours of the oils and oil combinations were also assessed using impregnated filter paper discs that were placed on the underside of the Petri dish lid at a distance of 8mm from the bacteria. The most inhibitory combinations of oils for each strain were used in a dressing model constructed using a four layers of dressings: the primary layer consisted of either Jelonet or TelfaClear with or without Flamazine; the second was a layer of gauze, the third a layer of Gamgee and the final layer was Crepe bandage. The oil combinations were placed in either the gauze or the Gamgee layer. This four-layered dressing was placed over the seeded agar plate, incubated for 24h at 37 degrees C and the zones of inhibition measured. All experiments were repeated on three separate occasions. No anti-bacterial effects were observed when Flamazine was smeared on the gauze in the dressing model. When Telfaclear was used as the primary layer in the dressing model compared to Jelonet, greater zones of inhibition were observed. A combination of Citricidal and geranium oil showed the greatest-anti-bacterial effects against MRSA, whilst a combination of geranium and tea tree oil was most active against the methicillin-sensitive S. aureus (Oxford strain). This study demonstrates the potential of essential oils and essential oil vapours as antibacterial agents and for use in the treatment of MRSA infection.
Hu, Yanmin; Liu, Alexander; Vaudrey, James; Vaiciunaite, Brigita; Moigboi, Christiana; McTavish, Sharla M.; Kearns, Angela; Coates, Anthony
Bacterial infections remain the leading killer worldwide which is worsened by the continuous emergence of antibiotic resistance. In particular, methicillin-sensitive (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) are prevalent and the latter can be difficult to treat. The traditional strategy of novel therapeutic drug development inevitably leads to emergence of resistant strains, rendering the new drugs ineffective. Therefore, rejuvenating the therapeutic potentials of existing antibiotics offers an attractive novel strategy. Plectasin, a defensin antimicrobial peptide, potentiates the activities of other antibiotics such as β-lactams, aminoglycosides and glycopeptides against MSSA and MRSA. We performed in vitro and in vivo investigations to test against genetically diverse clinical isolates of MSSA (n = 101) and MRSA (n = 115). Minimum inhibitory concentrations (MIC) were determined by the broth microdilution method. The effects of combining plectasin with β-lactams, aminoglycosides and glycopeptides were examined using the chequerboard method and time kill curves. A murine neutropenic thigh model and a murine peritoneal infection model were used to test the effect of combination in vivo. Determined by factional inhibitory concentration index (FICI), plectasin in combination with aminoglycosides (gentamicin, neomycin or amikacin) displayed synergistic effects in 76-78% of MSSA and MRSA. A similar synergistic response was observed when plectasin was combined with β-lactams (penicillin, amoxicillin or flucloxacillin) in 87–89% of MSSA and MRSA. Interestingly, no such interaction was observed when plectasin was paired with vancomycin. Time kill analysis also demonstrated significant synergistic activities when plectasin was combined with amoxicillin, gentamicin or neomycin. In the murine models, plectasin at doses as low as 8 mg/kg augmented the activities of amoxicillin and gentamicin in successful treatment of MSSA and MRSA infections. We
Introduction Staphylococcus aureus has emerged as a major nosocomial pathogen in the last decades and also represents the second most common pathogen isolated from patients in outpatient settings. Although methicillin-resistant S.aureus infections were traditionally limited to hospitals, community-associated cases of methicillin-resistant S.aureus infections have been reported. In our case, we observed an unexpected event during treatment. Case presentation A 60-year-old Caucasian man developed fever and multiple muscle and brain abscesses caused by Panton-Valentine leukocidin-negative community-associated methicillin-resistant S. aureus. Conclusion Although our patient was given antimicrobials active against the isolated methicillin-resistant S. aureus strain, it was only after the introduction of daptomycin that his skin, soft tissue and muscle lesions and also brain manifestations improved. PMID:22938025
Co-infection of methicillin-resistant Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus and extended spectrum β-lactamase producing Escherichia coli in bovine mastitis--three cases reported from India.
Bandyopadhyay, Samiran; Samanta, Indranil; Bhattacharyya, Debaraj; Nanda, Pramod Kumar; Kar, Debasish; Chowdhury, Jayanta; Dandapat, Premanshu; Das, Arun Kumar; Batul, Nayan; Mondal, Bimalendu; Dutta, Tapan Kumar; Das, Gunjan; Das, Bikash Chandra; Naskar, Syamal; Bandyopadhyay, Uttam Kumar; Das, Suresh Chandra; Bandyopadhyay, Subhasish
Emergence of antimicrobial resistance among bovine mastitis pathogens is the major cause of frequent therapeutic failure and a cause of concern for veterinary practitioners. This study describes intra-mammary infection of methicillin-resistant Staphylococcus epidermidis (MRSE), methicillin-resistant Staphylococcus aureus (MRSA) and extended spectrum β-lactamase (ESBL) producing Escherichia coli in two Holstein Friesian crossbred cows with subclinical mastitis and one non-descript cow with clinical mastitis in two different districts of West Bengal, India. In total, three MRSE, one MRSA and three ESBL producing E. coli were isolated from these cases. Both the crossbreds were detected with MRSE (HFSE1 and HFSE2) and ESBL producing E. coli (HFEC1 and HFEC2), whereas, simultaneous infection of three pathogens viz. MRSA (NDSA1), MRSE (NDSE1) and ESBL producing E. coli (NDEC1) was found in the non-descript cow. The methicillin-resistant isolates possessed mecA gene and exhibited resistance to various antibiotics such as amikacin, tetracycline and glycopeptides. The ESBL producers were positive for blaCTX-M and blaTEM genes; in addition, HFEC1 and HFEC2 were positive for blaSHV and possessed the genes for class I integron (int1), sulphonamide resistance (sul1), quinolone resistance (qnrS) and other virulence factors (papC, iucD and ESTA1). All the ESBL producers exhibited resistance to a variety of antibiotics tested including third- and fourth-generation cephalosporins and were also intermediately resistant to carbapenems. This is the first ever report on simultaneous occurrence of MRSE, MRSA and ESBL producing E. coli in bovine mastitis indicating a major concern for dairy industry and public health as well.
Bagcigil, Funda A; Moodley, Arshnee; Baptiste, Keith E; Jensen, Vibeke F; Guardabassi, Luca
beta-Lactams and macrolides are important antibiotics for treatment of staphylococcal infections in both humans and animals. The aim of the study was to investigate the occurrence, species distribution and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of dogs, horses, pigs, and cattle in Denmark. Nasal swabs were collected from a total of 400 animals, including 100 individuals of each species. Methicillin- and erythromycin-resistant staphylococci were isolated on selective media, identified by 16S rDNA sequencing, and typed by pulsed field gel electrophoresis (PFGE). Methicillin-resistant coagulase-negative staphylococci (MRCoNS) harbouring mecA were isolated from horses (50%) and dogs (13%), but not from food animals. The species identified were S. haemolyticus (n=21), S. vitulinus (n=19), S. sciuri (n=13), S. epidermidis (n=8), and S. warneri (n=2). mecA-mediated methicillin resistance in S. vitulinus was described for the first time. Methicillin-resistant S. aureus was not detected. PFGE analysis revealed the presence of specific MRCoNS clones in samples originating from the same veterinary hospital or equine farm. Erythromycin-resistant S. aureus (ERSA) was detected in 38% of pigs and all isolates harboured a constitutively expressed erm(C) gene. The vast majority (37/38) of pigs carrying ERSA originated from a farm characterized by frequent use of macrolides. Most ERSA isolates (28/38) displayed indistinguishable or closely related PFGE patterns, indicating clonal distribution within the farm. Based on the analysis of data on antimicrobial consumption, the occurrence of MRCoNS in companion animals and that of ERSA in pigs reflected national and local patterns of antimicrobial usage.
Chao, Guoxiang; Zhang, Xiaoping; Zhang, Xiaorong; Huang, Yao; Xu, Lan; Zhou, Liping; Yang, Weixia; Jiang, Yuan; Xue, Feng; Wu, Yantao
A diverse collection of 261 Staphylococcus aureus strains from human, animal, food, and environmental sources were tested for the presence and type of SCCmec elements, antibiotic susceptibility to various antibiotics, and non-ß-lactam antibiotic resistance genes. About 18.39% (48/261) of strains were methicillin-resistant S. aureus (MRSA) including 29.75% (36/121) human strains of which 29 strains were hospital-acquired MRSA (HA-MRSA) and 7 strains were community-associated MRSA (CA-MRSA) and 19.67% (12/61) animal strains that all were CA-MRSA strains. The percentage of CA-MRSA strains from animals was significantly higher than that from human (p<0.01). Most of MRSA strains and a part of methicillin-susceptible S. aureus (MSSA) strains harbored unique combinations of non-ß-lactamase genes aac(6')/aph(2″), aph(3')-III, ant (4',4″), ermA, ermC, mrsA, tetM, and tetK. Antibiotic resistance genes were detected more frequently in HA-MRSA strains than in CA-MRSA strains (p<0.01). MRSA strains and MSSA strains had 22 and 39 antibiotic profiles to 15 tested antibiotics, respectively. The resistant proportion was higher in HA-MRSA strains than in CA-MSSA strains for various antibiotics, as well as higher in MRSA strains than in MSSA strains. Animal MRSA reservoirs (particularly pigs and cows) might represent an important source of human CA-MRSA. CA-MRSA strains might acquire more different resistance genes gradually, depending on the selective pressure of antibiotics in different regions or environments. CA-MRSA is not yet endemic in China, but could be prevalent in future, contributing to its acquiring more resistance genes and huge animal sources. Infection with multidrug-resistant MSSA strains acquired from food, animal, and human sources might also become a significant problem for human medicine, which warrants further study.
Sarkar, Abdullah; Raji, Adeola; Garaween, Ghada; Soge, Olusegun; Rey-Ladino, Jose; Al-Kattan, Wael; Shibl, Atef; Senok, Abiola
Most Staphylococcus aureus infections occur in previously colonized persons who also act as reservoirs for continued dissemination. This study aimed to investigate the carriage of antimicrobial resistance and virulence markers in S. aureus isolates associated with nasal colonization. The study was conducted from December 2013-April 2014. Nasal swabs were collected and questionnaires administered to 97 medical students in Riyadh Saudi Arabia. Bacterial culture, identification and antimicrobial susceptibility testing were performed by conventional methods and chromogenic agar was used for methicillin resistant S. aureus (MRSA) screening. Molecular characterization of isolates was carried out using the StaphyType DNA microarray. Thirty two students (43%) had S. aureus nasal carriage (MSSA = 31; MRSA = 1). Seventeen clonal complexes (CC) were identified namely: CC15-MSSA (n = 5), CC1-MSSA-SCCfus (n = 4), CC8-MSSA (n = 3), CC22-MSSA (n = 3), CC25-MSSA (n = 3), CC101-MSSA (n = 2). Other CC found as single isolates were CC5-MSSA, CC6-MSSA, CC30-MSSA, CC45-MSSA, CC96-MSSA, CC188-MSSA, CC398-MSSA, CC942-MSSA/PVL+, CC1290-MSSA, ST2482-MSSA, CC80-MRSA-IV/PVL+. The CC1-SCCfus isolates harbored the Staphylococcal cassette chromosome (SCC) with ccrA-1; ccrB-1 and ccrB-3 genes plus the putative fusidic acid resistance marker Q6GD50. One MSSA isolate was genotyped as coagulase negative Staphylococcus spp with an irregular composite SCCmec element. Majority of the isolates harbored various virulence genes including the hemolysin, enterotoxin, and exfoliative genes as well as various adhesive protein producing genes. Although there was low carriage of MRSA, the MSSA isolates harbored various resistance and virulence genes including those usually seen in MRSA isolates. The presence of isolates with incomplete SCCmec elements plus putative resistance and virulence genes is of concern.
Frana, Timothy S; Beahm, Aleigh R; Hanson, Blake M; Kinyon, Joann M; Layman, Lori L; Karriker, Locke A; Ramirez, Alejandro; Smith, Tara C
In the last decade livestock-associated methicillin-resistant S. aureus (LA-MRSA) has become a public health concern in many parts of the world. Sequence type 398 (ST398) has been the most commonly reported type of LA-MRSA. While many studies have focused on long-term exposure experienced by swine workers, this study focuses on short-term exposures experienced by veterinary students conducting diagnostic investigations. The objectives were to assess the rate of MRSA acquisition and longevity of carriage in students exposed to pork farms and characterize the recovered MRSA isolates. Student nasal swabs were collected immediately before and after farm visits. Pig nasal swabs and environmental sponge samples were also collected. MRSA isolates were identified biochemically and molecularly including spa typing and antimicrobial susceptibility testing. Thirty (30) veterinary students were enrolled and 40 pork farms were visited. MRSA was detected in 30% of the pork farms and in 22% of the students following an exposure to a MRSA-positive pork farm. All students found to be MRSA-positive initially following farm visit were negative for MRSA within 24 hours post visit. Most common spa types recovered were t002 (79%), t034 (16%) and t548 (4%). Spa types found in pork farms closely matched those recovered from students with few exceptions. Resistance levels to antimicrobials varied, but resistance was most commonly seen for spectinomycin, tetracyclines and neomycin. Non-ST398 MRSA isolates were more likely to be resistant to florfenicol and neomycin as well as more likely to be multidrug resistant compared to ST398 MRSA isolates. These findings indicate that MRSA can be recovered from persons visiting contaminated farms. However, the duration of carriage was very brief and most likely represents contamination of nasal passages rather than biological colonization. The most common spa types found in this study were associated with ST5 and expands the range of livestock
Kardas-Sloma, Lidia; Boëlle, Pierre-Yves; Opatowski, Lulla; Guillemot, Didier; Temime, Laura
Interventions designed to reduce antibiotic consumption are under way worldwide. While overall reductions are often achieved, their impact on the selection of antibiotic-resistant selection cannot be assessed accurately from currently available data. We developed a mathematical model of methicillin-sensitive and methicillin-resistant Staphylococcus aureus (MSSA and MRSA) transmission inside and outside the hospital. A systematic simulation study was then conducted with two objectives: to assess the impact of antibiotic class-specific changes during an antibiotic reduction period and to investigate the interactions between antibiotic prescription changes in the hospital and the community. The model reproduced the overall reduction in MRSA frequency in French intensive-care units (ICUs) with antibiotic consumption in France from 2002 to 2003 as an input. However, the change in MRSA frequency depended on which antibiotic classes changed the most, with the same overall 10% reduction in antibiotic use over 1 year leading to anywhere between a 69% decrease and a 52% increase in MRSA frequency in ICUs and anywhere between a 37% decrease and a 46% increase in the community. Furthermore, some combinations of antibiotic prescription changes in the hospital and the community could act in a synergistic or antagonistic way with regard to overall MRSA selection. This study shows that class-specific changes in antibiotic use, rather than overall reductions, need to be considered in order to properly anticipate the impact of an antibiotic reduction campaign. It also highlights the fact that optimal gains will be obtained by coordinating interventions in hospitals and in the community, since the effect of an intervention in a given setting may be strongly affected by exogenous factors.
El-Adawy, Hosny; Ahmed, Marwa; Hotzel, Helmut; Monecke, Stefan; Schulz, Jochen; Hartung, Joerg; Ehricht, Ralf; Neubauer, Heinrich; Hafez, Hafez M
Methicillin-resistant Staphylococcus aureus (MRSA) is a major human health problem and recently, domestic animals are described as carriers and possible reservoirs. Twenty seven S. aureus isolates from five turkey farms (n = 18) and two broiler farms (n = 9) were obtained by culturing of choana and skin swabs from apparently healthy birds, identified by Taqman-based real-time duplex nuc-mecA-PCR and characterized by spa typing as well as by a DNA microarray based assay which covered, amongst others, a considerable number of antibiotic resistance genes, species controls, and virulence markers. The antimicrobial susceptibility profiles were tested by agar diffusion assays and genotypically confirmed by the microarray. Five different spa types (3 in turkeys and 2 in broilers) were detected. The majority of MRSA isolates (24/27) belonged to clonal complex 398-MRSA-V. The most frequently occurring spa types were accordingly t011, t034, and t899. A single CC5-MRSA-III isolated from turkey and CC398-MRSA with an unidentified/truncated SCCmec element in turkey and broiler were additionally detected. The phenotypic antimicrobial resistance profiles of S. aureus isolated from both turkeys and broilers against 14 different antimicrobials showed that all isolates were resistant to ampicillin, cefoxitin, oxacillin, doxycycline, and tetracycline. Moreover, all S. aureus isolated from broilers were resistant to erythromycin and azithromycin. All isolates were susceptible to gentamicin, chloramphenicol, sulphonamides, and fusidic acid. The resistance rate against ciprofloxacin was 55.6% in broiler isolates and 42.1% in turkey isolates. All tetracycline resistant isolates possessed genes tetK/M. All erythromycin-resistant broiler isolates carried ermA. Only one broiler isolate (11.1%) carried genes ermA, ermB, and ermC, while 55.6% of turkey isolates possessed ermA and ermB genes. Neither PVL genes (lukF/S-PV), animal-associated leukocidin (lukM and luk-P83) nor the gene encoding
El-Adawy, Hosny; Ahmed, Marwa; Hotzel, Helmut; Monecke, Stefan; Schulz, Jochen; Hartung, Joerg; Ehricht, Ralf; Neubauer, Heinrich; Hafez, Hafez M.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major human health problem and recently, domestic animals are described as carriers and possible reservoirs. Twenty seven S. aureus isolates from five turkey farms (n = 18) and two broiler farms (n = 9) were obtained by culturing of choana and skin swabs from apparently healthy birds, identified by Taqman-based real-time duplex nuc-mecA-PCR and characterized by spa typing as well as by a DNA microarray based assay which covered, amongst others, a considerable number of antibiotic resistance genes, species controls, and virulence markers. The antimicrobial susceptibility profiles were tested by agar diffusion assays and genotypically confirmed by the microarray. Five different spa types (3 in turkeys and 2 in broilers) were detected. The majority of MRSA isolates (24/27) belonged to clonal complex 398-MRSA-V. The most frequently occurring spa types were accordingly t011, t034, and t899. A single CC5-MRSA-III isolated from turkey and CC398-MRSA with an unidentified/truncated SCCmec element in turkey and broiler were additionally detected. The phenotypic antimicrobial resistance profiles of S. aureus isolated from both turkeys and broilers against 14 different antimicrobials showed that all isolates were resistant to ampicillin, cefoxitin, oxacillin, doxycycline, and tetracycline. Moreover, all S. aureus isolated from broilers were resistant to erythromycin and azithromycin. All isolates were susceptible to gentamicin, chloramphenicol, sulphonamides, and fusidic acid. The resistance rate against ciprofloxacin was 55.6% in broiler isolates and 42.1% in turkey isolates. All tetracycline resistant isolates possessed genes tetK/M. All erythromycin-resistant broiler isolates carried ermA. Only one broiler isolate (11.1%) carried genes ermA, ermB, and ermC, while 55.6% of turkey isolates possessed ermA and ermB genes. Neither PVL genes (lukF/S-PV), animal-associated leukocidin (lukM and luk-P83) nor the gene encoding
Kwon, Nam Hoon; Park, Kun Taek; Jung, Woo Kyung; Youn, Hwa Young; Lee, Yeonhee; Kim, So Hyun; Bae, Wonki; Lim, Ji Youn; Kim, Ji Yeon; Kim, Jun Man; Hong, Soon Keun; Park, Yong Ho
Methicillin resistant Staphylococcus aureus (MRSA) is one of the most important pathogens in human and veterinary hospitals. The isolation of MRSA from animals and foodstuffs has been reported with an increased incidence. However, methicillin (oxacillin) is not used in animal husbandry or in animal hospitals in Korea. In this study, three pre-MRSA and one silent mecA-carrying methicillin susceptible S. aureus (smMSSA) were isolated from retail chicken meat, and three MRSA were isolated from hospitalized dogs in Korea. The three pre-MRSA isolates were determined to have a staphylococcal cassette chromosome mec (SCCmec) type III, and the smMSSA isolate was not classified. The animal hospital isolates were found to contain SCCmec type II. Seven and 15 S. aureus isolated from hospitalized humans and bovine milk, respectively, were also examined in this study in order to determine the epidemiological origins of MRSA. Multilocus sequencing typing (MLST) revealed that the chicken meat and bovine milk isolates were closely related to the animal hospital isolates. The SCCmec characteristics and MLST analyses indicated the possibility of the human to animal transmission of MRSA. These results highlight the importance of identifying MRSA carriers as well as intercepting MRSA transmission because MRSA is becoming increasingly widespread without any plausible relationship with the use of methicillin (oxacillin).
Discrimination of methicillin-resistant Staphylococcus aureus from methicillin-susceptible Staphylococcus aureus or coagulase-negative staphylococci by detection of penicillin-binding protein 2 and penicillin-binding protein 2' using a bioluminescent enzyme immunoassay.
Shiga, Kazuki; Gomi, Keiko; Nishimura, Motoi; Watanabe, Masaharu; Nomura, Fumio; Kajiyama, Naoki
For the discrimination of methicillin-resistant S. aureus (MRSA) from methicillin-susceptible S. aureus (MSSA) or coagulase-negative staphylococci (CNS), we developed a bioluminescent enzyme immunoassay (BLEIA) for detecting penicillin-binding protein 2 (PBP2) and penicillin-binding protein 2' (PBP2') using biotinylated firefly luciferase. The BLEIA was able to detect recombinant PBP2 at 50 pg/ml and recombinant PBP2' at 500 pg/ml. PBP2 and PBP2' present in the membranes of S. aureus were extracted by acid and detergent treatment. The method was able to detect PBP2 or PBP2' extracted from 10(6) colony forming units of S. aureus because of efficient extraction and the high sensitivity of luciferase. In a study of clinical isolates previously characterized as either MRSA or MSSA by antibiotic susceptibility testing, all 34 specimens identified as MRSA were both PBP2 and PBP2' positive. The 34 MSSA specimens were PBP2 positive and PBP2' negative. Moreover, the BLEIA could detect PBP2' extracted from four species of methicillin-resistant CNS, but not PBP2 extracted from four species of methicillin-resistant and methicillin-susceptible CNS. This result suggested that PBP2 could be a unique marker for discrimination of S. aureus from CNS. A BLEIA that is able to detect PBP2 and PBP2' may be useful in clinical diagnostics.
Metwally, L; Gomaa, N; Hassan, R
We evaluated the utility of 2 methods for detection of methicillin-resistant Staphylococcus aureus (MRSA) directly from signal-positive blood culture bottles: loop-mediated isothermal amplification (LAMP) assay, and direct cefoxitin disk diffusion (DCDD) test using a 30 μg cefoxitin disk. In parallel, standard microbiological identification and oxacillin susceptibility testing with MecA PCR was performed. Of 60 blood cultures positive for Gram-positive cocci in clusters, LAMP (via detection of the FemA and MecA genes) showed 100% sensitivity and specificity for identification of MRSA/MSSA. When coagulase-negative staphylococci were tested, sensitivity for detection of methicillin resistance was 91.7% and specificity was 100%. DCDD along with direct tube coagulase assay detected only 80.6% of MRSA/MSSA. LAMP showed higher diagnostic accuracy although DCDD was more cost-effective and did not require additional reagents or supplies.
Gomez, Eric O; Jafary, Ahmar; Dever, Lisa L
Daptomycin has demonstrated clinical efficacy in the treatment of methicillin-resistant Staphylococcus aureus-associated bacteremia and right-sided infective endocarditis. Although daptomycin is not approved for treatment of gram-positive pneumonia, clinical evidence suggests that it may be effective therapy for S. aureus-associated septic pulmonary emboli (SPE). We present our clinical experience with the use of daptomycin in combination with rifampin in four patients with SPE in the absence of infective endocarditis. Three of the patients had a history of injection drug use; two of these patients also had soft-tissue infections. All patients had clinical resolution of their infections. Daptomycin and rifampin appear to have a role in the treatment of methicillin-resistant S. aureus bacteremia with SPE in the absence of infective endocarditis and should be considered in patients that have failed therapy with vancomycin.
Stewart, Audra; Dyamenahalli, Umesh; Greenberg, S Bruce; Drummond-Webb, Jonathan
We present the case of a 6-month-old previously healthy girl who presented with high fever, labored breathing, and an enlarged cardiac silhouette on her chest radiograph. Comprehensive evaluation discovered a ductus arteriosus aneurysm and pericardial effusion with methicillin-resistant Staphylococcus aureus bacteremia. Despite pericardiocentesis and appropriate intravenous antibiotics, there was rapid enlargement of the aneurysm and accumulation of echogenic material within the ductus arteriosus aneurysm. Infected aneurysm rupture was identified during emergency surgery. This infant also had vocal cord paresis, a likely complication of the surgery. The clinical course, diagnosis, and treatment of this patient are discussed. Infection of a ductus arteriosus or an infected ductal arteriosus aneurysm is a rare and potentially fatal clinical entity. In the era of increasing community-acquired methicillin-resistant S aureus infections, this is a diagnosis that requires a high index of suspicion.
Merchant, Chandni; Villanueva, Daphne-Dominique; Lalani, Ishan; Eng, Margaret; Kang, Yong
Staphylococcus schleiferi subsp. schleiferi is a coagulase-negative Staphylococcus which has been described as a pathogen responsible for various nosocomial infections including bacteremia, brain abscess, and infection of intravenous pacemakers. Recently, such bacteria have been described to be found typically on skin and mucosal surfaces. It is also believed to be a part of the preaxillary human flora and more frequently found in men. It is very similar in its pathogenicity with Staphylococcus aureus group and expresses a fibronectin binding protein. Literature on this pathogen reveals that it commonly causes otitis among dogs because of its location in the auditory meatus of canines. Also, it has strong association with pyoderma in dogs. The prime concern with this organism is the antibiotic resistance and relapse even after appropriate treatment. Very rarely, if any, cases have been reported about prostatic abscess (PA) with this microbe. Our patient had a history of recurrent UTIs and subsequent PA resulting in S. schleiferi bacteremia in contrast to gram negative bacteremia commonly associated with UTI. This organism was found to be resistant to methicillin, in spite of being negative for PBP2, which is a rare phenomenon and needs further studies. PMID:27092283
Chu, Ming; Zhang, Ming-Bo; Liu, Yan-Chen; Kang, Jia-Rui; Chu, Zheng-Yun; Yin, Kai-Lin; Ding, Ling-Yu; Ding, Ran; Xiao, Rong-Xin; Yin, Yi-Nan; Liu, Xiao-Yan; Wang, Yue-Dan
Berberine is an isoquinoline alkaloid widely used in the treatment of microbial infections. Recent studies have shown that berberine can enhance the inhibitory efficacy of antibiotics against clinical multi-drug resistant isolates of methicillin-resistant Staphylococcus aureus (MRSA). However, the underlying mechanisms are poorly understood. Here, we demonstrated that sub-minimum inhibitory concentrations (MICs) of berberine exhibited no bactericidal activity against MRSA, but affected MRSA biofilm development in a dose dependent manner within the concentration ranging from 1 to 64 μg/mL. Further study indicated that berberine inhibited MRSA amyloid fibrils formation, which consist of phenol-soluble modulins (PSMs). Molecular dynamics simulation revealed that berberine could bind with the phenyl ring of Phe19 in PSMα2 through hydrophobic interaction. Collectively, berberine can inhibit MRSA biofilm formation via affecting PSMs’ aggregation into amyloid fibrils, and thereby enhance bactericidal activity of antibiotics. These findings will provide new insights into the multiple pharmacological properties of berberine in the treatment of microbial-generated amyloid involved diseases.
Pastar, Irena; Nusbaum, Aron G.; Gil, Joel; Patel, Shailee B.; Chen, Juan; Valdes, Jose; Stojadinovic, Olivera; Plano, Lisa R.; Tomic-Canic, Marjana; Davis, Stephen C.
Understanding the pathology resulting from Staphylococcus aureus and Pseudomonas aeruginosa polymicrobial wound infections is of great importance due to their ubiquitous nature, increasing prevalence, growing resistance to antimicrobial agents, and ability to delay healing. Methicillin-resistant S. aureus USA300 is the leading cause of community-associated bacterial infections resulting in increased morbidity and mortality. We utilized a well-established porcine partial thickness wound healing model to study the synergistic effects of USA300 and P. aeruginosa on wound healing. Wound re-epithelialization was significantly delayed by mixed-species biofilms through suppression of keratinocyte growth factor 1. Pseudomonas showed an inhibitory effect on USA300 growth in vitro while both species co-existed in cutaneous wounds in vivo. Polymicrobial wound infection in the presence of P. aeruginosa resulted in induced expression of USA300 virulence factors Panton-Valentine leukocidin and α-hemolysin. These results provide evidence for the interaction of bacterial species within mixed-species biofilms in vivo and for the first time, the contribution of virulence factors to the severity of polymicrobial wound infections. PMID:23451098
Vijayamohan, N.; Nair, S. Pradeep
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a cause of increased morbidity and mortality in debilitated and immunocompromised patients. Aims and Objectives: To study the prevalence of MRSA among dermatology inpatients and elucidate its predisposing factors and antibiotic sensitivity pattern. Materials and Methods: This prospective study involved all the inpatients in the dermatology ward with suspected bacterial infections, both primary and secondary. Gram's stain, pus culture, and antibiotic sensitivity were the main tests done. Results: There were 61 patients in this study, with 36 males (59%) and 25 females (41%). The age group maximally affected was 41-50 years (21.6%). Vesiculobullous disorders (22, 36.1%) and eczemas (16, 26.2%) were the commonest dermatoses with secondary infection. Extremities (72.1%) and the trunk (26.2%) were the commonest sites involved. Diabetes mellitus (DM) was the commonest systemic illness (11.5%). Staphylococcus aureus was the commonest isolate, grown in 52 patients (85.2%) of whom 16 (30.80%) were MRSA positive. MRSA constituted 22.2% of the total bacterial isolates. All the MRSA isolates (100%) were resistant to cloxacillin and oxacillin, while all the isolates (100%) were sensitive to vancomycin and linezolid. Conclusions: The prevalence of MRSA in this study was high. It was most commonly seen in patients with vesiculobullous disorders. A high sensitivity of MRSA was found to amikacin. There was no relation between prolonged antibiotic therapy and MRSA in this study. PMID:25396125
Miyamoto, Tatsuro; Eguchi, Hiroshi; Tserennadmid, Ehkhmaa; Mitamura-Aizawa, Sayaka; Hotta, Fumika; Mitamura, Yoshinori
Purpose We report a case of methicillin-resistant Staphylococcus aureus (MRSA) keratitis after Descemet's stripping automated endothelial keratoplasty (DSAEK). Case Report An 87-year-old woman who had undergone a DSAEK 4 months previously was referred to Tokushima University Hospital with a diagnosis of infectious keratitis after DSAEK. A white abscess and infiltration in the inferior cornea of the right eye were observed. We started an empiric therapy using topical levofloxacin and chloramphenicol on the basis of the microscopic findings of the corneal scraping concurrently with cultivation of the cornea. Results A strain of MRSA was isolated from the corneal sample. Although the strain was susceptible to chloramphenicol, it was resistant to quinolone. The keratitis improved rapidly due to empiric therapy, and topical steroids could be resumed 6 days after initiation of the empiric therapy. Conclusions To our knowledge, this is the first case of MRSA keratitis, and the second case of bacterial keratitis, after DSAEK. MRSA keratitis can occur following uneventful DSAEK. The empiric therapy on the basis of results from a light microscopic examination of a Gram-stained corneal scraping and restarting topical steroids in the early stages of medication contributed to the good clinical course of this case. PMID:24348415
Joung, Dae-Ki; Mun, Su-Hyun; Choi, Sung-Hoon; Kang, Ok-Hwa; Kim, Sung-Bae; Lee, Young-Seob; Zhou, Tian; Kong, Ryong; Choi, Jang-Gi; Shin, Dong-Won; Kim, Youn-Chul; Lee, Dong-Sung; Kwon, Dong-Yeul
Oxyresveratrol (ORV) is a naturally occurring compound found in mulberries that exhibits a wide spectrum of biological activities. However, the underlying mechanism of the action of ORV against the methicillin-resistant S. aureus (MRSA) pathogen has not yet been reported. MRSA is multidrug-resistant, causing skin and other types of infections. The aim of the present study was to examine the antimicrobial activity of ORV and the underlying mechanism of its action on MRSA. The antibacterial activity of ORV was evaluated using a minimum inhibitory concentration (MIC) assay, and the mechanism of its antibacterial action on S. aureus was investigated using a combination of ORV with detergent, ATPase inhibitors and peptidoglycan (PGN). In addition, the survival characteristics and changes in MRSA morphology were monitored using transmission electron microscopy (TEM). The MIC value of ORV against all S. aureus strains was found to be 125 µg/ml. The optical density at 600 nm of each suspension treated using a combination of ORV with Triton X-100, N,N'-dicyclohexylcarbodiimide or sodium azide was reduced by 68.9–89.8% compared with the value upon treatment with ORV alone. In the ORV and PGN combination assay, direct binding of ORV with PGN from S. aureus was evident. Furthermore, TEM examination of MRSA treated with ORV showed alterations in septa formation. In conclusion, these results showed that ORV has a strong antibacterial effect against S. aureus, mainly by increasing membrane permeability and inhibiting ATPase when combined with other drugs. PMID:27588079
Chu, Ming; Zhang, Ming-bo; Liu, Yan-chen; Kang, Jia-rui; Chu, Zheng-yun; Yin, Kai-lin; Ding, Ling-yu; Ding, Ran; Xiao, Rong-xin; Yin, Yi-nan; Liu, Xiao-yan; Wang, Yue-dan
Berberine is an isoquinoline alkaloid widely used in the treatment of microbial infections. Recent studies have shown that berberine can enhance the inhibitory efficacy of antibiotics against clinical multi-drug resistant isolates of methicillin-resistant Staphylococcus aureus (MRSA). However, the underlying mechanisms are poorly understood. Here, we demonstrated that sub-minimum inhibitory concentrations (MICs) of berberine exhibited no bactericidal activity against MRSA, but affected MRSA biofilm development in a dose dependent manner within the concentration ranging from 1 to 64 μg/mL. Further study indicated that berberine inhibited MRSA amyloid fibrils formation, which consist of phenol-soluble modulins (PSMs). Molecular dynamics simulation revealed that berberine could bind with the phenyl ring of Phe19 in PSMα2 through hydrophobic interaction. Collectively, berberine can inhibit MRSA biofilm formation via affecting PSMs’ aggregation into amyloid fibrils, and thereby enhance bactericidal activity of antibiotics. These findings will provide new insights into the multiple pharmacological properties of berberine in the treatment of microbial-generated amyloid involved diseases. PMID:27103062
Cao, Huiliang; Qin, Hui; Zhao, Yaochao; Jin, Guodong; Lu, Tao; Meng, Fanhao; Zhang, Xianlong; Liu, Xuanyong
Since the use of systemic antibiotics for preventing acute biomaterial-associated infections (BAIs) may build up bacterial resistance and result in huge medical costs and unpredictable mortality, new precaution strategies are required. Here, it demonstrated that titanium armed with a nano-thick calcium oxide layer was effective on averting methicillin-resistant Staphylococcus aureus (MRSA) infections in rabbits. The calcium oxide layer was constructed by, firstly, injecting of metallic calcium into titanium via a plasma immersion ion implantation process, and then transforming the outer most surface into oxide by exposing to the atmosphere. Although the calcium oxide armed titanium had a relative low reduction rate (~74%) in growth of MRSA in vitro, it could markedly promote the osteogenic differentiation of bone marrow stem cells (BMSCs), restore local bone integration against the challenge of MRSA, and decrease the incidence of MRSA infection with a rate of 100% (compared to the titanium control). This study demonstrated for the first time that calcium, as one of the major elements in a human body, could be engineered to avert MRSA infections, which is promising as a safe precaution of disinfection for implantable biomedical devices. PMID:26899567
Wang, Zhaofei; Zheng, Panpan; Ji, Wenhui; Fu, Qiang; Wang, Hengan; Yan, Yaxian; Sun, Jianhe
Staphylococcus aureus (S. aureus) is a Gram-positive pathogen causing a variety of infections in humans and animals. Extensive use of antibiotics has led to the emergence of methicillin-resistant S. aureus (MRSA). As an alternative antibacterial agent against drug-resistant S. aureus, a lytic phage, designated SLPW, was isolated from fecal sewage in a pig farm. The SLPW was morphologically classified under Podoviridae and contains a double-stranded DNA genome. The genome of SLPW was 17,861 bp (29.35% G+C) containing 20 open reading frames and lacked regions encoding lysogeny-related integrase gene and cI repressor gene. Phage SLPW showed a broad host range and high efficiency of plating against various types of S. aureus. One-step growth curve showed a short latency period (10 min) and a long lytic period (120 min). Phage SLPW remained stable under a wide range of temperatures or pH and was almost unaffected in chloroform or ultraviolet light. Further, it efficiently lysed MRSA strains in vitro and in vivo. Intraperitoneal phage administration at 1 h post-infection cured the mice and reduced the bacterial expression of inflammatory cytokines in mice. Specifically, the phage SLPW displayed a wide antibacterial spectrum. It was therapeutically effective against intra-abdominal infection in mice harboring different multilocus sequence typing (MLST) types of S. aureus strains. Therefore, phage SLPW is a potential therapeutic agent against MRSA infections. PMID:27379064
Deguchi, K; Yokota, N; Koguchi, M; Suzuki, Y; Suzuki, K; Fukayama, S; Ishihara, R; Oda, S
Efficacies of combined uses of arbekacin(ABK) with other antibiotics against between 57 and 76 clinically isolated methicillin-resistant Staphylococcus aureus(MRSA) were examined. Other antibiotics tested were fosfomycin(FOM), 5 cephems(CEPs), 2 penicillins(PCs) and imipenem. Percentages of strains that grew at various combination of serially diluted drugs in a microdilution method were scored, and for each pair of drugs > or = MIC50 or > or = MIC90 was determined. FIC-indices were calculated at > or = MIC50. Bacterial growth was inhibited by low concentrations of ABK dose-dependently, while most of the strains tested were resistant to 9 other drugs. FIC-indices with ABK were lowest when used together with ampicillin (ABPC) (0.516) followed by combinations with cefotiam (CTM) and cefuzonam (CZON) (each, 0.53), and FOM (0.625). When combined with 0.5 microgram/ml of ABK, 0.5 microgram/ml of ABPC inhibited more than 50% of MRSA strains. CTM or CZON in combination with the same amount of ABK to 2 micrograms/ml inhibited more than 50% of MRSA. Thus ABPC was found to be the most efficient in combined use with ABK to inhibit clinical MRSA strains.
Cao, Huiliang; Qin, Hui; Zhao, Yaochao; Jin, Guodong; Lu, Tao; Meng, Fanhao; Zhang, Xianlong; Liu, Xuanyong
Since the use of systemic antibiotics for preventing acute biomaterial-associated infections (BAIs) may build up bacterial resistance and result in huge medical costs and unpredictable mortality, new precaution strategies are required. Here, it demonstrated that titanium armed with a nano-thick calcium oxide layer was effective on averting methicillin-resistant Staphylococcus aureus (MRSA) infections in rabbits. The calcium oxide layer was constructed by, firstly, injecting of metallic calcium into titanium via a plasma immersion ion implantation process, and then transforming the outer most surface into oxide by exposing to the atmosphere. Although the calcium oxide armed titanium had a relative low reduction rate (~74%) in growth of MRSA in vitro, it could markedly promote the osteogenic differentiation of bone marrow stem cells (BMSCs), restore local bone integration against the challenge of MRSA, and decrease the incidence of MRSA infection with a rate of 100% (compared to the titanium control). This study demonstrated for the first time that calcium, as one of the major elements in a human body, could be engineered to avert MRSA infections, which is promising as a safe precaution of disinfection for implantable biomedical devices.
Weickert, Martin O.
Insulin resistance has been proposed as the strongest single predictor for the development of Type 2 Diabetes (T2DM). Chronic oversupply of energy from food, together with inadequate physical activity, have been recognized as the most relevant factors leading to overweight, abdominal adiposity, insulin resistance, and finally T2DM. Conversely, energy reduced diets almost invariably to facilitate weight loss and reduce abdominal fat mass and insulin resistance. However, sustained weight loss is generally difficult to achieve, and distinct metabolic characteristics in patients with T2DM further compromise success. Therefore, investigating the effects of modulating the macronutrient composition of isoenergetic diets is an interesting concept that may lead to additional important insights. Metabolic effects of various different dietary concepts and strategies have been claimed, but results from randomized controlled studies and particularly from longer-term-controlled interventions in humans are often lacking. However, some of these concepts are supported by recent research, at least in animal models and short-term studies in humans. This paper provides an update of the current literature regarding the role of nutrition in the modulation of insulin resistance, which includes the discussion of weight-loss-independent metabolic effects of commonly used dietary concepts. PMID:24278690
Olischar, Monika; Hunt, Rod W; Daley, Andrew J; Clifford, Vanessa; Tingay, David G
Methicillin-resistant Staphylococcus aureus (MRSA) can cause serious infection among hospitalised patients. The emergence of community acquired strains of MRSA (CA-MRSA) increases the potential exposure of newborns. The high incidence of reported meningitis in neonates in large CA-MRSA outbreaks suggests increased virulence in the neonatal population. The authors describe a case of severe meningitis due to MRSA in a previously healthy neonate and include findings from amplitude-integrated electroencephalography (aEEG) and MRI. PMID:22802366
Salgueiro, Vivian Carolina; Azevedo, Milena Borgo; Iorio, Natalia Lopes Pontes; Amorim, Efigênia de Lourdes Teixeira; dos Santos, Kátia Regina Netto
To characterize 46 methicillin-resistant coagulase-negative staphylococci from Brazilian neonates, we investigated their SCCmec type, susceptibility and clonality. Staphylococcus epidermidis and Staphylococcus haemolyticus were the prevalent species. SCCmec types III or IV were detected in 53.3% S. epidermidis, whereas 63.6% S. haemolyticus were nontypeable. Despite the diversity, specific clones carried specific SCCmec elements, highlighting that effective typing can help in epidemiological analysis.
Verhulst, Carlo; Nonhoff, Claire; Nagant, Carole; Denis, Olivier; Kluytmans, Jan
Three chromogenic media, chromID MRSA SMART (SMART), chromID MRSA first generation (chromID), and Brilliance MRSA (OX2), were evaluated for methicillin-resistant Staphylococcus aureus (MRSA) screening using 1,220 samples. The sensitivity at 24 h was significantly better with the SMART agar (66.4%) than that with chromID agar (50.5%). Enrichment and incubation until 48 h are still needed for an optimal yield. PMID:26109446
Pape, John; Wadlin, Jill; Nachamkin, Irving
We evaluated the ability of BBL CHROMagar MRSA medium (Becton Dickinson, Sparks, MD) to identify methicillin-resistant Staphylococcus aureus (MRSA) directly upon subculture from positive blood culture bottles. There were 124 MRSA isolates recovered from blood cultures in the study. BBL CHROMagar MRSA medium was highly sensitive (97.6% [121/124] at 18 to 24 h of incubation and 100% [124/124] at 48 h) and 99.9% specific for identifying MRSA from positive blood cultures. PMID:16825383
Stefańska, Joanna; Antoszczak, Michał; Stępień, Karolina; Bartoszcze, Michał; Mirski, Tomasz; Huczyński, Adam
For the first time, a series of tertiary amides of polyether antibiotic-Salinomycin have been obtained and screened for their antibacterial activity against different strains of bacteria, including Bacillus anthracis and clinical methicillin-resistant Staphylococcus epidermidis (MRSE). Moreover, biofilm inhibition of MRSE and genotoxicity tests against Bacillus subtilis have been performed. Our studies show that Salinomycin and its some derivatives are active against tested bacteria and exhibited definitely bacteriostatic, not bactericidal activity.
Moreno, Luisa Z; Dutra, Mauricio C; Moreno, Marina; Ferreira, Thais SP; da Silva, Givago FR; Matajira, Carlos EC; Silva, Ana Paula S; Moreno, Andrea M
Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has been mainly related with pig farming, in Europe and North America, with the ST398 as the most commonly identified type of LA-MRSA. Here we present the draft genome of the first vancomycin-intermediate MRSA ST398/t9538 isolated from a swine presenting exudative epidermitis in Brazil. PMID:27759766
Bardiau, Marjorie; Yamazaki, Kazuko; Ote, Isabelle; Misawa, Naoaki; Mainil, Jacques G
The aim of this study was to explore the presence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in a collection of S. pseudintermedius strains isolated from dogs and cats with dermatitis in Japan and to compare their genotypic and phenotypic characteristics. Clonal relationships were determined by pulse field gel electrophoresis (PFGE), staphylococcal chromosomal cassette mec (SCCmec) typing, and multilocus sequence typing (MLST). Biofilm formation assay was performed using safranin staining in microplates. Three virulence genes coding for S. intermedius exfoliative toxin and Panton-Valentine leukocidin (siet, lukS-PV and lukF-PV) were searched for in a collection of strains. Antimicrobial resistance against 15 antibiotics was studied by a disc diffusion method. Twenty-seven MRSP were isolated. According to PFGE results the isolates were not closely related except for a few strains. MLST showed that the strains belonged to five groups, ST71 and ST26 being the two most prevalent. Three types of SCCmec (II, II-III and V) were identified. All isolates were siet-positive but PVL-negative. Most strains (except for two) produced strong biofilm in tryptic soy broth with glucose. Seventy-eight percent of the isolates were resistant or intermediate to twelve or more antibiotics. Our study demonstrates that the ST71 lineage is widespread in Japan and that ST26 could represent an emerging lineage. Moreover, most of our strains are capable of forming strong biofilm and possess siet gene, two virulence characteristics that probably help the bacteria to persist and spread. Finally, our MRSP strains show a strong resistance profile to antibiotics commonly used in veterinary medicine.
Loncaric, Igor; Künzel, Frank; Licka, Theresia; Simhofer, Hubert; Spergser, Joachim; Rosengarten, Renate
The aim of this study was to investigate the antimicrobial resistance, resistance gene patterns and genetic relatedness of a collection of Austrian methicillin-resistant Staphylococcus aureus (MRSA) isolates from companion animals and horses. A total of 89 non-repetitive MRSA isolates collected during routine veterinary microbiological examinations from April 2004 to the end of 2012, and one isolate from 2013 were used for this study. The presence of mecA and other resistance genes was confirmed by PCR. Isolates were genotyped by spa typing, two multiple-locus variable-number tandem repeat analyses (MLVA) analyses, SCCmec typing and multilocus sequence typing (MLST). PCR targeting Panton-Valentine leukocidin (PVL) and detection of staphylococcal enterotoxins (SE), toxic shock syndrome toxin (TSST) was performed using PCR assays. Antimicrobial susceptibility testing was performed. Five sequence types (STs-ST398, ST254, ST22, ST5 and ST1), SCCmec types II, IVa, V, and non-type-abele, 8 spa-types (t003, t011, t036, t127, t386, t1348, and t4450), and two isolates could not be assigned, 21 MLVA-14Orsay types Multiplex-PCR MLVA (mMLVA) displayed 17 different MLVA types. The present study is the most comprehensive dealing with MRSA from Austrian companion animals and horses. The results confirm that MRSA ST398 is present in a wide range of animal species and is predominant especially in horses. In other companion animals it is unclear whether the infections with the different MRSA isolates investigated in the present study truly represents a rare phenomenon or may be an emerging problem in companion animals.
Ekambaram, Sanmuga Priya; Perumal, Senthamil Selvan; Balakrishnan, Ajay; Marappan, Nathiya; Gajendran, Sabari Srinivasan; Viswanathan, Vinodhini
Aim/Background: Medicinal plants have ability to resist microorganisms by synthesizing secondary metabolites such as phenols. Rosmarinic acid (RA) is a phenylpropanoid widely distributed in plants and well known as therapeutic and cosmetic agent. Methicillin-resistant Staphylococcus aureus (MRSA) which is resistant to all kinds of β-lactams, threatens even most potent antibiotics. To improve the efficiency of antibiotics against multi-drug resistant bacteria and to reduce the antibiotic dose, the antibacterial activity and the synergistic effect of RA with standard antibiotics against S. aureus and MRSA was investigated. Materials and Methods: Antibacterial activity of RA against S. aureus and a clinical isolate of MRSA was evaluated by agar well diffusion method. Minimum inhibitory concentration (MIC) of RA was determined by broth dilution method. Synergism of RA with various antibiotics against S. aureus and MRSA was studied by broth checkerboard method and time-kill kinetic assay. Effect of RA on microbial surface components recognizing adhesive matrix molecules (MSCRAMM’s) of S. aureus and MRSA was studied using sodium dodecyl sulfate - polyacrylamide gel electrophoresis. Results: MIC of RA was found to be 0.8 and 10 mg/ml against S. aureus and MRSA, respectively. RA was synergistic with vancomycin, ofloxacin, and amoxicillin against S. aureus and only with vancomycin against MRSA. The time-kill analysis revealed that synergistic combinations were a more effective than individual antibiotics. MSCRAMM’s protein expression of S. aureus and MRSA was markedly suppressed by RA + vancomycin combination rather than RA alone. Conclusion: The synergistic effects of RA with antibiotics were observed against S. aureus and MRSA. RA showed inhibitory effect on the surface proteins MSCRAMM’s. Even though RA was shown to exhibit a synergistic effect with antibiotics, the MIC was found to be higher. Thus, further studies on increasing the efficacy of RA can develop it
Oliveira, C J B; Tiao, N; de Sousa, F G C; de Moura, J F P; Santos Filho, L; Gebreyes, W A
The aim of this study was to investigate the phenotypic and genotypic diversity and anti-microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north-eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti-microbial susceptibility by Kirby-Bauer disc diffusion method. Confirmation of methicillin-resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP-2a latex agglutination. Clonal relatedness of isolates was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase-negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622-ST1625). The findings show that MRSA is prevalent in milk from semi-extensive dairy cows in north-eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm-to-table continuum is warranted.
Zoraghi, Roya; See, Raymond H; Axerio-Cilies, Peter; Kumar, Nag S; Gong, Huansheng; Moreau, Anne; Hsing, Michael; Kaur, Sukhbir; Swayze, Richard D; Worrall, Liam; Amandoron, Emily; Lian, Tian; Jackson, Linda; Jiang, Jihong; Thorson, Lisa; Labriere, Christophe; Foster, Leonard; Brunham, Robert C; McMaster, William R; Finlay, B Brett; Strynadka, Natalie C; Cherkasov, Artem; Young, Robert N; Reiner, Neil E
Novel classes of antimicrobials are needed to address the challenge of multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA). Using the architecture of the MRSA interactome, we identified pyruvate kinase (PK) as a potential novel drug target based upon it being a highly connected, essential hub in the MRSA interactome. Structural modeling, including X-ray crystallography, revealed discrete features of PK in MRSA, which appeared suitable for the selective targeting of the bacterial enzyme. In silico library screening combined with functional enzymatic assays identified an acyl hydrazone-based compound (IS-130) as a potent MRSA PK inhibitor (50% inhibitory concentration [IC50] of 0.1 μM) with >1,000-fold selectivity over human PK isoforms. Medicinal chemistry around the IS-130 scaffold identified analogs that more potently and selectively inhibited MRSA PK enzymatic activity and S. aureus growth in vitro (MIC of 1 to 5 μg/ml). These novel anti-PK compounds were found to possess antistaphylococcal activity, including both MRSA and multidrug-resistant S. aureus (MDRSA) strains. These compounds also exhibited exceptional antibacterial activities against other Gram-positive genera, including enterococci and streptococci. PK lead compounds were found to be noncompetitive inhibitors and were bactericidal. In addition, mutants with significant increases in MICs were not isolated after 25 bacterial passages in culture, indicating that resistance may be slow to emerge. These findings validate the principles of network science as a powerful approach to identify novel antibacterial drug targets. They also provide a proof of principle, based upon PK in MRSA, for a research platform aimed at discovering and optimizing selective inhibitors of novel bacterial targets where human orthologs exist, as leads for anti-infective drug development.
Bæk, Kristoffer T.; Gründling, Angelika; Mogensen, René G.; Thøgersen, Louise; Petersen, Andreas; Paulander, Wilhelm
Methicillin-resistant Staphylococcus aureus (MRSA) has acquired the mecA gene encoding a peptidoglycan transpeptidase, penicillin binding protein 2a (PBP2a), which has decreased affinity for β-lactams. Quickly spreading and highly virulent community-acquired (CA) MRSA strains recently emerged as a frequent cause of infection in individuals without exposure to the health care system. In this study, we found that the inactivation of the components of the ClpXP protease substantially increased the β-lactam resistance level of a CA-MRSA USA300 strain, suggesting that the proteolytic activity of ClpXP controls one or more pathways modulating β-lactam resistance. These pathways do not involve the control of mecA expression, as the cellular levels of PBP2a were unaltered in the clp mutants. An analysis of the cell envelope properties of the clpX and clpP mutants revealed a number of distinct phenotypes that may contribute to the enhanced β-lactam tolerance. Both mutants displayed significantly thicker cell walls, increased peptidoglycan cross-linking, and altered composition of monomeric muropeptide species compared to those of the wild types. Moreover, changes in Sle1-mediated peptidoglycan hydrolysis and altered processing of the major autolysin Atl were observed in the clp mutants. In conclusion, the results presented here point to an important role for the ClpXP protease in controlling cell wall metabolism and add novel insights into the molecular factors that determine strain-dependent β-lactam resistance. PMID:24867990
Pohlentz, Gottfried; Xia, Guoqing; Hussain, Muzaffar; Foster, Simon; Peters, Georg
Most bacterial glycoproteins identified to date are virulence factors of pathogenic bacteria, i.e. adhesins and invasins. However, the impact of protein glycosylation on the major human pathogen Staphylococcus aureus remains incompletely understood. To study protein glycosylation in staphylococci, we analyzed lysostaphin lysates of methicillin-resistant Staphylococcus aureus (MRSA) strains by SDS-PAGE and subsequent periodic acid-Schiff’s staining. We detected four (>300, ∼250, ∼165, and ∼120 kDa) and two (>300 and ∼175 kDa) glycosylated surface proteins with strain COL and strain 1061, respectively. The ∼250, ∼165, and ∼175 kDa proteins were identified as plasmin-sensitive protein (Pls) by mass spectrometry. Previously, Pls has been demonstrated to be a virulence factor in a mouse septic arthritis model. The pls gene is encoded by the staphylococcal cassette chromosome (SCC)mec type I in MRSA that also encodes the methicillin resistance-conferring mecA and further genes. In a search for glycosyltransferases, we identified two open reading frames encoded downstream of pls on the SCCmec element, which we termed gtfC and gtfD. Expression and deletion analysis revealed that both gtfC and gtfD mediate glycosylation of Pls. Additionally, the recently reported glycosyltransferases SdgA and SdgB are involved in Pls glycosylation. Glycosylation occurs at serine residues in the Pls SD-repeat region and modifying carbohydrates are N-acetylhexosaminyl residues. Functional characterization revealed that Pls can confer increased biofilm formation, which seems to involve two distinct mechanisms. The first mechanism depends on glycosylation of the SD-repeat region by GtfC/GtfD and probably also involves eDNA, while the second seems to be independent of glycosylation as well as eDNA and may involve the centrally located G5 domains. Other previously known Pls properties are not related to the sugar modifications. In conclusion, Pls is a glycoprotein and Pls glycosyl
Soloaga, R; Corso, A; Gagetti, P; Faccone, D; Galas, M
Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen that has emerged over the last four decades, causing both nosocomial and community-acquired infections. Rapid and accurate detection of methicillin resistance in S. aureus is important for the use of appropriate antimicrobial therapy and for the control of nosocomial spread of MRSA strains. We evaluated the efficiency of conventional methods for detection of methicillin resistance such as the disk diffusion, agar dilution, oxacillin agar screen test, and the latex agglutination test MRSA-Screen latex, in 100 isolates of S. aureus, 79 mecA positive and 21 mecA negative. The MRSA-Screen latex (Denka Seiken, Niigata, Japón), is a latex agglutination method that detects the presence of PLP-2a, product of mecA gene in S. aureus. The PCR of the mecA gene was used as the "gold standard" for the evaluation of the different methods tested. The percentages of sensitivity and specificity were as follows: disk difusión 97 and 100%, agar dilution 97 and 95%, oxacillin agar screen test 100 and 100%, and MRSA-Screen latex, 100 and 100 %. All methods presented high sensitivity and specificity, but MRSA-Screen latex had the advantage of giving a reliable result, equivalent to PCR, in only 15 minutes.
Zouhir, Abdelmajid; Taieb, Malek; Lamine, Mohamed Ashraf; Cherif, Ammar; Jridi, Taoufik; Mahjoubi, Basma; Mbarek, Sarra; Fliss, Ismail; Nefzi, Adel; Sebei, Khaled; Ben Hamida, Jeannette
Staphylococcus aureus and methicillin-resistant S. aureus are major pathogens. The antimicrobial peptides and essential oils (EOs) display narrow- or broad-spectrum activity against bacteria including these strains. A centralized resource, such as a database, designed specifically for anti-S. aureus/anti-methicillin-resistant S. aureus antimicrobial peptides and EOs is therefore needed to facilitate the comprehensive investigation of their structure/activity associations and combinations. The database ANTISTAPHYBASE is created to facilitate access to important information on antimicrobial peptides and essential peptides against methicillin-resistant S. aureus and S. aureus. At the moment, the database contains 596 sequences of antimicrobial peptides produced by diverse organisms and 287 essential oil records. It permits a quick and easy search of peptides based on their activity as well as their general, physicochemical properties and literature data. These data are very useful to perform further bioinformatic or chemometric analysis and would certainly be useful for the development of new drugs for medical use. The ANTISTAPHYBASE database is freely available at: https://www.antistaphybase.com/ .
Abdel-moein, K A; El-Hariri, M; Samir, A
Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) has emerged to be a pathogen of public health burden causing infections with significant concern. This study was conducted to investigate methicillin-resistant Staphylococcus aureus (MRSA) infection in pet dogs and cats as an emerging zoonosis that could be disseminated in the community. A total of 184 (nasal, oral, ear and wound) swabs were collected from 70 pet dogs and 48 pet cats, whereas 50 nasal and oral swabs were collected from 28 apparently healthy companion persons in intimate contact with pets and without history of hospitalization. All samples were cultured for the isolation and identification of Staphylococcus aureus using selective media, biochemical and serological tests, while isolates were identified as MRSA after antimicrobial susceptibility testing and determination of the MIC. PCR was applied using specific primers to confirm MRSA. Three MRSA isolates have been recovered from two dogs of 70 (2.9%) and one isolate from 28 examined persons (3.6%), while none of the examined cats yielded MRSA. Furthermore, we found that two MRSA isolates recovered from one diseased dog seemed to be hospital-associated methicillin-resistant Staphylococcus aureus (HA-MRSA), whereas the other dog isolate as well as the human isolate were considered as community-acquired (CA-MRSA). The occurrence of MRSA in apparently healthy and/or diseased pet dogs makes it an emerging veterinary pathogen which could be considered a public health burden if it is disseminated in our community outside hospitals.
Djoudi, Ferhat; Bonura, Celestino; Touati, Abdelaziz; Aléo, Aurora; Benallaoua, Said; Mammina, Caterina
Genetic mechanisms of methicillin resistance are still relevant in staphylococci. The aims of this study are to assess the possible exchanges of staphylococcal cassette chromosome mec (SCCmec) among isolates of methicillin-resistant staphylococci (MRS) and to check for known or new mutations in mecA DNA. A total of 35 MRS non-repetitive isolates were recovered, including 20 Staphylococcushaemolyticus, 7 Staphylococcusaureus, 4 Staphylococcussciuri, 2 Staphylococcussaprophyticus and 1 isolate each of Staphylococcusxylosus and Staphylococcuslentus. Only 16 of the 35 strains were assigned to known SCCmec types: 7 SCCmec VII, 6 SCCmec IV and 3 SCCmec III, with possible horizontal transfer of the SCCmec VII from methicillin-resistant S. haemolyticus to methicillin-susceptible S. aureus. mecA gene sequencing in ten selected isolates allowed description of nine punctual mutations, seven of which were reported for the first time. The most frequent mutation was G246E, identified in isolates of methicillin-resistant S. aureus, S. sciuri, S. saprophyticus and S. lentus. These results emphasized the high degree of genetic diversity of SCCmec element in MRS and describe new missense mutations in mecA, which might be important in understanding the evolution of methicillin and new β-lactam resistance.
Kim, Eu Suk; Kim, Hong Bin; Kim, Gayeon; Kim, Kye-Hyung; Park, Kyung-Hwa; Lee, Shinwon; Choi, Young Hwa; Yi, Jongyoun; Kim, Chung Jong; Song, Kyoung-Ho; Choe, Pyoeng Gyun; Kim, Nam-Joong; Lee, Yeong-Seon; Oh, Myoung-Don
Successful empirical therapy of Staphylococcus aureus infections requires the ability to predict methicillin resistance. Our aim was to identify predictors of methicillin resistance in community-onset (CO) invasive S. aureus infections. Sixteen hospitals across Korea participated in this study from May to December 2012. We prospectively included cases of S. aureus infection in which S. aureus was isolated from sterile clinical specimens ≤ 72 hours after hospitalization. Clinical and epidemiological data were gathered and compared in methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) cases. Community-associated (CA) infections were defined as in previous studies. In total, there were 786 cases of community-onset S. aureus infection, 102 (13.0%) of which were CA-MRSA. In addition to known risk factors, exposure to 3rd generation cephalosporins in the past 6 months [odds ratio (OR), 1.922; 95% confidence interval (CI), 1.176-3.142] and close contact with chronically ill patients in the past month (OR, 2.647; 95% CI, 1.189-5.891) were independent risk factors for MRSA infection. However, no clinical predictors of CA-MRSA were identified. Methicillin resistance, CO infection, and appropriateness of empirical antibiotics were not significantly related to 30-day mortality. MRSA infection should be suspected in patients recently exposed to 3rd generation cephalosporins or chronically-ill patients. There were no reliable predictors of CA-MRSA infection, and mortality was not affected by methicillin resistance.
Cui, Lanqing; Wang, Yang; Li, Yun; He, Tao; Schwarz, Stefan; Ding, Yujing; Shen, Jianzhong; Lv, Yuan
Four methicillin-resistant coagulase-negative staphylococci (MRCoNS), one Staphylococcus haemolyticus and three Staphylococcus cohnii, from infections of humans collected via the Ministry of Health National Antimicrobial Resistance Surveillance Net (Mohnarin) program in China were identified as linezolid-resistant. These four isolates were negative for the 23S rRNA mutations, but positive for the gene cfr. Mutations in the gene for the ribosomal protein L3, which resulted in the amino acid exchanges Gly152Asp and Tyr158Phe, were identified in S. haemolyticus 09D279 and S. cohnii NDM113, respectively. In each isolate, the cfr gene was located on a plasmid of ca. 35.4 kb, as shown by S1 nuclease pulsed-field gel electrophoresis and Southern blotting experiments. This plasmid was indistinguishable from the previously described plasmid pSS-02 by its size, restriction pattern, and a sequenced 14-kb cfr-carrying segment. Plasmid pSS-02 was originally identified in staphylococci isolated from pigs. This is the first time that a cfr-carrying plasmid has been detected in MRCoNS obtained from intensive care patients in China. Based on the similarities to the cfr-carrying plasmid pSS-02 from porcine coagulase-negative staphylococci, a transmission of this cfr-carrying plasmid between staphylococci from pigs and humans appears to be likely.
Zhao, Zhiwei; Yan, Rong; Yi, Xuan; Li, Jingling; Rao, Jiaming; Guo, Zhengqing; Yang, Yanmei; Li, Weifeng; Li, Yong-Qiang; Chen, Chunying
Despite numerous advanced imaging and sterilization techniques available nowadays, the sensitive in vivo diagnosis and complete elimination of drug-resistant bacterial infections remain big challenges. Here we report a strategy to design activatable theranostic nanoprobes against methicillin-resistant Staphylococcus aureus (MRSA) infections. This probe is based on silica nanoparticles coated with vancomycin-modified polyelectrolyte-cypate complexes (SiO2-Cy-Van), which is activated by an interesting phenomenon of bacteria-responsive dissociation of the polyelectrolyte from silica nanoparticles. Due to the aggregation of hydrophobic cypate fluorophores on silica nanoparticles to induce ground-state quenching, the SiO2-Cy-Van nanoprobes are nonfluorescent in aqueous environments. We demonstrate that MRSA can effectively pull out the vancomycin-modified polyelectrolyte-cypate complexes from silica nanoparticles and draw them onto their own surface, changing the state of cypate from off (aggregation) to on (disaggregation) and leading to in vitro MRSA-activated near-infrared fluorescence (NIRF) and photothermal elimination involving bacterial cell wall and membrane disruption. In vivo experiments show that this de novo-designed nanoprobe can selectively enable rapid (4 h postinjection) NIRF imaging with high sensitivity (10(5) colony-forming units) and efficient photothermal therapy (PTT) of MRSA infections in mice. Remarkably, the SiO2-Cy-Van nanoprobes can also afford a long-term tracking (16 days) of the development of MRSA infections, allowing real-time estimation of bacterial load in infected tissues and further providing a possible way to monitor the efficacy of antimicrobial treatment. The strategy of bacteria-activated polyelectrolyte dissociation from nanoparticles proposed in this work could also be used as a general method for the design and fabrication of bacteria-responsive functional nanomaterials that offer possibilities to combat drug-resistant
Cázares-Domínguez, Vicenta; Cruz-Córdova, Ariadnna; Ochoa, Sara A.; Escalona, Gerardo; Arellano-Galindo, José; Rodríguez-Leviz, Alejandra; Hernández-Castro, Rigoberto; López-Villegas, Edgar O.; Xicohtencatl-Cortes, Juan
Methicillin-resistant Staphylococcus aureus (MRSA) is an important opportunistic pathogen that causes both healthcare- and community-acquired infections. An increase in the incidence of these infections may lead to a substantial change in the rate of vancomycin usage. Incidence of reduced susceptibility to vancomycin has been increasing worldwide for the last few years, conferring different levels of resistance to vancomycin as well as producing changes in the cell wall structure. The aim of the present study was to determine the effect of vancomycin on cell wall thickening in clinical isolates of vancomycin-tolerant (VT) MRSA obtained from pediatric patients. From a collection of 100 MRSA clinical isolates from pediatric patients, 12% (12/100) were characterized as VT-MRSA, and from them, 41.66% (5/12) exhibited the heterogeneous vancomycin-intermediate S. aureus (hVISA) phenotype. Multiplex-PCR assays revealed 66.66% (8/12), 25% (3/12), and 8.33% (1/12) of the VT-MRSA isolates were associated with agr group II, I, and III polymorphisms, respectively; the II-mec gene was amplified from 83.3% (10/12) of the isolates, and the mecIVa gene was amplified from 16.66% (2/12) of the isolates. Pulsed field electrophoresis (PFGE) fingerprint analysis showed 62% similarity among the VT-MRSA isolates. Thin transverse sections analyzed by transmission electron microscopy (TEM) revealed an average increase of 24 nm (105.55%) in the cell wall thickness of VT-MRSA compared with untreated VT-MRSA isolates. In summary, these data revealed that the thickened cell walls of VT-MRSA clinical isolates with agr type II and SCCmec group II polymorphisms are associated with an adaptive resistance to vancomycin. PMID:25793280
Liu, Xiaowei; Li, Zhan; Wang, Xiaodong; Chen, Yujuan; Wu, Fengbo; Men, Ke; Xu, Ting; Luo, Yan; Yang, Li
Infections caused by multidrug-resistant bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA), have become a public threat; therefore, development of new antimicrobial drugs or strategies is urgently required. In this study, a new antibacterial peptide DP7-C (Chol-suc-VQWRIRVAVIRK-NH2) and DP7-C-modified azithromycin (AZT)-loaded liposomes (LPs) are developed for the treatment of MRSA infection, and it was found that DP7-C inserted into the LP lipid bilayer not only functioned as a carrier to encapsulate the antibiotic AZT but also synergized the antibacterial effect of the encapsulated AZT. In vitro assays showed that DP7-C-modified LPs possessed sustained drug release profile and immune regulatory effect and did not show obvious cytotoxicity in mammal cells, but they did not possess direct antibacterial activity in vitro. In vivo studies revealed that DP7-C-modified LPs did not exhibit obvious side effects or toxicity in mice but were able to significantly reduce the bacterial counts in an MRSA-infectious mouse model and possessed high antibacterial activity. In particular, DP7-C-modified AZT-loaded LPs showed more positive therapeutic effects than either DP7-C-modified blank LPs or nonmodified AZT-loaded LPs treatment alone. Molecular mechanism studies demonstrated that DP7-C formulations effectively upregulated the production of anti-inflammatory cytokines and chemokines without inducing harmful immune response, suggesting that DP7-C was synergistic with AZT against the bacterial infection by activating the innate immune response. Most importantly, although DP7-C activated the innate immune response, it did not possess direct antibacterial activity in vitro, indicating that DP7-C did not possess the potential to induce bacteria resistance. The findings indicate that DP7-C-modified AZT-loaded LPs developed in this study have a great potential required for the clinical treatment of MRSA infections. PMID:28008253
Cázares-Domínguez, Vicenta; Cruz-Córdova, Ariadnna; Ochoa, Sara A; Escalona, Gerardo; Arellano-Galindo, José; Rodríguez-Leviz, Alejandra; Hernández-Castro, Rigoberto; López-Villegas, Edgar O; Xicohtencatl-Cortes, Juan
Methicillin-resistant Staphylococcus aureus (MRSA) is an important opportunistic pathogen that causes both healthcare- and community-acquired infections. An increase in the incidence of these infections may lead to a substantial change in the rate of vancomycin usage. Incidence of reduced susceptibility to vancomycin has been increasing worldwide for the last few years, conferring different levels of resistance to vancomycin as well as producing changes in the cell wall structure. The aim of the present study was to determine the effect of vancomycin on cell wall thickening in clinical isolates of vancomycin-tolerant (VT) MRSA obtained from pediatric patients. From a collection of 100 MRSA clinical isolates from pediatric patients, 12% (12/100) were characterized as VT-MRSA, and from them, 41.66% (5/12) exhibited the heterogeneous vancomycin-intermediate S. aureus (hVISA) phenotype. Multiplex-PCR assays revealed 66.66% (8/12), 25% (3/12), and 8.33% (1/12) of the VT-MRSA isolates were associated with agr group II, I, and III polymorphisms, respectively; the II-mec gene was amplified from 83.3% (10/12) of the isolates, and the mecIVa gene was amplified from 16.66% (2/12) of the isolates. Pulsed field electrophoresis (PFGE) fingerprint analysis showed 62% similarity among the VT-MRSA isolates. Thin transverse sections analyzed by transmission electron microscopy (TEM) revealed an average increase of 24 nm (105.55%) in the cell wall thickness of VT-MRSA compared with untreated VT-MRSA isolates. In summary, these data revealed that the thickened cell walls of VT-MRSA clinical isolates with agr type II and SCCmec group II polymorphisms are associated with an adaptive resistance to vancomycin.
Wang, Xiaoguang; Ouyang, Lin; Luo, Lingfei; Liu, Jiqian; Song, Chiping; Li, Cuizhen; Yan, Hongjing; Wang, Ping
Methicillin-resistant Staphylococcus aureus (MRSA) strains are now common both in the health care setting and in the community. Active surveillance is critical for MRSA control and prevention. Specimens of patients (200 patients with 1119 specimens) as well as medical staff and hospital setting (1000 specimens) were randomly sampled in a level 2 hospital in Shanghai from September 2011 to August 2012. Isolation, cultivation and identification of S. aureus were performed. Totally, 67 S. aureus strains were isolated. 32 S. aureus strains were isolated from patient samples; 13 (13/32, 40.6%) of the 32 S. aureus isolates were MRSA; sputum sample and patients in the department of general internal medicine were the most frequent specimen and patient group for S. aureus strains isolation. Remaining 35 S. aureus strains were isolated from the medical staff and hospital setting; 20 (20/35, 57.1%) of the 35 S. aureus isolates were MRSA; specimens sampled from doctors and nurses’ hands and nose and hospital facilities were the most frequent samples to isolate S. aureus. Resistant and virulent genes detection showed that, all 33 MRSA strains were mecA positive which accounts for 49.3% of the 67 S. aureus strains; 38 isolates were Panton-Valentine leukocidin (PVL) gene positive which accounts for 56.7% of the 67 S. aureus strains; and 17 (17/67, 25.4%) isolates are mecA and PVL genes dual positive. Multidrug-resistant strains of MRSA and PVL positive S. aureus are common in patients, medical staff and hospital setting, the potential health threat is worthy of our attention. PMID:28030828
Wang, Xiaoguang; Ouyang, Lin; Luo, Lingfei; Liu, Jiqian; Song, Chiping; Li, Cuizhen; Yan, Hongjing; Wang, Ping
Methicillin-resistant Staphylococcus aureus (MRSA) strains are now common both in the health care setting and in the community. Active surveillance is critical for MRSA control and prevention. Specimens of patients (200 patients with 1119 specimens) as well as medical staff and hospital setting (1000 specimens) were randomly sampled in a level 2 hospital in Shanghai from September 2011 to August 2012. Isolation, cultivation and identification of S. aureus were performed. Totally, 67 S. aureus strains were isolated. 32 S. aureus strains were isolated from patient samples; 13 (13/32, 40.6%) of the 32 S. aureus isolates were MRSA; sputum sample and patients in the department of general internal medicine were the most frequent specimen and patient group for S. aureus strains isolation. Remaining 35 S. aureus strains were isolated from the medical staff and hospital setting; 20 (20/35, 57.1%) of the 35 S. aureus isolates were MRSA; specimens sampled from doctors and nurses' hands and nose and hospital facilities were the most frequent samples to isolate S. aureus. Resistant and virulent genes detection showed that, all 33 MRSA strains were mecA positive which accounts for 49.3% of the 67 S. aureus strains; 38 isolates were Panton-Valentine leukocidin (PVL) gene positive which accounts for 56.7% of the 67 S. aureus strains; and 17 (17/67, 25.4%) isolates are mecA and PVL genes dual positive. Multidrug-resistant strains of MRSA and PVL positive S. aureus are common in patients, medical staff and hospital setting, the potential health threat is worthy of our attention.
Argudín, M. A.; Mendoza, M. C.; Méndez, F. J.; Martín, M. C.; Guerra, B.; Rodicio, M. R.
Molecular epidemiology studies have allowed the identification of the methicillin (meticillin)-resistant (MRSA) and methicillin-susceptible (MSSA) clonal complexes (CCs) and clones of Staphylococcus aureus circulating in a Spanish hospital recently. Of 81 isolates tested, 32.1% were MRSA. Most of them carried staphylococcal cassette chromosome mec (SCCmec) IVc (88.5%) and belonged to CC5 (88.5%; multilocus sequence typing types ST125 [mainly associated with spa type t067], ST5, and ST228). A higher diversity was found among MSSA isolates (67.9%). Eighty percent shared the genetic background of major MRSA lineages (CC5 [38.2%; ST125 and ST5], CC30 [25.5%; ST30], CC45 [14.5%; ST45 and ST47], and CC8 [1.8%; ST8]), but CC12, CC15, CC51, and CC59 were also detected. Many exotoxin genes were present in each of the 81 isolates, independent of whether they were involved in sepsis (11 to 22) or other types of infections (13 to 21), and they appeared in 73 combinations. The relevant data are that (i) all isolates were positive for hemolysin and leukotoxin genes (98.8% for lukED and 25.9% for lukPV); (ii) all contained an enterotoxin gene cluster (egc with or without seu), frequently with one or more genes encoding classical enterotoxins; (iii) about half were positive for tst and 95% were positive for exfoliatin-encoding genes (eta, etb, and/or etd); and (iv) the four agr groups were detected, with agrII (55.6%) and agrIII (23.5%) being the most frequent. Taken together, results of the present study suggest a frequent acquisition and/or loss of exotoxin genes, which may be mediated by efficient intralineage transfer of mobile genetic elements and exotoxin genes therein and by eventual breakage of interlineage barriers. PMID:19458176
Sergi, Simona; Donnarumma, Francesca; Mastromei, Giorgio; Goti, Emanuele; Nicoletti, Pierluigi; Pecile, Patrizia; Cecconi, Daniela; Mannino, Roberta; Fanci, Rosa; Bosi, Alberto; Bartolozzi, Benedetta; Casalone, Enrico
In this study we report the results of analysis of 253 isolates of Staphylococcus aureus (132 methicillin [meticillin]-resistant S. aureus [MRSA] isolates and 121 methicillin-susceptible S. aureus [MSSA] isolates) from 209 patients admitted to 18 high-risk wards of six hospitals located in Florence, Italy, over an 8-month period during which a program of epidemiological surveillance of hospital-acquired infections was conducted. The majority (69%) of the 87 reported S. aureus infections were caused by MRSA. No outbreak events have been reported. All the isolates were typed by amplified fragment length polymorphism (AFLP), and AFLP profiles were analyzed in order to define similarity groups. The discriminatory power of AFLP is very high with MSSA (Simpson index of diversity [D], 95.9%), whereas its resolution capability with MRSA (D, 44.7%) is hampered by the well-known high clonality of these populations (the main MRSA group accounted for 74% of the MRSA isolates). Combining AFLP, improved by visual inspection of polymorphisms, with multiplex PCR greatly increases MRSA resolution (D, 85.5%), resolving the MRSA population to a level that is one of the highest reported in the literature. Widespread and sporadic clones of MSSA and MRSA were identified, and their diffusion in the different hospitals and wards over the surveillance period was studied. The understanding of MSSA and MRSA population structures should be the starting point for the design of a more rational surveillance program for S. aureus species, maximizing benefits and reducing the cost of infection control strategies.
Alt, Volker; Kirchhof, Kristin; Seim, Florian; Hrubesch, Isabelle; Lips, Katrin S; Mannel, Henrich; Domann, Eugen; Schnettler, Reinhard
New strategies to decrease infection rates in cementless arthroplasty are needed, especially in the context of the growing incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections. The purpose of this study was to investigate the antimicrobial activity of a rifampicin-fosfomycin coating against methicillin-sensitive Staphylococcus aureus (MSSA) and MRSA in a rabbit infection prophylaxis model. Uncoated or rifampicin-fosfomycin-coated K-wires were inserted into the intramedullary canal of the tibia in rabbits and contaminated with an inoculation dose of 10(5) or 10(6) colony-forming units of MSSA EDCC 5055 in study 1 and MRSA T6625930 in study 2, respectively. After 28days the animals were killed and clinical, histological and microbiological assessment, including pulse-field gel electrophoresis, was conducted. Positive culture growth in agar plate testing and/or clinical signs and/or histological signs were defined positive for infection. Statistical evaluation was performed using Fisher's exact test. Both studies showed a statistically significant reduction of infection rates for rifampicin-fosfomycin-coated implants compared to uncoated K-wires (P=0.015). In both studies none of the 12 animals that were treated with a rifampicin-fosfomycin-coated implant showed clinical signs of infection or a positive agar plate testing result. In both studies, one animal of the coating group showed the presence of sporadic bacteria with concomitant inflammatory signs in histology. The control groups in both studies exhibited an infection rate of 100% with clear clinical signs of infection and positive culture growth in all animals. In summary, the rifampicin-fosfomycin-coating showed excellent antimicrobial activity against both MSSA and MRSA, and therefore warrants further clinical testing.
Sanchini, A; Del Grosso, M; Villa, L; Ammendolia, M G; Superti, F; Monaco, M; Pantosti, A
Panton-Valentine leukocidin (PVL) is the hallmark of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) but can also be found in methicillin-susceptible S. aureus (MSSA) sharing pathogenic and epidemiological characteristics of CA-MRSA. PVL is encoded by two co-transcribed genes that are carried by different staphylococcal bacteriophages. We applied an extended PCR-based typing scheme for the identification of two morphological groups (elongated-head group and icosahedral-head group I phages) and specific PVL phage types in S. aureus isolates recovered in Italy. We examined 48 PVL-positive isolates (25 MSSA and 23 MRSA) collected from different hospital laboratories from April 2005 to May 2011. spa typing, multilocus sequence typing and staphylococcal cassette chromosome mec typing were applied to categorize the isolates. Phage typeability was 48.0% in MSSA and 91.3% in MRSA, highlighting the limitation of the PCR typing scheme when applied to PVL-positive MSSA. Five different PVL phages and two variants of a known phage were detected, the most prevalent being ΦSa2usa, recovered in 15 out of 48 (31.2%) isolates, and carried by both MSSA and MRSA belonging to CC8 and CC5. The recently described ΦTCH60 was recovered in four isolates. A PVL phage (ΦSa119) from an ST772 MRSA, that was not detected using the previous typing scheme, was sequenced, and new primers were designed for the identification of the icosahedral-head group II PVL phages present in ST772 and ST59 MRSA. A comprehensive PVL-phage typing can contribute to the understanding of the epidemiology and evolution of PVL-positive MSSA and MRSA.
Peyrani, P; Allen, M; Seligson, D; Roberts, C; Chen, A; Haque, N; Zervos, M; Wiemken, T; Harting, J; Christensen, D; Ramirez, R
Methicillin-resistant Staphylococcus aureus (MRSA) USA-300 strains have emerged as an important cause of community-acquired infections. These strains have been recognized as an etiology of osteomyelitis but data on their incidence and outcomes are limited. We retrospectively studied the incidence and clinical outcomes of MRSA USA-300 osteomyelitis in patients at the University of Louisville Hospital and the Henry Ford Health System between January 2007 and March 2008. Pulsed-field gel electrophoresis was used to determine USA type. Clinical outcomes were defined as management success versus failure at 12 months. Chi-square tests, Fisher exact tests, and Mann-Whitney tests were used to compare patient characteristics on the basis of clinical outcomes and USA type. Of the 50 patients with MRSA osteomyelitis, 27 (54%) had the USA-300 strain. Clinical failure was identified in 22% (6/27) of the patients with MRSA USA-300 and in 30% (7/23) of the patients with MRSA non-USA-300 osteomyelitis (P = .509). Our results showed that MRSA USA-300 is a significant etiology of MRSA osteomyelitis. With current surgical and medical management, outcomes of patients with MRSA USA-300 osteomyelitis are similar to those of patients with MRSA non-USA-300 osteomyelitis.
Neethirajan, Suresh; DiCicco, Matthew
The influence of fosfomycin on methicillin-resistant Staphylococcus pseudintermedius (MRSP) as the target cell was studied by atomic force microscopy (AFM). Nanoscale images of the effects of the antibiotic fosfomycin on this gram-positive bacterium's cell were obtained in situ without fixing agents. Our study has demonstrated substantial morphological and topographical differences between the control and fosfomycin-treated MRSP cells. The AFM investigations further revealed the rough surface morphology and a 30 % shrinkage in size of the fosfomycin-treated cell and the leakage of cytoplasmic components from the cell. The damage of cell membrane integrity and the cell surface degradation as observed elaborates the antibacterial activity of fosfomycin. The AFM image analysis also reveals that the fosfomycin inhibits cell division, and prevents the adhesion on the surface discouraging the biofilm attachment. The micro-titre plate assay results conform to the atomic force microscopy image analysis. This is the first visual demonstration of the effect of fosfomycin on MRSP cells.
Yang, Hee-Young; Suh, Jin-Tae; Lee, Hee-Joo
To evaluate methicillin-resistant S. aureus (MRSA) detection methods, we compared (a) mannitol salt agar with cefoxitin (MSA-FX), (b) MRSASelect agar (Bio-Rad), (c) MRSA ID (bioMerieuex), and (d) CHROMagar MRSA (BD Diagnostics) as selective media for culturing nasal swab specimens collected from intensive care unit (ICU) patients and healthcare personnel. A total of 99 (17.1%) cases of MRSA were recovered from 578 specimens. Four (5.5%) cases were identified from healthcare personnel and 95 (18.8%) were from ICU patients. The sensitivity of MSA-FX, MRSASelect, MRSA ID, and CHROMagar MRSA was 83.8, 87.9, 80.8, and 84.8% after 18 hr; 92.9, 94.9, 90.9, and 91.9% after 24 hr; and 96.0, 100, 99.0, and 99.0% after 48 hr, respectively. The specificity of MSA-FX, MRSASelect, MRSA ID, and CHROMagar MRSA was 97.7, 99.0, 98.7. and 99.8% after 18 hr; 97.1, 98.5, 98.1, and 99.5% after 24 hr; and 95.2, 97.7, 97.9, and 99.0% after 48 hr, respectively. In conclusion, all four media showed good results after the 24 hr readings, but MRSA ID and CHROMagar MRSA media required readings at 48 hr due to increased sensitivity at this time point.
Kiem, Sungmin; Schentag, Jerome J
Although antibiotics whose epithelial lining fluid (ELF) concentrations are reported high tend to be preferred in treatment of pneumonia, measurement of ELF concentrations of antibiotics could be misled by contamination from lysis of ELF cells and technical errors of bronchoalveolar lavage (BAL). In this review, ELF concentrations of anti-methicillin resistant Staphylococcus aureus (MRSA) antibiotics were interpreted considering above confounding factors. An equation used to explain antibiotic diffusion into CSF (cerebrospinal fluid) was adopted: ELF/free serum concentration ratio = 0.96 + 0.091 × ln (partition coefficient / molecular weight(1/2)). Seven anti-MRSA antibiotics with reported ELF concentrations were fitted to this equation to see if their ELF concentrations were explainable by the penetration capacity only. Then, outliers were modeled under the assumption of varying contamination from lysed ELF cells (test range 0-10% of ELF volume). ELF concentrations of oritavancin, telavancin, tigecycline, and vancomycin were well described by the diffusion equation, with or without additional impact from cell lysis. For modestly high ELF/free serum concentration ratio of linezolid, technical errors of BAL should be excluded. Although teicoplanin and iclaprim showed high ELF/free serum ratios also, their protein binding levels need to be cleared for proper interpretation. At the moment, it appears very premature to use ELF concentrations of anti-MRSA antibiotics as a relevant guide for treatment of lung infections by MRSA.
Nowrouzian, Forough L; Karami, Nahid; Welinder-Olsson, Christina; Ahrén, Christina
Methicillin-resistant Staphylococcus aureus (MRSA) has widely spread to all parts of the world. For surveillance and effective infection control molecular typing is required. We have evaluated the utility of virulence gene determination as a complementary tool for epidemiological typing of MRSA in relation to spa-typing and pulsed-field gel electrophoresis (PFGE). We assessed 63 community-acquired MRSA (CA-MRSA) isolates detected in the West part of Sweden for 30 virulence factor genes (VF) and agr allele variations by serial polymerase chain reaction (PCR) assays. These isolates belonged to sequence types (ST) 8, 80, 45 and 30 as classified by multilocus sequence typing. The isolates in each spa-type and PFGE-type were examined over an extended time-period and constituted a varying number of PFGE-subtypes (5-14) and spa-types (3-11) within four major PFGE types. Each ST had a unique VF profile. For isolates within a major PFGE type showing high diversity both in PFGE subtypes and spa the VF profile varied as well in contrast to those with low diversity where no alterations were seen. Thus, the accuracy of each typing method does not only vary by the method per se but is rather dependent on the genetic repertoire of the typed strains and genes evaluated. For strains demonstrating high diversity VF typing may be a useful complement in the epidemiological investigations, and may highlight the accurate discriminatory power of spa or PFGE typing.
Over the last 50 years methicillin-resistant S. aureus (MRSA) spread globally. Vancomycin is still the most recommended antibiotic for MRSA-infections. Teicoplanin is an alternative glycopeptide with longer elimination half-life. Telavancin is a more recently developed derivative of vancomycin with similar clinical efficacy as vancomycin. It is not recommended for treatment of patients with renal insufficiency. Nephrotoxicity limits the therapeutic use of glycopeptide antibiotics. The oxazolidinone linezolid exhibits similar to superior therapeutic efficacy. Hematologic controls are necessary during treatment with this antibacterial agent. Neurotoxic effects have been observed mainly in patients who received prolonged linezolid treatment. Attention must be paid to possible interactions with concomitantly given drugs acting on the serotonergic system. New therapeutic options arise with ceftaroline, the first β-lactam antibiotic with activity against MRSA. However, controlled clinical trials with pulmonary MRSA infections have not been conducted with ceftaroline. Daptomycin, a lipopeptide, and tigecycline, a glycylcyclin are active in vitro against MRSA as well, but are also not indicated in pulmonary MRSA infections. These antibiotics show in an exemplary manner that antibacterial activity in vitro is an important prerequisite, but relevant data for a therapeutic decision should be derived from randomized controlled clinical double-blind trials.
Jiamboonsri, Pimsumon; Pithayanukul, Pimolpan; Bavovada, Rapepol; Chomnawang, Mullika T
Plant extracts are a valuable source of novel antibacterial compounds to combat pathogenic isolates of methicillin-resistant Staphylococcus aureus (MRSA), a global nosocomial infection. In this study, the alcoholic extract from Thai mango (Mangifera indica L. cv. 'Fahlun') seed kernel extract (MSKE) and its phenolic principles (gallic acid, methyl gallate and pentagalloylglucopyranose) demonstrated potent in vitro antibacterial activity against Staphylococcus aureus and 19 clinical MRSA isolates in studies of disc diffusion, broth microdilution and time-kill assays. Electron microscopy studies using scanning electron microscopy and transmission electron microscopy revealed impaired cell division and ultra-structural changes in bacterial cell morphology, including the thickening of cell walls, of microorganisms treated with MSKE; these damaging effects were increased with increasing concentrations of MSKE. MSKE and its phenolic principles enhanced and intensified the antibacterial activity of penicillin G against 19 clinical MRSA isolates by lowering the minimum inhibitory concentration by at least 5-fold. The major phenolic principle, pentagalloylglucopyranose, was demonstrated to be the major contributor to the antibacterial activity of MSKE. These results suggest that MSKE may potentially be useful as an alternative therapeutic agent or an adjunctive therapy along with penicillin G in the treatment of MRSA infections.
Wolters, Manuel; Rohde, Holger; Maier, Thomas; Belmar-Campos, Cristina; Franke, Gefion; Scherpe, Stefanie; Aepfelbacher, Martin; Christner, Martin
Early detection of outbreaks of methicillin-resistant Staphylococcus aureus (MRSA) and initiation of adequate infection control measures are important objectives in hospital hygiene. To reach these goals, prompt determination of epidemiologic relatedness of clinical MRSA isolates is essential. Genetic typing methods like pulsed-field gel electrophoresis (PFGE), spa typing, or multilocus sequence typing (MLST) have a high discriminatory power, however, these methods are time consuming and cost intensive. The aim of this study was to investigate the potential of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for discrimination of major MRSA lineages. By analysis of mass spectra from 25 representative MRSA isolates belonging to the 5 major hospital-acquired (HA) MRSA clonal complexes (CC5, CC8, CC22, CC30, CC45; deduced from spa typing), reproducible spectrum differences were observed at 13 characteristic m/z values allowing robust discrimination of the clonal complexes. When 60 independent clinical MRSA isolates were tested for the presence or absence of the 13 characteristic MALDI-TOF MS peaks, 15 different profiles (MALDI types) could be detected. Hierarchical clustering of the MALDI types showed high concordance with the clonal complexes. Our results suggest that MALDI-TOF MS has the potential to become a valuable first-line tool for inexpensive and rapid typing of MRSA in infection control.
Lee, Young-Seob; Lee, Dae-Young; Kim, Yeon Bok; Lee, Sang-Won; Cha, Seon-Woo; Park, Hong-Woo; Kim, Geum-Soog; Kwon, Dong-Yeul; Lee, Min-Ho; Han, Sin-Hee
Shikonin (SKN), a highly liposoluble naphthoquinone pigment isolated from the roots of Lithospermum erythrorhizon, is known to exert antibacterial, wound-healing, anti-inflammatory, antithrombotic, and antitumor effects. The aim of this study was to examine SKN antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). The SKN was analyzed in combination with membrane-permeabilizing agents Tris and Triton X-100, ATPase inhibitors sodium azide and N,N′-dicyclohexylcarbodiimide, and S. aureus-derived peptidoglycan; the effects on MRSA viability were evaluated by the broth microdilution method, time-kill test, and transmission electron microscopy. Addition of membrane-permeabilizing agents or ATPase inhibitors together with a low dose of SKN potentiated SKN anti-MRSA activity, as evidenced by the reduction of MRSA cell density by 75% compared to that observed when SKN was used alone; in contrast, addition of peptidoglycan blocked the antibacterial activity of SKN. The results indicate that the anti-MRSA effect of SKN is associated with its affinity to peptidoglycan, the permeability of the cytoplasmic membrane, and the activity of ATP-binding cassette (ABC) transporters. This study revealed the potential of SKN as an effective natural antibiotic and of its possible use to substantially reduce the use of existing antibiotic may also be important for understanding the mechanism underlying the antibacterial activity of natural compounds. PMID:26265924
Dakheel, Khulood Hamid; Abdul Rahim, Raha; Hun, Tan Geok
Twenty-five methicillin-resistant Staphylococcus aureus (MRSA) isolates were characterized by staphylococcal protein A gene typing and the ability to form biofilms. The presence of exopolysaccharides, proteins, and extracellular DNA and RNA in biofilms was assessed by a dispersal assay. In addition, cell adhesion to surfaces and cell cohesion were evaluated using the packed-bead method and mechanical disruption, respectively. The predominant genotype was spa type t127 (22 out of 25 isolates); the majority of isolates were categorized as moderate biofilm producers. Twelve isolates displayed PIA-independent biofilm formation, while the remaining 13 isolates were PIA-dependent. Both groups showed strong dispersal in response to RNase and DNase digestion followed by proteinase K treatment. PIA-dependent biofilms showed variable dispersal after sodium metaperiodate treatment, whereas PIA-independent biofilms showed enhanced biofilm formation. There was no correlation between the extent of biofilm formation or biofilm components and the adhesion or cohesion abilities of the bacteria, but the efficiency of adherence to glass beads increased after biofilm depletion. In conclusion, nucleic acids and proteins formed the main components of the MRSA clone t127 biofilm matrix, and there seems to be an association between adhesion and cohesion in the biofilms tested. PMID:28078291
Lee, Bruce Y.; Singh, Ashima; David, Michael Z.; Bartsch, Sarah M.; Slayton, Rachel B.; Huang, Susan S.; Zimmer, Shanta M.; Potter, Margaret A.; Macal, Charles M.; Lauderdale, Diane S.; Miller, Loren G.; Daum, Robert S.
The economic impact of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) remains unclear. We developed an economic simulation model to quantify the costs associated with CA-MRSA infection from the societal and third-party payer perspectives. A single CA-MRSA case costs third-party payers $2,277 – $3,200 and society $7,070 – $20,489, depending on patient age. In the United States (US), CA-MRSA imposes an annual burden of $478 million - 2.2 billion on third-party payers and $1.4 billion - 13.8 billion on society, depending on the CA-MRSA definitions and incidences. The US jail system and Army may be experiencing annual total costs of $7 – 11 million ($6 – 10 million direct medical costs) and $15 – 36 million ($14 – 32 million), respectively. Hospitalization rates and mortality are important cost drivers. CA-MRSA confers a substantial economic burden to third-party payers and society, with CA-MRSA-attributable productivity losses being major contributors to the total societal economic burden. Although decreasing transmission and infection incidence would decrease costs, even if transmission were to continue at present levels, early identification and appropriate treatment of CA-MRSA infections before they progress could save considerable costs. PMID:22712729
Non, Lemuel; Kosmin, Aaron
Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is a rare cause of necrotising fasciitis (NF), and is usually not fulminant as in group A Streptococcus (GAS), the archetypal aetiology. We report an unusually fulminant case of NF by CA-MRSA in an immunocompetent patient. A 52-year-old man presented to the emergency department with 1 week of progressive left thigh pain and swelling. The patient had ecchymoses, bullae and hypoesthesia of the involved skin, and CT scan revealed extensive fascial oedema. He was immediately started on broad spectrum antibiotics. Within 12 h of presentation, he underwent surgical debridement. Despite aggressive supportive care, the patient died less than 24 h after presentation. MRSA, with an antibiogram suggestive of a community-acquired strain, was recovered from intraoperative specimens and admission blood cultures. This case underscores that CA-MRSA, while rarely reported, can cause a fulminant presentation of NF similar to GAS in immunocompetent patients. PMID:25824286
Duerden, Brian; Fry, Carole; Johnson, Alan P.; Wilcox, Mark H.
Methicillin-resistant Staphylococcus aureus (MRSA) blood stream infection (BSI) is a major healthcare burden in some but not all healthcare settings, and it is associated with 10%–20% mortality. The introduction of mandatory reporting in England of MRSA BSI in 2001 was followed in 2004 by the setting of target reductions for all National Health Service hospitals. The original national target of a 50% reduction in MRSA BSI was considered by many experts to be unattainable, and yet this goal has been far exceeded (∼80% reduction with rates still declining). The transformation from endemic to sporadic MRSA BSI involved the implementation of serial national infection prevention directives, and the deployment of expert improvement teams in organizations failed to meet their improvement trajectory targets. We describe and appraise the components of the major public health infection prevention campaign that yielded major reductions in MRSA infection. There are important lessons and opportunities for other healthcare systems where MRSA infection remains endemic. PMID:26380336
Wang, Li; Zhang, Han-Bo; Chen, Qian; Liu, Hua; Tang, Xun; Jin, Tao; Zhu, Chong-Tao; Li, Fu-Bing; Sun, Lin-Hui; Xu, Xin-Ming; Xu, Yong-Qing
Infection by methicillin-resistant Staphylococcus aureus (MRSA) is a life-threatening condition, and formation of biofilms can lead to treatment failure in a clinical setting. The aim of this study was to demonstrate the in vivo bactericidal effects of a combination of vancomycin (VAN) and fosfomycin (FOS) against MRSA in a rat carboxymethyl cellulose-pouch biofilm model. The results of the time-kill assay showed that the combination therapy was capable of killing at low minimal inhibitory concentrations (MIC) (½× MIC VAN +1× MIC FOS and 1× MIC VAN + 1× MIC FOS). In the in vivo study, a synergistically bactericidal effect was observed when using the combination therapy on MRSA embedded in the mature biofilm model. In comparison with the untreated control group and the groups receiving either VAN or FOS alone, the rats treated with combination therapy had lower MRSA colony counts in exudates from the pouch, lower white blood cell and neutrophil counts, and C-reactive protein (CRP) in peripheral blood. Furthermore, histological analysis of the pouch wall indicated combination therapy resulted in disappearance of biofilm-like structures, marked decrease in necrosis, and formation of granular tissue. In conclusion, the combination of VAN with FOS had a synergistic bactericidal effect on chronic MRSA infection embedded in biofilm, providing an alternative approach to treating this condition. PMID:25551618
Sneha, S; Venishetty, Shanthan; Seshadri, Shubha; Mukhopadhyay, Chiranjay
Infective endocarditis by Methicillin Resistant Staphylococcus aureus (MRSA) bacteraemia is a common association and carries a high mortality. However, rising Minimum Inhibitory Concentrations (MICs) for vancomycin amongst MRSA strains is an emerging threat which carries poor prognosis and higher mortality. Here, we report a case of 41-year-old young non-addict gentleman presenting with fever of 3 days duration following recovery from an acute kidney injury necessitating haemodialysis. Systemic examination revealed signs of mitral regurgitation and left sided cerebellar signs. Laboratory investigations revealed anaemia, MRSA bacteraemia and transthoracic echocardiogram showed a mitral valve vegetation. He was diagnosed with MRSA mitral valve endocarditis with acute left cerebellar infarct. He was initially treated with intravenous vancomycin based on MIC at the dose of one gram every twelfth hourly. As the MRSA strain was shown to demonstrate a vancomycin creep phenomemon along with worsening clinical condition of the patient, treatment was changed to intravenous daptomycin at dose of 9mg/kg as per blood culture and sensitivity pattern which was administered for 6 weeks. Following initiation of daptomycin patient became afebrile and vegetations reduced in size and finally disappeared. PMID:28208915
Sizemore, Elizabeth Nicole; Rivas, Kenya Maria; Valdes, Jose; Caballero, Joshua
A geriatric patient status post intraabdominal surgery presented with persistent diarrhoea and heavy intestinal methicillin-resistant Staphylococcus aureus (MRSA) growth after multiple courses of antibiotic therapy. Additionally, swab cultures of the anterior nares tested positive for MRSA. In order to impede infection and prevent future complications, the patient received a 10-day course of vancomycin oral solution 250 mg every 6 h, 15-day course of Saccharomyces boulardii 250 mg orally twice daily and a 5-day course of topical mupirocin 2% twice daily intranasally. Diarrhoea ceased during therapy and repeat cultures 11 days after initiating therapy demonstrated negative MRSA growth from the stool and nares. Further repeat cultures 5 months later revealed negative MRSA growth in the stools and minimal MRSA growth in the nares. Overall, enteral vancomycin and probiotics successfully eradicated MRSA infection without intestinal recurrence. Although the results were beneficial treating MRSA diarrhoea for our patient, these agents remain highly controversial. PMID:22847566
Chihara, Shingo; Hayden, Mary K.; Minogue-Corbett, Eileen; Singh, Kamaljit
The ability to rapidly differentiate coagulase-negative staphylococcus (CoNS) from Staphylococcus aureus and to determine methicillin resistance is important as it affects the decision to treat empiric antibiotic selection. The objective of this study was to evaluate CHROMagar S. aureus and CHROMagar MRSA (Becton Dickinson) for rapid identification of Staphylococcus spp. directly from blood cultures. Consecutive blood culture bottles (BacT Alert 3D SA and SN, bioMérieux) growing gram-positive cocci in clusters were evaluated. An aliquot was plated onto CHROMagar MRSA (C-MRSA) and CHROMagar S. aureus (C-SA) plates, which were read at 12 to 16 hours. C-SA correctly identified 147/147 S. aureus (100% sensitivity); 2 CoNS were misidentified as S. aureus (98% specificity). C-MRSA correctly identified 74/77 MRSA (96% sensitivity). None of the MSSA isolates grew on C-MRSA (100% specificity). In conclusion, CHROMagar is a rapid and sensitive method to distinguish MRSA, MSSA, and coagulase-negative Staphylococcus and may decrease time of reporting positive results. PMID:20016679
Visutthi, Monton; Srimanote, Potjanee; Voravuthikunchai, Supayang Piyawan
Rhodomyrtone from a medicinal plant species, Rhodomyrtus tomentosa, is a challenged effective agent against Gram-positive bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). The present study was undertaken to provide insight into MRSA extracellular protein expression following rhodomyrtone treatment. Secreteomic approach was performed on a representative clinical MRSA isolate exposing to subinhibitory concentration rhodomyrtone (0.174 μg/ml). The identified extracellular proteins of a response of MRSA to rhodomyrtone treated condition were both suppressed and overexpressed. Staphylococcal antigenic proteins, immunodominant antigen A (IsaA) and staphylococcal secretory antigen (SsaA) involved in cell wall hydrolysis were downregulated after the treatment. The results suggested that rhodomyrtone may interfere with WalK/WalR (YycG/YycF) system. Other enzymes such as lipase precursor and another lipase, glycerophosphoryl diester phosphodiesterase, were absent. In contrast, cytoplasmic proteins such as SpoVG and glycerol phosphate lipoteichoic acid synthase, and ribosomal proteins were found in the treated sample. Appearance of several cytoplasmic proteins in the treated culture supernatant revealed that the bacterial cell wall biosynthesis was disturbed. This finding provides a proteomic mapping of extracellular proteins after rhodomytone treatment. Extensive investigation is required for this natural compound as it has a great potency as an alternative anti-MRSA drug.
Turk, Ryen; Singh, Ameet; Rousseau, Joyce; Weese, J Scott
Methicillin resistant Staphylococcus pseudintermedius (MRSP) is an important canine pathogen that has been shown to produce biofilm in vitro. Biofilm production may be an important virulence factor and methods to eliminate biofilm-associated infections are required. An enzyme of Aggregatibacter actinomycetemcomitans, DispersinB, has been shown to degrade the extracellular matrix in the biofilm of a variety of bacteria including Staphylococcus aureus and Staphylococcus epidermidis. The objective of this study was to determine the effect of DispersinB on MRSP biofilm production and eradication in vitro. A quantitative microtitre plate assay was used to assess the impact of DispersinB on 30 MRSP isolates from dogs. While DispersinB did not have any effect on MRSP growth (P=0.98), it reduced biofilm formation (P=0.0002) and degraded established biofilm (P=0.0001). These data indicate that in vivo study of the effect of this enzyme in indicated to determine if it may be a useful treatment option for MRSP biofilm-associated infections.
Karasartova, Djursun; Cavusoglu, Zeynep Burcin; Turegun, Buse; Ozsan, Murat T; Şahin, Fikret
Bacteriophages play an important role in the pathogenicity of Staphylococcus aureus (S. aureus) either by carrying accessory virulence factors or several superantigens. Despite their importance, there are not many studies showing the actual distribution of the virulence genes carried by the prophages obtained from the clinically isolated Staphylococcus. In this study, we investigated prophages obtained from methicillin-resistant S. aureus (MRSA) strains isolated from hospital- and community-associated (HA-CA) infections for the virulence factors. In the study, 43 phages isolated from 48 MRSA were investigated for carrying toxin genes including the sak, eta, lukF-PV, sea, selp, sek, seg, seq chp, and scn virulence genes using polymerase chain reaction (PCR) and Southern blot. Restriction fragment length polymorphism was used to analyze phage genomes to investigate the relationship between the phage profiles and the toxin genes' presence. MRSA strains isolated from HA infections tended to have higher prophage presence than the MRSA strains obtained from the CA infections (97% and 67%, respectively). The study showed that all the phages with the exception of one phage contained one or more virulence genes in their genomes with different combinations. The most common toxin genes found were sea (83%) followed by sek (77%) and seq (64%). The study indicates that prophages encode a significant proportion of MRSA virulence factors.
David, Michael Z.; Daum, Robert S.
Summary: Staphylococcus aureus is an important cause of skin and soft-tissue infections (SSTIs), endovascular infections, pneumonia, septic arthritis, endocarditis, osteomyelitis, foreign-body infections, and sepsis. Methicillin-resistant S. aureus (MRSA) isolates were once confined largely to hospitals, other health care environments, and patients frequenting these facilities. Since the mid-1990s, however, there has been an explosion in the number of MRSA infections reported in populations lacking risk factors for exposure to the health care system. This increase in the incidence of MRSA infection has been associated with the recognition of new MRSA clones known as community-associated MRSA (CA-MRSA). CA-MRSA strains differ from the older, health care-associated MRSA strains; they infect a different group of patients, they cause different clinical syndromes, they differ in antimicrobial susceptibility patterns, they spread rapidly among healthy people in the community, and they frequently cause infections in health care environments as well. This review details what is known about the epidemiology of CA-MRSA strains and the clinical spectrum of infectious syndromes associated with them that ranges from a commensal state to severe, overwhelming infection. It also addresses the therapy of these infections and strategies for their prevention. PMID:20610826
Woon, Colin YL; Patel, Kushal R; Goldberg, Benjamin A
Gluteal compartment syndrome (GCS) is a rare condition. We present a case of gluteal muscle strain with hematoma formation, methicillin-resistant Staphylococcus aureus (MRSA) superinfection, leading to acute GCS, rhabdomyolysis and acute kidney injury. This combination of diagnoses has not been reported in the literature. A 36-year-old Caucasian male presented with buttock pain, swelling and fever after lifting weights. Gluteal compartment pressure was markedly elevated compared with the contralateral side. Investigations revealed elevated white blood cell, erythrocyte sedimentation rate, C-reactive protein, creatine kinase, creatinine and lactic acid. Urinalysis was consistent with myoglobinuria. Magnetic resonance imaging showed increased T2 signal in the gluteus maximus and a central hematoma. Cultures taken from the emergency debridement and fasciotomy revealed MRSA. He had repeat, debridement 2 d later, and delayed primary closure 3 d after. GCS is rare and must be suspected when patients present with pain and swelling after an inciting event. They are easily diagnosed with compartment pressure monitoring. The treatment of gluteal abscess and compartment syndrome is the same and involves rapid surgical debridement. PMID:27190761
Zhang, Hong; Li, Haiqing; Liu, Yan; Li, Qingyan; Bi, Yufang; Fang, Guiqing
The aim of the study was to investigate the characteristic function of the upregulated effects of miR-7 in methicillin-resistant Staphylococcus aureus (MRSA). After separating the MRSA in clinic, the expression of miR-7 mRNA was tested by reverse transcription polymerase chain reaction. The overexpression, inhibition of miR-7, and control group were established by plasmid in vitro. Following transfection of the bacterial strain, the effect of β-lactam antibiotics in minimum inhibitory concentration (MIC) was observed using the microporous dilution method, and antibacterial effects in vitro were observed using the dynamic growth curve method. The expression of miR-7 in sensitive MRSA was upregulated distinctly, with significant difference (P<0.05). MIC and the number of bacteria in the miR-7 overexpression group significantly increased while the inhibition group decreased prominently, with significant difference (P<0.05). The control and null plasmid groups revealed no significant difference. In conclusion, miR-7 upregulated the antimicrobial activity of MRSA, and the intervention of its expression may become a possible antibacterial target. PMID:28101153
Bühlmann, Manuela; Bögli-Stuber, Katja; Droz, Sara; Mühlemann, Kathrin
PCR tests for the rapid and valid detection of methicillin-resistant Staphylococcus aureus (MRSA) are now available. We evaluated the costs associated with contact screening for MRSA carriage in a tertiary-care hospital with low MRSA endemicity. Between 1 October 2005 and 28 February 2006, 232 patients were screened during 258 screening episodes (644 samples) for MRSA carriage by GenoType MRSA Direct (Hain Lifescience GmbH, Nehren, Germany). Conventional culture confirmed all PCR results. According to in-house algorithms, 34 of 258 screening episodes (14.7%) would have qualified for preemptive contact isolation, but such isolation was not done upon negative PCR results. MRSA carriage was detected in 4 (1.5%) of 258 screening episodes (i.e., in four patients), of which none qualified for preemptive contact isolation. The use of PCR for all 258 screening episodes added costs (in Swiss francs [CHF]) of CHF 104,328.00 and saved CHF 38,528.00 (for preemptive isolation). The restriction of PCR screening to the 34 episodes that qualified for preemptive contact isolation and screening all others by culture would have lowered costs for PCR to only CHF 11,988.00, a savings of CHF 38,528.00. Therefore, PCR tests are valuable for the rapid detection of MRSA carriers, but high costs require the careful evaluation of their use. In patient populations with low MRSA endemicity, the broad use of PCR probably is not cost-effective.
Efstathopoulos, Nicolas; Giamarellos-Bourboulis, Evangellos; Kanellakopoulou, Kyriaki; Lazarettos, Ioannis; Giannoudis, Peter; Frangia, Konstantina; Magnissalis, Evangellos; Papadaki, Maria; Nikolaou, Vassilios S
The authors examined the effectiveness of the local anti-microbial treatment on methicillin resistant Staphylococcus aureus (MRSA) experimental osteomyelitis. Thirty-six rabbits with chronic MRSA osteomyelitis of the right femur were treated with local grepafloxacin delivery system prepared by a mixture of acrylic bone cement (polymethyl methacrylate, PMMA) plus 4% grepafloxacin. Osteomyelitis was induced by inoculating MRSA (100 microl of cultured bacteria; 10(7)) and the local insertion of a needle, serving as a foreign body, at the upper third of the femur. The course of the infection was followed by clinical, radiographic and microbiological examination. In the third week, all animals were re-operated, needles were removed, and antibiotic containing acrylic cement was implanted. Thereafter, one control and five treated animals were sacrificed per week, within 6 weeks. Osteomyelitis was found in all rabbits. In vitro grepafloxacin levels remained high throughout the 6 weeks of the experiment. Histologically tissue reaction against the cement was not observed. Osteomyelitis lesions and bone structure were progressively repaired after cement implantation. Biomechanical analysis showed no significant influence on the mechanical properties of acrylic cement due to grepafloxacin. The above mixture could prove to be an important supplementary method for the treatment of bone infections. Such a system could replace the use of gentamycin PMMA beads in the treatment of patients with chronic osteomyelitis due to MRSA. Furthermore, the proposed method could be used as a spacer after removal septic loosened prostheses in combination with systemic administration of antibiotics.
Gücükoğlu, Ali; Çadirci, Özgür; Terzi, Göknur; Kevenk, T Onur; Alişarli, Mustafa
The aim of this study was to determine the prevalence of enterotoxigenic and methicillin-resistant Staphylococcus aureus in ice creams. After culture-based identification of isolates, the presence of 16S rRNA and nuc was confirmed by mPCR. S. aureus was identified in 18 of 56 fruity (32.1%), 4 of 32 vanilla (12.5%), and 1 of 12 chocolate (8.3%) ice creams. S. aureus was identified as 38 isolates in 23 ice cream samples by culture-based techniques, but only 35 isolates were confirmed by PCR as S. aureus. To determine the enterotoxigenic properties of PCR-confirmed S. aureus isolates, a toxin detection kit was used (SET RPLA®). Of the 12 enterotoxigenic S. aureus isolates, 9 SEB (75%), 1 SED (8.3%), 1 SEB+SED (8.3%), and 1 SEA+SEB+SED (8.3%) expressing isolates were found. The presence of enterotoxin genes (sea, seb, sed) was identified in 13 (37.1%) out of 35 isolates by the mPCR technique. In the ice cream isolates, the sea, seb, and sed genes were detected: 1 sea (7.6%), 9 seb (69.2%), 1 sed (7.6%), 1 seb+sed (7.6%), and 1 sea+seb+sed (7.6%), respectively. The sec gene was not detected in any of these isolates. One of the 35 (2.8%) S. aureus strain was mecA positive.
Kuusela, P; Hildén, P; Savolainen, K; Vuento, M; Lyytikäinen, O; Vuopio-Varkila, J
Seventy-nine methicillin-resistant Staphylococcus aureus (MRSA) strains, isolated during 1980 to 1990, were classified as MRSA Aggl- (14 strains) and MRSA Aggl+ (65 strains) strains on the basis of test results in slide agglutination assays designed to detect fibrinogen-binding protein (clumping factor) and protein A on the staphylococcal surface. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that lysostaphin digests of MRSA Aggl- strains contained a high-molecular-weight protein which was not detected in digests of MRSA Aggl+ strains. Immunization of rabbits with an MRSA Aggl- strain produced an antiserum which agglutinated all MRSA Aggl- strains and also 64 of 65 MRSA Aggl+ strains. Only 1 of 68 coagulase-negative staphylococci showed agglutination in this assay. The anti-MRSA Aggl- antiserum reacted mainly with a 230-kDa staphylococcal surface protein but also with a 175-kDa protein, probably formed by proteolysis of the former and a few slightly smaller proteins. These could not be immunologically detected in lysostaphin digests of MRSA Aggl+ strains. Purified antibodies reacting with the 230-kDa protein agglutinated all MRSA Aggl- strains, indicating that the protein is located on the surfaces of staphylococci. The results suggest a tentative role for the 230-kDa protein or its fragments as a novel target to develop more efficient rapid identification methods for S. aureus, including MRSA. Images PMID:8126170
Hall, David L
The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in nursing home residents now averages 20-35%. This includes both numerous asymptomatic mostly unidentified carriers, and the occasional patient with an active infection. Among the most common sites for positive MRSA colonization are the nares and mouth (saliva). Ohio State University (OSU) dental students perform routine restorative dental care onsite in local nursing homes using portable equipment including handpieces that can generate aerosols. Using a series of cultured test swabs and plates, this pilot study suggests that protection for both dental health care personnel and patients are provided by the following: 1. universal barrier precautions (for example, gloves, gowns, masks, hats, facial shields, glasses), 2. surface disinfectants, 3. pre-op 0.12% chlorhexidene mouth rinses, 4. high volume evacuation, 5. perioral skin scrubs. Additional infection control methods, techniques and equipment were evaluated and compared including rubber dam isolation, hand excavation and bond technique, high-speed air turbine and electric "high" speed handpiece. There was no indication of a special tendency or heightened ability of MRSA to aerosolize.
Blanc, D. S.; Struelens, M. J.; Deplano, A.; De Ryck, R.; Hauser, P. M.; Petignat, C.; Francioli, P.
To determine the stability of pulsed-field gel electrophoresis (PFGE) patterns of methicillin-resistant Staphylococcus aureus in the nosocomial setting, we analyzed isolates from long-term carriers (>1 month) and from patients involved in well-defined nosocomial epidemics. The number of fragment differences between the first isolate and subsequent isolates in long-term carriers showed a bimodal distribution, with one group having 0 to 6 fragment differences and the other group having 14 to 24 fragment differences. The PFGE patterns of isolates involved in epidemics also presented a similar bimodal distribution of the number of fragment differences. Typing these isolates with another molecular method (inter-IS256 PCR) showed that isolates of the first group (i.e., with 1 to 6 fragment differences) were clonally related, whereas the second group (with 14 to 24 fragment differences) could be considered genetically different. Among long-term carriers with clonally related isolates, 74 of 84 (88%) of consecutive isolates showed indistinguishable patterns, whereas 10 of 84 (12%) showed related patterns differing by one to six fragments. Moreover, the frequency of apparition of related patterns is higher when the time between the first and the subsequent isolate is longer. During seven nosocomial epidemics lasting from 1 to 15 months, only 2 of 120 isolates (1.7%) showed a pattern which was different, although related, from the predominant one involved in each of these outbreaks. PMID:11574553
Lee, Young-Seob; Kang, Ok-Hwa; Choi, Jang-Gi; Oh, You-Chang; Chae, Hee-Sung; Kim, Jong Hak; Park, Hyun; Sohn, Dong Hwan; Wang, Zheng-Tao; Kwon, Dong-Yeul
The antimicrobial killing activity toward methicillin-resistant Staphylococcus aureus (MRSA) has been a serious emerging global issue. New effective antimicrobials and/or new approaches to settle this issue are urgently needed. The oriental herb, Alpinia officinarum, has been used in Korea for several hundreds of years to treat various infectious diseases. As it is well known, one of the active constituents of Alpinia officinarum is galangin. Against the 17 strains, the minimum inhibitory concentrations (MICs) of galangin (GAL) were in the range of 62.5 ~ 125 microg/ml, and the MICs of gentamicin (GEN) ranged from 1.9 microg/ml to 2,000 microg/ml. The fractional inhibitory concentrations (FICs) of GAL, in combination with GEN, against 3 test strains were 0.4, 3.9, and 250 microg/ml, and were all 15.62 microg/ml in GEN. The FIC index showed marked synergism in the value range of 0.19 to 0.25. By determining time-kill curves, also confirmed the low synergism of the GAL and GEN combination against 4 h, 8 h, 12 h, and 24 h cultured MRSA. The time-kill study results indicated a low synergistic effect against 3 test strains. Thus, the mixture of GAL and GEN could lead to the development of new combination antibiotics against MRSA infection.
Shafer, W M; Iandolo, J J
Biophysical and genetic analysis of staphylococcal enterotoxin B (SEB) synthesis in 16 methicillin-resistant (Mecr) Staphylococcus aureus isolates demonstrated that the toxin gene (entB) can occupy either a plasmid or chromosomal locus. Biophysical analysis of the plasmid deoxyribonucleic acid content of these strains by agarose gel electrophoresis revealed the presence of a 1.15-megadalton plasmid in six isolates that appears to contain the entB gene. Genetic manipulation of SEB synthesis by transduction and elimination procedures demonstrated that this plasmid is critical for enterotoxigenesis. Nevertheless, the majority of the Mecr SEB+ isolates (62.5%) analyzed in this investigation were found to lack the 1.15-megadalton plasmid. In at least two of these strains (COL and 57-dk), transduction and elimination procedures showed that entB was chromosomal. Genetic studies involving strains harboring either a plasmid or chromosomal entB gene demonstrated that toxin synthesis was coeliminated with mec. However, analysis of the entB and mec loci by transformation or transduction showed that the genes are not closely linked. On the other hand, transduction of entB, regardless of the donor, was observed when both mec and the Tcr plasmid were jointly cotransduced. This finding suggests that, during transduction, a transient association between entB, mec, and the Tcr plasmid may exist. Images PMID:259057
Van den Eede, A; Hermans, K; Van den Abeele, A; Floré, K; Dewulf, J; Vanderhaeghen, W; Crombé, F; Butaye, P; Gasthuys, F; Haesebrouck, F; Martens, A
Given the significance of methicillin-resistant Staphylococcus aureus (MRSA) infections for both horses and staff in equine veterinary hospitals, protocols are required to minimise the risk of nosocomial transmission, including the screening of the skin and nasal chambers of equine patients for evidence of infection. The objective of this study was to clarify the potential existence and extent of MRSA on the skin of horses requiring long-term hospitalisation (≥ 6 months). Thirty such horses were sampled at eight different locations on their skin and from their nasal chambers. MRSA was isolated from 12 animals (40%), with all sample sites testing positive on at least one occasion. Organisms were most frequently detected in the nasal chambers (relative sensitivity, 83.3%; 34.5% positive horses; isolation rate 33.3%). Skin presence was found in 30% of animals with the highest isolation rates found at the carpus (16.7%), neck, withers and croup (13.3% each). To achieve a relative screening sensitivity of >90%, at least one skin site was required in addition to nasal sampling. This evidence of skin as well as nasal reservoirs of MRSA in long-term hospitalised horses should facilitate the design of effective screening and containment protocols.
Calderwood, Michael S.; Desjardins, Christopher A.; Sakoulas, George; Nicol, Robert; DuBois, Andrea; Delaney, Mary L.; Kleinman, Ken; Cosimi, Lisa A.; Feldgarden, Michael; Onderdonk, Andrew B.; Birren, Bruce W.; Platt, Richard; Huang, Susan S.
Background. Methicillin-resistant Staphylococcus aureus (MRSA) colonization predicts later infection, with both host and pathogen determinants of invasive disease. Methods. This nested case-control study evaluates predictors of MRSA bacteremia in an 8–intensive care unit (ICU) prospective adult cohort from 1 September 2003 through 30 April 2005 with active MRSA surveillance and collection of ICU, post-ICU, and readmission MRSA isolates. We selected MRSA carriers who did (cases) and those who did not (controls) develop MRSA bacteremia. Generating assembled genome sequences, we evaluated 30 MRSA genes potentially associated with virulence and invasion. Using multivariable Cox proportional hazards regression, we assessed the association of these genes with MRSA bacteremia, controlling for host risk factors. Results. We collected 1578 MRSA isolates from 520 patients. We analyzed host and pathogen factors for 33 cases and 121 controls. Predictors of MRSA bacteremia included a diagnosis of cancer, presence of a central venous catheter, hyperglycemia (glucose level, >200 mg/dL), and infection with a MRSA strain carrying the gene for staphylococcal enterotoxin P (sep). Receipt of an anti-MRSA medication had a significant protective effect. Conclusions. In an analysis controlling for host factors, colonization with MRSA carrying sep increased the risk of MRSA bacteremia. Identification of risk-adjusted genetic determinants of virulence may help to improve prediction of invasive disease and suggest new targets for therapeutic intervention. PMID:24041793
Bhargava, Kanika; Zhang, Yifan
This study was to understand the extent of methicillin-resistant coagulase-negative staphylococci (MRCoNS) serving as a mecA reservoir in retail meat. MRCoNS were isolated from retail meat (beef, chicken, and turkey) in Detroit and characterized by sodA gene sequencing for species identification, staphylococcal cassette chromosome mec (SCCmec) typing, and pulsed-field gel electrophoresis (PFGE). Unique MRCoNS isolates recovered from 25 meat samples were comprised of Staphylococcus sciuri (n = 13), Staphylococcus fleuretti (n = 4), Staphylococcus lentus (n = 3), Staphylococcus epidermidis (n = 2), Staphylococcus vitulinus (n = 1), Staphylococcus saprophyticus (n = 1) and Staphylococcus pasteuri (n = 1). Heterogeneous and composite SCCmec types, including I, III, IV, V, I + V and III + V were identified in 16 isolates. Same SCCmec types were recovered in different staphylococcal species and meat sources. Indistinguishable PFGE patterns were also observed in S. sciuri isolated from beef, chicken, and turkey, and with different SCCmec types. In conclusion, multiple CoNS species can serve as reservoirs for mecA. In addition to the clonal transmission of MRCoNS in meat, horizontal occurrence of SCCmec is observed in staphylococcal species.
Perencevich, Eli N.; David, Michael Z.
In the United States, methicillin-resistant Staphylococcus aureus (MRSA) with the USA300 pulsed-field gel electrophoresis type causes most community-associated MRSA infections and is an increasingly common cause of health care–associated MRSA infections. USA300 probably emerged during the early 1990s. To assess the spatiotemporal diffusion of USA300 MRSA and USA100 MRSA throughout the United States, we systematically reviewed 354 articles for data on 33,543 isolates, of which 8,092 were classified as USA300 and 2,595 as USA100. Using the biomedical literature as a proxy for USA300 prevalence among genotyped MRSA samples, we found that USA300 was isolated during 2000 in several states, including California, Texas, and midwestern states. The geographic mean center of USA300 MRSA then shifted eastward from 2000 to 2013. Analyzing genotyping studies enabled us to track the emergence of a new, successful MRSA type in space and time across the country. PMID:26484389
Moodley, Arshnee; Stegger, Marc; Ben Zakour, Nouri L; Fitzgerald, J Ross; Guardabassi, Luca
A putative staphylococcal protein A (spa) gene was discovered in the genome of Staphylococcus pseudintermedius and used for developing a species-specific spa typing protocol. Thirty-one clinical methicillin-resistant S. pseudintermedius (MRSP) isolates from dogs and cats in four countries were characterized by spa typing, pulsed-field gel electrophoresis (PFGE) and staphylococcal cassette chromosome (SCCmec) typing. The results indicated the occurrence of two MRSP clones that acquired distinct SCCmec elements in Europe (t02, PFGE type A, SCCmec type III,) and California (t06, PFGE type B, SCCmec type V). Sequence analysis of mecA revealed the occurrence of four alleles (mecA1 to mecA4), which correlated with the geographical origin of the isolates and enabled discrimination of two distinct subtypes within the European clone. The newly developed spa typing method appeared to be a promising tool for easy and rapid typing of MRSP, either alone or in combination with SCCmec and mecA typing for fine-structure epidemiological analysis.
Dakheel, Khulood Hamid; Abdul Rahim, Raha; Neela, Vasantha Kumari; Al-Obaidi, Jameel R; Hun, Tan Geok; Yusoff, Khatijah
Twenty-five methicillin-resistant Staphylococcus aureus (MRSA) isolates were characterized by staphylococcal protein A gene typing and the ability to form biofilms. The presence of exopolysaccharides, proteins, and extracellular DNA and RNA in biofilms was assessed by a dispersal assay. In addition, cell adhesion to surfaces and cell cohesion were evaluated using the packed-bead method and mechanical disruption, respectively. The predominant genotype was spa type t127 (22 out of 25 isolates); the majority of isolates were categorized as moderate biofilm producers. Twelve isolates displayed PIA-independent biofilm formation, while the remaining 13 isolates were PIA-dependent. Both groups showed strong dispersal in response to RNase and DNase digestion followed by proteinase K treatment. PIA-dependent biofilms showed variable dispersal after sodium metaperiodate treatment, whereas PIA-independent biofilms showed enhanced biofilm formation. There was no correlation between the extent of biofilm formation or biofilm components and the adhesion or cohesion abilities of the bacteria, but the efficiency of adherence to glass beads increased after biofilm depletion. In conclusion, nucleic acids and proteins formed the main components of the MRSA clone t127 biofilm matrix, and there seems to be an association between adhesion and cohesion in the biofilms tested.
Jang, Hyoyeong; Lim, Soo Hyeon; Choi, Jae Sue; Park, Youmie
Silver nanoparticles (AgNPs) were synthesized using Artemisia capillaris extracts in the absence and presence of stabilizers, and the antibacterial activities of the AgNPs against methicillin-resistant Staphylococcus aureus (MRSA) were determined by a minimum inhibitory concentration (MIC) assay. Two stabilizers, cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS), were utilized during the one-step synthesis to increase the colloidal stability of the AgNPs. The extract of A. capillaris was used as a reducing agent to convert silver ions into AgNPs. Sharp surface plasmon resonance bands in the range of 423-426 nm were observed in the UV-Visible spectra of the AgNPs, which indicated that the AgNPs were predominantly spherical shaped. Their average diameter, which ranged from 15.11 to 16.54 nm, was measured from high-resolution transmission electron microscopy images. Remarkably, the CTAB-stabilized AgNPs exhibited greater antibacterial activity (2- to 8-fold increase, MIC 15.6-62.5 μg/mL) against MRSA compared with the extract, AgNPs prepared in the absence of stabilizers, and SDS-stabilized AgNPs. The results indicate that CTAB-stabilized AgNPs prepared using plant extracts as reducing agents are very promising for the development of novel antibacterial agents.
Kraushaar, Britta; Fetsch, Alexandra
Staphylococcus aureus is an important food-borne pathogen due to the ability of enterotoxigenic strains to produce staphylococcal enterotoxins (SEs) in food. Methicillin-resistant S. aureus (MRSA) is also an important pathogen for humans, causing severe and hard to treat diseases in hospitals and in the community due to its multiresistance against antimicrobials. In particular, strains harbouring genes encoding for the Panton-Valentine leukocidin (PVL) toxin are of concern from a public health perspective as they are usually capable of causing severe skin and soft tissue infections (sSSTIs) and occasionally necrotizing pneumonia which is associated with high mortality. This is the first report on the detection of MRSA with genes encoding for PVL in wild boar meat. Among the 28 MRSA isolated from wild boar meat in the course of a national monitoring programme in Germany, seven harboured PVL-encoding genes. Six of the isolates were identical according to the results of spa-, MLST-, microarray- and PFGE-typing. They could be assigned to the epidemic MRSA clone USA300. Epidemiological investigations revealed that people handling the food were the most likely common source of contamination with these MRSA. These findings call again for suitable hygienic measures at all processing steps of the food production chain. The results of the study underline that monitoring along the food chain is essential to closely characterise the total burden of MRSA for public health.
Nickerson, Emma K.; Wuthiekanun, Vanaporn; Kumar, Varun; Amornchai, Premjit; Wongdeethai, Nattavut; Chheng, Kheng; Chantratita, Narisara; Putchhat, Hor; Thaipadungpanit, Janjira; Day, Nicholas P.; Peacock, Sharon J.
We previously described the first reported isolation of methicillin-resistant Staphylococcus aureus (MRSA) (a case series of pediatric community-associated MRSA infections) in Cambodia. We define the rate of pediatric MRSA carriage in the same population and characterize the associated bacterial genotypes by using pulsed-field gel electrophoresis and multilocus sequence typing. A prospective cohort study of MRSA carriage conducted over one month at the Angkor Hospital for Children, Siem Reap, Cambodia, identified MRSA carriage in 87 (3.5%) of 2,485 children who came to the outpatient department, and 6 (4.1%) of 145 inpatients, including at least two with cases of nosocomial acquisition. Genotyping of all 93 MRSA isolates resolved 5 genotypes. Most (91%) isolates were assigned to sequence type 834. Only 28 (32%) of 87 MRSA carriers identified in the outpatient department had no history of recent healthcare contact. The study findings have important implications for healthcare in a setting where diagnostic microbiology and access to antimicrobial drugs with efficacy against MRSA are limited. PMID:21292906
Background Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) was first reported in remote regions of Western Australia and is now the predominant MRSA isolated in the state. The objective of this study is to determine the genetic relatedness of Western Australian CA-MRSA clones within different multilocus sequence type (MLST) clonal clusters providing an insight into the frequency of S. aureus SCCmec acquisition within a region. Results The CA-MRSA population in Western Australia is genetically diverse consisting of 83 unique pulsed-field gel electrophoresis strains from which 46 MLSTs have been characterised. Forty five of these sequence types are from 18 MLST clonal clusters and two singletons. While SCCmec IV and V are the predominant SCCmec elements, SCCmec VIII and several novel and composite SCCmec elements are present. The emergence of MRSA in diverse S. aureus clonal clusters suggests horizontal transmission of the SCCmec element has occurred on multiple occasions. Furthermore DNA microarray and spa typing suggests horizontal transfer of SCCmec elements has also occurred within the same CC. For many single and double locus variant CA-MRSA clones only a few isolates have been detected. Conclusions Although multiple CA-MRSA clones have evolved in the Western Australian community only three clones have successfully adapted to the Western Australian community environment. These data suggest the successful evolution of a CA-MRSA clone may not only depend on the mobility of the SCCmec element but also on other genetic determinants. PMID:21955438
Kim, Dong Hee; Kim, Hye Kyung; Kim, Kang Min; Kim, Chun Kyu; Jeong, Min Ho; Ko, Chan Young; Moon, Kyung Ho; Kang, Jae Seon
The principal objective of this study was to evaluate the antibacterial activities of macrolactin A (MA) and 7-O-succinyl macrolactin A (SMA) generated from Bacillus polyfermenticus KJS-2 against vancomycin-resistant enterococci (VREs) and methicillin-resistant Staphylococcus aureus. The minimal inhibitory concentrations (MICs) of MA and SMA against VREs were 16 and 2∼1 μg/mL, respectively, and the MICs of MA and SMA against methicillin-resistant Staphylococcus aureus were 2 and < 0.25 μg/mL, respectively. Their MIC values were comparable or superior to those of teicoplanin. In evaluating the inhibitory effects of intestinal VRE colonization in mice, the oral MA and SMA effected a rapid inhibition of intestinal VRE colonization in mice, and the intraperitoneal SMA also inhibited VRE colonization, whereas intraperitoneal MA did not.
McMahon, T.J.; Basso, T.S.; Rummel, S.R.
Shunt resistance of cells in photovoltaic modules can affect module power output and could indicate flawed manufacturing processes and reliability problems. The authors describe a two-terminal diagnostic method to directly measure the shunt resistance of individual cells in a series-connected module non-intrusively, without deencapsulation. Peak power efficiency vs. light intensity was measured on a 12-cell, series-connected, single crystalline module having relatively high cell shunt resistances. The module was remeasured with 0.5-, 1-, and 2-ohm resistors attached across each cell to simulate shunt resistances of several emerging technologies. Peak power efficiencies decreased dramatically at lower light levels. Using the PSpice circuit simulator, they verified that cell shunt and series resistances can indeed be responsible for the observed peak power efficiency vs. intensity behavior. They discuss the effect of basic cell diode parameters, i.e., shunt resistance, series resistance, and recombination losses, on PV module performance as a function of light intensity.
Short communication: Prevalence of methicillin resistance in coagulase-negative staphylococci and Staphylococcus aureus isolated from bulk milk on organic and conventional dairy farms in the United States.
Cicconi-Hogan, K M; Belomestnykh, N; Gamroth, M; Ruegg, P L; Tikofsky, L; Schukken, Y H
The objective of this study was to evaluate the presence of methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus spp. in bulk tank milk samples from 288 organic and conventional dairy farms located in New York, Wisconsin, and Oregon from March 2009 to May 2011. Due to recent publications reporting the presence mecC (a mecA homolog not detected by traditional mecA-based PCR methods), a combination of genotypic and phenotypic approaches was used to enhance the recovery of methicillin-resistant organisms from bulk tank milk. In total, 13 isolates were identified as methicillin resistant: Staph. aureus (n=1), Staphylococcus sciuri (n=5), Staphylococcus chromogenes (n=2), Staphylococcus saprophyticus (n=3), Staphylococcus agnetis (n=1), and Macrococcus caseolyticus (n=1). The single methicillin-resistant Staph. aureus isolate was identified from an organic farm in New York, for an observed 0.3% prevalence at the farm level. The methicillin-resistant coagulase-negative staphylococci prevalence was 2% in the organic population and 5% in the conventional population. We did not identify mecC in any of the isolates from our population. Of interest was the relatively high number of methicillin-resistant Staph. sciuri recovered, as the number of isolates from our study was considerably higher than those recovered from other recent studies that also assessed milk samples. Our research suggests that the presence of a potential methicillin-resistant Staphylococcus reservoir in milk, and likely the dairy farm population in the United States, is independent of the organic or conventional production system.
Mukherjee, Nabanita; Sulaiman, Irshad M; Banerjee, Pratik
Indoor skin-contact surfaces of public fitness centers may serve as reservoirs of potential human transmission of methicillin-resistant Staphylococcus aureus (MRSA). We found a high prevalence of multidrug-resistant (MDR) MRSA of clonal complex 59 lineage harboring a variety of extracellular toxin genes from surface swab samples collected from inanimate surfaces of fitness centers in the Memphis metropolitan area, Tennessee. Our findings underscore the role of inanimate surfaces as potential sources of transmission of MDR MRSA strains with considerable genetic diversity.
Bandara, Aloka B; Zuo, Ziwei; Ramachandran, Siddharth; Ritter, Alfred; Heflin, James R; Inzana, Thomas J
Methicillin-resistance among Staphylococcus species is a major health problem in hospitals, communities, and animals. There is a need for culture-free diagnostic assays that can be carried out rapidly, and maintain a high degree of sensitivity and specificity. To address this need an ionic self-assembled multilayer (ISAM) film was deposited on the surface of a long-period grating (LPG) optical fiber by immersion alternately in poly-allylamine hydrochloride and in poly-1-[p-(3'-carboxy-4'-hydroxyphenylazo) benzenesulfonamido]-1,2-ethandiyl (PCBS), resulting in terminal carboxyl groups on the LPG-ISAM. The terminal carboxyl groups were covalently conjugated to monoclonal antibodies (MAb) specific to penicillin-binding-protein 2a of methicillin resistant (MR) staphylococci. After exposure of the LPG-ISAM to 10(2) colony forming units (CFU)/ml of MR S. aureus (MRSA) for 50 min., light transmission was reduced by 19.7%. In contrast, after exposure to 10(6) CFU/ml of methicillin-sensitive S. aureus (MSSA) attenuation of light transmission was less than 1.8%. Exposure of the LPG-ISAM to extracts of liver, lungs, or spleen from mice infected with MRSA attenuated light transmission by 11.7-73.5%. In contrast, exposure of the biosensor to extracts from MSSA-infected mice resulted in 5.6% or less attenuation of light transmission. When the sensor was tested with 36 strains of MR staphylococci, 15 strains of methicillin-sensitive staphylococci, 10 strains of heterologous genera (all at 10(4) CFU/ml), or tissue samples from mice infected with MRSA, there was complete agreement between MR and non-MR bacteria determined by antibiotic susceptibility testing and the biosensor assay when the cutoff value for attenuation of light transmission was 6.3%. Thus, the biosensor described has the potential to detect MR staphylococci in clinical samples with a high degree of sensitivity and specificity.
Zhang, Hongwei; Ma, Luyao; Ma, Lina; Hua, Marti Z; Wang, Shuo; Lu, Xiaonan
Methicillin-resistant Staphylococcus aureus (MRSA) is considered as one of the leading causes of food poisonings worldwide. Due to the high prevalence and extensive challenges in clinical treatment, a rapid and accurate detection method is required to differentiate MRSA from other S. aureus isolated from foods. Since the methicillin resistance of S. aureus is due to the acquisition of the mecA gene from staphylococcal chromosome cassette, the presence of the mecA gene is interpreted as a marker for the identification of MRSA. In this study, a low-cost lateral flow immunoassay (LFI) strip was used to detect the mecA amplicons subsequent to polymerase chain reaction (PCR). The specificity of this PCR-LFI assay was tested between MRSA and methicillin-susceptive S. aureus. Both the test line and control line were shown up on the LFI strip for MRSA, whereas only the control line developed for methicillin-susceptive S. aureus. The detection limit of PCR-LFI assay was 20fg for genomic DNA (100 times more sensitive than gel electrophoresis) and 2×10(0)CFU per 100g of pork products after enrichment at 37°C for 48h. The total detection time of using LFI was 3min, which was faster than the conventional electrophoresis (~45min). With the performance of PCR-LFI, 7 out of 42 S. aureus isolates were identified to be MRSA from imported pork products, which was consistent to the standardized minimum inhibitory concentration assay. This mecA-based PCR-LFI strip can be used for rapid and accurate detection of MRSA isolated from commercial pork products.
de Dios Caballero, Juan; Pastor, María Dolores; Vindel, Ana; Máiz, Luis; Yagüe, Genoveva; Salvador, Carme; Cobo, Marta; Morosini, María-Isabel; Cantón, Rafael
Resistance to linezolid (LZD) in methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients with cystic fibrosis (CF) is due mainly to ribosomal mutations. We report on four CF patients with LZD-resistant MRSA bronchopulmonary infections by strains carrying the cfr gene. Strains from one patient also harbored the G2576U mutation (23S rRNA) and the G139R substitution (L3 protein). All strains belonged to the epidemic clone ST125 MRSA IVc. Our results support the monitoring of LZD resistance emergence in CF and non-CF MRSA isolates. PMID:26666940
de Dios Caballero, Juan; Pastor, María Dolores; Vindel, Ana; Máiz, Luis; Yagüe, Genoveva; Salvador, Carme; Cobo, Marta; Morosini, María-Isabel; del Campo, Rosa; Cantón, Rafael
Resistance to linezolid (LZD) in methicillin-resistant Staphylococcus aureus (MRSA) isolates from patients with cystic fibrosis (CF) is due mainly to ribosomal mutations. We report on four CF patients with LZD-resistant MRSA bronchopulmonary infections by strains carrying the cfr gene. Strains from one patient also harbored the G2576U mutation (23S rRNA) and the G139R substitution (L3 protein). All strains belonged to the epidemic clone ST125 MRSA IVc. Our results support the monitoring of LZD resistance emergence in CF and non-CF MRSA isolates.
Rasool, Muhammad Hidayat; Yousaf, Rukhsana; Siddique, Abu Baker; Saqalein, Muhammad; Khurshid, Mohsin
Background In recent years, antibiotic resistance has been indicated as a paramount threat to public health. The use of bacteriophages appears to be a safer alternative for the control of bacterial infections. Objectives The present study aims to explore sewage water for the presence of indigenous bacteriophages, and to investigate their antibacterial potential against Methicillin-resistant Staphylococcus aureus (MRSA). Methods Bacterial isolates were first collected and identified from pus samples taken from the surgical and burn units using standard microbiological procedures. A cefoxitin disk screen test was then used and interpreted according to the clinical laboratory standards institute (CLSI) guidelines for the detection of MRSA. The sewage samples were processed and the phages enriched using S. aureus as a host organism. Turbid and clear plaques of different sizes were isolated using an overlay method, purified, and then enumerated by means of a dilution method. Results The phages exhibited good lytic activity against MRSA when tested in-vitro, and the highest activity was attained within three to six hours of phage infection. The isolated phage pq/48 was also found efficient in decreasing the bacterial count during an in-vivo trial in rabbits. A protein analysis using SDS-PAGE revealed 10 proteins of between 20 kDa and 155 kDa in size. Conclusions The overall results indicated that bacteriophages isolated from sewage exhibited excellent lytic activity against MRSA strains. In conclusion, bacteriophages can be further characterized and appear to be a promising candidate for phage therapy against MRSA in the future. PMID:27942361
Himsworth, Chelsea G.; Miller, Ruth R.; Montoya, Vincent; Hoang, Linda; Romney, Marc G.; Al-Rawahi, Ghada N.; Kerr, Thomas; Jardine, Claire M.; Patrick, David M.; Tang, Patrick; Weese, J. Scott
Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of multi-drug-resistant infections in people, particularly indigent populations. MRSA can be transmitted between people and domestic animals, but the potential for transmission between people and commensal pests, particularly rodents, had not been investigated. The objective of this study was to identify the presence and characterize the ecology of MRSA in rats (Rattus spp.) from in an impoverished, inner-city neighborhood. Oropharyngeal swabs were collected from rats trapped in 33 city blocks and one location within the adjacent port. Bacterial culture was performed and MRSA isolates were characterized using a variety of methods, including whole-genome sequencing (WGS). The ecology of MRSA in rats was described using phylogenetic analysis, geospatial analysis, and generalized linear mixed models. MRSA was identified 22 of 637 (3.5%) rats tested, although prevalence varied from 0 – 50% among blocks. Isolates belonged to 4 clusters according to WGS, with the largest cluster (n = 10) containing isolates that were genetically indistinguishable from community-acquired USA300 MRSA strains isolated from people within the study area. MRSA strains demonstrated both geographic clustering and dispersion. The odds of an individual rat carrying MRSA increased with increased body fat (OR = 2.53, 95% CI = 1.33 – 4.82), and in the winter (OR = 5.29, 95% CI = 1.04 – 26.85) and spring (OR = 5.50, 95% CI = 1.10 – 27.58) compared to the fall. The results show that urban rats carried the same MRSA lineages occurring in local human and/or animal populations, supporting recent transmission from external sources. MRSA carriage was influenced by season, most likely as a result of temporal variation in rat behavior and rat-human interactions. PMID:24498421
Mine, Yoshiko; Nakasone, Isamu; Yamamoto, Yuichi; Utani, Atsushi; Yamane, Nobuhisa; Uezato, Hiroshi; Takahashi, Kenzo
Panton-Valentine leukocidin (PVL) is a pore-forming cytotoxin that is produced by Staphylococcus aureus closely associated with skin and soft-tissue infections (SSTI). PVL-positive S. aureus strains have been identified worldwide, including in the USA; however, few studies have reported the presence of these strains in Japan. In this study, we prospectively investigated the prevalence of PVL in S. aureus strains from outpatients presenting with SSTI in Okinawa and characterized the PVL-positive S. aureus strains by polymerase chain reaction (PCR) and multilocus sequence typing (MLST). From 2008-2010, 499 clinical samples were obtained from 497 people. S. aureus was identified in 274 samples, and 36% (99 of 274) were methicillin-resistant S. aureus (MRSA). Seventeen (6.2%) PVL-positive S. aureus strains were detected by PCR, and 12 of the 17 PVL-positive strains were MRSA. Most PVL-positive S. aureus caused furuncles or carbuncles. Nine of the 17 PVL-positive isolates had an ST8 MRSA genotype and most harbored SCCmec type IVa and the arcA gene of the arginine catabolic mobile element, which is identical to the USA300 clone prevalent in the USA. PVL-positive S. aureus strains were more likely to be resistant to erythromycin (65%) and levofloxacin (53%). PVL-positive S. aureus strains have emerged and are spreading as a causative pathogen for SSTI in Okinawa.
Rodríguez-Mirones, Cristina; Acosta, Felix; Icardo, Jose M.; Martínez-Martínez, Luis; Ramos-Vivas, José
Ceftaroline (CPT) is a novel cephalosporin with in vitro activity against Staphylococcus aureus. Ceftaroline exhibits a level of binding affinity for PBPs in S. aureus including PBP2a of methicillin-resistant S. aureus (MRSA). The aims of this study were to investigate the morphological, physiological and molecular responses of MRSA clinical strains and MRSA biofilms to sub-MICs (1/4 and 1/16 MIC) of ceftaroline by using transmission, scanning and confocal microscopy. We have also used quantitative Real-Time PCR to study the effect of sub-MICs of ceftaroline on the expression of the staphylococcal icaA, agrA, sarA and sasF genes in MRSA biofilms. In one set of experiments, ceftaroline was able to inhibit biofilm formation in all strains tested at MIC, however, a strain dependent behavior in presence of sub-MICs of ceftaroline was shown. In a second set of experiments, destruction of preformed biofilms by addition of ceftaroline was evaluated. Ceftaroline was able to inhibit biofilm formation at MIC in all strains tested but not at the sub-MICs. Destruction of preformed biofilms was strain dependent because the biofilm formed by a matrix-producing strain was resistant to a challenge with ceftaroline at MIC, whereas in other strains the biofilm was sensitive. At sub-MICs, the impact of ceftaroline on expression of virulence genes was strain-dependent at 1/4 MIC and no correlation between ceftaroline-enhanced biofilm formation and gene regulation was established at 1/16 MIC. Our findings suggest that sub-MICs of ceftaroline enhance bacterial attachment and biofilm formation by some, but not all, MRSA strains and, therefore, stress the importance of maintaining effective bactericidal concentrations of ceftaroline to fight biofilm-MRSA related infections. PMID:26800524
Himsworth, Chelsea G; Miller, Ruth R; Montoya, Vincent; Hoang, Linda; Romney, Marc G; Al-Rawahi, Ghada N; Kerr, Thomas; Jardine, Claire M; Patrick, David M; Tang, Patrick; Weese, J Scott
Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of multi-drug-resistant infections in people, particularly indigent populations. MRSA can be transmitted between people and domestic animals, but the potential for transmission between people and commensal pests, particularly rodents, had not been investigated. The objective of this study was to identify the presence and characterize the ecology of MRSA in rats (Rattus spp.) from in an impoverished, inner-city neighborhood. Oropharyngeal swabs were collected from rats trapped in 33 city blocks and one location within the adjacent port. Bacterial culture was performed and MRSA isolates were characterized using a variety of methods, including whole-genome sequencing (WGS). The ecology of MRSA in rats was described using phylogenetic analysis, geospatial analysis, and generalized linear mixed models. MRSA was identified 22 of 637 (3.5%) rats tested, although prevalence varied from 0 - 50% among blocks. Isolates belonged to 4 clusters according to WGS, with the largest cluster (n = 10) containing isolates that were genetically indistinguishable from community-acquired USA300 MRSA strains isolated from people within the study area. MRSA strains demonstrated both geographic clustering and dispersion. The odds of an individual rat carrying MRSA increased with increased body fat (OR = 2.53, 95% CI = 1.33-4.82), and in the winter (OR = 5.29, 95% CI = 1.04-26.85) and spring (OR = 5.50, 95% CI = 1.10-27.58) compared to the fall. The results show that urban rats carried the same MRSA lineages occurring in local human and/or animal populations, supporting recent transmission from external sources. MRSA carriage was influenced by season, most likely as a result of temporal variation in rat behavior and rat-human interactions.
Bessa, Lucinda J; Palmeira, Andreia; Gomes, Ana S; Vasconcelos, Vitor; Sousa, Emília; Pinto, Madalena; Martins da Costa, Paulo
The extensive use of antimicrobials is leaving medicine with few effective therapeutic options to treat many infections due to the fact that many organisms developed resistance to commonly used drugs. It is therefore pertinent to search not only for new antimicrobials but also for compounds able to restore or potentiate the activity of existing antibiotics. We have screened a library consisting of 40 (thio)xanthone derivatives for antibacterial activity and possible synergistic effects when used in combination with antibiotics. Nine out of the 40 compounds exhibited antibacterial activity against Gram-positive bacteria. Two xanthone derivatives, 1-formyl-4-hydroxy-3-methoxy (7), 2-formyl-3-hydroxy-4-methoxyxanthone (8) and the thioxanthone derivative 1-((2-(diethylamino)ethyl)amino)-4-propoxythioxanthone (10) and its hydrochloride form 13, showed activity against a methicillin-resistant Staphylococcus aureus (MRSA) isolate with minimum inhibitory concentration (MIC) values lower than 256 μg/ml. Thioxanthone 10 demonstrated antibacterial activity and also synergy when combined with ampicillin and oxacillin against MRSA. Additionally, thioxanthone 1-(piperidin-1-yl)-4-propoxythioxanthone (9), despite not having antibacterial activity presented remarkable synergy with oxacillin against MRSA; the MIC of tioxanthone 9 and oxacillin when both were in combination were 128 and 8 μg/ml, respectively. Thioxanthones 9 and 10 were also found to be synergistic when both were combined. Subsequently, docking simulations between thioxanthones 9 and 10 and the allosteric domain of penicillin-binding protein 2A (PBP2A) were undertaken in AutoDock Vina. Both compounds had the ability to bind with an allosteric domain of PBP2A, which may explain their synergy with oxacillin. These two thioxanthone derivatives with different profiles may be promising tools for restoring the activity of oxacillin against MRSA.
Buultjens, Andrew H.; Giulieri, Stefano; Owusu-Mireku, Evelyn; Aboagye, Samuel Y.; Baines, Sarah L.; Gonçalves da Silva, Anders; Howden, Benjamin P.; Pluschke, Gerd; Yeboah-Manu, Dorothy
Background The emergence and evolution of community-acquired methicillin resistant Staphylococcus aureus (CA-MRSA) strains in Africa is poorly understood. However, one particular MRSA lineage called ST88, appears to be rapidly establishing itself as an “African” CA-MRSA clone. In this study, we employed whole genome sequencing to provide more information on the genetic background of ST88 CA-MRSA isolates from Ghana and to describe in detail ST88 CA-MRSA isolates in comparison with other MRSA lineages worldwide. Methods We first established a complete ST88 reference genome (AUS0325) using PacBio SMRT sequencing. We then used comparative genomics to assess relatedness among 17 ST88 CA-MRSA isolates recovered from patients attending Buruli ulcer treatment centres in Ghana, three non-African ST88s and 15 other MRSA lineages. Results We show that Ghanaian ST88 forms a discrete MRSA lineage (harbouring SCCmec-IV [2B]). Gene content analysis identified five distinct genomic regions enriched among ST88 isolates compared with the other S. aureus lineages. The Ghanaian ST88 isolates had only 658 core genome SNPs and there was no correlation between phylogeny and geography, suggesting the recent spread of this clone. The lineage was also resistant to multiple classes of antibiotics including β-lactams, tetracycline and chloramphenicol. Discussion This study reveals that S. aureus ST88-IV is a recently emerging and rapidly spreading CA-MRSA clone in Ghana. The study highlights the capacity of small snapshot genomic studies to provide actionable public health information in resource limited settings. To our knowledge this is the first genomic assessment of the ST88 CA-MRSA clone. PMID:28265515
Street, Cale N.; Pedigo, Lisa; Gibbs, Aaron; Loebel, Nicolas G.
The nosocomial infection rate has increased dramatically due to emergence of antibiotic resistant bacterial strains such as methicillin resistant Staphylococcus aureus (MRSA). The primary anatomical site of MRSA colonization is the anterior nares, and this reservoir represents a primary vector of transmission from non-infected carriers to susceptible individuals. Antimicrobial photodynamic therapy (aPDT) has been used successfully for topical disinfection in the oral cavity. The aim of this study was to evaluate the utility of aPDT for nasal MRSA decolonization at the preclinical and clinical level. The nasal aPDT system consists of a 670 nm diode laser fibre-optically coupled to a disposable light diffusing tip, used to activate a methylene blue based photosensitizer formulation. Preclinical testing was done both in a custom nasal reservoir model and on human skin cultures colonized on the epithelial surface with MRSA. Human clinical testing was performed by clinicians in regions in which the system is approved by the regulatory authority. In vitro testing demonstrated that aPDT eradicated planktonic MRSA in an energy and photosensitizer concentration dependent manner. Furthermore, aPDT eliminated sustained colonization of MRSA on cultured human epithelial surfaces, an effect that was sustained over multiple days post-treatment. In preliminary human testing, aPDT eradicated MRSA completely from the nose with total treatment times <10 minutes. aPDT is effective against MRSA when used topically in the nose. Energy dose and photosensitizer parameters have been optimized for the nasal environment. Controlled clinical studies are currently underway to further evaluate safety and efficacy.
Cohen, Philip R
A healthy 19-year-old black man without any methicillin-resistant Staphylococcus aureus risk factors developed axillary boils after he began lifting weights at the university gym in Houston, TX. He presented with a large tender erythematous fluctuant abscess of his right axillae and a superiorly located smaller painful red indurated nodule; the surrounding cellulitis extended into the adjacent tissue (Figure). The abscess was incised, drained, and cultured. Empiric treatment with cephalexin 500 mg q.i.d. was given for 7 days. The culture grew methicillin-resistant S. aureus. Susceptibility testing of the S. aureus isolate was performed by Laboratory Corp. of America (Houston, TX); the Vitek system (Biomerieux, Hazelwood, MO) was used, and the specimen was incubated for 8 hours. Confirmation of methicillin resistance was performed using a methicillin-resistant S. aureus plate and the specimen was incubated for 24 hours. In addition to resistance to methicillin, the bacterial isolate was also resistant to ciprofloxacin, erythromycin, and penicillin. The S. aureus strain had intermediate susceptibility to levofloxacin and was susceptible to clindamycin, gentamicin, rifampin, tetracycline, trimethoprim/sulfamethoxazole, and vancomycin. The infection persisted and the antibiotic was changed to double strength trimethoprim/sulfamethoxazole, taken twice daily for 15 days. In addition, topical care included lesional and intranasal application of mupirocin 2% ointment and daily cleaning of the area with 10% povidone-iodine liquid soap. The skin infection completely resolved without recurrence within 2 weeks.
Vincze, Szilvia; Paasch, Angela; Walther, Birgit; Ruscher, Claudia; Lübke-Becker, Antina; Wieler, Lothar H; Barbara, Kohn
A case of a dog with a long-term inflammatory skin disorder due to infection with methicillin-resistant Staphylococcus pseudintermedius (MRSP) is described. After initial diagnostics of MRSP, follow-up swabs of the dog (nose, skin) were taken twice after four and seven weeks. MRSP was constantly isolated from the skin and once from the nose. Since infected humans might be a source of reinfection, the owners of the dog were screened (nasal) three times during their pet's therapy. Thereby, the male owner was found to be colonized with MRSP once in the first sampling round. Comparative typing of all MRSP-isolates by pulsed-field gel electrophoresis (PFGE), SCCmec typing, multilocus sequence typing (MLST), spa typing, PCR-detection of the leukotoxin encoding operon (LukI) and the Staphylococcus intermedius-exfoliative toxin (SIET) as well as antimicrobial resistance profiling by broth microdilution revealed that all five MRSP isolates from the dog and the single isolate from the owner were indistinguishable by any of the applied methods. All isolates were assigned to a certain strain, a multidrug-resistant MRSP belonging to sequence type (ST) 71, spa type (t)05, harbouring SCCmecIII as well as the genes encoding LukI and SIET. In this case, a number of reasons might have contributed to therapy failure and re-infection, respectively (e. g. contact to other MRSP-colonized dogs, contact to MRSP-colonized humans, refusal to clip the dog's fur). In addition, MRSP-contaminated objects or surfaces in the household, which were difficult to disinfect or simply not considered as a potential source of MRSP, might have served as a source of re-infection. These results envision the possibility of a dog-to-human transmission of MRSP and the relevance of this aspect as a potential source of re-infection in cases of bacterial-supported long-term skin disorders in canine patients. First cases of MRSP infections in humans have been described only recently. However, the general
Rivera, Ana Maria; Boucher, Helen W
Gram-positive bacteria cause a broad spectrum of disease in immunocompetent and immunocompromised hosts. Despite increasing knowledge about resistance transmission patterns and new antibiotics, these organisms continue to cause significant morbidity and mortality, especially in the health care setting. Methicillin-resistant Staphylococcus aureus poses major problems worldwide as a cause of nosocomial infection and has emerged as a cause of community-acquired infections. This change in epidemiology affects choices of empirical antibiotics for skin and skin-structure infections and community-acquired pneumonia in many settings. Throughout the world, the treatment of community-acquired pneumonia and other respiratory tract infections caused by penicillin-resistant Streptococcus pneumoniae has been complicated by resistance to β-lactam and macrolide antibacterial drugs. Vancomycin-resistant enterococci are a major cause of infection in the hospital setting and remain resistant to treatment with most standard antibiotics. Treatment of diseases caused by resistant gram-positive bacteria requires appropriate use of available antibiotics and stewardship to prolong their effectiveness. In addition, appropriate and aggressive infection control efforts are vital to help prevent the spread of resistant pathogens.
Majcherczyk, Paul A; McKenna, Therese; Moreillon, Philippe; Vaudaux, Pierre
To explore the discriminatory power of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for detecting subtle differences in isogenic isolates, we tested isogenic strains of Staphylococcus aureus differing in their expression of resistance to methicillin or teicoplanin. More important changes in MALDI-TOF MS spectra were found with strains differing in methicillin than in teicoplanin resistance. In comparison, very minor or no changes were recorded in pulsed-field gel electrophoresis profiles or peptidoglycan muropeptide digest patterns of these strains, respectively. MALDI-TOF MS might be useful to detect subtle strain-specific differences in ionizable components released from bacterial surfaces and not from their peptidoglycan network.
Kadlec, Kristina; Schwarz, Stefan; Goering, Richard V; Weese, J Scott
Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged in a remarkable manner as an important problem in dogs and cats. However, limited molecular epidemiological information is available. The aims of this study were to apply direct repeat unit (dru) typing in a large collection of well-characterized MRSP isolates and to use dru typing to analyze a collection of previously uncharacterized MRSP isolates. Two collections of MRSP isolates from dogs and cats were included in this study. The first collection comprised 115 well-characterized MRSP isolates from North America and Europe. The data for these isolates included multilocus sequence typing (MLST) and staphylococcal protein A gene (spa) typing results as well as SmaI macrorestriction patterns after pulsed-field gel electrophoresis (PFGE). The second collection was a convenience sample of 360 isolates from North America. The dru region was amplified by PCR, sequenced, and analyzed. For the first collection, the discriminatory indices of the typing methods were calculated. All isolates were successfully dru typed. The discriminatory power for dru typing (D = 0.423) was comparable to that of spa typing (D = 0.445) and of MLST (D = 0.417) in the first collection. Occasionally, dru typing was able to further discriminate between isolates that shared the same spa type. Among all 475 isolates, 26 different dru types were identified, with 2 predominant types (dt9a and dt11a) among 349 (73.4%) isolates. The results of this study underline that dru typing is a useful tool for MRSP typing, being an objective, standardized, sequence-based method that is relatively cost-efficient and easy to perform.
Schwarz, Stefan; Goering, Richard V.; Weese, J. Scott
Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged in a remarkable manner as an important problem in dogs and cats. However, limited molecular epidemiological information is available. The aims of this study were to apply direct repeat unit (dru) typing in a large collection of well-characterized MRSP isolates and to use dru typing to analyze a collection of previously uncharacterized MRSP isolates. Two collections of MRSP isolates from dogs and cats were included in this study. The first collection comprised 115 well-characterized MRSP isolates from North America and Europe. The data for these isolates included multilocus sequence typing (MLST) and staphylococcal protein A gene (spa) typing results as well as SmaI macrorestriction patterns after pulsed-field gel electrophoresis (PFGE). The second collection was a convenience sample of 360 isolates from North America. The dru region was amplified by PCR, sequenced, and analyzed. For the first collection, the discriminatory indices of the typing methods were calculated. All isolates were successfully dru typed. The discriminatory power for dru typing (D = 0.423) was comparable to that of spa typing (D = 0.445) and of MLST (D = 0.417) in the first collection. Occasionally, dru typing was able to further discriminate between isolates that shared the same spa type. Among all 475 isolates, 26 different dru types were identified, with 2 predominant types (dt9a and dt11a) among 349 (73.4%) isolates. The results of this study underline that dru typing is a useful tool for MRSP typing, being an objective, standardized, sequence-based method that is relatively cost-efficient and easy to perform. PMID:26378275
Background Nasal colonization with methicillin-resistant Staphylococcus aureus (MRSA) is a well defined risk factor for subsequent bacteremia and death in various groups of patients, but its impact on outcome in patients receiving long-term hemodialysis (HD) is under debate. Methods This prospective interventional cohort study (performed 2004 to 2010) enrolled 289 HD outpatients of an urban dialysis-unit. Nasal swab cultures for MRSA were performed in all patients upon first admission, at transfer from another dialysis facility or readmission after hospitalisation. Nasal MRSA carriers were treated in a separate ward and received mupirocin nasal ointment. Concomitant extra-nasal MRSA colonization was treated with 0.2% chlorhexidine mouth rinse (throat) or octenidine dihydrochloride containing antiseptic soaps and 2% chlorhexidine body washes (skin). Clinical data and outcome of carriers and noncarriers were systematically analyzed. Results The screening approach identified 34 nasal MRSA carriers (11.7%). Extra-nasal MRSA colonization was observed in 11/34 (32%) nasal MRSA carriers. History of malignancy and an increased Charlson Comorbidity Index were significant predictors for nasal MRSA carriers, whereas traditional risk factors for MRSA colonization or markers of inflammation or malnutrition were not able to discriminate. Kaplan-Meier analysis demonstrated significant survival differences between MRSA carriers and noncarriers. Mupirocin ointment persistently eliminated nasal MRSA colonization in 26/34 (73.5%) patients. Persistent nasal MRSA carriers with failure of this eradication approach had an extremely poor prognosis with an all-cause mortality rate >85%. Conclusions Nasal MRSA carriage with failure of mupirocin decolonization was associated with increased mortality despite a lack of overt clinical signs of infection. Further studies are needed to demonstrate whether nasal MRSA colonization represents a novel predictor of worse outcome or just another
Althaqafi, Abdulhakeem O; Matar, Madonna J; Moghnieh, Rima; Alothman, Adel F; Alenazi, Thamer H; Farahat, Fayssal; Corman, Shelby; Solem, Caitlyn T; Raghubir, Nirvana; Macahilig, Cynthia; Haider, Seema; Stephens, Jennifer M
Objectives The objective of this study is to describe the real-world treatment patterns and burden of suspected or confirmed methicillin-resistant Staphylococcus aureus (MRSA) pneumonia in Saudi Arabia and Lebanon. Methods A retrospective chart review study evaluated 2011–2012 data from hospitals in Saudi Arabia and Lebanon. Patients were included if they had been discharged with a diagnosis of MRSA pneumonia, which was culture proven or suspected based on clinical criteria. Hospital data were abstracted for a random sample of patients to capture demographics (eg, age and comorbidities), treatment patterns (eg, timing and use of antimicrobials), hospital resource utilization (eg, length of stay), and clinical outcomes (eg, clinical status at discharge and mortality). Descriptive results were reported using frequencies or proportions for categorical variables and mean and standard deviation for continuous variables. Results Chart-level data were collected for 93 patients with MRSA pneumonia, 50 in Saudi Arabia and 43 in Lebanon. The average age of the patients was 56 years, and 60% were male. The most common comorbidities were diabetes (39%), congestive heart failure (30%), coronary artery disease (29%), and chronic obstructive pulmonary disease (28%). Patients most frequently had positive cultures from pulmonary (87%) and blood (27%) samples. All isolates were sensitive to vancomycin, teicoplanin, and linezolid, and only one-third of the isolates tested were sensitive to ciprofloxacin. Beta-lactams (inactive therapy for MRSA) were prescribed 21% of the time across all lines of therapy, with 42% of patients receiving first-line beta-lactams. Fifteen percent of patients did not receive any antibiotics that were considered to be MRSA active. The mean hospital length of stay was 32 days, and in-hospital mortality was 30%. Conclusion The treatment for MRSA pneumonia in Saudi Arabia and Lebanon may be suboptimal with inactive therapy prescribed a substantial proportion
Lebeaux, David; Barbier, François; Angebault, Cécile; Benmahdi, Lahcene; Ruppé, Etienne; Felix, Benjamin; Gaillard, Kevin; Djossou, Félix; Epelboin, Loïc; Dupont, Claire; Renard, Magaly; Peroz, Gilles; Vandenesch, François; Wolff, Michel; Andremont, Antoine
Nasal carriage of methicillin-resistant coagulase-negative staphylococci (MR-CoNS) is highly prevalent in community subjects, but its dynamic has been little investigated. Nasal swabbing was performed in 2006 and 2008 in 154 Amerindians living isolated in French Guiana. MR-CoNS strains were identified and characterized by non-β-lactam susceptibility testing and staphylococcal cassette chromosome mec element (SCCmec) typing, characterizing the associations of ccr and mec gene complex allotypes, and for MR Staphylococcus epidermidis (MRSE), multilocus variable number of tandem repeats analysis (MLVA) was used. The impact of sociodemographic and medical characteristics on the persistence of MR-CoNS carriage was assessed by bivariate analysis. Prevalence of MR-CoNS carriage was 50.6% in 2006 and 46.8% in 2008. The 274 MR-CoNS isolates, including S. epidermidis (n = 89, 62 MLVA patterns), Staphylococcus haemolyticus (n = 78), and Staphylococcus hominis (n = 72), exhibited 41 distinct ccr and mec gene complex associations. Persistent carriage (in 2006 and 2008), intermittent carriage (either in 2006 or 2008), and noncarriage were documented in 25.3, 47.4, and 27.3% of the participants, respectively. Persistent carriage of a given MRSE isolate was rarely observed (n = 8 isolates). Furthermore, no epidemiological factor, including antibiotic exposure, was associated with persistent carriage. The high diversity of MRSE clones and their ccr and mec gene complex associations contrasted with the high carriage rates in this isolated community, which might reflect the occurrence of SCCmec rearrangement and the generation of new MR-CoNS strains. PMID:22064532
Rasschaert, G; Vanderhaeghen, W; Dewaele, I; Janez, N; Huijsdens, X; Butaye, P; Heyndrickx, M
This study evaluates the multiple-locus variable-number tandem-repeat assay (MLVA) and pulsed-field gel electrophoresis (PFGE) when using restriction enzymes BstZI, SacII, and ApaI to fingerprint a diverse collection of methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) sequence type 398 (ST398) isolates. These isolates had been characterized previously by multilocus sequence typing, spa typing, and staphylococcal cassette chromosome mec (SCCmec) typing. Typeability and discriminatory power were analyzed, and the concordance between the various methods was determined. All MRSA ST398 isolates were typeable by the MLVA and PFGE using BstZI, SacII, and ApaI. With each method, the MRSA ST398 isolates formed a separate group from the two non-ST398 MRSA strains. PFGE, performed with any of the three restriction enzymes, had the most discriminatory power, followed by MLVA, spa typing, and SCCmec typing. The MLVA showed the highest concordance with PFGE using ApaI and spa typing. As further expressed by the Wallace coefficient, the MLVA type was poorly predicted by spa typing, whereas the spa type was well predicted by MLVA. PFGE, using a combination of all three restriction enzymes, had the highest concordance with the MLVA but had a low probability of being predicted by MLVA. PFGE, using a combination of all three restriction enzymes, was able to predict SCCmec type and MLVA type completely and had a high probability of predicting spa type. Both the MLVA and PFGE could be used to discriminate among the MRSA ST398 isolates. Although the MLVA is a faster technique, PFGE had more discriminatory power than the MLVA, especially when a combination of restriction enzymes was used.
Johnson, Amanda; Nava-Hoet, Rocio C.; Bateman, Shane; Hillier, Andrew; Dyce, John; Gebreyes, Wondwossen A.; Wittum, Thomas E.
Abstract Concurrent to reports of zoonotic and nosocomial transmission of methicillin-resistant Staphylococcus aureus (MRSA) in veterinary settings, recent evidence indicates that the environment in veterinary hospitals may be a potential source of MRSA. The present report is a cross-sectional study to determine the prevalence of MRSA on specific human and animal contact surfaces at a large veterinary hospital during a nonoutbreak period. A total of 156 samples were collected using Swiffers® or premoistened swabs from the small animal, equine, and food animal sections. MRSA was isolated and identified by pre-enrichment culture and standard microbiology procedures, including growth on Mueller-Hinton agar supplemented with NaCl and oxacillin, and by detection of the mecA gene. Staphylococcal chromosome cassette mec (SCCmec) typing and pulsed-field gel electrophoresis profile were also determined. MRSA was detected in 12% (19/157) of the hospital environments sampled. The prevalence of MRSA in the small animal, equine, and food animal areas were 16%, 4%, and 0%, respectively. Sixteen of the MRSA isolates from the small animal section were classified as USA100, SCCmec type II, two of which had pulsed-field gel electrophoresis pattern that does not conform to any known type. The one isolate obtained from the equine section was classified as USA500, SCCmec type IV. The molecular epidemiological analysis revealed a very diverse population of MRSA isolates circulating in the hospital; however, in some instances, multiple locations/surfaces, not directly associated, had the same MRSA clone. No significant difference was observed between animal and human contact surfaces in regard to prevalence and type of isolates. Surfaces touched by multiple people (doors) and patients (carts) were frequently contaminated with MRSA. The results from this study indicate that MRSA is present in the environment even during nonoutbreak periods. This study also identified specific surfaces in
Popovich, Kyle J.; Snitkin, Evan; Green, Stefan J.; Aroutcheva, Alla; Hayden, Mary K.; Hota, Bala; Weinstein, Robert A.
Background. In a community, it is unknown what factors account for transmission of methicillin-resistant Staphylococcus aureus (MRSA). We integrated whole genome sequencing (WGS) and epidemiologic data to identify factors associated with MRSA transmission networks in an urban community. Methods. WGS was performed on colonizing USA300 MRSA isolates from 74 individuals within 72 hours of admission to a public hospital in Chicago, IL. Single nucleotide variants (SNVs) were used to reconstruct the phylogeny of sequenced isolates, and epidemiologic data was overlaid to identify factors associated with transmission networks. Results. The maximum within-patient SNV difference for an individual with multisite colonization was 41 SNVs, with no systematic divergence among body sites. We observed a minimum of 7 SNVs and maximum of 153 SNVs between isolates from different individuals. We identified 4 pairs of individuals whose isolates were within 40 SNVs of each other. Putting our isolates in the context of previously sequenced USA300 isolates from other communities, we identified a 13-member group and two 4-member groups that represent samples from putative local transmission networks. Individuals in these groups were more likely to be African American, to be human immunodeficiency virus–infected, to reside in high detainee release areas, and to be current users of illicit drugs. Conclusions. Using WGS, we observed potential transmission networks in an urban community and that certain epidemiologic factors were associated with inclusion in these networks. Future work with contact tracing and advanced molecular diagnostics may allow for identification of MRSA “epicenters” in the community where interventions can be targeted. PMID:26347509
Popovich, Kyle J.; Hota, Bala; Aroutcheva, Alla; Kurien, Lisa; Patel, Janki; Lyles-Banks, Rosie; Grasso, Amanda E.; Spec, Andrej; Beavis, Kathleen G.; Hayden, Mary K.; Weinstein, Robert A.
Background. The epidemic of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has had a disproportionate impact on patients with human immunodeficiency virus (HIV). Methods. We evaluated CA-MRSA colonization burden (number of colonized sites per total number sampled) among HIV-infected and HIV-negative inpatients within 72 hours of hospitalization. From March 2011 through April 2012, we obtained cultures from nasal and extranasal sites (throat, axilla, inguinal, perirectal, and chronic wound if present) and collected risk factor data. Results. Of 745 patients (374 HIV-infected, 371 HIV-negative), 15.7% were colonized with CA-MRSA at any site: 20% of HIV and 11% of HIV-negative patients (relative prevalence = 1.8, P = .002). HIV-infected patients had a higher prevalence of nasal, extranasal, and exclusive extranasal colonization as well as higher colonization burden. Perirectal and inguinal areas were the extranasal sites most frequently colonized, and 38.5% of colonized patients had exclusive extranasal colonization. Seventy-three percent of isolates were identified as USA300. Among HIV-infected patients, male sex, younger age, and recent incarceration were positively associated whereas Hispanic ethnicity was negatively associated with higher colonization burden. Among HIV-negative patients, temporary housing (homeless, shelter, or substance abuse center) was the only factor associated with higher colonization burden. Predictors of USA300 included HIV, younger age, illicit drug use, and male sex; all but 1 colonized individual with current or recent incarceration carried USA300. Conclusions. HIV-infected patients were more likely to have a higher CA-MRSA colonization burden and carry USA300. In certain populations, enhanced community and outpatient-based infection control strategies may be needed to prevent CA-MRSA cross-transmission and infection. PMID:23325428
Kaur, Sandeep; Harjai, Kusum; Chhibber, Sanjay
Phage therapy presents an alternative approach against the emerging methicillin-resistant Staphylococcus aureus (MRSA) threat. Some of the problems encountered during isolation of MRSA phages include the high prevalence of enteric phages in natural sources, nonspecific absorption of viable phage, and the formation of pinpoint or tiny plaques. The phage isolated in this study, MR-5, also formed tiny plaques against its host S. aureus ATCC 43300 (MRSA), making its detection and enumeration difficult. An improved method of increasing the plaque size of MRSA phage by incorporating sublethal concentrations of three different classes of antibiotics (inhibitors of protein synthesis) in the classical double-layer agar (DLA) method was investigated. The β-lactam and quinolone antibiotics commonly employed in earlier studies for increasing the plaque size did not show any significant effect on the plaque size of isolated MR-5 phage. Linezolid (oxazolidinone class), tetracycline, and ketolide antibiotics brought significant enhancements (3 times the original size) in the plaque size of MR-5 phage. Prior treatment with these antibiotics resulted in significant reductions in the time of adsorption and the latent period of MR-5 phage. To rule out whether the action of linezolid (which brought the maximum increase in plaque size) was specific for a single phage only, its effect on the plaque size of seven other S. aureus-specific phages was also assessed. Significant enhancements in the plaque size of these phages were observed. These results indicate that this modification can therefore safely be incorporated in the traditional DLA overlay method to search for new MRSA-virulent phages.
Orellana, Robert C; Hoet, Armando E; Bell, Christopher; Kelley, Christina; Lu, Bo; Anderson, Sarah E; Stevenson, Kurt B
The objective was to determine the nasal carriage prevalence of methicillin-resistant Staphylococcus aureus (MRSA) among emergency medical service (EMS) personnel and the associated risk factors. A cross-sectional study was conducted among Ohio EMS personnel randomly sampled from 84 urban and rural agencies. Surveys assessing demographics, occupational history, health, cohabitation status, and hygiene practice were collected with nasal swabs from those who enrolled. Survey weight adjusted analysis was performed (1) to estimate MRSA nasal carriage prevalence of Ohio EMS providers, and (2) to identify variables associated with MRSA. MRSA was detected in 4.6% (13/280) EMS personnel sampled. After employing a survey-weighted analysis the following risk factors associated with MRSA carriage were identified: those who did not practice frequent hand hygiene after glove use (OR, 10.51; 95% CI, 2.54-43.45; P = 0.0012), living with someone with a recent staphylococcal infection (OR, 9.02; 95% CI, 1.03-78.98; P = 0.0470), and individuals with low frequency of hand washing (< 8 times per shift) (OR, 4.20; 95% CI 1.02-17.27; P = 0.0468). An additional risk factor identified through the logistic regression analysis on the study population was EMS workers with an open wound or skin infection (OR, 6.75; 95% CI, 1.25-36.36; P = 0.0262). However, this was not significant in the survey-weighted analysis. The high prevalence of MRSA in Ohio EMS personnel is both an occupational hazard and patient safety concern. Implementing methods to reinforce CDC guidelines for proper hygiene could decrease MRSA found in the EMS setting. Previous literature suggests that a reduction in MRSA colonization can lead to decreases in transmission and improved health for both patients and personnel.
Maeda, Yasunori; Loughrey, Anne; Earle, J A Philip; Millar, B Cherie; Rao, Juluri R; Kearns, Angela; McConville, Ogie; Goldsmith, Colin E; Rooney, Paul J; Dooley, James S G; Lowery, Colm J; Snelling, William J; McMahon, Ann; McDowell, David; Moore, John E
Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has now been described globally, as a clinically significant pathogen, particularly associated with skin and soft tissue infections, including abscesses, cellulitis and furunculosis. The recent emergence of CA-MRSA combined with its predominant presentation associated with skin and soft tissue infection, the previous literature indicating honey as an effective treatment of healthcare-associated HA-MRSA-related wound infection, as well as honey's ease of topical application, make the current study timely and of interest to healthcare practitioners involved with wound management. Although previous studies have examined the antimicrobial activity of honey against HA-MRSA, such data are limited regarding the activity of honey against this emerging type of MRSA. CA-MRSA (n=6 isolates), was examined for its susceptibility to natural honey (n=3 honey produced from bees in Northern Ireland and one commercial French honey). Results demonstrated that all honey was able to reduce the cultural count of all CA-MRSA from approximately 10(6) colony-forming units (cfus) (mean = 6.46 log10 cfu/g) to none detectable within 24h of co-culture of separate CA-MRSA organisms individually with all four-honey types examined. Subsequent non-selective enrichment of honey demonstrated that inoculated honey remained positive for CA-MRSA until 72h postinoculation, after which point no culturable organisms could be detected. This study demonstrated that, in vitro, these natural products had an antimicrobial activity against the CA-MRSA organisms tested. Further studies are now required to demonstrate if this antimicrobial activity has any clinical application.
Chen, Yi-Guo; Zhang, Yong; Deng, Lin-Qiang; Chen, Hui; Zhang, Yu-Juan; Zhou, Nan-Jin; Yuan, Keng; Yu, Li-Zhi; Xiong, Zhang-Hua; Gui, Xiao-Mei; Yu, Yan-Rong; Wu, Xiao-Mu; Min, Wei-Ping
The spread of methicillin-resistant Staphylococcus aureus (MRSA) is a critical health issue that has drawn greater attention to the potential use of immunotherapy. Toll-like receptor 2 (TLR2), a pattern recognition receptor, is an essential component in host innate defense system against S. aureus infection. However, little is known about the innate immune response, specifically TLR2 activation, against MRSA infection. Here, we evaluate the protective effect and the mechanism of MRSA murine pneumonia after pretreatment with Pam3CSK4, a TLR2 agonist. We found that the MRSA-pneumonia mouse model, pretreated with Pam3CSK4, had reduced bacteria and mortality in comparison to control mice. As well, lower protein and mRNA levels of TNF-α, IL-1β and IL-6 were observed in lungs and bronchus of the Pam3CSK4 pretreatment group. Conversely, expression of anti-inflammatory cytokine IL-10, but not TGF-β, increased in Pam3CSK4-pretreated mice. Our additional studies showed that CXCL-2 and CXCL1, which are necessary for neutrophil recruitment, were less evident in the Pam3CSK4-pretreated group compared to control group, whereas the expression of Fcγ receptors (FcγⅠ/Ⅲ) and complement receptors (CR1/3) increased in murine lungs. Furthermore, we found that increased survival and improved bacterial clearance were not a result of higher levels of neutrophil infiltration, but rather a result of enhanced phagocytosis and bactericidal activity of neutrophils in vitro and in vivo as well as increased robust oxidative activity and release of lactoferrin. Our cumulative findings suggest that Pam3CSK4 could be a novel immunotherapeutic candidate against MRSA pneumonia. PMID:26974438
Verhegghe, Marijke; Crombé, Florence; Luyckx, Kaat; Haesebrouck, Freddy; Butaye, Patrick; Herman, Lieve; Heyndrickx, Marc; Rasschaert, Geertrui
Since the first description of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA), a high prevalence was observed in pigs. At present, questions remain about the transmission of LA-MRSA to the general human population through pork. The objectives of the present study were to determine the prevalence of LA-MRSA in Belgian pork and to determine the role of the pork production chain and butcheries in transmission of LA-MRSA to the human population. Pig meat samples (chops, bacon, minced pork, ribs, forelimbs, and ears; n = 137) originating from four butcheries (A through D) were spread plated on ChromID MRSA plates both before and after overnight enrichment culture. Suspect colonies were confirmed using a MRSA-specific triplex PCR assay and a CC398-specific PCR assay. The isolates (n = 147) were further characterized by SCCmec typing, multiple-locus variable number tandem repeat analysis, and antimicrobial susceptibility testing, a selection of isolates were subjected to pulsed-field gel electrophoresis and spa typing. Direct plating revealed a MRSA prevalence of 8%. After enrichment, MRSA was isolated from 98 (72%) of 137 samples of which the majority were from rib, ear, and forelimb. The majority (97%) of obtained isolates belonged to CC398, the main LA-MRSA type. A high level of genetic diversity was noted among the isolates from one butchery. Thirty antimicrobial susceptibility profiles were found; 13 and 9% of the isolates had Cip-Tet-Tri and Gen-Kan-Tet-Tob-Tri profiles, respectively. These results indicate the importance of enrichment for MRSA detection of pork. The observed genetic diversity of the isolates indicated that the pork production chain can be considered a source of multiple MRSA types that could be transmitted to the human population through cross-contaminated meat.
Abdallah, Soad A; Al-Asfoor, Khulood K; Salama, Mona F; Al-Awadi, Bashayer M
Background: Since the early nineties, a new methicillin-resistant Staphylococcus aureus (MRSA) has existed in a form correlating with community health personnel. Community-acquired MRSA (CA-MRSA) could be differentiated from healthcare-associated MRSA (HA-MRSA) microbiologically, epidemiologically, and molecularly. Aims: To determine the prevalence, risk factors of MRSA infections in community and hospital. Settings: The incidence and risk factors for CA-MRSA and HA-MRSA among patients of medical, surgical, and pediatrics wards and ICU at a Kuwaiti teaching hospital between 1 March 2011 and 30 November 2011 were studied. Materials and Methods: Cultures for MRSA were taken from nasal (nostril), groin, axilla, wound, sputum, or throat, and the inguinal area in all enrolled patients upon admission. All preserved isolates were examined for their susceptibility to different types of antibiotics. Results and Conclusion: A total of 71 MRSA patients admitted to different hospital wards were examined. Among these patients, 52 (73.2%) were carriers of MRSA before they were admitted to the hospital. Nineteen patients (26.8%) were found to have acquired MRSA during their stay in the hospital. Twenty-nine patients (40.8%) were given mupirocin local skin antibiotic. Binomial and the t-test (paired) were used to compare the prevalence of CA-MRSA and HA-MRSA; significant correlation (P < 0.05) between the type of MRSA and different wards, sites, and lengths of hospital stay was found. The level of serum albumin that is routinely measured at hospital admission is a predictor to MRSA infection. This study suggests that S. aureus and MRSA should become a national priority for disease control to avoid outbreaks. PMID:23599613
Chen, Michelle M. S.; Boardman, Wayne S. J.; Brown, Melissa H.
Background Infection with methicillin-resistant staphylococci (MRS) can be life-threatening in humans and its presence in animals is a cause for public health concern. The aim of this study was to measure the prevalence of MRS in captive and free-ranging wallabies over a 16-month period in South Australia, Australia. Materials and methods Eighty-nine purified staphylococcal isolates recovered from 98 captive and free-ranging wallabies' anterior nasal swabs were used in this study. All isolates were tested for the presence of the mecA, mecA1, and mecC genes. Multiplex PCR-directed SCCmec-typing, ccrB-typing, and determination of the minimal inhibitory concentration of oxacillin were performed on mec-positive isolates. Results and discussion In total, 11 non-Staphylococcus aureus MRS were isolated from 7 out of 98 animals, corresponding to a 7.1% carriage rate. The SCCmec types I, III, and V were identified by multiplex PCR and sequencing of the ccrB gene. This is the first report of MRS carriage in both captive and free-ranging wallabies in Australia. These data demonstrate a low prevalence of MRS and no association between wallaby captivity status and MRS carriage could be assigned. These animals may act as a reservoir for the exchange of genetic elements between staphylococci. Furthermore, the mecA genes of animal isolates were identical to that found in human MRS strains and thus the possibility of zoonotic transfer must be considered. PMID:27193419
Abdulgader, Shima M.; Shittu, Adebayo O.; Nicol, Mark P.; Kaba, Mamadou
Methicillin-resistant Staphylococcus aureus (MRSA) infections are a serious global problem, with considerable impact on patients and substantial health care costs. This systematic review provides an overview on the clonal diversity of MRSA, as well as the prevalence of Panton-Valentine leukocidin (PVL)-positive MRSA in Africa. A search on the molecular characterization of MRSA in Africa was conducted by two authors using predefined terms. We screened for articles published in English and French through to October 2014 from five electronic databases. A total of 57 eligible studies were identified. Thirty-four reports from 15 countries provided adequate genotyping data. CC5 is the predominant clonal complex in the healthcare setting in Africa. The hospital-associated MRSA ST239/ST241-III [3A] was identified in nine African countries. This clone was also described with SCCmec type IV [2B] in Algeria and Nigeria, and type V [5C] in Niger. In Africa, the European ST80-IV [2B] clone was limited to Algeria, Egypt and Tunisia. The clonal types ST22-IV [2B], ST36-II [2A], and ST612-IV [2B] were only reported in South Africa. No clear distinctions were observed between MRSA responsible for hospital and community infections. The community clones ST8-IV [2B] and ST88-IV [2B] were reported both in the hospital and community settings in Angola, Cameroon, Gabon, Ghana, Madagascar, Nigeria, and São Tomé and Príncipe. The proportion of PVL-positive MRSA carriage and/or infections ranged from 0.3 to 100% in humans. A number of pandemic clones were identified in Africa. Moreover, some MRSA clones are limited to specific countries or regions. We strongly advocate for more surveillance studies on MRSA in Africa. PMID:25983721
Perona, Paul J; Johnson, Aaron J; Perona, John P; Issa, Kimona; Kapadia, Bhaveen H; Bonutti, Peter M; Mont, Michael A
Periprosthetic infections with methicillin-resistant Staphylococcus aureus (MRSA) can be particularly burdensome and difficult to eradicate. One of the measures that infection control officers have emphasized in our hospitals has been the use of various hand sanitizers throughout the hospital. Our objective was to determine the level of growth inhibition of common hand sanitizers and surgical scrub solutions that are used to prevent the spread of community-acquired strains of MRSA. Various hospital and surgical agents (n = 13) were applied to community-acquired MRSA bacteria that had been cultured on agar plates. These different commercially available solutions were incubated for 48 h, and the plates were assessed to determine the level of growth inhibition (0, 25, 75, or 100%). The negative control was a test in which no agent was added to the MRSA culture, while a positive control tested 100% alcohol. Eight of the solutions tested had 100% growth inhibition, four solutions had partial growth inhibition effects, and one solution did not inhibit MRSA. Of the solutions with alcohol, the 62% solution did not kill MRSA, while the 80% solution only inhibited MRSA. Both the 95 and 100% alcohol solutions had 100% growth inhibition. Of the two surgical scrub solutions, only the one with iodine had 100% growth inhibition, whereas the solution with chloroxylenol (PCMX 3%) had only partial growth inhibition. This study suggests that the solutions with high levels of alcohol, chlorhexidine, or iodine appear to better kill MRSA and might best be used to prevent the spread of community-acquired MRSA in both the hospital and the surgical environment.
Christopher, Solomon; Verghis, Rejina Mariam; Antonisamy, Belavendra; Sowmyanarayanan, Thuppal Varadachari; Brahmadathan, Kootallur Narayanan; Kang, Gagandeep; Cooper, Ben Symons
Background Methicillin-resistant Staphylococcus aureus (MRSA) is a global pathogen and an important but seldom investigated cause of morbidity and mortality in lower and middle-income countries where it can place a major burden on limited resources. Quantifying nosocomial transmission in resource-poor settings is difficult because molecular typing methods are prohibitively expensive. Mechanistic statistical models can overcome this problem with minimal cost. We analyse the transmission dynamics of MRSA in a hospital in south India using one such approach and provide conservative estimates of the organism's economic burden. Methods and Findings Fifty months of MRSA infection data were collected retrospectively from a Medical Intensive Care Unit (MICU) in a tertiary hospital in Vellore, south India. Data were analysed using a previously described structured hidden Markov model. Seventy-two patients developed MRSA infections and, of these, 49 (68%) died in the MICU. We estimated that 4.2% (95%CI 1.0, 19.0) of patients were MRSA-positive when admitted, that there were 0.39 MRSA infections per colonized patient month (0.06, 0.73), and that the ward-level reproduction number for MRSA was 0.42 (0.08, 2.04). Anti-MRSA antibiotic treatment costs alone averaged $124/patient, over three times the monthly income of more than 40% of the Indian population. Conclusions Our analysis of routine data provides the first estimate of the nosocomial transmission potential of MRSA in India. The high levels of transmission estimated underline the need for cost-effective interventions to reduce MRSA transmission in hospital settings in low and middle income countries. PMID:21750700
Yilmaz, Ayhan; Kaleta, Erhard Franz
Two commercially available preparations from the disinfectants list of the German Veterinary Association (Deutsche Veterinärmedizinische Gesellschaft e. V. (DVG)) were tested against a methicillin-resistant Staphylococcus aureus (MRSA) field isolate in a concentration as recommended in the list for disinfection of bacteria. The "Guidelines of DVG for testing of disinfection methods and chemical disinfectants" were used as methodical base. Initially, the examinations for determination of the minimal inhibitory concentrations (MIC) and suitable inactivation substances were carried out. The bactericidal effect of both commercial preparations--a formic acid preparation (55 g per 100 g) and a preparation containing 21 g glutaraldhyde plus 17 g per 100 g formaldehyde--as well as reference disinfectants (37% formaldehyde and 99.5% phenol) were examined in suspension and carrier tests. The suspension tests were performed without and with protein load (20% FBS). Pieces of linden wood were used as carriers. The recommendation of DVG for both Venno Vet 1 super and M&enno Veterinär B neu as bactericide (except of bacterial spores) is 30 min (for preventive disinfections) as well as 120 min contact (for specific disinfections) of 1% concentration. The results show that the disinfection of MRSA do not require higher concentrations and longer contact times as recommended in the disinfectant list of DVG for both commercial disinfectants. Both products were effective in suspension tests in a 1% concentration and within five minutes independent of protein load. In contrast, 1% phenol (reference disinfectant for suspension tests) needed 30 min reaction time without protein load as well as 60 min with protein load for complete inactivation of the MRSA field strain. In carrier tests, a 30 min contact of 1% concentration of both commercial disinfectants and 3% concentration of formalin (reference disinfectant for carrier tests) was sufficient for complete disinfection.
Hoet, Armando E; Johnson, Amanda; Nava-Hoet, Rocio C; Bateman, Shane; Hillier, Andrew; Dyce, John; Gebreyes, Wondwossen A; Wittum, Thomas E
Concurrent to reports of zoonotic and nosocomial transmission of methicillin-resistant Staphylococcus aureus (MRSA) in veterinary settings, recent evidence indicates that the environment in veterinary hospitals may be a potential source of MRSA. The present report is a cross-sectional study to determine the prevalence of MRSA on specific human and animal contact surfaces at a large veterinary hospital during a nonoutbreak period. A total of 156 samples were collected using Swiffers(®) or premoistened swabs from the small animal, equine, and food animal sections. MRSA was isolated and identified by pre-enrichment culture and standard microbiology procedures, including growth on Mueller-Hinton agar supplemented with NaCl and oxacillin, and by detection of the mecA gene. Staphylococcal chromosome cassette mec (SCCmec) typing and pulsed-field gel electrophoresis profile were also determined. MRSA was detected in 12% (19/157) of the hospital environments sampled. The prevalence of MRSA in the small animal, equine, and food animal areas were 16%, 4%, and 0%, respectively. Sixteen of the MRSA isolates from the small animal section were classified as USA100, SCCmec type II, two of which had pulsed-field gel electrophoresis pattern that does not conform to any known type. The one isolate obtained from the equine section was classified as USA500, SCCmec type IV. The molecular epidemiological analysis revealed a very diverse population of MRSA isolates circulating in the hospital; however, in some instances, multiple locations/surfaces, not directly associated, had the same MRSA clone. No significant difference was observed between animal and human contact surfaces in regard to prevalence and type of isolates. Surfaces touched by multiple people (doors) and patients (carts) were frequently contaminated with MRSA. The results from this study indicate that MRSA is present in the environment even during nonoutbreak periods. This study also identified specific surfaces in a
Biber, A; Parizade, M; Taran, D; Jaber, H; Berla, E; Rubin, C; Rahav, G; Glikman, D; Regev-Yochay, G
Data on community-associated (CA) methicillin-resistant Staphylococcus aureus (MRSA) in Israel are scarce. The objective of this study was to characterize the major CA-MRSA clones in Israel. All clinical MRSA isolates detected in the community during a period of 2.5 years (2011-2013) from individuals insured by a major health maintenance organization in Israel were collected, with additional data from medical records. Antibiotic susceptibility patterns and staphylococcal chromosomal cassette mec (SCCmec) typing were determined. SCCmec IV and V isolates were further typed by pulsed-field gel electrophoresis (PFGE), spa typing, and detection of a panel of toxin genes. MRSA were detected in 280 patients, mostly from skin infections. Patients with SCCmec IV (n = 120, 43 %) were younger (p < 0.0001) and reported less contact with healthcare facilities. Almost all isolates were trimethoprim-sulfamethoxazole susceptible (98 %). spa-CC032, a typical nosocomial MRSA clone, accounted for 28 % of SCCmec IV. The two major CA-MRSA clones were t008 USA300 (13 %) and t991 (10 %); t991 was isolated mainly from children (75 %), was Panton-Valentine leukocidin (PVL) negative but eta-positive, and was typically susceptible to most antibiotic groups. PVL-positive strains (n = 31) included mainly USA300 (52 %) and t019 (13 %). While multiple genetic lineages were evident among community-onset MRSA in Israel, approximately 20 % are typical CA-MRSA clones, mainly USA300 and a local clone, t991.
Haeili, Mehri; Moore, Casey; Davis, Christopher J C; Cochran, James B; Shah, Santosh; Shrestha, Tej B; Zhang, Yaofang; Bossmann, Stefan H; Benjamin, William H; Kutsch, Olaf; Wolschendorf, Frank
Macrophages take advantage of the antibacterial properties of copper ions in the killing of bacterial intruders. However, despite the importance of copper for innate immune functions, coordinated efforts to exploit copper ions for therapeutic interventions against bacterial infections are not yet in place. Here we report a novel high-throughput screening platform specifically developed for the discovery and characterization of compounds with copper-dependent antibacterial properties toward methicillin-resistant Staphylococcus aureus (MRSA). We detail how one of the identified compounds, glyoxal-bis(N4-methylthiosemicarbazone) (GTSM), exerts its potent strictly copper-dependent antibacterial properties on MRSA. Our data indicate that the activity of the GTSM-copper complex goes beyond the general antibacterial effects of accumulated copper ions and suggest that, in contrast to prevailing opinion, copper complexes can indeed exhibit species- and target-specific activities. Based on experimental evidence, we propose that copper ions impose structural changes upon binding to the otherwise inactive GTSM ligand and transfer antibacterial properties to the chelate. In turn, GTSM determines target specificity and utilizes a redox-sensitive release mechanism through which copper ions are deployed at or in close proximity to a putative target. According to our proof-of-concept screen, copper activation is not a rare event and even extends to already established drugs. Thus, copper-activated compounds could define a novel class of anti-MRSA agents that amplify copper-dependent innate immune functions of the host. To this end, we provide a blueprint for a high-throughput drug screening campaign which considers the antibacterial properties of copper ions at the host-pathogen interface.
Woeste, G; Zapletal, C; Wullstein, C; Golling, M; Bechstein, W O
The prevalence of methicillin-resistant Staphylococus aureus (MRSA) has increased worldwide and MRSA has emerged as an important cause of sepsis in cirrhotic patients and liver transplant recipients. In this retrospective study, the prevalence of MRSA colonization and its influence on infections following orthotopic liver transplantation (OLT) was investigated. From August, 2002 until November, 2004, 66 primary cadaver OLT were performed for adult recipients. Antibody induction used Daclizumab (n = 49) or ATG (n = 14). Maintenance immunosuppression consisted of tacrolimus and steroids, with 30 patients receiving mycophenolate mofetil and 4, rapamune. For perioperative anti-infectious prophylaxis cefotaxime, metronidazole, and tobramycin were administered for 48 hours. The preoperatively performed routine swabs revealed MRSA colonization in 12 of 66 (18.2%) patients. The stage of cirrhosis was equivalent for MRSA(-) patients according to Child score. The mean MELD score was significantly higher for MRSA(+) patients (24.3 versus 18.7, P = .036). More MRSA(+) patients were hospitalized at the time of transplantation (14/54 versus 8/12, P = .018). The incidence of posttransplant infections was not significantly different among the two groups. Within the first year 7 of 66 (10.6%) patients died: 3 of 12 (25%) MRSA(+) and 4 of 54 (7.4%) MRSA(-). The 1-year survival was lower in the MRSA(+) group (74.1% versus 94.1%). In conclusion, this study did not show that an MRSA-positive carrier status implies an increased risk for septic complications following OLT. Mortality was increased for MRSA(+), but failed to show a significant difference. A significantly higher MELD score and pretransplant hospitalization for MRSA(+) patients may contribute to the higher mortality and reflect sicker patients.
Lima, Danielle F; Cohen, Renata WF; Rocha, Géssica A; Albano, Rodolpho M; Marques, Elizabeth A; Leão, Robson S
Alarmingly, the isolation of methicillin-resistant Staphylococcus aureus (MRSA) has been increasing among patients with cystic fibrosis (CF). During a previous molecular characterisation of MRSA isolates obtained from patients with CF from Rio de Janeiro, Brazil, one isolate was identified as the ST398 clone, a livestock-associated (LA) MRSA. In this study, we report the draft genome sequence of an LA-MRSA ST398 clone isolated from a patient with CF. PMID:28076471
Lima, Danielle F; Cohen, Renata Wf; Rocha, Géssica A; Albano, Rodolpho M; Marques, Elizabeth A; Leão, Robson S
Alarmingly, the isolation of methicillin-resistant Staphylococcus aureus (MRSA) has been increasing among patients with cystic fibrosis (CF). During a previous molecular characterisation of MRSA isolates obtained from patients with CF from Rio de Janeiro, Brazil, one isolate was identified as the ST398 clone, a livestock-associated (LA) MRSA. In this study, we report the draft genome sequence of an LA-MRSA ST398 clone isolated from a patient with CF.
de Kraker, Marlieke E A; Wolkewitz, Martin; Davey, Peter G; Koller, Walter; Berger, Jutta; Nagler, Jan; Icket, Claudine; Kalenic, Smilja; Horvatic, Jasminka; Seifert, Harald; Kaasch, Achim J; Paniara, Olga; Argyropoulou, Athina; Bompola, Maria; Smyth, Edmond; Skally, Mairead; Raglio, Annibale; Dumpis, Uga; Kelmere, Agita Melbarde; Borg, Michael; Xuereb, Deborah; Ghita, Mihaela C; Noble, Michelle; Kolman, Jana; Grabljevec, Stanko; Turner, David; Lansbury, Louise; Grundmann, Hajo
Antimicrobial resistance is threatening the successful management of nosocomial infections worldwide. Despite the therapeutic limitations imposed by methicillin-resistant Staphylococcus aureus (MRSA), its clinical impact is still debated. The objective of this study was to estimate the excess mortality and length of hospital stay (LOS) associated with MRSA bloodstream infections (BSI) in European hospitals. Between July 2007 and June 2008, a multicenter, prospective, parallel matched-cohort study was carried out in 13 tertiary care hospitals in as many European countries. Cohort I consisted of patients with MRSA BSI and cohort II of patients with methicillin-susceptible S. aureus (MSSA) BSI. The patients in both cohorts were matched for LOS prior to the onset of BSI with patients free of the respective BSI. Cohort I consisted of 248 MRSA patients and 453 controls and cohort II of 618 MSSA patients and 1,170 controls. Compared to the controls, MRSA patients had higher 30-day mortality (adjusted odds ratio [aOR] = 4.4) and higher hospital mortality (adjusted hazard ratio [aHR] = 3.5). Their excess LOS was 9.2 days. MSSA patients also had higher 30-day (aOR = 2.4) and hospital (aHR = 3.1) mortality and an excess LOS of 8.6 days. When the outcomes from the two cohorts were compared, an effect attributable to methicillin resistance was found for 30-day mortality (OR = 1.8; P = 0.04), but not for hospital mortality (HR = 1.1; P = 0.63) or LOS (difference = 0.6 days; P = 0.96). Irrespective of methicillin susceptibility, S. aureus BSI has a significant impact on morbidity and mortality. In addition, MRSA BSI leads to a fatal outcome more frequently than MSSA BSI. Infection control efforts in hospitals should aim to contain infections caused by both resistant and susceptible S. aureus.
Many methicillin-resistant Staphylococcus aureus (MRSA) strains are multidrug-resistant; consequently, infectious diseases involving MRSA are recognized as troublesome diseases not only in human health care but also in animal health care. A bovine mastitis case caused by MRSA isolates of the New York/Japan clone (NJC), which occurred in Japan in 2005, was monitored in the current study. Isolates of the NJC are typical of hospital-acquired MRSA in Japan. The genetic backgrounds of these strains differ from those of bovine-associated S. aureus, which are typically of clonal complex (CC)97, CC705, and CC133. Moreover, the NJC isolates in this bovine outbreak possessed a β-hemolysin-converting bacteriophage and an immune evasion cluster, as found in the NJC isolates from humans, so it is possible that this clone was introduced into the dairy herd by a human carrier. Most bovine intramammary infections (IMIs) caused by the NJC isolates in our study were asymptomatic, and obvious clinical signs were recognized in only the first 3 infected cows. Of a total of 78 cows, 31 cows were MRSA carriers, and these carrier cows were detected by testing the milk of all lactating cows at 1-month intervals. These S. aureus carrier cows were culled or the infected quarter was dried off and no longer milked. Both IMI and mastitis caused by MRSA were completely eradicated after 5 months. Genotyping data suggested that exchanging of the staphylococcal cassette chromosome mec (the determining factor in methicillin resistance) occurred easily between MRSA and methicillin-sensitive S. aureus in the udders of carrier cows. This case study demonstrates an effective procedure against the spread of MRSA in a dairy herd, and highlights the risk of emergence of new MRSA strains in a dairy herd.
Roy, S; Barman, T K; Hossain, M A; Paul, S K; Haque, N; Ahmed, S; Nasreen, S A; Hossain, M S; Sarkar, S R; Kubayashi, N; Laskar, N
Infections caused by Staphylococcus aureus were treated by methicillin, but about 95% of S. aureus has been resistance to methicillin, both in the community and hospitals and are increasing day by day. MRSA produces altered penicillin binding protein, PBP2a, due to the expression of mecA gene. Some strains of both the MRSA and MSSA carry PVL gene. This cross sectional observational study was conducted to detect the molecular-characterization of methicillin resistant Staphylococcus aureus (MRSA) and carried out in the Department of Microbiology, Mymensingh Medical College from July 2014 to December 2015. Clinical samples for this study were wound swab, pus, exudates from diabetic ulcer and burn ulcer, aural swab, blood and urine which were collected from three tertiary care hospitals such as from MMCH, BIRDEM hospital and SSMCH. Standard microbiological procedure & biochemical tests were carried out to detect S. aureus. Oxacillin disk diffusion method (ODDM) was done by Kirby-Bauer disk diffusion method. Out of a total 109 culture positive samples 69 isolates of S. aureus were selected for the study. Among the 69 isolates 33, 27 and 09 were from MMCH, BIRDEM hospital and SSMCH respectively. Among the 69 isolates, 17(24.6%) and 52(75.3%) were distinguished as MRSA and MSSA respectively by ODDM. In contrast, detection of presence and absence of mecA gene by PCR identified 20(28.9%) and 49(71.01%) isolates as MRSA and MSSA respectively. Multiplex PCR was performed by standard protocol with specific primers for detection of 16S rRNA gene for Staphylococcus, nuc gene for Staphylococcus aureus, mecA gene for MRSA, PVL gene as a virulence factor and ACME-arc gene for worldwide spreading USA 300 MRSA clone. The PVL gene were detected in 3 out of 20 MRSA (15%) and 19 out of 49 MSSA (38.7%) and the ACME- arc gene was not found in any isolates. All of the S. aureus (MRSA and MSSA) isolates were sensitive to Vancomycin and Gentamicin. All MRSA isolates (100%) showed resistance
Tanner, Joshua; Lin, Ying; Kornblum, John; Herzig, Carolyn T. A.; Bystritsky, Rachel; Uhlemann, Anne-Catrin
Inmates of Rikers Island jail potentially introduce Staphylococcus aureus into New York State prisons upon transfer. In this study, methicillin-resistant Staphylococcus aureus isolates (n = 452), collected from infected inmates (2009 to 2013), were characterized. spa type t008 was the predominant clone identified, accounting for 82.3% of the isolates, with no evidence of mupirocin or chlorhexidine resistance. PMID:24899033
Yadav, Mukesh Kumar; Chae, Sung-Won; Im, Gi Jung; Chung, Jae-Woo; Song, Jae-Jun
Background Inhibition and eradication of Staphylococcus aureus biofilms with conventional antibiotic is difficult, and the treatment is further complicated by the rise of antibiotic resistance among staphylococci. Consequently, there is a need for novel antimicrobials that can treat biofilm-related infections and decrease antibiotics burden. Natural compounds such as eugenol with anti-microbial properties are attractive agents that could reduce the use of conventional antibiotics. In this study we evaluated the effect of eugenol on MRSA and MSSA biofilms in vitro and bacterial colonization in vivo. Methods and Results Effect of eugenol on in vitro biofilm and in vivo colonization were studied using microtiter plate assay and otitis media-rat model respectively. The architecture of in vitro biofilms and in vivo colonization of bacteria was viewed with SEM. Real-time RT-PCR was used to study gene expression. Check board method was used to study the synergistic effects of eugenol and carvacrol on established biofilms. Eugenol significantly inhibited biofilms growth of MRSA and MSSA in vitro in a concentration-dependent manner. Eugenol at MIC or 2×MIC effectively eradicated the pre-established biofilms of MRSA and MSSA clinical strains. In vivo, sub-MIC of eugenol significantly decreased 88% S. aureus colonization in rat middle ear. Eugenol was observed to damage the cell-membrane and cause a leakage of the cell contents. At sub-inhibitory concentration, it decreases the expression of biofilm-and enterotoxin-related genes. Eugenol showed a synergistic effect with carvacrol on the eradication of pre-established biofilms. Conclusion/Major Finding This study demonstrated that eugenol exhibits notable activity against MRSA and MSSA clinical strains biofilms. Eugenol inhibited biofilm formation, disrupted the cell-to-cell connections, detached the existing biofilms, and killed the bacteria in biofilms of both MRSA and MSSA with equal effectiveness. Therefore, eugenol may
Yabuki, Hiroshi; Tabata, Toshiharu; Sugawara, Takafumi; Fukaya, Ken; Fujimura, Shigefumi
A case is 48-year-old man who had a history of Blalock-Taussig shunt and the radical operation for Fallot's tetralogy, had been performed cavernostomy and fenestration operation for aspergilloma of left upper lobe in the previous hospital due to control blood spitting. Although the contents of the abscess cavity were removed, the opened cavity was again infected by methicillin-resistant Staphylococcus aureus (MRSA) and he was referred to our hospital. The plombage of free omental flap with vascular anastomosis was performed. He has been well without any symptoms or recurrence of empyema for 6 years after surgery.
Cavrić, Gordana; Ilić, Diana; Njers, Kristina; Prkacin, Ingrid; Hamp, Dubravka Bartolek
We present a case of a 50-year-old man with advanced liver cirrhosis and native valve infective endocarditis caused by methicillin-resistant Staphylococcus epidermidis. Bacterial infections are one of the most common complications of liver cirrhosis, but reports of infective endocarditis in patients with liver cirrhosis are relatively rare. Because of vulnerability of patients with advanced cirrhosis for developing infections, it is necessary to pay attention to the pathogens that are sometimes considered contamination and actively seek for the seat of infection, even in less expected areas (e.g., native heart valves without a history of heart disease).
Fujimura, Shigeru; Sato, Tetsuro; Hayakawa, Sachiko; Kawamura, Masato; Furukawa, Emiko; Watanabe, Akira
In vitro efficacy of combined eradication therapy with clarithromycin and daptomycin against biofilm-formed methicillin-resistant Staphylococcus aureus on the orthopedic titanium devices was evaluated. The bactericidal effect of this antibiotic was investigated by a re-culture test, the scanning electron microscopy, and fluorescence microscopy using a double-staining dyes. Clarithromycin decreased the amount to half in 24 h. Although MRSA biofilms were not eradicated with clarithromycin or daptomycin alone, clarithromycin combined with daptomycin was useful to sterilize titanium devices within 72 h. This in vitro study showed that combined treatment with clarithromycin plus daptomycin is useful to eradicate staphylococcal biofilms formed on orthopedic devices.
Baldwin, L N; Lowe, A D
We report a case of community acquired methicillin resistant Staphylococcus aureus pneumonia. The causative organism was positive for the toxin Panton-Valentine Leukocidin. This resulted in a severe pneumonia requiring a prolonged stay on our intensive care unit. This infection is becoming more common in the United Kingdom. It can cause a far more aggressive illness than the hospital acquired infection with a high mortality if it becomes an invasive infection. The Department of Health has recently produced interim guidelines for its treatment which we have also reviewed.
Adnalizawati, A. Siti Noor; Nazlina, I.; Yaacob, W. A.
The in vitro activity of methanolic extract from Phyllanthus columnaris stem bark was studied against Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300 and MRSA BM1 (clinical strain) using time-kill curves in conjunction with scanning and transmission electron microscopy. The extract showed more markedly bactericidal activity in MRSA BM1 clinical strain within less than 4 h by 6.25-12.5 mg/mL and within 6 h by 1.56 mg/mL. Scanning electron microscopy of MRSA BM1 revealed distortion of cell whilst transmission electron microscopy revealed disruption in cell wall division.
Adnalizawati, A. Siti Noor; Nazlina, I.; Yaacob, W. A.
The in vitro activity of methanolic extract from Phyllanthus columnaris stem bark was studied against Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300 and MRSA BM1 (clinical strain) using time-kill curves in conjunction with scanning and transmission electron microscopy. The extract showed more markedly bactericidal activity in MRSA BM1 clinical strain within less than 4 h by 6.25-12.5 mg/mL and within 6 h by 1.56 mg/mL. Scanning electron microscopy of MRSA BM1 revealed distortion of cell whilst transmission electron microscopy revealed disruption in cell wall division.
Udumula, Venkatareddy; Endres, Jennifer L; Harper, Caleb N; Jaramillo, Lee; Zhong, Haizhen A; Bayles, Kenneth W; Conda-Sheridan, Martin
Community-associated methicillin resistant Staphylococcus aureus (CA-MRSA) has become a severe health concern because of its treatment difficulties. Herein, we report the synthesis and biological evaluation of two phenazine natural products and a series of phenazines that show promising activities against MRSA with MIC values in the low micromolar range. Basic studies revealed that these compounds are bacteriostatic agents. The most active compound also displayed promising IC50 values against HaCat cells. Finally, a QSAR model was developed to understand the key structural features of the molecules.
Duim, Birgitta; Fluit, Ad C; Carney, Jennifer; van Nes, Arie; Wagenaar, Jaap A
Methicillin-resistant Staphylococcus aureus (MRSA) is an important colonizer in animals and an opportunistic pathogen in humans. In humans, MRSA can cause infections that might be difficult to treat because of antimicrobial resistance. The use of bacteriophages has been suggested as a potential approach for the control of MRSA colonization to minimize the—often occupational—exposure of humans. The aim of this study was to assess the efficacy of bacteriophage treatment on porcine nasal colonization with MRSA in vitro, in vivo, and ex vivo. The effectiveness of a bacteriophage combination of phage K*710 and P68 was assessed in vitro by incubating them with MRSA V0608892/1 (ST398) measuring the OD600 hourly. To study the in vivo effect, bacteriophages were administered in a gel developed for human application, which contain 109 plaque-forming units (pfu)/mL (K and P68 in a 19.25:1 ratio) for 5 days to piglets (N = 8) that were experimentally colonized with the MRSA strain. Eight piglets experimentally colonized were used as a negative control. The MRSA strain was also used to colonize porcine nasal mucosa explants and bacteriophages were applied to assess the ex vivo efficacy of treatment. Bacteriophages were effective in vitro. In vivo, sixteen piglets were colonized with MRSA but the number of CFU recovered after the application of the bacteriophages in 8 piglets was not reduced compared to the control animals (approx. 105 CFU/swab). In the ex vivo model, 108 CFU were used to establish colonization with MRSA; a reduction of colonization was not observed after application of bacteriophages. However, application of mupirocin both in vivo and ex vivo resulted in a near eradication of MRSA. In conclusion: i) The MRSA strain was killed in the presence of the bacteriophages phage K*710 and P68 in vitro. ii) Bacteriophages did not reduce porcine nasal colonization in vivo or ex vivo. Physiological in vivo and ex vivo conditions may explain these observations. Efficacy
Maraqa, Nizar F.; Cook, Robert L.; Johnson, Judith A.; Bailey, Christine; Wheeler, Sarah; Nolan, David; Rathore, Mobeen H.; Morris, J. Glenn; Salemi, Marco
Despite infection prevention efforts, neonatal intensive care unit (NICU) patients remain at risk of Methicillin-resistant Staphylococcus aureus (MRSA) infection. Modes of transmission for healthcare-associated (HA) and community-associated (CA) MRSA remain poorly understood and may vary by genotype, hindering the development of effective prevention and control strategies. From 2008–2010, all patients admitted to a level III NICU were screened for MRSA colonization, and all available isolates were spa-typed. Spa-type t008, the most prevalent CA- genotype in the United States, spa-type t045, a HA- related genotype, and a convenience sample of strains isolated from 2003–2011, underwent whole-genome sequencing and phylodynamic analysis. Patient risk factors were compared between colonized and noncolonized infants, and virulence and resistance genes compared between spa-type t008 and non-t008 strains. Epidemiological and genomic data were used to estimate MRSA importations and acquisitions through transmission reconstruction. MRSA colonization was identified in 9.1% (177/1940) of hospitalized infants and associated with low gestational age and birth weight. Among colonized infants, low gestational age was more common among those colonized with t008 strains. Our data suggest that approximately 70% of colonizations were the result of transmission events within the NICU, with the remainder likely to reflect importations of “outside” strains. While risk of transmission within the NICU was not affected by spa-type, patterns of acquisition and importation differed between t008 and t045 strains. Phylodynamic analysis showed the effective population size of spa-type t008 has been exponentially increasing in both community and hospital, with spa-type t008 strains possessed virulence genes not found among t045 strains; t045 strains, in contrast, appeared to be of more recent origin, with a possible hospital source. Our data highlight the importance of both intra
Normanno, Giovanni; Dambrosio, Angela; Lorusso, Vanessa; Samoilis, Georgios; Di Taranto, Pietro; Parisi, Antonio
Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen present in the hospital environment (HA-MRSA), in the community (CA-MRSA) and in livestock, including pigs (LA-MRSA). MRSA may enter the human food chain during slaughtering and may infect humans coming into direct contact with pigs or pork products. This study aimed to determine the prevalence and characteristics of MRSA isolated from pigs and workers at industrial abattoirs in southern Italy. A total of 215 pig nasal swabs were screened for the presence of MRSA using PCR. An MRSA isolate was detected from each mecA/nuc PCR-positive sample and characterized by spa-typing, Multi-Locus Sequence Typing, SCC-mec and Panton-Valentine Leukocidin (PVL), and also tested for the production of staphylococcal enterotoxins (SEs). Eighty-one MRSA isolates (37.6%) were obtained from the 215 pig nasal swabs; 37 of these isolates were further characterized, and showed 18 different spa-types and 8 different STs. The most frequently recovered STs were ST398 (CC398-t034, t011, t899, t1939 - 43.2%) followed by ST8 (CC8-t008, t064, t2953, t5270 - 24.3%) and ST1 (CC1-t127, t174, t2207 - 10.8%). Nine MRSA isolates were obtained from the 113 human swabs; the isolates showed 5 different spa-types and 5 different STs, including the novel ST2794 (t159). The most representative STs recovered were ST1 (CC1-t127) and ST398 (CC398-t034) (33.3%). None of the MRSA isolates showed the ability to produce SEs and PVL and all resulted resistant to two or more classes of antimicrobials. This study shows the great genetic diversity of MRSA strains in slaughtered pigs and in abattoir employees in Italy, and clearly demonstrates the need for improved hygiene standards to reduce the risk of occupational and food-borne infection linked to the handling/consumption of raw pork containing MRSA.
Cunningham-Oakes, Edward; Soren, Odel; Moussa, Caroline; Rathor, Getika; Liu, Yingjun; Coates, Anthony; Hu, Yanmin
Infections caused by methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) are prevalent. MRSA infections are difficult to treat and there are no new classes of antibiotics produced to the market to treat infections caused by the resistant bacteria. Therefore, using antibiotic enhancers to rescue existing classes of antibiotics is an attractive strategy. Nordihydroguaiaretic acid (NDGA) is an antioxidant compound found in extracts from plant Larrea Tridentata. It exhibits antimicrobial activity and may target bacterial cell membrane. Combination efficacies of NDGA with many classes of antibiotics were examined by chequerboard method against 200 clinical isolates of MRSA and MSSA. NDGA in combination with gentamicin, neomycin, and tobramycin was examined by time-kill assays. The synergistic combinations of NDGA and aminoglycosides were tested in vivo using a murine skin infection model. Calculations of the fractional inhibitory concentration index (FICI) showed that NDGA when combined with gentamicin, neomycin, or tobramycin displayed synergistic activities in more than 97% of MSSA and MRSA, respectively. Time kill analysis demonstrated that NDGA significantly augmented the activities of these aminoglycosides against MRSA and MSSA in vitro and in murine skin infection model. The enhanced activity of NDGA resides on its ability to damage bacterial cell membrane leading to accumulation of the antibiotics inside bacterial cells. We demonstrated that NDGA strongly revived the therapeutic potencies of aminoglycosides in vitro and in vivo. This combinational strategy could contribute major clinical implications to treat antibiotic resistant bacterial infections. PMID:26579101
Casero, Carina; Estévez-Braun, Ana; Ravelo, Angel G; Demo, Mirta; Méndez-Álvarez, Sebastián; Machín, Félix
Currently, there is a pressing need for novel antibacterial agents against drug-resistant bacteria, especially those which have been common in our communities and hospitals, such as methicillin-resistant Staphylococcus aureus (MRSA). The South American plant Achyrocline satureioides ("Marcela") has been widely used in traditional medicine for a number of diseases, including infections. Several crude extracts from this plant have shown good antimicrobial activities in vitro. In the search for the active principle(s) that confers these antimicrobial activities, we have processed the dichloromethane extract from the aerial parts of the plant. One of the isolated compounds showed extraordinary antibacterial activities against a set of clinically relevant Gram-positive strains that widely differ in their antibiogram profiles. This compound was identified as achyrofuran on the basis of its spectroscopic and physical data. We determined the MIC to be around 0.1 μM (0.07 μg/ml) for the reference methicillin-resistant and vancomycin-intermediate S. aureus strain NRS402. Moreover, nanomolar concentrations of achyrofuran killed 10(6) bacteria within 12 h. Based on the presence of the 2,2'-biphenol core, we further studied whether achyrofuran killed bacteria through a mechanism of action similar to that reported for the naturally occurring antibiotic MC21-A. Indeed, we found that achyrofuran was not bacteriolytic by itself although it greatly compromised membrane impermeability as determined by increased SYTOX Green uptake.
Bozic, Dragana D.; Milenkovic, Marina; Ivkovic, Branka; Cirkovic, Ivana
Biofilm formation and adherence of bacteria to host tissue are one of the most important virulence factors of methicillin-resistant strains of Staphylococcus aureus (MRSA). The number of resistant strains is seriously increasing during the past years and bacteria have become resistant, not only to methicillin, but also to other commonly used antistaphylococcal antibiotics. There is a great need for discovering a novel antimicrobial agent for the treatment of staphylococcal infections. One of the most promising groups of compounds appears to be chalcones. In present study we evaluated the in vitro effect of three newly synthesized chalcones: 1,3- Bis-(2-hydroxy-phenyl)-propenone, 3-(3-Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone and 3-(4-Hydroxy-phenyl)-1-(2-hydroxy-phenyl)-propenone on glycocalyx production, biofilm formation and adherence to human fibronectin of clinical isolates and laboratory control strain of MRSA (ATCC 43300). Subinhibitory concentrations of the tested compounds reduced the production of glycocalyx, biofilm formation and adherence to human fibronectin of all MRSA strains. Inhibition of biofilm formation was dose dependent and the most effective was 1,3- Bis-(2-hydroxy-phenyl)-propenone. In our study we demonstrated that three newly-synthesized chalcones exhibited significant effect on adherence and biofilm formation of MRSA strains. Chalcones may be considered as promising new antimicrobial agents that can be used for prevention of staphylococcal infections or as adjunct to antibiotics in conventional therapy. PMID:24948943
Mbah, Andreas N; Isokpehi, Raphael D
Resistance to methicillin by Staphylococcus aureus is a persistent clinical problem worldwide. A mechanism for resistance has been proposed in which methicillin resistant Staphylococcus aureus (MRSA) isolates acquired a new protein called β -lactam inducible penicillin binding protein (PBP-2'). The PBP-2' functions by substituting other penicillin binding proteins which have been inhibited by β -lactam antibiotics. Presently, there is no structural and regulatory information on PBP-2' protein. We conducted a complete structural and functional regulatory analysis of PBP-2' protein. Our analysis revealed that the PBP-2' is very stable with more hydrophilic amino acids expressing antigenic sites. PBP-2' has three striking regulatory points constituted by first penicillin binding site at Ser25, second penicillin binding site at Ser405, and finally a single metallic ligand binding site at Glu657 which binds to Zn(2+) ions. This report highlights structural features of PBP-2' that can serve as targets for developing new chemotherapeutic agents and conducting site direct mutagenesis experiments.
Technical Education Research Center, Waco, TX.
This concept module on resistance is one of thirteen modules that provide a flexible, laboratory-based physics instructional package designed to meet the specialized needs of students in two-year, postsecondary technical schools. Each of the thirteen concept modules discusses a single physics concept and how it is applied to each energy system.…
Metsemakers, Willem-Jan; Emanuel, Noam; Cohen, Or; Reichart, Malka; Potapova, Inga; Schmid, Tanja; Segal, David; Riool, Martijn; Kwakman, Paulus H S; de Boer, Leonie; de Breij, Anna; Nibbering, Peter H; Richards, R Geoff; Zaat, Sebastian A J; Moriarty, T Fintan
Implant-associated bone infections caused by antibiotic-resistant pathogens pose significant clinical challenges to treating physicians. Prophylactic strategies that act against resistant organisms, such as methicillin-resistant Staphylococcus aureus (MRSA), are urgently required. In the present study, we investigated the efficacy of a biodegradable Polymer-Lipid Encapsulation MatriX (PLEX) loaded with the antibiotic doxycycline as a local prophylactic strategy against implant-associated osteomyelitis. Activity was tested against both a doxycycline-susceptible (doxy(S)) methicillin-susceptible S. aureus (MSSA) as well as a doxycycline-resistant (doxy(R)) methicillin-resistant S. aureus (MRSA). In vitro elution studies revealed that 25% of the doxycycline was released from the PLEX-coated implants within the first day, followed by a 3% release per day up to day 28. The released doxycycline was highly effective against doxy(S) MSSA for at least 14days in vitro. A bolus injection of doxycycline mimicking a one day release from the PLEX-coating reduced, but did not eliminate, mouse subcutaneous implant-associated infection (doxy(S) MSSA). In a rabbit intramedullary nail-related infection model, all rabbits receiving a PLEX-doxycycline-coated nail were culture negative in the doxy(S) MSSA-group and the surrounding bone displayed a normal physiological appearance in both histological sections and radiographs. In the doxy(R) MRSA inoculated rabbits, a statistically significant reduction in the number of culture-positive samples was observed for the PLEX-doxycycline-coated group when compared to the animals that had received an uncoated nail, although the reduction in bacterial burden did not reach statistical significance. In conclusion, the PLEX-doxycycline coating on titanium alloy implants provided complete protection against implant-associated MSSA osteomyelitis, and resulted in a significant reduction in the number of culture positive samples when challenged with a
Meeker, Daniel G.; Beenken, Karen E.; Mills, Weston B.; Loughran, Allister J.; Spencer, Horace J.; Lynn, William B.
We used in vitro and in vivo models of catheter-associated biofilm formation to compare the relative activity of antibiotics effective against methicillin-resistant Staphylococcus aureus (MRSA) in the specific context of an established biofilm. The results demonstrated that, under in vitro conditions, daptomycin and ceftaroline exhibited comparable activity relative to each other and greater activity than vancomycin, telavancin, oritavancin, dalbavancin, or tigecycline. This was true when assessed using established biofilms formed by the USA300 methicillin-resistant strain LAC and the USA200 methicillin-sensitive strain UAMS-1. Oxacillin exhibited greater activity against UAMS-1 than LAC, as would be expected, since LAC is an MRSA strain. However, the activity of oxacillin was less than that of daptomycin and ceftaroline even against UAMS-1. Among the lipoglycopeptides, telavancin exhibited the greatest overall activity. Specifically, telavancin exhibited greater activity than oritavancin or dalbavancin when tested against biofilms formed by LAC and was the only lipoglycopeptide capable of reducing the number of viable bacteria below the limit of detection. With biofilms formed by UAMS-1, telavancin and dalbavancin exhibited comparable activity relative to each other and greater activity than oritavancin. Importantly, ceftaroline was the only antibiotic that exhibited greater activity than vancomycin when tested in vivo in a murine model of catheter-associated biofilm formation. These results emphasize the need to consider antibiotics other than vancomycin, most notably, ceftaroline, for the treatment of biofilm-associated S. aureus infections, including by the matrix-based antibiotic delivery methods often employed for local antibiotic delivery in the treatment of these infections. PMID:27401574
Fernandez, Silvina; Murzicato, Sofía; Sandoval, Orlando; Fernández-Canigia, Liliana; Mollerach, Marta
Community-acquired methicillin-resistant Staphylococcus aureus is the first cause of skin and soft tissue infections, but can also produce severe diseases such as bacteremia, osteomyelitis and necrotizing pneumonia. Some S. aureus lineages have been described in cases of necrotizing pneumonia worldwide, usually in young, previously healthy patients. In this work, we describe a fatal case of necrotizing pneumonia due to community-acquired methicillin-resistant S. aureus clone ST30-SCCmecIVc-spat019-PVL positive in an immunocompetent adult patient.
Sedighi, Iraj; Moez, Hoda Jaefari; Alikhani, Mohammad Yousef
Nasal colonization with community acquired methicillin resistant Staphylococcus aureus (CA-MRSA) is being increasingly reported, especially in places where people are in close contact and in reduced hygiene, such as day-care centers. In this study we investigated the frequency of MRSA colonization and their antibiotic susceptibility patterns in 1-6 years old children of day-care centers in Hamadan, West of Iran.Five hundred nasal swabs were collected from children of 27 day-care centers that had no risk factors for colonization by S. aureus. The specimens were cultured for isolation of S. aureus by standard methods. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. For evaluation of the frequency of erythromycin induced clindamycin resistance, disk approximation test (D-test) was applied.Totally, 148 (29.6%) children were colonized by S. aureus. Out of 260 male, 94 (36.2%) and of 240 female, 54 (22.5%) cases were nasal carriers of S. aureus (P value = 0.001). Six (4.1%) of the 148 S. aureus isolated from children were MRSA strains. None of MRSA and methicillin susceptible S. aureus (MSSA) was resistant to vancomycin and clindamycin. Three of the 6 strains of MRSA and 7 (4.9%) of the 142 MSSA strains were resistant to erythromycin, and D-test was positive in all of them.We conclude that the rate of colonization by S. aureus is high in children attending day-care centers but colonization with MRSA is not common in our areas. Clindamycin or trimethoprim-sulfamethoxazol could be used in mild to moderataly severe diseases caused by CA-MRSA. However, if the CA-MRSA isolates are erythromycin resistant, D-test should be carried out for detection of inducible clindamycin resistance.
Gaur, Rashmi; Gupta, Vivek Kumar; Singh, Pooja; Pal, Anirban; Darokar, Mahendra Padurang; Bhakuni, Rajendra Singh
Isoliquiritigenin (ISL) and liquiritigenin (LTG) are structurally related flavonoids found in a variety of plants. Discovery of novel antimicrobial combinations for combating methicillin-resistant Staphylococcus aureus (MRSA) infections is of vital importance in the post-antibiotic era. The present study was taken to explore the in vitro and in vivo combination effect of LTG and ISL with β-lactam antibiotics (penicillin, ampicillin and oxacillin) against mec A-containing strains of MRSA. Minimum inhibitory concentration (MIC) of both LTG and ISL exhibited significant anti-MRSA activity (50-100 µg/mL) against clinical isolates of MRSA. The result of in vitro combination study showed that ISL significantly reduced MIC of β-lactam antibiotics up to 16-folds [∑ fractional inhibitory concentration (FIC) 0.312-0.5], while LTG reduced up to 8-folds (∑FIC 0.372-0.5). Time kill kinetics at graded MIC combinations (ISL/LTG + β-lactam) indicated 3.27-9.79-fold and 2.59-3.48-fold reduction in the growth of clinical isolates of S. aureus respectively. In S. aureus-infected Swiss albino mice model, combination of ISL with oxacillin significantly (p < 0.05, p < 0.01, p < 0.001) lowered the systemic microbial burden in blood, liver, kidney, lung and spleen tissues in comparison with ISL, oxacillin alone as well as untreated control. Considering its synergistic antibacterial effect, we suggest both ISL and LTG as promising compounds for the development of novel antistaphylococcal combinations. Copyright © 2016 John Wiley & Sons, Ltd.
Claeys, Kimberly C.; Lagnf, Abdalhamid M.; Hallesy, Jessica A.; Compton, Matthew T.; Gravelin, Alison L.; Davis, Susan L.
Vancomycin remains the mainstay treatment for methicillin-resistant Staphylococcus aureus (MRSA) infections, including pneumonia. There is concern regarding the emergence of vancomycin tolerance, caused by heterogeneous vancomycin-intermediate S. aureus (hVISA), and subsequent vancomycin treatment failure. Pneumonia is associated with high morbidity and mortality, especially with delays in appropriate therapy. This study evaluated the clinical outcomes of patients with hVISA pneumonia compared to those with vancomycin-susceptible S. aureus (VSSA) pneumonia. A retrospective cohort of patients with MRSA pneumonia from 2005 to 2014 was matched at a ratio of 2:1 VSSA to hVISA infections to compare patient characteristics, treatments, and outcomes. hVISA was determined by the 48-h population analysis profile area under the curve. Characteristics between VSSA and hVISA infections were compared by univariate analysis and multivariable logistic regression analysis to determine independent risk factors of inpatient mortality. Eighty-seven patients were included, representing 29 hVISA and 58 VSSA cases of pneumonia. There were no significant differences in demographics or baseline characteristics. Sequential organ failure assessment (SOFA) scores were a median of 7 (interquartile ratio [IQR], 5 to 8) in hVISA patients and 5 (IQR, 3 to 8) in VSSA (P = 0.092) patients. Inpatient mortality was significantly higher in hVISA patients (44.8% versus 24.1%; P = 0.049). Predictors of inpatient mortality upon multivariable regression were SOFA score (adjusted odds ratio [aOR], 1.36; 95% confidence interval [CI], 1.08 to 1.70), Panton-Valentine leukocidin (PVL) positivity (aOR, 6.63; 95% CI, 1.79 to 24.64), and hVISA phenotype (aOR, 3.95; 95% CI, 1.18 to 13.21). Patients with hVISA pneumonia experienced significantly higher inpatient mortality than those with VSSA pneumonia. There is a need to consider the presence of vancomycin heteroresistance in pneumonia caused by MRSA in order to
Nillius, Dorothea; von Müller, Lutz; Wagenpfeil, Stefan; Klein, Renate; Herrmann, Mathias
Background Multiresistant organisms pose a threat for patients and care recipients. Control interventions need to be tailored to region, the type of institution considered, and risk factors. The German state of Saarland is ideally suited to study colonisation epidemiology throughout its various health and care institutions. After conclusion of a large admission prevalence study in acute care hospitals, we now performed a methicillin-resistant Staphylococcus aureus (MRSA) point prevalence study in Saarland long term care facilities (LTCF), allowing for a direct comparison with respect of MRSA prevalence and associated risk factors between these two institutional types located within a confined region. Methodology and Principal Findings Of all LTCF of the region, 65/136 participated in the study performed between 09/2013 and 07/2014. Overall, complete microbiological specimen and questionnaires of 2,858 of 4,275 (66.8%) LTCF residents were obtained. 136/2,858 (4.8%) screened residents revealed MRSA carrier status. Multivariate risk factor analysis yielded ulcer/deep soft tissue infection, urinary tract catheter, and MRSA history with multiple MRSA decolonisation cycles to be independently associated with MRSA carrier status. Conclusion As already known from previous studies, colonisation with MRSA is common in LTCF residents even in an area with relatively low MRSA prevalence. This found prevalence can now be related to the acute care admission prevalence (2.2%) as well as to the admission prevalence in acute care geriatric departments (7.6%). The common clonal attribution (spa type) of MRSA isolates prevalent in the LTCF population as well as in the acute care admission population points towards a close relationship between both types of institutions. However, the ostensible absence of risk factors such as “previous hospitalisation” in conjunction with newly identified factors such as “multiple decolonisation cycles” refers to MRSA colonisation risks
Wang, Min; Zheng, Yi; Mediavilla, Jose. R.; Chen, Liang; Kreiswirth, Barry. N.; Song, Yajun; Yang, Ruifu; Du, Hong
Methicillin-resistant Staphylococcus aureus (MRSA), is one of the most prevalent clinical pathogens isolated from hospital settings, and has increasingly identified in community settings. In China, the SCCmecIII-ST239 strains are disseminated in different geographic regions, accounting for >75% of all MRSA isolates in some national studies. Here we characterized 150 non-duplicate MRSA isolates collected from February 2012 to May 2013 in a tertiary hospital in Suzhou, Eastern China, to explore the molecular epidemiology. All isolates were characterized by spa typing, SCCmec typing, and detection of genes encoding Panton-Valentine leukocidin (PVL) and toxic shock syndrome toxin (TSST-1). Representative genotypes were also subjected to multilocus sequence typing (MLST). Antibiotic susceptibility testing was performed using BD Phoenix™ Automated Microbiology System. Molecular typing identified 11 clonal complex (CC) and 28 spa types, with the CC5-spa t002 (29.3%) and CC239-spa t037 (14.7%) being the most prevalent. SCCmec types II, III, IV, and V were identified in 33.3, 21.3, 23.3, and 21.3% of all isolates, respectively. PVL genes (lukF/S-PV) were detected in 11.3% of all isolates and from 6 CCs (5, 8, 59, 88, 239, and 398). The TSST-1 gene (tst) was detected in 18.0% of the all isolates, predominantly in CC5 (96.3%). All the tst-1-positve CC5 isolates were spa t002. Eighteen patients died within 30 days of hospitalization, and the in-hospital 30-day mortality was 12.0%. Multivariable analysis showed that 60 years old (odds ratio [OR] = 7.2, P = 0.026), cancer diagnosis (OR = 9.6, P = 0.022), and MRSA isolate carriage of tst-1 (OR = 62.5, P < 0.001) were independent factors associated with 30-day mortality. Our study revealed unique MRSA dissemination patterns in our hospital in comparison to those of other regions in China. The finding that tst-1-positive CC5 strains were associated with higher mortality highlights the need for strict infection control measures in
Medina, G; Egea, A L; Otth, C; Otth, L; Fernández, H; Bocco, J L; Wilson, M; Sola, C
Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen of public health importance. In Chile, the Cordobes/Chilean clone was the predominant healthcare-associated MRSA (HA-MRSA) clone in 1998. Since then, the molecular epidemiological surveillance of MRSA has not been performed in Southern Chile. We aimed to investigate the molecular epidemiology of HA-MRSA infections in Southern Chile to identify the MRSA clones involved, and their evolutionary relationships with epidemic international MRSA lineages. A total of 303 single inpatient isolates of S. aureus were collected in the Valdivia County Hospital (2007-2008), revealing 33% (100 MRSA/303) prevalence for HA-MRSA infections. The SCCmec types I and IV were identified in 97% and 3% of HA-MRSA, respectively. All isolates lacked the pvl genes. A random sample (n = 29) of all MRSA was studied by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), SCCmec subtyping, agr and spa typing, and virulence genes profiling. PFGE analysis revealed the predominance (89%, 26/29) of pulsotype A and three additional pulsotypes, designated H1, I33, and G1. Pulsotype A (ST5-SCCmecI-spa-t149) is clonally related to the Cordobes/Chilean clone. Pulsotype H1 (ST5-SCCmecIVNT-spa-t002) is genetically related to the Pediatric clone (ST5-SCCmecIV). Pulsotype I33 (ST5-SCCmecIVc-spa-t002) is clonally related by PFGE to the community-associated MRSA (CA-MRSA) clone spread in Argentina, I-ST5-IVa-PVL(+). The G1 pulsotype (ST8-SCCmecIVc-spa-t024) is clonally related to the epidemic USA300 CA-MRSA. Here, we demonstrate the stability of the Cordobes/Chilean clone over time as the major HA-MRSA clone in Southern Chile. The identification of two CA-MRSA clones might suggest that these clones have entered into the healthcare setting from the community. These results emphasize the importance of the local surveillance of MRSA infections in the community and hospital settings.
Conceição, Teresa; Coelho, Céline; Santos Silva, Isabel; de Lencastre, Hermínia; Aires-de-Sousa, Marta
Although the nosocomial prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Angola is over 60% and one of the highest in Africa, the extent of MRSA in the community is unknown. To fill this gap, we conducted a hospital-based study in which 158 children attending the emergency ward and ambulatory services of a pediatric hospital in Luanda, the capital of Angola, were screened for S. aureus nasal colonization. Overall, 70 (44.3%) individuals were colonized with S. aureus, of which 20 (28.6%) carried MRSA, resulting in a prevalence of 12.7% (20/158) of MRSA in the population screened. Molecular characterization by pulsed-field gel electrophoresis (PFGE), spa typing, multilocus sequence typing, and SCCmec typing distributed the isolates into two major MRSA clones and one dominant methicillin-susceptible S. aureus (MSSA) lineage, corresponding to the main clones circulating in hospitals in Luanda. The MRSA isolates mainly belonged to clones A (PFGE type A, spa type t105, ST5-IVa-65%) and B (PFGE B, t3869, ST88-IVa-30%), while MSSA isolates mainly belonged to clone L (PFGE type L, t861, ST508-42%). S. aureus isolates showed resistance to penicillin (96%), rifampin (87%), and trimethoprim-sulfamethoxazole (21%). In conclusion, the prevalence of MRSA among children in the community in Luanda is high and seems to originate from hospitals, warranting continuous monitoring and implementation of additional infection control measures.
Singh Lubana, Sandeep; Alfishawy, Mostafa; Singh, Navdeep; Brennessel, Debra J.
Acupuncture is one of the oldest medical procedures in the world and originated in China about 2,000 years ago. Acupuncture is a form of complementary medicine and has gained popularity worldwide in the last few decades. It is mainly used for the treatment of chronic pain. Acupuncture is usually considered a safe procedure but has been reported to cause serious complications including death. It has been associated with transmission of many viruses and bacteria. Two cases of Methicillin-Resistant Staphylococcus aureus have been reported recently following acupuncture therapy. We are reporting a case of a 57-year-old Korean female who developed vertebral osteomyelitis and intraspinal and paraspinal abscesses as a complication of acupuncture. Blood cultures, skin lesion culture, and body fluid culture yielded Methicillin-Resistant Staphylococcus aureus (MRSA). Good anatomical and medical knowledge, good hygiene standards, and proper acupuncture techniques should be followed to prevent the complications. Acupuncturists should consistently review the infection control guidelines to acupuncture. This case should raise awareness of such condition and hazards of presumably benign procedures such as acupuncture. PMID:26257786
Necula, Bogdan S; Fratila-Apachitei, Lidy E; Zaat, Sebastian A J; Apachitei, Iulian; Duszczyk, Jurek
The aim of this study was the synthesis of a porous TiO(2)-Ag composite coating and assessment of its in vitro bactericidal activity against methicillin-resistant Staphylococcus aureus. The coating was produced by plasma electrolytic oxidation of Ti-6Al-7Nb medical alloy in a calcium acetate/calcium glycerophosphate electrolyte bearing Ag nanoparticles. Following oxidation, the surface of the titanium substrate was converted into the corresponding oxide (TiO(2)) bearing Ca and P species from the electrolyte. In addition, Ag was detected associated with particles present in the oxide layers. The coatings revealed a porous interconnected structure with pores up to 3 microm in size, a threefold increase in roughness and improved wettability relative to the non-oxidized specimens. The composite TiO(2)-Ag coating showed complete killing of methicillin-resistant S. aureus within 24h in all culture conditions, whereas a 1000-fold increase in bacterial numbers was recorded with the ground titanium specimens and the samples oxidized in the absence of Ag nanoparticles.
Hill-Cawthorne, Grant A; Hudson, Lyndsey O; El Ghany, Moataz Fouad Abd; Piepenburg, Olaf; Nair, Mridul; Dodgson, Andrew; Forrest, Matthew S; Clark, Taane G; Pain, Arnab
Clinical laboratories are increasingly using molecular tests for methicillin-resistant Staphylococcus aureus (MRSA) screening. However, primers have to be targeted to a variable chromosomal region, the staphylococcal cassette chromosome mec (SCCmec). We initially screened 726 MRSA isolates from a single UK hospital trust by recombinase polymerase amplification (RPA), a novel, isothermal alternative to PCR. Undetected isolates were further characterised using multilocus sequence, spa typing and whole genome sequencing. 96% of our tested phenotypically MRSA isolates contained one of the six orfX-SCCmec junctions our RPA test and commercially available molecular tests target. However 30 isolates could not be detected. Sequencing of 24 of these isolates demonstrated recombinations within the SCCmec element with novel insertions that interfered with the RPA, preventing identif