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Sample records for aureus clone st228

  1. Molecular characterization of resistance to Rifampicin in an emerging hospital-associated Methicillin-resistant Staphylococcus aureus clone ST228, Spain

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    Liñares Josefina

    2010-03-01

    Full Text Available Abstract Background Methicillin-resistant S. aureus (MRSA has been endemic in Hospital Universitari de Bellvitge, Barcelona, since 1990. During the 1990-95 period the Iberian clone (ST-247; SCCmec-I was dominant. Isolates of clonal complex 5 (ST-125; SCCmec-IV gradually replaced the Iberian clone from 1996 to 2003. A new multiresistant MRSA phenotype showing rifampicin resistance emerged in 2004 and rapidly increased from 25% in 2004 to 45% in 2006. The aims of this study were i the molecular characterisation of rifampicin resistant MRSA isolates, ii the study of the rifampicin resistance expression by disk diffusion, microdilution and E-test, and iii the analysis of the rpoB gene mutations involved in rifampicin resistance. Results A sample of representative 108 rifampicin-resistant MRSA isolates belonged to a single PFGE genotype, ST-228, SCCmec type I and spa type t041. Of 108 isolates, 104 (96% had a low-level rifampicin resistance (MICs, 2 to 4 mg/L and 4 a high-level rifampicin resistance (MICs, 128 - ≥ 256 mg/L. Disk diffusion and E-test methods failed to identify a low-level rifampicin resistance in 20 and 12 isolates, respectively. A low-level rifampicin resistance was associated with amino acid substitution 481His/Asn in the beta-subunit of RNA polymerase. Isolates with a high-level rifampicin resistance carried additional mutations in the rpoB gene. Conclusions The emergence of MRSA clone ST228-SCCmecI, related to the Southern Germany clone, involved a therapeutical challenge for treating serious MRSA infections. Decreased susceptibility to rifampicin in MRSA strains of ST228-SCCmecI was associated with one or two specific mutations in the rpoB gene. One fifth of isolates with low-level rifampicin-resistance were missed by the diffusion methods.

  2. Missense mutations in PBP2A Affecting ceftaroline susceptibility detected in epidemic hospital-acquired methicillin-resistant Staphylococcus aureus clonotypes ST228 and ST247 in Western Switzerland archived since 1998.

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    Kelley, William L; Jousselin, Ambre; Barras, Christine; Lelong, Emmanuelle; Renzoni, Adriana

    2015-04-01

    The development and maintenance of an arsenal of antibiotics is a major health care challenge. Ceftaroline is a new cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA); however, no reports concerning MRSA ceftaroline susceptibility have been reported in Switzerland. We tested the in vitro activity of ceftaroline against an archived set of 60 MRSA strains from the University Hospital of Geneva collected from 1994 to 2003. Our results surprisingly revealed ceftaroline-resistant strains (MIC, >1 μg/ml in 40/60 strains; EUCAST breakpoints, susceptible [S], ≤1 μg/ml; resistant [R], >1 μg/ml) were present from 1998 to 2003. The detected resistant strains predominantly belonged to sequence type 228 (ST228) (South German clonotype) but also to ST247 (Iberian clonotype). A sequence analysis of these strains revealed missense mutations in the penicillin-binding protein 2A (PBP2A) allosteric domain (N146K or E239K and N146K-E150K-G246E). The majority of our ST228 PBP2A mutations (N146K or E150K) were distinct from ST228 PBP2A allosteric domain mutations (primarily E239K) recently described for MRSA strains collected in Thailand and Spain during the 2010 Assessing Worldwide Antimicrobial Resistance Evaluation (AWARE) global surveillance program. We also found that similar allosteric domain PBP2A mutations (N146K) correlated with ceftaroline resistance in an independent external ST228 MRSA set obtained from the nearby University Hospital of Lausanne, Lausanne, Switzerland, collected from 2003 to 2008. Thus, ceftaroline resistance was observed in our archived strains (including two examples of an MIC of 4 µg/ml for the Iberian ST247 clonotype with the triple mutation N146K/E150K/G246E), at least as far back as 1998, considerably predating the commercial introduction of ceftaroline. Our results reinforce the notion that unknown parameters can potentially exert selective pressure on PBP2A that can subsequently modulate ceftaroline

  3. Factors contributing to epidemic MRSA clones replacement in a hospital setting.

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    Rossella Baldan

    Full Text Available The mechanisms governing the epidemiology dynamics and success determinants of a specific healthcare-associated methicillin-resistant S. aureus (HA-MRSA clone in hospital settings are still unclear. Important epidemiological changes have occurred in Europe since 2000 that have been related to the appearance of the ST22-IV clone. Between 2006 and 2010, we observed the establishment of the ST22-IV clone displacing the predominant Italian clone, ST228-I, in a large Italian university hospital. To investigate the factors associated with a successful spread of epidemic MRSA clones we studied the biofilm production, the competitive behavior in co-culture, the capacity of invasion of the A549 cells, and the susceptibility to infection in a murine model of acute pneumonia of the two major HA-MRSA clones, ST22-IV and ST228-I. We showed that persistence of ST22-IV is associated with its increased biofilm production and capacity to inhibit the growth of ST228-I in co-culture. Compared to ST228-I, ST22-IV had a significantly higher capacity to invade the A549 cells and a higher virulence in a murine model of acute lung infection causing severe inflammation and determining death in all the mice within 60 hours. On the contrary, ST228-I was associated with mice survival and clearance of the infection. ST22-IV, compared with ST228-I, caused a higher number of persistent, long lasting bacteremia. These data suggest that ST22-IV could have exploited its capacity to i increase its biofilm production over time, ii maintain its growth kinetics in the presence of a competitor and iii be particularly invasive and virulent both in vitro and in vivo, to replace other well-established MRSA clones, becoming the predominant European clone.

  4. Pneumonia and new methicillin-resistant Staphylococcus aureus clone.

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    Garnier, Fabien; Tristan, Anne; François, Bruno; Etienne, Jerome; Delage-Corre, Manuella; Martin, Christian; Liassine, Nadia; Wannet, Wim; Denis, François; Ploy, Marie-Cécile

    2006-01-01

    Necrotizing pneumonia caused by Staphylococcus aureus strains carrying the Panton-Valentin leukocidin gene is a newly described disease entity. We report a new fatal case of necrotizing pneumonia. An S. aureus strain with an agr1 allele and of a new sequence type 377 was recovered, representing a ne

  5. Epidemiology of Staphylococcus aureus in Italy: First nationwide survey, 2012.

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    Campanile, Floriana; Bongiorno, Dafne; Perez, Marianna; Mongelli, Gino; Sessa, Laura; Benvenuto, Sabrina; Gona, Floriana; Varaldo, Pietro E; Stefani, Stefania

    2015-12-01

    A 3-month epidemiological study to determine the prevalence and antibiotic resistance of Staphylococcus aureus nosocomial infections was performed in 52 centres throughout Italy in 2012. A total of 21,873 pathogens were analysed. The prevalence of S. aureus among all nosocomial pathogens isolated in that period was 11.6% (n=2541), whilst the prevalence of methicillin-resistant S. aureus (MRSA) among the S. aureus was 35.8% (n=910). All tested antimicrobials demonstrated ≥92.2% susceptibility against methicillin-susceptible S. aureus, with the exception of clindamycin (89.7%) and erythromycin (84.2%). Among MRSA, percentages of resistance ranged from 12.6% to >39% for tetracycline, rifampicin, clindamycin and gentamicin; higher percentages were found for erythromycin (65.4%) and fluoroquinolones (72.3-85.8%). Overall, the glycopeptide minimum inhibitory concentration (MIC) distribution showed that 58.3% of strains possessed MICs of 1-2mg/L and few strains were linezolid- or daptomycin-resistant. Molecular characterisation was performed on 102 MRSA selected from Northern, Central and Southern regions. Five major clones were found: Italian/ST228-I (t001-t023-t041-t1686-t3217), 33.3%; USA500/ST8-IV (t008), 17.6%; E-MRSA15/ST22-IVh (t020-t025-t032-t223), 16.7%; USA100/ST5-II (t002-t653-t1349-t2164-t3217-t388), 14.7%; and Brazilian/ST239/241-III (t030-t037), 3.9%. Five PVL-positive CA-MRSA isolates, belonging to USA300 and minor clones, were also identified. In conclusion, this first nationwide surveillance study showed that in Italy, S. aureus infections accounted for 11.6% of all nosocomial infections; MRSA accounted for approximately one-third of the S. aureus isolates and these were multidrug-resistant organisms. Five major MRSA epidemic clones were observed and were inter-regionally distributed, with ST228-SCCmecI becoming predominant.

  6. Prevalence of Adhesion and Regulation of Biofilm-Related Genes in Different Clones of Staphylococcus aureus

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    Salman Sahab Atshan

    2012-01-01

    Full Text Available Clinical information about genotypically different clones of biofilm-producing Staphylococcus aureus is largely unknown. We examined whether different clones of methicillin-sensitive and methicillin-resistant S. aureus (MSSA and MRSA differ with respect to staphylococcal microbial surface components recognizing adhesive matrix molecules (MSCRAMMs in biofilm formation. The study used 60 different types of spa and determined the phenotypes, the prevalence of the 13 MSCRAMM, and biofilm genes for each clone. The current investigation was carried out using a modified Congo red agar (MCRA, a microtiter plate assay (MPA, polymerase chain reaction (PCR, and reverse transcriptase polymerase chain reaction (RT-PCR. Clones belonging to the same spa type were found to have similar properties in adheringto thepolystyrene microtiter plate surface. However, their ability to produce slime on MCRA medium was different. PCR experiments showed that 60 clones of MSSA and MRSA were positive for 5 genes (out of 9 MSCRAMM genes. icaADBC genes were found to be present in all the 60 clones tested indicating a high prevalence, and these genes were equally distributed among the clones associated with MSSA and those with MRSA. The prevalence of other MSCRAMM genes among MSSA and MRSA clones was found to be variable. MRSA and MSSA gene expression (MSCRAMM and icaADBC was confirmed by RT-PCR.

  7. Livestock Origin for a Human Pandemic Clone of Community-Associated Methicillin-Resistant Staphylococcus aureus

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    Spoor, Laura E.; McAdam, Paul R.; Weinert, Lucy A.;

    2013-01-01

    ABSTRACT The importance of livestock as a source of bacterial pathogens with the potential for epidemic spread in human populations is unclear. In recent years, there has been a global increase in community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections of healthy...... of emergent clones of methicillin-resistant Staphylococcus aureus (MRSA) that originated in livestock and switched to humans, followed by host-adaptive evolution and epidemic spread in global human populations. Our findings demonstrate that livestock can act as a reservoir for the emergence of new human...

  8. Severe invasive methicillin-resistant S. aureus (USA300 clone infection in an Italian adolescent

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    Piero Valentini

    2009-12-01

    Full Text Available This report describes an uncommon presentation of invasive community-acquired methicillin-resistant S. aureus (CA-MRSA infection in an immunocompetent adolescent without any other risk factor, characterized by septicaemia, meningitis, necrotising pneumonia and deep venous thrombosis (DVT. Successful treatment was performed with linezolid, rifampicin and low-molecular-weight heparin (LMWH. MRSA molecular typing revealed the presence of Panton-Valentine leukocidin (PVL gene, and a genetic background identical to USA300 clone, an emerging aggressive CA-MRSA strain in USA and Europe.

  9. Cloning and Expression of Nano Body Gene against Enterotoxin B of Staphylococcus Aureus

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    Zahra Tavassoli

    2017-02-01

    Full Text Available Background & Objectives: Staphylococcus aureus bacteria causes many different diseases by secretion of various enterotoxins. Therefore, it is necessary to develop ways that facilitate the detection of enterotoxins. Nowadays, immunochemical methods which are based on monoclonal antibody technology are used. The heavy chain antibodies that are called VHH or Nano body were found in blood serum of the Camelidae family. The unique properties of this antibody such as their binding to small molecules like toxins make them attractive candidates for the development of immunodiagnostic tests. The present study was done to achieve a VHH molecules against Staphylococcus enterotoxin B. Materials & Methods: Freighting phage library for isolate private Nano bodies against enterotoxin B was done in previous works. Next, pCANTAB 5E vector that consists VHH, extracted from E.coli bacteria strain xl1blue, and after doing PCR process with relative primers, sub cloning in pET21a(+ as an expression vector with cut sites NdeI and XhoI was done. Transformation in E.coli bacteria strain BL21(DE3 was done. Then, the cells effected with IPTG and producing time, and other terms were optimized. Finally, the expression of the protein with SDS-PAGE and western blot techniques was evaluated. Result: For proving cloning of nano body gene in pET21a (+ vector, nucleotide sequence of gene was analyzed, and transforming to E.coli bacteria strain BL21(DE3 was successful. After inspiration, active protein in cell was seen by SDS-PAGE technique and proved by western blot. Conclusion: cloning, sub cloning, and nonabody expression were surveyed in this research. Production of this protein can help to develop new therapeutic methods and produce vaccine against enterotoxin B of Staphylococcus aureus

  10. Characterization of the epidemic European fusidic acid-resistant impetigo clone of Staphylococcus aureus.

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    O'Neill, A J; Larsen, A R; Skov, R; Henriksen, A S; Chopra, I

    2007-05-01

    Resistance to the antibiotic fusidic acid in European strains of Staphylococcus aureus causing impetigo has increased in recent years. This increase appears to have resulted from clonal expansion of a strain we have designated the epidemic European fusidic acid-resistant impetigo clone (EEFIC), which carries the fusidic acid resistance determinant fusB on its chromosome. To understand better the properties of the EEFIC responsible for its success, we have performed detailed phenotypic and genotypic characterization of this clone. Molecular typing revealed the EEFIC to be ST123, spa type t171, and agr type IV and therefore unrelated to earlier prevalent fusB(+) strains found in the United Kingdom. EEFIC strains exhibited resistance to fusidic acid, penicillin, and, in some cases, erythromycin, which are all used in the treatment of impetigo. PCR analysis of the EEFIC and complete DNA sequencing of the 39.3 Kb plasmid it harbors identified genes encoding several toxins previously implicated in impetigo (exfoliative toxins A and B and EDIN-C). The location of fusB was mapped on the chromosome and found to be associated with a novel 16.6-kb genomic island integrated downstream of groEL. Although this element is related to classical staphylococcal pathogenicity islands, it does not encode any known virulence factors and consequently has been designated SaRI(fusB) (for "S. aureus resistance island carrying fusB").

  11. Outbreak in newborns of methicillin-resistant Staphylococcus aureus related to the sequence type 5 Geraldine clone.

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    Leroyer, Camille; Lehours, Philippe; Tristan, Anne; Boyer, Frederique; Marie, Veronique; Elleau, Christophe; Nolent, Paul; Venier, Anne-Gaelle; Brissaud, Olivier; de Barbeyrac, Bertille; Megraud, Francis; Rogues, Anne-Marie

    2016-02-01

    We describe the first nosocomial outbreak of a toxic shock syndrome-positive methicillin-resistant Staphylococcus aureus (MRSA) sequence type 5 Geraldine clone. Infection control interventions that are usually successful were implemented to control the outbreak. Spread of this virulent MRSA strain highlights the need to be vigilant to MRSA antibiotic susceptibilities.

  12. A field guide to pandemic, epidemic and sporadic clones of methicillin-resistant Staphylococcus aureus.

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    Stefan Monecke

    Full Text Available In recent years, methicillin-resistant Staphylococcus aureus (MRSA have become a truly global challenge. In addition to the long-known healthcare-associated clones, novel strains have also emerged outside of the hospital settings, in the community as well as in livestock. The emergence and spread of virulent clones expressing Panton-Valentine leukocidin (PVL is an additional cause for concern. In order to provide an overview of pandemic, epidemic and sporadic strains, more than 3,000 clinical and veterinary isolates of MRSA mainly from Germany, the United Kingdom, Ireland, France, Malta, Abu Dhabi, Hong Kong, Australia, Trinidad & Tobago as well as some reference strains from the United States have been genotyped by DNA microarray analysis. This technique allowed the assignment of the MRSA isolates to 34 distinct lineages which can be clearly defined based on non-mobile genes. The results were in accordance with data from multilocus sequence typing. More than 100 different strains were distinguished based on affiliation to these lineages, SCCmec type and the presence or absence of PVL. These strains are described here mainly with regard to clinically relevant antimicrobial resistance- and virulence-associated markers, but also in relation to epidemiology and geographic distribution. The findings of the study show a high level of biodiversity among MRSA, especially among strains harbouring SCCmec IV and V elements. The data also indicate a high rate of genetic recombination in MRSA involving SCC elements, bacteriophages or other mobile genetic elements and large-scale chromosomal replacements.

  13. A field guide to pandemic, epidemic and sporadic clones of methicillin-resistant Staphylococcus aureus.

    LENUS (Irish Health Repository)

    Monecke, Stefan

    2011-04-01

    In recent years, methicillin-resistant Staphylococcus aureus (MRSA) have become a truly global challenge. In addition to the long-known healthcare-associated clones, novel strains have also emerged outside of the hospital settings, in the community as well as in livestock. The emergence and spread of virulent clones expressing Panton-Valentine leukocidin (PVL) is an additional cause for concern. In order to provide an overview of pandemic, epidemic and sporadic strains, more than 3,000 clinical and veterinary isolates of MRSA mainly from Germany, the United Kingdom, Ireland, France, Malta, Abu Dhabi, Hong Kong, Australia, Trinidad & Tobago as well as some reference strains from the United States have been genotyped by DNA microarray analysis. This technique allowed the assignment of the MRSA isolates to 34 distinct lineages which can be clearly defined based on non-mobile genes. The results were in accordance with data from multilocus sequence typing. More than 100 different strains were distinguished based on affiliation to these lineages, SCCmec type and the presence or absence of PVL. These strains are described here mainly with regard to clinically relevant antimicrobial resistance- and virulence-associated markers, but also in relation to epidemiology and geographic distribution. The findings of the study show a high level of biodiversity among MRSA, especially among strains harbouring SCCmec IV and V elements. The data also indicate a high rate of genetic recombination in MRSA involving SCC elements, bacteriophages or other mobile genetic elements and large-scale chromosomal replacements.

  14. Single-nucleotide polymorphism genotyping identifies a locally endemic clone of methicillin-resistant Staphylococcus aureus.

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    Ulrich Nübel

    Full Text Available We developed, tested, and applied a TaqMan real-time PCR assay for interrogation of three single-nucleotide polymorphisms that differentiate a clade (termed 't003-X' within the radiation of methicillin-resistant Staphylococcus aureus (MRSA ST225. The TaqMan assay achieved 98% typeability and results were fully concordant with DNA sequencing. By applying this assay to 305 ST225 isolates from an international collection, we demonstrate that clade t003-X is endemic in a single acute-care hospital in Germany at least since 2006, where it has caused a substantial proportion of infections. The strain was also detected in another hospital located 16 kilometers away. Strikingly, however, clade t003-X was not found in 62 other hospitals throughout Germany nor among isolates from other countries, and, hence, displayed a very restricted geographical distribution. Consequently, our results show that SNP-typing may be useful to identify and track MRSA clones that are specific to individual healthcare institutions. In contrast, the spatial dissemination pattern observed here had not been resolved by other typing procedures, including multilocus sequence typing (MLST, spa typing, DNA macrorestriction, and multilocus variable-number tandem repeat analysis (MLVA.

  15. Bovine mastitis outbreak in Japan caused by methicillin-resistant Staphylococcus aureus New York/Japan clone.

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    Hata, Eiji

    2016-05-01

    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.

  16. Rapid Emergence and Evolution of Staphylococcus aureus Clones Harboring fusC-Containing Staphylococcal Cassette Chromosome Elements.

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    Baines, Sarah L; Howden, Benjamin P; Heffernan, Helen; Stinear, Timothy P; Carter, Glen P; Seemann, Torsten; Kwong, Jason C; Ritchie, Stephen R; Williamson, Deborah A

    2016-04-01

    The prevalence of fusidic acid (FA) resistance amongStaphylococcus aureusstrains in New Zealand (NZ) is among the highest reported globally, with a recent study describing a resistance rate of approximately 28%. Three FA-resistantS. aureusclones (ST5 MRSA, ST1 MSSA, and ST1 MRSA) have emerged over the past decade and now predominate in NZ, and in all three clones FA resistance is mediated by thefusCgene. In particular, ST5 MRSA has rapidly become the dominant MRSA clone in NZ, although the origin of FA-resistant ST5 MRSA has not been explored, and the genetic context offusCin FA-resistant NZ isolates is unknown. To better understand the rapid emergence of FA-resistantS. aureus, we used population-based comparative genomics to characterize a collection of FA-resistant and FA-susceptible isolates from NZ. FA-resistant NZ ST5 MRSA displayed minimal genetic diversity and represented a phylogenetically distinct clade within a global population model of clonal complex 5 (CC5)S. aureus In all lineages,fusCwas invariably located within staphylococcal cassette chromosome (SCC) elements, suggesting that SCC-mediated horizontal transfer is the primary mechanism offusCdissemination. The genotypic association offusCwithmecAhas important implications for the emergence of MRSA clones in populations with high usage of fusidic acid. In addition, we found thatfusCwas colocated with a recently described virulence factor (tirS) in dominant NZS. aureusclones, suggesting a fitness advantage. This study points to the likely molecular mechanisms responsible for the successful emergence and spread of FA-resistantS. aureus.

  17. Molecular characterization and clonal diversity of meticillin-resistant Staphylococcus aureus isolated from the community in Spain: emergence of clone sequence type 72.

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    Potel, C; Rey, S; Otero, S; Rubio, J; Álvarez, M

    2016-08-01

    Sequence type 72 meticillin-resistant Staphylococcus aureus (ST72 MRSA) was recently detected in our hospital. Although in Europe this clone is rarely isolated, it is the leading cause of community-associated MRSA infections in Korea, spreading also into hospitals, where it has also emerged as the main MRSA clone recovered from raw meat. We studied MRSA isolated from outpatients in Spain during a nine-year period. More than 70% of the isolates belonged to predominant clones found in hospitals. There was a significant increase in the ST72 prevalence. It appears that boundaries of dominance among MRSA clones have become blurred, demanding continuous surveillance.

  18. Hospital clones of methicillin-resistant Staphylococcus aureus are carried by medical students even before healthcare exposure

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    Ido Orlin

    2017-01-01

    Full Text Available Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA strains are prevalent in healthcare and the community. Few studies have examined MRSA carriage among medical students. The aim of this study is to examine Staphylococcus aureus (SA carriage, and particular MRSA, over time in cohort medical students Methods Prospective collection of nasal swabs from medical students in Israel and assessment of SA carriage. Three samples were taken per student in preclinical and clinical parts of studies. Antibiotic susceptibilities were recorded and MRSA typing was performed by staphylococcal cassette chromosome mec (SCCmec types, Panton Valentine Leukocidin (PVL encoding genes, and spa types. Clonality was assessed by pulsed-field gel electrophoresis. Results Among 58 students, SA carriage rates increased from 33% to 38% to 41% at baseline (preclinical studies, 13 and 19 months (clinical studies, respectively (p = 0.07. Methicillin-susceptible SA (MSSA carriage increased in the clinical studies period (22 to 41%, p = 0.01. Overall, seven students (12% carried 13 MRSA isolates. MRSA isolates were PVL negative and were characterized as SCCmecII-t002, SCCmecIV-t032, or t12435 with untypable SCCmec. MRSA carriage during the pre-clinical studies was evident in 4/7 students. Two students carried different MRSA clones at various times and persistent MRSA carriage was noted in one student. Simultaneous carriage of MRSA and MSSA was not detected. Conclusions MSSA carriage increased during the clinical part of studies in Israeli medical students. Compared with previous reports, higher rates of MRSA carriage were evident. MRSA strains were genotypically similar to Israeli healthcare-associated clones; however, carriage occurred largely before healthcare exposure, implying community-acquisition of hospital strains.

  19. Staphylococcus aureus eye infections in two Indian hospitals: emergence of ST772 as a major clone

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    Nadig S

    2012-01-01

    Full Text Available Savitha Nadig1, Nithya Velusamy2, Prajna Lalitha2, Sarita Kar3, Savitri Sharma3, Gayathri Arakere11Society for Innovation and Development, Indian Institute of Science, Bengaluru, Karnataka, 2Aravind Eye Hospital, Madurai, Tamil Nadu, 3LV Prasad Eye Institute, Bhubaneswar, Odisha, IndiaPurpose: The purpose of this study was to perform molecular characterization of Staphylococcus aureus isolates causing a variety of eye infections from two major eye care hospitals in India.Methods: Twenty-four isolates from Aravind Eye Hospital, Madurai, India, and nine isolates from LV Prasad Eye Institute, Bhubaneswar, India, representing severe to nonsevere eye infections like microbial keratitis to lacrimal sac abscess, were characterized. Staphylococcal cassette chromosome mec typing, multilocus sequence typing, accessory gene regulator typing, staphylococcal protein A typing, and pulsed field gel electrophoresis were used, along with determination of the presence of Panton–Valentine leucocidin toxin and endotoxin gene cluster among each sequence type.Results: The majority of eye infections, both severe and nonsevere, were caused by sequence type (ST772, positive for the Panton–Valentine leucocidin gene, and carrying methicillin-resistant staphylococcal cassette chromosome mec type V cassette (22/33, 67%. Some of the other sequence types that caused severe eye infections were ST1 (9%, 5 (3%, 72 (6%, 88 (3%, 121 (3%, and 672 (3%. This is the first report of the presence of ST1 and 88 in India.Conclusion: Although the number of isolates included in this study was small, most of the eye infections were caused by community-associated S. aureus where patients had no history of hospitalization or treatment in the past year. In the case of six severe infections, patients were admitted for surgeries and there is probability of hospital infection. In addition, only methicillin-resistant S. aureus isolates carrying staphylococcal cassette chromosome mec type V were

  20. Phylogeographic variation in recombination rates within a global clone of Methicillin-Resistant Staphylococcus aureus (MRSA)

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    Castillo-Ramirez, Santiago; Corander, Jukka; Marttinen, Pekka;

    2012-01-01

    that the rapid global dissemination of a single pathogenic bacterial clone results in local variation in measured recombination rates. Possible explanatory variables include the size and time since emergence of each defined sub-population (as determined by the sampling frame), variation in transmission dynamics...... and demonstrate that the rate of recombination varies significantly between phylogeographic sub-groups representing independent introductions from Europe. These differences are most striking when mobile non-core genes are included, but remain apparent even when only considering the stable core genome...

  1. Interspecies spread of Staphylococcus aureus clones among companion animals and human close contacts in a veterinary teaching hospital. A cross-sectional study in Greece.

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    Drougka, Eleanna; Foka, Antigoni; Koutinas, Christos K; Jelastopulu, Eleni; Giormezis, Nikolaos; Farmaki, Ourania; Sarrou, Styliani; Anastassiou, Evangelos D; Petinaki, Efthimia; Spiliopoulou, Iris

    2016-04-01

    Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) prevalence among companion animals and veterinary personnel (VP) was investigated. Strains' molecular characteristics were evaluated in order to assess S. aureus transmission. Specimens (224) from colonized and infected sites of 102 animals (92 dogs, 10 cats) and 18 VP were collected during 2012 and 2013. Antibiotic susceptibility was performed by the disk diffusion method and Etest. mecA, mecC, tst (toxic shock syndrome toxin) and lukF/lukS-PV (Panton-Valentine leukocidin, PVL) genes were investigated by PCR. Genotypes were identified by Multi Locus Sequence Typing (MLST), Staphylococcal Cassette Chromosome mec (SCCmec), accessory gene regulator group (agr), spa and Pulsed Field Gel Electrophoresis (PFGE). S. aureus prevalence among pets and VP was 36.3% (37/102) and 38.9% (7/18), respectively. Younger companion animals, those living in rural areas, having a disease upon admission or Coagulase-negative staphylococci co-carriage showed significantly higher prevalence of S. aureus isolation (panimals and VP. Companion animals harbor PVL-positive clones constituting a possible source for transmission to humans.

  2. Complete genome sequence of Staphylococcus aureus strain M1, a unique t024-ST8-IVa Danish methicillin-resistant S. aureus clone

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    Larner-Svensson, Hanna; Worning, Peder; Bartels, Mette

    2013-01-01

    We report the genome sequence, in five contigs, of a methicillin-resistant Staphylococcus aureus isolate designated M1. This clinical isolate was from the index patient of a methicillin-resistant Staphylococcus aureus (MRSA) outbreak in Copenhagen, Denmark, that started in 2003. This strain is se...... is sequence type 8 (ST8), spa type t024, and staphylococcal cassette chromosome mec element (SCCmec) type IVa....

  3. The dissemination of ST80-SCCmec-IV community-associated methicillin resistant Staphylococcus aureus clone in Kuwait hospitals

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    Sarkhoo Eiman

    2010-11-01

    Full Text Available Abstract Background Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA is a global healthcare problem. The purpose of this study was to characterize CA-MRSA clones and their distribution in Kuwait hospitals. Methods In total, 135 CA-MRSA isolates, carrying the SCCmec IV or V genetic elements, isolated in eight hospitals were characterized using antibiogram, pulsed-field gel electrophoresis, multilocus sequence typing, and carriage of genes for Panton-Valentine Leukocidin (PVL, capsular polysaccharides types (cap 5 and 8, accessory genes regulators (agr, Staphylococcal enterotoxins (SE and toxic shock syndrome toxin 1 (tst. Results They were susceptible to vancomycin, teicoplanin and linezolid but resistant to kanamycin (62%, fusidic acid (42.2%, tetracycline (39.3%, erythromycin and clindamycin (21.5%, gentamicin (5.9%, streptomycin (6.7%, trimethoprim (5.9%, mupirocin (6.6% and cadmium acetate (82.2%. They consisted of 10 pulsotypes with the majority belonging to PFGE type I (51.1%, type II (22.2%, type IV (13.3% and type III (3.7%. They belonged to 10 sequence types (ST comprising ST80 (51.1%, ST30 (22.2%, ST5 (14.1%, ST1 (4.45, ST6 (3.7%, ST88 (1.5%, ST834 (1.5%, ST8 (0.7%, ST46 (0.7% and ST950 (0.7%. Genes for PVL, cap 8, cap 5 and agr III, agr I and agr II were detected in 61.5%, 77.3%, 20.7% and 62.2%, 17% and 8.1% of the isolates respectively. Nine (6.7% isolates contained tst while 103 isolates were positive for SE genes with sei (63.0%, seg (41.5% and sed (29.6% as the common SE genes. Conclusions ST80-SCCmecIV was the most common CA-MRSA clone in Kuwait hospitals presenting new challenges for infection control.

  4. Cloning

    Science.gov (United States)

    ... copies of whole animals Therapeutic cloning, which creates embryonic stem cells. Researchers hope to use these cells to grow healthy tissue to replace injured or diseased tissues in the human body. NIH: National Human Genome Research Institute

  5. Epidemiology of methicillin-resistant and -susceptible Staphylococcus aureus in Luanda, Angola: first description of the spread of the MRSA ST5-IVa clone in the African continent.

    Science.gov (United States)

    Conceição, Teresa; Coelho, Céline; Santos-Silva, Isabel; de Lencastre, Hermínia; Aires-de-Sousa, Marta

    2014-10-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a major human pathogen worldwide, and although surveillance studies are available in the most developed countries, data from Angola are inexistent. In June 2012, 295 inpatients and 199 healthcare workers from three hospitals in Luanda, Angola were nasal swabbed for S. aureus and MRSA carriage. A total of 117 individuals (23.7%) were S. aureus nasal carriers, out of which 68 (58.1%) were colonized with MRSA. The majority of the MRSA isolates (74%) belonged to a single clonal lineage, pulsed-field gel electrophoresis (PFGE) A-ST5-IVa associated with three spa types (spa types t105/t311/t11657), followed by PFGE C-ST88-IVa (spa types t186/t325/t786/t1951/t3869) (n=9; 12%); the other 11 MRSA isolates were representatives of 4 additional lineages. Almost half (49%) of the methicillin-susceptible S. aureus (MSSA) isolates belonged to three major clones: PFGE B-ST508 (spa types t050/t861/t1346/t1574/t2626/t12218), PFGE D-ST45 (spa types t939/t11656), and PFGE E-ST30 (spa types t1202/t9118). MSSA isolates presented a high variability of virulence factors, including Panton-Valentine leukocidine (7.9%). MRSA carriage in Luanda is considerably high, and the major clone corresponds to a worldwide epidemic lineage, so far scarcely reported in Africa. Additional infection control measures in this metropolis are mandatory for a global MRSA control.

  6. Lack of association between genotypes and haematogenous seeding infections in a large cohort of patients with methicillin-resistant Staphylococcus aureus bacteraemia from 21 Spanish hospitals.

    Science.gov (United States)

    Gasch, O; Camoez, M; Dominguez, M A; Padilla, B; Pintado, V; Almirante, B; Martín-Gandul, C; López-Medrano, F; de Gopegui, E Ruiz; Ramón Blanco, J; García-Pardo, G; Calbo, E; Horcajada, J P; Granados, A; Jover-Sáenz, A; Dueñas, C; Pujol, M

    2014-04-01

    There is increasing concern regarding the association between certain methicillin-resistant Staphylococcus aureus (MRSA) genotypes and poor clinical outcome. To assess this issue, a large cohort of 579 subjects with MRSA bacteraemia was prospectively followed from June 2008 to December 2009, in 21 hospitals in Spain. Epidemiology, clinical data, therapy, and outcome were recorded. All MRSA strains were analysed in a central laboratory. Presence of a haematogenous seeding infection was the dependent variable in an adjusted logistic regression model. Of the 579 patients included in the study, 84 (15%) had haematogenous seeding infections. Microdilution vancomycin median MIC (IQR) was 0.73 (0.38-3) mg/L. Most MRSA isolates (n = 371; 67%) belonged to Clonal Complex 5 (CC5) and carried an SCCmec element type IV and agr type 2. Isolates belonging to ST8-agr1-SCCmecIV, ST22-agr1-SCCmecIV and ST228-agr2-SCCmecI--a single locus variant of ST5--accounted for 8%, 9% and 9% of the isolates, respectively. After adjusting by clinical variables, any of the clones was associated with increased risk of haematogenous seeding infections. Higher vancomycin MIC was not identified as an independent risk factor, either. In contrast, persistent bacteraemia (OR 4.2; 2.3-7.8) and non-nosocomial acquisition (3.0; 1.7-5.6) were associated with increased risk.

  7. Detection of ST772 Panton-Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus (Bengal Bay clone and ST22 S. aureus isolates with a genetic variant of elastin binding protein in Nepal

    Directory of Open Access Journals (Sweden)

    R.H. Pokhrel

    2016-05-01

    Full Text Available Genetic characteristics were analysed for recent clinical isolates of methicillin-resistant and -susceptible Staphylococcus aureus (MRSA and MSSA respectively in Kathmandu, Nepal. MRSA isolates harbouring Panton-Valentine leukocidin (PVL genes were classified into ST1, ST22 and ST88 with SCCmec-IV and ST772 with SCCmec-V (Bengal Bay clone, while PVL-positive MSSA into ST22, ST30 and ST772. ST22 isolates (PVL-positive MRSA and MSSA, PVL-negative MRSA possessed a variant of elastin binding protein gene (ebpS with an internal deletion of 180 bp, which was similar to that reported for ST121 S. aureus previously outside Nepal. Phylogenetic analysis indicated that the ebpS variant in ST22 might have occurred independently of ST121 strains. This is the first report of ST772 PVL-positive MRSA in Nepal and detection of the deletion variant of ebpS in ST22 S. aureus.

  8. Molecular Characterization of Methicillin Resistant Staphylococcus aureus Strains Isolated from Intensive Care Units in Iran: ST22-SCCmec IV/t790 Emerges as the Major Clone.

    Directory of Open Access Journals (Sweden)

    Mehdi Goudarzi

    Full Text Available The emergence of methicillin-resistant Staphylococcus aureus (MRSA in different patient populations is a major public health concern. This study determined the prevalence and distribution of circulating molecular types of MRSA in hospitalized patients in ICU of hospitals in Tehran.A total of 70 MRSA isolates were collected from patients in eight hospitals. Antimicrobial resistance patterns were determined using the disk diffusion method. The presence of toxin encoding genes and the vancomycin resistance gene were determined by PCR. The MRSA isolates were further analyzed using multi-locus sequence, spa, SCCmec, and agr typing.The MRSA prevalence was 93.3%. Antimicrobial susceptibility testing revealed a high resistance rate (97.1% to ampicillin and penicillin. The rate of resistance to the majority of antibiotics tested was 30% to 71.4%. Two isolates belonging to the ST22-SCCmec IV/t790 clone (MIC ≥ 8 μg/ml had intermediate resistance to vancomycin. The majority of MRSA isolates (24.3% were associated with the ST22-SCCmec IV/t790 clone; the other MRSA clones were ST859-SCCmec IV/t969 (18.6%, ST239-SCCmec III/t037 (17.1%, and ST291-SCCmec IV/t030 (8.6%.The circulating MRSA strains in Iranian hospitals were genetically diverse with a relatively high prevalence of the ST22-SCCmec IV/t790 clone. These findings support the need for future surveillance studies on MRSA to better elucidate the distribution of existing MRSA clones and detect emergence of new MRSA clones.

  9. The dominant Australian community-acquired methicillin-resistant Staphylococcus aureus clone ST93-IV [2B] is highly virulent and genetically distinct.

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    Kyra Y L Chua

    Full Text Available Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA USA300 has spread rapidly across North America, and CA-MRSA is also increasing in Australia. However, the dominant Australian CA-MRSA strain, ST93-IV [2B] appears distantly related to USA300 despite strikingly similar clinical and epidemiological profiles. Here, we compared the virulence of a recent Australian ST93 isolate (JKD6159 to other MRSA, including USA300, and found that JKD6159 was the most virulent in a mouse skin infection model. We fully sequenced the genome of JKD6159 and confirmed that JKD6159 is a distinct clone with 7616 single nucleotide polymorphisms (SNPs distinguishing this strain from all other S. aureus genomes. Despite its high virulence there were surprisingly few virulence determinants. However, genes encoding α-hemolysin, Panton-Valentine leukocidin (PVL and α-type phenol soluble modulins were present. Genome comparisons revealed 32 additional CDS in JKD6159 but none appeared to encode new virulence factors, suggesting that this clone's enhanced pathogenicity could lie within subtler genome changes, such as SNPs within regulatory genes. To investigate the role of accessory genome elements in CA-MRSA epidemiology, we next sequenced three additional Australian non-ST93 CA-MRSA strains and compared them with JKD6159, 19 completed S. aureus genomes and 59 additional S. aureus genomes for which unassembled genome sequence data was publicly available (82 genomes in total. These comparisons showed that despite its distinctive genotype, JKD6159 and other CA-MRSA clones (including USA300 share a conserved repertoire of three notable accessory elements (SSCmecIV, PVL prophage, and pMW2. This study demonstrates that the genetically distinct ST93 CA-MRSA from Australia is highly virulent. Our comparisons of geographically and genetically diverse CA-MRSA genomes suggest that apparent convergent evolution in CA-MRSA may be better explained by the rapid

  10. Cloning of type 8 capsule genes and analysis of gene clusters for the production of different capsular polysaccharides in Staphylococcus aureus.

    Science.gov (United States)

    Sau, S; Lee, C Y

    1996-04-01

    Eleven serotypes of capsular polysaccharide from Staphylococcus aureus have been reported. We have previously cloned a cluster of type 1 capsule (cap1) genes responsible for type 1 capsular polysaccharide biosynthesis in S. aureus M. To clone the type 8 capsule (cap8) genes, a plasmid library of type 8 strain Becker was screened with a labelled DNA fragment containing the cap1 genes under low-stringency conditions. One recombinant plasmid containing a 14-kb insert was chosen for further study and found to complement 14 of the 18 type 8 capsule-negative (Cap8-) mutants used in the study. Additional library screening, subcloning, and complementation experiments showed that all of the 18 Cap8- mutants were complemented by DNA fragments derived from a 20.5-kb contiguous region of the Becker chromosome. The mutants were mapped into six complementation groups, indicating that the cap8 genes are clustered. By Southern hybridization analyses under high-stringency conditions, we found that DNA fragments containing the cap8 gene cluster show extensive homology with all 17 strains tested, including type 1 strains. By further Southern analyses and cloning of the cap8-related homolog from strain M, we show that strain M carries an additional capsule gene cluster different from the cap1 gene cluster. In addition, by using DNA fragments containing different regions of the cap8 gene cluster as probes to hybridize DNA from different strains, we found that the central region of the cap8 gene cluster hybridizes only to DNAs from certain strains tested whereas the flanking regions hybridize to DNAs of all strains tested. Thus, the cap8 gene clusters and its closely related homologs are likely to have organizations similar to those of the encapsulation genes of other bacterial systems.

  11. Insights into alpha-hemolysin (Hla) evolution and expression among Staphylococcus aureus clones with hospital and community origin

    DEFF Research Database (Denmark)

    Tavares, Ana; Nielsen, Jesper B; Boye, Kit;

    2014-01-01

    in the RNAIII binding site were not associated to hla expression. Although expression rates of hla were in general strain-specific, we observed CA clones showed significantly higher hla expression (p = 0.003) when compared with HA clones. CONCLUSION: We propose that the hla gene has evolved together...

  12. ST2249-MRSA-III: a second major recombinant methicillin-resistant Staphylococcus aureus clone causing healthcare infection in the 1970s.

    Science.gov (United States)

    Nimmo, G R; Steen, J A; Monecke, S; Ehricht, R; Slickers, P; Thomas, J C; Appleton, S; Goering, R V; Robinson, D A; Coombs, G W

    2015-05-01

    Typing of healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) from Australia in the 1970s revealed a novel clone, ST2249-MRSA-III (CC45), present from 1973 to 1979. This clone was present before the Australian epidemic caused by the recombinant clone, ST239-MRSA-III. This study aimed to characterize the genome of ST2249-MRSA-III to establish its relationship to other MRSA clones. DNA microarray analysis was conducted and a draft genome sequence of ST2249 was obtained. The recombinant structure of the ST2249 genome was revealed by comparisons to publicly available ST239 and ST45 genomes. Microarray analysis of genomic DNA of 13 ST2249 isolates showed gross similarities with the ST239 chromosome in a segment around the origin of replication and with ST45 for the remainder of the chromosome. Recombination breakpoints were precisely determined by the changing pattern of nucleotide polymorphisms in the genome sequence of ST2249 isolate SK1585 compared with ST239 and ST45. One breakpoint was identified to the right of oriC, between sites 1014 and 1065 of the gene D484_00045. Another was identified to the left of oriC, between sites 1185 and 1248 of D484_01632. These results indicate that ST2249 inherited approximately 35.3% of its chromosome from an ST239-like parent and 64.7% from an ST45-like parent. ST2249-MRSA-III resulted from a major recombination between parents that resemble ST239 and ST45. Although only limited Australian archival material is available, the oldest extant isolate of ST2249 predates the oldest Australian isolate of ST239 by 3 years. It is therefore plausible that these two recombinant clones were introduced into Australia separately.

  13. New patterns of methicillin-resistant Staphylococcus aureus (MRSA) clones, community-associated MRSA genotypes behave like healthcare-associated MRSA genotypes within hospitals, Argentina.

    Science.gov (United States)

    Egea, Ana L; Gagetti, Paula; Lamberghini, Ricardo; Faccone, Diego; Lucero, Celeste; Vindel, Ana; Tosoroni, Dario; Garnero, Analía; Saka, Hector A; Galas, Marcelo; Bocco, José L; Corso, Alejandra; Sola, Claudia

    2014-11-01

    Methicillin-resistant Staphylococcus aureus (MRSA) burden is increasing worldwide in hospitals [healthcare-associated (HA)-MRSA] and in communities [community-associated (CA)-MRSA]. However, the impact of CA-MRSA within hospitals remains limited, particularly in Latin America. A countrywide representative survey of S. aureus infections was performed in Argentina by analyzing 591 clinical isolates from 66 hospitals in a prospective cross-sectional, multicenter study (Nov-2009). This work involved healthcare-onset infections-(HAHO, >48 hospitalization hours) and community-onset (CO) infections [including both, infections (HACO) in patients with healthcare-associated risk-factors (HRFs) and infections (CACO) in those without HRFs]. MRSA strains were genetically typed as CA-MRSA and HA-MRSA genotypes (CA-MRSAG and HA-MRSAG) by SCCmec- and spa-typing, PFGE, MLST and virulence genes profile by PCR. Considering all isolates, 63% were from CO-infections and 55% were MRSA [39% CA-MRSAG and 16% HA-MRSAG]. A significantly higher MRSA proportion among CO- than HAHO-S. aureus infections was detected (58% vs 49%); mainly in children (62% vs 43%). The CA-MRSAG/HA-MRSAG have accounted for 16%/33% of HAHO-, 39%/13% of HACO- and 60.5%/0% of CACO-infections. Regarding the epidemiological associations identified in multivariate models for patients with healthcare-onset CA-MRSAG infections, CA-MRSAG behave like HA-MRSAG within hospitals but children were the highest risk group for healthcare-onset CA-MRSAG infections. Most CA-MRSAG belonged to two major clones: PFGE-type N-ST30-SCCmecIVc-t019-PVL(+) and PFGE-type I-ST5-IV-SCCmecIVa-t311-PVL(+) (45% each). The ST5-IV-PVL(+)/ST30-IV-PVL(+) clones have caused 31%/33% of all infections, 20%/4% of HAHO-, 43%/23% of HACO- and 35%/60% of CACO- infections, with significant differences by age groups (children/adults) and geographical regions. Importantly, an isolate belonging to USA300-0114-(ST8-SCCmecIVa-spat008-PVL(+)-ACME(+)) was detected

  14. Two distinct clones of methicillin-resistant Staphylococcus aureus (MRSA) with the same USA300 pulsed-field gel electrophoresis profile: a potential pitfall for identification of USA300 community-associated MRSA

    DEFF Research Database (Denmark)

    Larsen, Anders Rhod; Goering, Richard; Stegger, Marc;

    2009-01-01

    Analysis of methicillin-resistant Staphylococcus aureus (MRSA) characterized as USA300 by pulsed-field gel electrophoresis identified two distinct clones. One was similar to community-associated USA300 MRSA (ST8-IVa, t008, and Panton-Valentine leukocidin positive). The second (ST8-IVa, t024...

  15. Molecular characterization of methicillin-resistant Panton-valentine leukocidin positive staphylococcus aureus clones disseminating in Tunisian hospitals and in the community

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    Mariem Ben Jomàa-Jemili

    2013-01-01

    Full Text Available Abstract Background The spread of MRSA strains at hospitals as well as in the community are of great concern worldwide. We characterized the MRSA clones isolated at Tunisian hospitals and in the community by comparing them to those isolated in other countries. Results We characterized 69 MRSA strains isolated from two Tunisian university hospitals between the years 2004-2008. Twenty-two of 28 (79% community-associated MRSA (CA-MRSA strains and 21 of 41 (51% healthcare-associated MRSA (HA-MRSA strains were PVL-positive. The PVL-positive strains belonged to predicted founder group (FG 80 in MLST and carried either type IVc SCCmec or nontypeable SCCmec that harbours the class B mec gene complex. In contrast, very diverse clones were identified in PVL-negative strains: three FGs (5, 15, and 22 for HA-MRSA strains and four FGs (5, 15, 45, and 80 for CA-MRSA strains; and these strains carried the SCCmec element of either type I, III, IVc or was nontypeable. The nucleotide sequencing of phi7401PVL lysogenized in a CA-MRSA strain JCSC7401, revealed that the phage was highly homologous to phiSA2mw, with nucleotide identities of more than 95%. Furthermore, all PVL positive strains were found to carry the same PVL phage, since these strains were positive in two PCR studies, identifying gene linkage between lukS and mtp (major tail protein and the lysogeny region, both of which are in common with phi7401PVL and phiSa2mw. Conclusions Our experiments suggest that FG80 S. aureus strains have changed to be more virulent by acquiring phi7401PVL, and to be resistant to β-lactams by acquiring SCCmec elements. These novel clones might have disseminated in the Tunisian community as well as at the Tunisian hospitals by taking over existing MRSA clones.

  16. Sequence type 72 community-associated meticillin-resistant Staphylococcus aureus emerged as a predominant clone of nasal colonization in newly admitted patients.

    Science.gov (United States)

    Park, S Y; Chung, D R; Yoo, J R; Baek, J Y; Kim, S H; Ha, Y E; Kang, C-I; Peck, K R; Lee, N Y; Song, J-H

    2016-08-01

    Current knowledge of community-associated (CA) meticillin-resistant Staphylococcus aureus (MRSA) carriage in hospitalized patients is incomplete. Genotypic characteristics of 637 nasal MRSA isolates from newly admitted patients in South Korea were investigated. Sequence type (ST) 72 accounted for 52.1%, 46.3%, and 52.8% of the isolates during the periods of 2007-2008, 2009-2010, and 2013-2014, respectively. Instead of classic MRSA clones responsible for healthcare-associated infections, including ST5 and ST239, MRSA with community genotype ST72 was the predominant strain in newly admitted patients regardless of age and home province of the patients. Active strategies are needed to prevent healthcare-associated infection by CA-MRSA.

  17. An uncommon presentation for a severe invasive infection due to methicillin-resistant Staphylococcus aureus clone USA300 in Italy: a case report

    Directory of Open Access Journals (Sweden)

    Tronci Mirella

    2008-04-01

    Full Text Available Abstract Background Methicillin resistant Staphylococcus aureus (MRSA has been considered for many years a typical nosocomial pathogen. Recently MRSA has emerged as a frequent cause of infections in the community. More commonly, community-acquired (CA-MRSA is a cause of infections of the skin and soft-tissues, but life-threatening infections such as necrotizing pneumonia and sepsis can occasionally occur. Case presentation This report describes an uncommon presentation of invasive CA-MRSA infection in an adolescent without known risk factors. The presentation was typical for bacterial meningitis, but the clinical findings also revealed necrotizing pneumonia. Following the development of deep venous thrombosis, the presence of an inherited trombophilic defect (factor V Leiden was detected. The patient was successfully treated with an antibiotic combination including linezolid and with anticoagulant therapy. CA-MRSA was isolated from both cerebrospinal fluid and blood. The isolates were resistant to oxacillin and other beta-lactam antibiotics and susceptible to the other antibiotics tested including erythromycin. Molecular typing revealed that the strains contained the Panton-Valentine leukocidin genes and type IV SCCmec, and were ST8, spa type t008, and agr type 1. This genetic background is identical to that of the USA300 clone. Conclusion This report highlights that meningitis can be a new serious presentation of CA-MRSA infection. CA-MRSA strains with the genetic background of the USA300 clone are circulating in Italy and are able to cause severe infections.

  18. Methicillin-resistant Staphylococcus aureus in hospitals in Tbilisi, the Republic of Georgia, are variants of the Brazilian clone

    DEFF Research Database (Denmark)

    Bartels, M.D.; Nanuashvili, A.; Boye, K.;

    2008-01-01

    The purpose of this study was to characterise methicillin-resistant Staphylococcus aureus (MRSA) isolates from the Republic of Georgia, part of the former Soviet Union. Thirty-two non-duplicate MRSA isolates were collected in the period from May 2006 to February 2007. The patient data were analysed...... and the isolates were characterised by staphylococcal protein A (spa) typing, staphylococcal chromosome cassette mec (SCCmec) typing, multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and the detection of Panton-Valentine leukocidin (PVL) genes. Only two closely related spa types were...

  19. 金黄色葡萄球菌FnBP配体结合区基因的克隆及其原核表达%Cloning and Prokaryotic Expression of FnBP Ligand Binding Gene of Staphylococcus aureus

    Institute of Scientific and Technical Information of China (English)

    尹荣兰; 杨正涛; 张艳晶; 刘辉; 刘珊; 杨琦; 曹永国; 张乃生

    2008-01-01

    [Objective] The study aimed to clone the FnBP ligand binding gene of Staphylococcus aureus and run prokaryotic expression by constructing a prokaryotic expression vector. [Method] The gene encoding FnBP ligand binding gene was amplified from S.aureus chromosomal DNA by PCR technique. After T-A cloning, plasmid pMD18- FnBP was constructed. pMD18- FnBP and pET28a(+)were digested by BamH Ⅰ and EcoR Ⅰ double enzymes, then the purified FnBP ligand binding gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-FnBP was thus constructed. The constructed plasmid pET28a-FnBP was transformed into Escherichia coli BL21(DE3) competent cells. The bacterium was induced by IPTG and the expressed products were analyzed by SDS-PAGE and Western blot. [Result] The gene fragment with the length of 370 bp was amplified by PCR approach. One approximately 30 kD exogenous protein was observed in SDS-PAGE analysis. Western blot analysis indicates the protein has antigenicity of S.aureus. [Conclusion] The FnBP ligand binding gene of S.aureus was successfully cloned and expressed in prokaryotic cells.

  20. Genome-wide single nucleotide polymorphism-based assay for high-resolution epidemiological analysis of the methicillin-resistant Staphylococcus aureus hospital clone EMRSA-15.

    Science.gov (United States)

    Holmes, A; McAllister, G; McAdam, P R; Hsien Choi, S; Girvan, K; Robb, A; Edwards, G; Templeton, K; Fitzgerald, J R

    2014-02-01

    The EMRSA-15 clone is a major cause of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infections in the UK and elsewhere but existing typing methodologies have limited capacity to discriminate closely related strains, and are often poorly reproducible between laboratories. Here, we report the design, development and validation of a genome-wide single nucleotide polymorphism (SNP) typing method and compare it to established methods for typing of EMRSA-15. In order to identify discriminatory SNPs, the genomes of 17 EMRSA-15 strains, selected to represent the breadth of genotypic and phenotypic diversity of EMRSA-15 isolates in Scotland, were determined and phylogenetic reconstruction was carried out. In addition to 17 phylogenetically informative SNPs, five binary markers were included to form the basis of an EMRSA-15 genotyping assay. The SNP-based typing assay was as discriminatory as pulsed-field gel electrophoresis, and significantly more discriminatory than staphylococcal protein A (spa) typing for typing of a representative panel of diverse EMRSA-15 strains, isolates from two EMRSA-15 hospital outbreak investigations, and a panel of bacteraemia isolates obtained in healthcare facilities in the east of Scotland during a 12-month period. The assay is a rapid, and reproducible approach for epidemiological analysis of EMRSA-15 clinical isolates in Scotland. Unlike established methods the DNA sequence-based method is ideally suited for inter-laboratory comparison of identified genotypes, and its flexibility lends itself to supplementation with additional SNPs or markers for the identification of novel S. aureus strains in other regions of the world.

  1. Antimicrobial activity of ceftaroline against methicillin-resistant Staphylococcus aureus (MRSA) isolates collected in 2013-2014 at the Geneva University Hospitals.

    Science.gov (United States)

    Andrey, D O; François, P; Manzano, C; Bonetti, E J; Harbarth, S; Schrenzel, J; Kelley, W L; Renzoni, A

    2017-02-01

    Ceftaroline is a broad-spectrum antibiotic with activity against methicillin-resistant Staphylococcus aureus (MRSA) strains. Ceftaroline susceptibility of an MRSA set archived between 1994 and 2003 in the Geneva University Hospitals detected a high percentage (66 %) of ceftaroline resistance in clonotypes ST228 and ST247 and correlated with mutations in PBP2a. The ceftaroline mechanism of action is based on the inhibition of PBP2a; thus, the identification of PBP2a mutations of recently circulating clonotypes in our institution was investigated. We analyzed ceftaroline susceptibility in MRSA isolates (2013 and 2014) and established that resistant strains correlated with PBP2a mutations and specific clonotypes. Ninety-six MRSA strains were analyzed from independent patients and were isolated from blood cultures (23 %), deep infections (38.5 %), and superficial (skin or wound) infections (38.5 %). This sample showed a ceftaroline minimum inhibitory concentration (MIC) range between 0.25 and 2 μg/ml and disk diameters ranging from 10 to 30 mm, with a majority of strains showing diameters ≥20 mm. Based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, 76 % (73/96) of isolates showed susceptibility to ceftaroline. Nevertheless, we still observed 24 % (23/96) of resistant isolates (MIC = 2 μg/ml). All resistant isolates were assigned to clonotype ST228 and carried the N146K mutation in PBP2a. Only two ST228 isolates showed ceftaroline susceptibility. The decreasing percentage of ceftaroline-resistant isolates in our hospital can be explained by the decline of ST228 clonotype circulating in our hospital since 2008. We present evidence that ceftaroline is active against recent MRSA strains from our hospital; however, the presence of PBP2a variants in particular clonotypes may affect ceftaroline efficacy.

  2. Epidemiological features, resistance genes, and clones among community-onset methicillin-resistant Staphylococcus aureus (CO-MRSA) isolates detected in northern Spain.

    Science.gov (United States)

    González-Domínguez, María; Seral, Cristina; Sáenz, Yolanda; Salvo, Soledad; Gude, María José; Porres-Osante, Nerea; Torres, Carmen; Castillo, Francisco Javier

    2012-12-01

    Twenty-nine community-onset methicillin-resistant Staphylococcus aureus (CO-MRSA) isolates were prospectively selected according to epidemiological criteria among 374 MRSA isolates collected in our laboratory during 2009-2010 in order to determine which community-associated MRSA (CA-MRSA) and healthcare-associated MRSA (HA-MRSA) clones are circulating in the community in northern Spain. PVL genes were detected in 5 strains (17.2%) that belonged to SCCmec type IV or V and to the agr group I (ST8 and ST2050), agr group II (ST121), and agr group III (ST30 and ST852). These strains were isolated from patients with different clinical manifestations such as urinary tract infection, abscess, or pneumonia, and most of them belonged to emergency department patients with no history of visits to General Practitioners (GPs) in the year before the isolation. We considered that the prevalence of CA-MRSA in community-onset isolates was low (17.2%). A high proportion of the CO-MRSA strains (58.6%) were ST125-MRSA-IVc (CC5), responsible for most of the infections caused by HA-MRSA strains in Spain. This endemic clone is also circulating in the community of northern Spain as we could demonstrate in this study. Antimicrobial resistance was found in spa type t067 isolates linked to the presence of ant(4')-Ia and msr(A). Most of the CO-MRSA isolates in this study corresponded to spa types more associated to the hospital environment, suggesting the interchange of genetic lineages of MRSA among community and hospital niches.

  3. Genotypic and phenotypic characterization of methicillin-resistant Staphylococcus aureus (MRSA) clones with high-level mupirocin resistance.

    Science.gov (United States)

    González-Domínguez, María; Seral, Cristina; Potel, Carmen; Sáenz, Yolanda; Álvarez, Maximiliano; Torres, Carmen; Castillo, Francisco Javier

    2016-06-01

    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.

  4. A timescale for evolution, population expansion, and spatial spread of an emerging clone of methicillin-resistant Staphylococcus aureus

    DEFF Research Database (Denmark)

    Nübel, Ulrich; Dordel, Janina; Kurt, Kevin;

    2010-01-01

    at 269 genetic loci (118,804 basepairs) within an international isolate collection, we ascertained extremely low diversity among European ST225 isolates, indicating that a recent population bottleneck had preceded the expansion of this clone. In contrast, US isolates were more divergent, suggesting......Due to the lack of fossil evidence, the timescales of bacterial evolution are largely unknown. The speed with which genetic change accumulates in populations of pathogenic bacteria, however, is a key parameter that is crucial for understanding the emergence of traits such as increased virulence...... they represent the ancestral population. While diversity was low, however, our results demonstrate that the short-term evolutionary rate in this natural population of MRSA resulted in the accumulation of measurable DNA sequence variation within two decades, which we could exploit to reconstruct its recent...

  5. Dissemination of multiple MRSA clones among community-associated methicillin-resistant Staphylococcus aureus infections from Japanese children with impetigo.

    Science.gov (United States)

    Hisata, Ken; Ito, Teruyo; Matsunaga, Nobuaki; Komatsu, Mitsutaka; Jin, Jingxun; Li, Shanshuang; Watanabe, Shinya; Shimizu, Toshiaki; Hiramatsu, Keiichi

    2011-10-01

    The proportion of MRSA strains that cause skin and soft infections has recently increased. In 3 months we have characterized 17 MRSA strains isolated from children with impetigo at a Japanese hospital. Seventeen MRSA strains belonged to 7 clones defined by clonal complex (CC) in MLST genotype and type of SCCmec, which were rarely identified among healthcare-associated MRSA: CC 91-SCCmecIIb (4 strains); CC91-SCCmecIIn (2 strains); CC91-SCCmecIVa (2 strains); CC91-SCCmecV (4 strains); CC88-SCCmecIVg (3 strains); CC1-SCCmecIVc (1 strain); and CC5-SCCmecIVn (1 strain). Although one strain belonged to CC5, which has been commonly identified in healthcare-associated MRSA, it did not carry type II SCCmec, but carried type IV SCCmec. Fourteen of the 17 strains carried exfoliative toxin a or b gene, and none carried Panton-Valentine leukocidine gene. Furthermore, we determined the entire nucleotide sequences of two type V SCCmec elements carried by strains JCSC5952, a CC91 strain, and TSGH17, a Taiwanese CC59 strain. The structure of SCCmecJCSC5952 was more than 99% homologous in nucleotide identity with those of Taiwanese PVL-positive ST59 MRSA strains TSGH17 and PM1, which were designated as type V (5C2&5). Identification of multiple MRSA clones distinct from those disseminating at the hospital suggests that MRSA strains might be emerging in the community from MSSA strains by acquiring SCCmec elements on various occasions. Carriage of the similar type V(5C2&5) SCCmec element by strains of distinct genetic backgrounds, CC91 and CC59, suggested horizontal transfer of the SCCmec element.

  6. Methicillin-resistant Staphylococcus aureus carrying SCCmec type II was more frequent than the Brazilian endemic clone as a cause of nosocomial bacteremia.

    Science.gov (United States)

    Caiaffa-Filho, Helio Hehl; Trindade, Priscila A; Gabriela da Cunha, Paula; Alencar, Cecilia Salete; Prado, Gladys V B; Rossi, Flavia; Levin, Anna S

    2013-08-01

    Fifty consecutive MRSA blood isolates were evaluated: 30(60%) carried SCCmec type II (single PFGE clone; sequence type 5 or ST105); 12 (26%), IV; 5 (10%), III; 3 (6%), I. Brazilian endemic clone, carrying SCCmec type III, has been the main nosocomial clone in Brazil; however, this study showed that a clone carrying type II predominated.

  7. Evolution of methicillin-resistant Staphylococcus aureus towards increasing resistance

    DEFF Research Database (Denmark)

    Strommenger, Birgit; Bartels, Mette Damkjær; Kurt, Kevin

    2014-01-01

    To elucidate the evolutionary history of Staphylococcus aureus clonal complex (CC) 8, which encompasses several globally distributed epidemic lineages, including hospital-associated methicillin-resistant S. aureus (MRSA) and the highly prevalent community-associated MRSA clone USA300....

  8. Characteristics of hospital patients colonized with livestock-associated meticillin-resistant Staphylococcus aureus (MRSA) CC398 versus other MRSA clones

    NARCIS (Netherlands)

    Köck, R; Siam, K; Al-Malat, S; Christmann, J; Schaumburg, F; Becker, K; Friedrich, A W

    2011-01-01

    Meticillin-resistant Staphylococcus aureus (MRSA) associated with the clonal complex (CC) 398 has emerged among livestock and humans exposed to these animals. MRSA CC398 has so far contributed relatively little to spread of MRSA and the burden of disease in the healthcare setting. This study aimed t

  9. Cloning and Occurrence of czrC, a Gene Conferring Cadmium and Zinc Resistance in Methicillin-Resistant Staphylococcus aureus CC398 Isolates

    NARCIS (Netherlands)

    Cavaco, L. M.; Hasman, H.; Stegger, M.; Andersen, P. S.; Skov, R.; Fluit, A. C.; Ito, T.; Aarestrup, F. M.

    2010-01-01

    We recently reported a phenotypic association between reduced susceptibility to zinc and methicillin resistance in Staphylococcus aureus CC398 isolates from Danish swine (F. M. Aarestrup, L. M. Cavaco, and H. Hasman, Vet. Microbiol. 142: 455-457, 2009). The aim of this study was to identify the gene

  10. Molecular characterisation of Panton–Valentine leucocidin-producing methicillin-resistant Staphylococcus aureus clones isolated from the main hospitals in Taif, KSA

    Directory of Open Access Journals (Sweden)

    E M Eed

    2016-01-01

    Full Text Available Introduction: Panton–Valentine leucocidin (PVL is a bicomponent pore-forming cytolytic toxin encoded by the lukF-PV and lukS-PV genes. Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA may carry the pvl genes which may be related to increased disease severity. This study aimed to characterise the PVL-producing MRSA recovered from different Taif Hospitals, Saudi Arabia. Methods: The study included 45 hospital-acquired-MRSA (HA-MRSA and 26 CA-MRSA strains which were identified from 445 S. aureus strains isolated from different clinical samples. MRSA strains were identified by standard oxacillin salt agar screening procedure and by the detection of the mecA gene by the polymerase chain reaction (PCR. Detection of the S. aureus-specific femA, mecA and pvl genes was performed by multiplex PCR. PCR-restriction fragment length polymorphism (PCR-RFLP analysis was done for coagulase (coa gene. Results: The staphylococcal cassette chromosome mec types of the 45 HA-MRSA strains were Type I (n = 24, Type II (n = 7 and Type III (n = 14 whereas the 26 CA-MRSA strains were Type IV (n = 14, Type V (n = 11 and one isolate was non-typeable. All the HA-MRSA and six CA-MRSA strains were PVL-negative PCR-RFLP analysis of coa gene showed that PVL-positive MRSA (n = 20 isolates showed six different patterns, and five patterns were shared by PVL-positive methicillin-susceptible S. aureus (MSSA. The eighth pattern was the most frequent in both MRSA and MSSA. Conclusion: PVL is more frequent among CA-MRSA than MSSA. All the HA-MRSA and 25% of CA-MRSA strains were negative for PVL. The pvl gene was related to the severity of infection but not related to coa gene RFLP pattern.

  11. Clonal types and antimicrobial resistance profiles of methicillin-resistant Staphylococcus aureus isolates from hospitals in south Brazil Tipos de clones e perfis de resistência antimicrobiana de Staphylococcus aureus resistentes à meticilina isolados de hospitais no sul do Brasil

    Directory of Open Access Journals (Sweden)

    Leandro Reus Rodrigues Perez

    2008-06-01

    Full Text Available In the present study were evaluated the DNA macrorestriction profile and SCCmec types for nine multi-resistant MRSA selected. Also antimicrobial susceptibility testing by disk diffusion method was evaluated for 68 MRSA isolates against 12 antimicrobial agents. The isolates were recovered from blood culture collected from hospitalized patients in three hospitals of Porto Alegre, Brazil. PFGE and PCR for mecA and SCCmec I, II, III, IV types genes were done on selected nine isolates with susceptibility only to vancomycin, teicoplanin and linezolid. Two clone profiles, with five subtypes, were demonstrated among multi-resistant MRSA analyzed. Eight isolates showed harbor SCCmec type III and one isolate was not typeable. The knowledge of SCCmec type, clone and antimicrobial profiles among S. aureus is essential mainly to prevention and control of dissemination of the antimicrobial resistance.No presente estudo foram avaliados o perfil de macrorrestrição do DNA e tipos de SCCmec para nove MRSA multirresistentes selecionados. Além disso, susceptibilidade a 12 agentes antimicrobianos pelo teste de disco-difusão foi avaliada para 68 isolados de MRSA. Os isolados foram obtidos de hemoculturas de pacientes hospitalizados de três hospitais de Porto Alegre, Brasil. PFGE e PCR para detecção do gene mecA e para os tipos genéticos SCCmec I, II, III e IV foram realizados em nove isolados selecionados que apresentaram susceptibilidade somente a vancomicina, teicoplanina e linezolida. Dois perfis clonais, com cinco subtipos, foram demonstrados entre os isolados analisados. Oito isolados apresentaram SCCmec tipo III e um isolado não foi caracterizado quanto ao tipo de SCCmec. O conhecimento do tipo de SCCmec bem como dos perfis clonais e de susceptibilidade aos antimicrobianos entre isolados de S. aureus é essencial, principalmente, para a prevenção e controle da disseminação da resistência antimicrobiana.

  12. Nasal carriage of a single clone of community-acquired methicillin-resistant Staphylococcus aureus among kindergarten attendees in northern Taiwan

    Directory of Open Access Journals (Sweden)

    Lee Shih-Yi

    2007-06-01

    Full Text Available Abstract Background: To evaluate the prevalence and microbiological characterization of community-acquired (CA methicillin-resistant Staphylococcus aureus (MRSA nasal carriage in a kindergarten. Methods: Point prevalence study. Nasal swabs were collected from healthy children younger than 7 years of age who were attending a kindergarten in Taipei, Taiwan. A parent questionnaire regarding MRSA risk factors was administered simultaneously. All CA-MRSA colonization isolates were archived for subsequent antimicrobial susceptibility and molecular typing. Results: Of the 68 children who participated in the study, 17 (25% had S. aureus isolated from nasal swabs. Nine (13.2% of the 68 children had CA-MRSA carriage, and none of them had any identified risk factors. Antimicrobial susceptibility testing revealed all of the 9 CA-MRSA colonization isolates had uniformly high resistance (100% to both clindamycin and erythromycin, the macrolide-lincosamide-streptogramin-constitutive phenotype and the ermB gene. Pulsed-field gel electrophoresis revealed 8 (88.9% of 9 CA-MRSA colonization isolates were genetically related and multilocus sequence typing revealed all isolates had sequence type 59. All of the colonization isolates carried the staphylococcal cassette chromosome mec type IV, but none were positive for the Panton-Valentine leukocidin genes. Conclusion: The results of this study suggest that a single predominant CA-MRSA colonization strain featuring high clindamycin resistance circulated in this kindergarten. Additionally, due to the established transmissibility of colonization isolates, the high prevalence of nasal carriage of CA-MRSA among healthy attendees in kindergartens may indicate the accelerated spread of CA-MRSA in the community.

  13. Cloning and gene construction analysis of β-actin in Oreochromis aureus%奥利亚罗非鱼β-actin基因的克隆与结构分析

    Institute of Scientific and Technical Information of China (English)

    胡海彦; 贾永义; 唐永凯

    2011-01-01

    β-actin在真核细胞的生理过程中起着重要作用,其序列具有高度保守性,是一种管家基因.通过RT-PCR方法克隆出奥利亚罗非鱼(Oreochromis aureus)β-actin的部分cDNA序列,其长度为424 bp,翻译成138个氨基酸,计算的蛋白质分子量为15.5 ku.氨基酸同源性分析显示,奥利亚罗非鱼β-actin与真鲷(Pagrus major)、斑马鱼(Danio rerio)、青鳝(Oryzias latipes)、龙溪鳝(Rivulus marmoratus)的相似性最高,为99.3%;与鲫(Carassius auratus)、红鳍东方鲍(Takifugu rubripes)等其它鱼的相似性也较高,为97.8%~98.6%.此外,还克隆出了奥利亚罗非鱼β-actin相应的DNA序列,共619 bp.cDNA和DNA的序列比对显示克隆出的奥利亚罗非鱼β-actin含有2个内含子,这为将来设计β-actin荧光定量PCR引物以及测定其在不同组织中的表达量变化打下基础.%Eukaryotic β-actin gene plays an important role in physiological process and its sequence is highly conservative as a housekeeping gene. The partial cDNA encoding β-actin in Oreochromis aureus was isolated using RT-PCR. The cDNA isolated was 424 bp encoding 138 amino acids with a calculated molecular weight of 15.5 ku. The comparison analysis of the deduced amino acid sequence of β-actin in O. aureus showed high similarity of 99.3% with those of Pagrus major, Danio rerio, Oryzias latipes, Rivulus marmoratus, and 97.8% ~ 98.6% homology with those of other fishes, such as Carassius auratus, Takifugu rubripes. The corresponding DNA sequence of O. aureus β-actin in size of 619 bp was also cloned. By comparing the sequences of cDNA and DNA, it was found that β-actin gene in O. aureus consisted of two introns. The present study provided some useful information in designing the β-actin primers for real time RT-PCR and evaluating the β-actin expression level for different tissues.

  14. Dissemination of methicillin-resistant Staphylococcus aureus SCCmec type IV and SCCmec type V epidemic clones in a tertiary hospital: challenge to infection control.

    Science.gov (United States)

    Dhawan, B; Rao, C; Udo, E E; Gadepalli, R; Vishnubhatla, S; Kapil, A

    2015-01-01

    Two-hundred MRSA strains from inpatients with healthcare-associated (HA) and 100 MRSA strains from outpatients with community-associated (CA) skin and soft tissue infections (SSTIs) were tested for antimicrobial susceptibility, staphylococcal cassette chromosome mec (SCCmec) typing, Panton-Valentine leucocidin (PVL) toxin, seh and arcA genes. Based on SCCmec typing, HA-MRSA isolates were further divided into HA-SCCmec I/II/III MRSA and HA-SCCmec IV/V MRSA, and CA-MRSA isolates into CA-SCCmec I/II/III MRSA and CA-SCCmec IV/V MRSA. SCCmec types were further characterized by pulsed-field gel electrophoresis, spa typing and multi-locus sequence typing. Seventy-five (37·5%) HA-MRSA isolates and 83/100 CA-MRSA isolates were SCCmec IV/V genotype. HA-SCCmec IV/V MRSA was associated with malignancy (P = 0·03) and bone fractures (P = 0·02) compared to CA-SCCmec IV/V MRSA. HA-SCCmec IV/V MRSA was associated with PVL gene carriage compared to HA-SCCmec I/II/III MRSA (P IV (EMRSA-15), ST772-MRSA-V, and ST36-MRSA-IV and ST239:EMRSA-I:III were the major clones identified. Our study documents the emergence of SCCmec IV and SCCmec V MRSA clones in an Indian hospital.

  15. Staphylococcus aureus spa type t437

    DEFF Research Database (Denmark)

    Glasner, C; Pluister, G; Westh, H;

    2015-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) belonging to the multilocus sequence type clonal complex 59 (MLST CC59) is the predominant community-associated MRSA clone in Asia. This clone, which is primarily linked with the spa type t437, has so far only been reported in low numbers among...

  16. Cloning and Expression of lsdD gene from Staphylococcus aureus%金黄色葡萄球菌IsdD基因的克隆与表达

    Institute of Scientific and Technical Information of China (English)

    马金柱; 宋佰芬; 王北艳; 曹宁; 于立权; 崔玉东; 朱战波

    2011-01-01

    Staphylococcusaureus causes a wide spectrum of human and animal diseases,including skin and soft-tissue infections and infectious endocarditis. IsdD gene is one of the important members of iron-regulated surface determinant (Isd) system from S. Aureus. According to the sequence of IsdD gene published in GenBank,a pair of specific primers was designed and synthesized. Using the total genome DNA from S. Aureus Wood46 strain as template,IsdD coding sequences were amplified by PCR method. The PCR product which had been digested by Nco Ⅰ and Xho Ⅰ restriction endonucleases was inserted into pET32a( + ) vector subsequently. The recombinant plasmids, designated as pET32a( + )-IsdD, were transformed into E. Coli BL21. The sequence analysis results showed the homologies of both nucleoside sequence of target gene and its deduced amino acid sequence to compare with standard sequence of IsdD could reach above 98%. SDS-PAGE analysis results indicated the recombinant IsdD protein of 61. 3 ku was successfully expressed by E. Coli BL21 induced with IPTG. The results obtained in this report provide important basis for further study on structure and function of the IsdD protein.%金黄色葡萄球菌是一种重要的人畜致病菌,IsdD属于金黄色葡萄球菌Isd系统中的重要成员.本研究根据GenBank报道的金黄色葡萄球菌IsdD基因序列设计一对特异性引物,以金黄色葡萄球菌基因组DNA为模板扩增出目的片段Is-dD,将其用Nco Ⅰ和Xho Ⅰ双酶切后连接到载体pET32a(+)上,构建重组质粒pET32a(+)-IsdD.序列分析结果显示,目的基因序列与网上的IsdD序列相比核苷酸序列和氨基酸序列同源性均在98%以上.将鉴定正确的重组质粒转化到大肠杆菌E.coli BL21中并成功诱导表达出大小为61.3 ku的蛋白,这为下一步研究IsdD蛋白的结构和功能奠定了良好基础.

  17. Nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) among Swiss veterinary health care providers: detection of livestock- and healthcare-associated clones.

    Science.gov (United States)

    Wettstein Rosenkranz, K; Rothenanger, E; Brodard, I; Collaud, A; Overesch, G; Bigler, B; Marschall, J; Perreten, V

    2014-07-01

    We screened a total of 340 veterinarians (including general practitioners, small animal practitioners, large animal practitioners, veterinarians working in different veterinary services or industry), and 29 veterinary assistants for nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP) at the 2012 Swiss veterinary annual meeting. MRSA isolates (n = 14) were detected in 3.8 % (95 % CI 2.1 - 6.3 %) of the participants whereas MRSP was not detected. Large animal practitioners were carriers of livestock-associated MRSA (LA-MRSA) ST398-t011-V (n = 2), ST398-t011-IV (n = 4), and ST398-t034-V (n = 1). On the other hand, participants working with small animals harbored human healthcare-associated MRSA (HCA-MRSA) which belonged to epidemic lineages ST225-t003-II (n = 2), ST225-t014-II (n = 1), ST5-t002-II (n = 2), ST5-t283-IV (n = 1), and ST88-t186-IV (n = 1). HCA-MRSA harbored virulence factors such as enterotoxins, β-hemolysin converting phage and leukocidins. None of the MRSA isolates carried Panton-Valentine leukocidin (PVL). In addition to the methicillin resistance gene mecA, LA-MRSA ST398 isolates generally contained additional antibiotic resistance genes conferring resistance to tetracycline [tet(M) and tet(K)], trimethoprim [dfrK, dfrG], and the aminoglycosides gentamicin and kanamycin [aac(6')-Ie - aph(2')-Ia]. On the other hand, HCA-MRSA ST5 and ST225 mainly contained genes conferring resistance to the macrolide, lincosamide and streptogramin B antibiotics [erm(A)], to spectinomycin [ant(9)-Ia], amikacin and tobramycin [ant(4')-Ia], and to fluoroquinolones [amino acid substitutions in GrlA (S84L) and GyrA (S80F and S81P)]. MRSA carriage may represent an occupational risk and veterinarians should be aware of possible MRSA colonization and potential for developing infection or for transmitting these strains. Professional exposure to animals should be reported upon hospitalization and before medical

  18. Genotyping of methicillin-resistant Staphylococcus aureus in the Sultan Qaboos University Hospital, Oman reveals the dominance of Panton–Valentine leucocidin-negative ST6-IV/t304 clone

    Directory of Open Access Journals (Sweden)

    E.E. Udo

    2014-07-01

    Full Text Available The objective of this study was to determine the prevalence and distribution of methicillin-resistant Staphylococcus aureus (MRSA genotypes circulating at a tertiary hospital in the Sultanate of Oman. A total of 79 MRSA isolates were obtained from different clinical samples and investigated using antibiogram, pulsed-field gel electrophoresis (PFGE, staphylococcal chromosome cassette mec (SCCmec, Spa typing and multilocus sequence typing (MLST. The isolates were susceptible to linezolid, vancomycin, teicoplanin, tigecycline and mupirocin but were resistant to tetracycline (30.4%, erythromycin (26.6%, clindamycin (24.1%, trimethoprim (19.0%, ciprofloxacin (17.7%, fusidic acid (15.2% and gentamicin (12.7%. Molecular typing revealed 19 PFGE patterns, 26 Spa types and 21 sequence types. SCCmec-IV (86.0% was the dominant SCCmec type, followed by SCCmec-V (10.1%. SCCmec-III (2.5% and SCCmec-II (1.3% were less common. ST6-IV/t304 (n = 30 and ST1295-IV/t690 (n = 12 were the dominant genotypes followed by ST772-V/t657 (n = 5, ST30-IV/t019/t021 (n = 5, ST22-IV/t852 (n = 4, ST80-IV/t044 (n = 3 and 18 single genotypes that were isolated sporadically. On the basis of SCCmec typing and MLST, 91.2% of the isolates were classified as community-associated MRSA and 8.8% of the isolates (consisting of four ST22-IV/t852, one ST239-III/t632, one ST5-III/t311 and one ST5-II/t003 were classified as healthcare-associated MRSA. The study has revealed the dominance of a Panton–Valentine leucocidin-negative ST6-IV/t304 clone and provided insights into the distribution of antibiotic resistance in MRSA at the tertiary hospital in Oman. It also highlights the importance of surveillance in detecting the emergence of new MRSA clones in a healthcare facility.

  19. Molecular cloning and analysis of enterotoxin gene from wild-type Staphylococcus aureus%野生型金黄色葡萄球菌肠毒素A基因的克隆与分析

    Institute of Scientific and Technical Information of China (English)

    邵奉公; 傅磊; 高晗; 杨滔; 郭焜鹏; 佘宇佳; 张海韵; 陈鸿鹏

    2012-01-01

    根据A、B、C和D肠毒素基因的序列设计特异引物,采用聚合酶链式反应(PCR)方法对食品中的金黄色葡萄球菌进行检测和鉴定肠毒素基因型,并进一步克隆肠毒素基因.结果表明,待检食品中的金黄色葡萄球菌的肠毒素基因为A型,序列长为774 bp,编码257个氨基酸残基,同其他肠毒素A高度相似,等电点为8.85,是一种膜结合蛋白,功能结构域和三位结构预测分析发现此蛋白含有肠毒素家族结构域和肠毒素C家族结构域.%A gene from food sample was identified and cloned by PCR method with special primers for A,B,C and D enterotoxin genes in the study. The results of bioinformaiics analysis suggested that the gene was a member of A enterotoxin gene family and was 774 bp in length,encoding 257 ammo acids,the deduced protein shared high homology with other enterotoxins from Genbanki the pi was 8. 85, and anchored in the membrane after secreted from the cell. The second structure and 3D prediction analysis indicated that the deduced protein contained Stap-Strp-toxin superfamily and Stap-Strp-tox C superiamily which composed the function domains of Stapkytococcus aureus.

  20. Why Clone?

    Science.gov (United States)

    ... How might cloning be used in medicine? Cloning animal models of disease Much of what researchers learn ... issue of the genetic reshuffling that happensduring sexual reproduction and simply clone our drug-producing cow. Cloning ...

  1. Molecular Epidemiology of Staphylococcus aureus among Patients with Skin and Soft Tissue Infections in Two Chinese Hospitals

    Directory of Open Access Journals (Sweden)

    Fei-Fei Gu

    2016-01-01

    Conclusions: The livestock ST398 was the most common clone among patients with S. aureus SSTIs in Jiangsu Province, China. Surveillance and further studies on the important livestock ST398 clone in human infections are necessarily requested.

  2. Staphylococcus aureus 'Down Under': contemporary epidemiology of S. aureus in Australia, New Zealand, and the South West Pacific.

    Science.gov (United States)

    Williamson, D A; Coombs, G W; Nimmo, G R

    2014-07-01

    The clinical and molecular epidemiology of Staphylococcus aureus disease has changed considerably over the past two decades, particularly with the emergence and spread of community-associated methicillin-resistant S. aureus (CA-MRSA) clones. Indeed, some of the first global descriptions of CA-MRSA were from remote indigenous communities in Western Australia, and from Pacific Peoples in New Zealand. The epidemiology of S. aureus infections in the South West Pacific has several unique features, largely because of the relative geographical isolation and unique indigenous communities residing in this region. In particular, a number of distinct CA-MRSA clones circulate in Australia and New Zealand, such as sequence type (ST) 93 methicillin-resistant S. aureus (MRSA) (Queensland clone) and clonal complex 75 S. aureus (Staphylococcus argenteus) in Australia, and ST30 MRSA (Southwest Pacific clone) in New Zealand. In addition, there is a disproportionate burden of S. aureus disease in indigenous paediatric populations, particularly in remote Aboriginal communities in Australia, and in Pacific Peoples and Maori in New Zealand. In this review, we provide a contemporary overview of the clinical and molecular epidemiology of S. aureus disease in the South West Pacific region, with a particular focus on features distinct to this region.

  3. What is Cloning?

    Science.gov (United States)

    Donate Home Cloning What is Cloning What is Cloning Clones are organisms that are exact genetic copies. ... clones made through modern cloning technologies. How Is Cloning Done? Many people first heard of cloning when ...

  4. Quantum cloning

    OpenAIRE

    Scarani, Valerio; Iblisdir, Sofyan; Gisin, Nicolas; Acin, Antonio

    2005-01-01

    The impossibility of perfectly copying (or cloning) an arbitrary quantum state is one of the basic rules governing the physics of quantum systems. The processes that perform the optimal approximate cloning have been found in many cases. These "quantum cloning machines" are important tools for studying a wide variety of tasks, e.g. state estimation and eavesdropping on quantum cryptography. This paper provides a comprehensive review of quantum cloning machines (both for discrete-dimensional an...

  5. New epidemiology of Staphylococcus aureus infection in Africa.

    Science.gov (United States)

    Schaumburg, F; Alabi, A S; Peters, G; Becker, K

    2014-07-01

    Research on African Staphylococcus aureus has been largely neglected in the past, despite the cultural and geographical diversity in Africa, which has a significant impact on the epidemiology of this pathogen. The polarity between developed urban societies and remote rural populations (e.g. Pygmies), combined with close contact with animals (e.g. livestock and domestic animals, and wildlife), makes the epidemiology of S. aureus on the African continent unique and fascinating. Here, we try to draw an epidemiological picture of S. aureus colonization and infection in Africa, and focus on the wide spread of Panton-Valentine leukocidin-positive isolates, the emergence of the hypervirulent methicillin-resistant S. aureus (MRSA) clone USA300, and the dissemination of the typical African clone MRSA sequence type 88.

  6. Staphylococcus aureus and Pregnancy

    Science.gov (United States)

    Staphylococcus aureus and Pregnancy In every pregnancy, a woman starts out with a 3-5% chance of having a ... This sheet talks about whether exposure to staphylococcus aureus may increase the risk for birth defects over ...

  7. Genomics of Natural Populations of Staphylococcus aureus.

    Science.gov (United States)

    Fitzgerald, J Ross; Holden, Matthew T G

    2016-09-01

    Staphylococcus aureus is a major human pathogen and an important cause of livestock infections. The first S. aureus genomes to be published, 15 years ago, provided the first view of genome structure and gene content. Since then, thousands of genomes from a wide array of strains from different sources have been sequenced. Comparison of these sequences has resulted in broad insights into population structure, bacterial evolution, clone emergence and expansion, and the molecular basis of niche adaptation. Furthermore, this information is now being applied clinically in outbreak investigations to inform infection control measures and to determine appropriate treatment regimens. In this review, we summarize some of the broad insights into S. aureus biology gained from the analysis of genomes and discuss future directions and opportunities in this dynamic field of research.

  8. Staphylococcus aureus toxins.

    Science.gov (United States)

    Otto, Michael

    2014-02-01

    Staphylococcus aureus is a dangerous pathogen that causes a variety of severe diseases. The virulence of S. aureus is defined by a large repertoire of virulence factors, among which secreted toxins play a preeminent role. Many S. aureus toxins damage biological membranes, leading to cell death. In particular, S. aureus produces potent hemolysins and leukotoxins. Among the latter, some were recently identified to lyse neutrophils after ingestion, representing an especially powerful weapon against bacterial elimination by innate host defense. Furthermore, S. aureus secretes many factors that inhibit the complement cascade or prevent recognition by host defenses. Several further toxins add to this multi-faceted program of S. aureus to evade elimination in the host. This review will give an overview over S. aureus toxins focusing on recent advances in our understanding of how leukotoxins work in receptor-mediated or receptor-independent fashions.

  9. Academic Cloning.

    Science.gov (United States)

    Sikula, John P.; Sikula, Andrew F.

    1980-01-01

    The authors define "cloning" as an integral feature of all educational systems, citing teaching practices which reward students for closely reproducing the teacher's thoughts and/or behaviors and administrative systems which tend to promote like-minded subordinates. They insist, however, that "academic cloning" is not a totally…

  10. Differential Analysis of the Nasal Microbiome of Pig Carriers or Non-Carriers of Staphylococcus aureus

    DEFF Research Database (Denmark)

    Espinosa-Gongora, Carmen; Larsen, Niels; Schonning, Kristian

    2016-01-01

    Staphylococcus aureus is presently regarded as an emerging zoonotic agent due to the spread of specific methicillin-resistant S. aureus (MRSA) clones in pig farms. Studying the microbiota can be useful for the identification of bacteria that antagonize such opportunistic veterinary and zoonotic p...

  11. First report of the predominance of clonal complex 398 Staphylococcus aureus strains in osteomyelitis complicating diabetic foot ulcers: a national French study.

    Science.gov (United States)

    Senneville, E; Brière, M; Neut, C; Messad, N; Lina, G; Richard, J-L; Sotto, A; Lavigne, J-P

    2014-04-01

    Staphylococcus aureus is the most common pathogen cultured from diabetic foot infection including diabetic foot osteomyelitis. This French multicentre study determined the genetic content of S. aureus isolated from 157 consecutive cases admitted to 12 diabetic foot centres between 2008 and 2011. We describe for the first time the emergence of the CC398 methicillin-susceptible S. aureus clone, the main clone in diabetic foot osteomyelitis, and its tropism for bone. This clone spreads to humans from an animal source through its intrinsic virulence. This adaptation of S. aureus isolates looks to be a worrisome problem and should be carefully monitored.

  12. Human Cloning

    Science.gov (United States)

    2006-07-20

    genes, for example, has led to new treatments developed by the biotechnology industry for diseases such as diabetes and hemophilia. In the context of...or imposed a moratorium. The legislation was opposed by a number of medical organizations, the biotechnology industry and many scientists and was not...cloning by FDA.36 They find little evidence to support FDA’s position that cloned human embryos are “drugs.” However, the biotechnology industry and the

  13. USA300 Methicillin-resistant Staphylococcus aureus in Cuba

    Directory of Open Access Journals (Sweden)

    Hopman Joost

    2012-01-01

    Full Text Available Abstract Background Methicillin-resistant Staphylococcus aureus is an increasing problem in the Caribbean. We investigated the molecular epidemiology of MRSA isolates on Cuba. Findings The predominant clone was of the spa type t149, followed by community-associated MRSA USA300. Conclusions We report the first molecular typing results of MRSA isolates from Cuba.

  14. The nosocomial transmission rate of animal-associated ST398 meticillin-resistant Staphylococcus aureus

    NARCIS (Netherlands)

    Bootsma, M.C.J.; Wassenberg, M.W.M.; Trapman, J.P.; Bonten, M.J.M.

    2010-01-01

    The global epidemiology of meticillin-resistant Staphylococcus aureus (MRSA) is characterized by different clonal lineages with different epidemiological behaviour. There are pandemic hospital clones (hospital-associated (HA-)MRSA), clones mainly causing community-acquired infections (community-asso

  15. Molecular cloning.

    Science.gov (United States)

    Lessard, Juliane C

    2013-01-01

    This protocol describes the basic steps involved in conventional plasmid-based cloning. The goals are to insert a DNA fragment of interest into a receiving vector plasmid, transform the plasmid into E. coli, recover the plasmid DNA, and check for correct insertion events.

  16. Diversity of mechanisms conferring resistance to β-lactams among OXA-23-producing Acinetobacter baumannii clones.

    Science.gov (United States)

    Cardoso, Juliana Provasi; Cayô, Rodrigo; Girardello, Raquel; Gales, Ana Cristina

    2016-05-01

    A total of 31 unrelated OXA-23-producing Acinetobacter baumannii strains isolated from 14 hospitals located in distinct Brazilian regions were evaluated in this study. These isolates were grouped into 12 different sequence types (STs), of which 7 had unique allelic sequences (ST188, ST189, ST190, ST191, ST192, ST228, and ST299). Most isolates belonged to the clonal complex CC79 followed by CC15 and CC1. Only polymyxin B and minocycline showed good activity against the OXA-23-producing A. baumannii clones. The ISAba1 upstream blaOXA-23, blaOXA-51-like, or ampC was found in 100%, 54.8%, and 77.4% of the isolates, respectively. High resistance rates to ceftazidime and cefotaxime were observed among those isolates possessing ISAba1 upstream ampC, in contrast to those isolates that did not carry this configuration. Moreover, a ≥2 Log2 decrease in the MICs of meropenem and ceftazidime was observed in the presence of phenyl-arginine-β-naphthylamide for 80.6% and 54.8% of isolates, respectively. Overexpression of the adeB was observed in 61.3% of isolates, particularly among those isolates belonging to the ST1 (CC1). It was also verified that ompW was down-regulated in all isolates belonging to the ST15 (CC15). On the other hand, carO and omp33-36 genes were overexpressed in 48.4% and 58.1% of the isolates, respectively. In this study, we show that overexpression of AdeABC system could significantly contribute for resistance to meropenem and ceftazidime among OXA-23-producing A. baumannii clones in Brazil, demonstrating the complexity involved in the β-lactam resistance in such isolates.

  17. Monoclonal Antibody Targeting Staphylococcus aureus Surface Protein A (SasA) Protect Against Staphylococcus aureus Sepsis and Peritonitis in Mice.

    Science.gov (United States)

    Yang, Yilong; Qian, Mengying; Yi, Shaoqiong; Liu, Shuling; Li, Bing; Yu, Rui; Guo, Qiang; Zhang, Xiaopeng; Yu, Changming; Li, Jianmin; Xu, Junjie; Chen, Wei

    2016-01-01

    Epidemic methicillin-resistant Staphylococcus aureus (MRSA) imposes an increasing impact on public health. Due to multi-antibiotics resistance in MRSA strains, there is an urgent need to develop novel therapeutics such as effective monoclonal antibodies (mAbs) against MRSA infections. Staphylococcus aureus surface protein A (SasA), a large surface-located protein (~240 kDa), is one of MSCRAMMs (microbial surface components recognizing adhesive matrix molecules) and a potential target for immunotherapeutic approaches against S. aureus infections. In the present study, we analyzed the sequence of SasA with bioinformatics tools and generated a protective monoclonal antibody (2H7) targeting the conserved domain of SasA. 2H7 was shown to recognize wild-type S. aureus and promote opsonophagocytic killing of S. aureus. In both sepsis and peritoneal infection models, prophylactic administration of 2H7 improved the survival of BALB/c mice challenged by S. aureus strain USA300 and ST239 (prevalent MRSA clones in North America and Asian countries, respectively) and enhanced bacterial clearance in kidneys. Additionally, 2H7 prophylaxis prevented the formation of intraperitoneal abscess in a murine model of peritoneal infection and therapeutic administration of 2H7 showed protective efficacy in a murine sepsis model. Our results presented here provide supporting evidences that an anti-SasA mAb might be a potential component in an antibody-based immunotherapeutic treatment of MRSA infections.

  18. Phosphorylation controls the functioning of Staphylococcus aureus isocitrate dehydrogenase--favours biofilm formation.

    Science.gov (United States)

    Prasad, U Venkateswara; Vasu, D; Yeswanth, S; Swarupa, V; Sunitha, M M; Choudhary, A; Sarma, P V G K

    2015-01-01

    Isocitrate dehydrogenase (IDH) gene from Staphylococcus aureus ATCC12600 was cloned, sequenced and characterized (HM067707). PknB site was observed in the active site of IDH; thus, it was predicted as IDH may be regulated by phosphorylation. Therefore, in this study, PknB, alkaline phosphatase III (SAOV 2675) and IDH genes (JN695616, JN645811 and HM067707) of S. aureus ATCC12600 were over expressed from clones PV 1, UVPALP-3 and UVIDH 1. On passing the cytosloic fractions through nickel metal chelate column, pure enzymes were obtained. Phosphorylation of pure IDH by PknB resulted in the complete loss of activity and was restored upon dephosphorylation with SAOV 2675 which indicated that phosphorylation and dephosphorylation regulate IDH activity in S. aureus. Further, when S. aureus ATCC12600 was grown in BHI broth, decreased IDH activity and increased biofilm units were observed; therefore, this regulation of IDH alters redox status in this pathogen favouring biofilm formation.

  19. High Nasal Carriage Rate of Staphylococcus aureus Containing Panton-Valentine leukocidin- and EDIN-Encoding Genes in Community and Hospital Settings in Burkina Faso

    Science.gov (United States)

    Ouedraogo, Abdoul-Salam; Dunyach-Remy, Catherine; Kissou, Aimée; Sanou, Soufiane; Poda, Armel; Kyelem, Carole G.; Solassol, Jérôme; Bañuls, Anne-Laure; Van De Perre, Philippe; Ouédraogo, Rasmata; Jean-Pierre, Hélène; Lavigne, Jean-Philippe; Godreuil, Sylvain

    2016-01-01

    The objectives of the present study were to investigate the rate of S.aureus nasal carriage and molecular characteristics in hospital and community settings in Bobo Dioulasso, Burkina Faso. Nasal samples (n = 219) were collected from 116 healthy volunteers and 103 hospitalized patients in July and August 2014. Samples were first screened using CHROMagar Staph aureus chromogenic agar plates, and S. aureus strains were identified by mass spectrometry. Antibiotic susceptibility was tested using the disk diffusion method on Müller-Hinton agar. All S. aureus isolates were genotyped using DNA microarray. Overall, the rate of S. aureus nasal carriage was 32.9% (72/219) with 29% in healthy volunteers and 37% in hospital patients. Among the S. aureus isolates, only four methicillin-resistant S. aureus (MRSA) strains were identified and all in hospital patients (3.9%). The 72 S. aureus isolates from nasal samples belonged to 16 different clonal complexes, particularly to CC 152-MSSA (22 clones) and CC1-MSSA (nine clones). Two clones were significantly associated with community settings: CC1-MSSA and CC45-MSSA. The MRSA strains belonged to the ST88-MRSA-IV or the CC8-MRSA-V complex. A very high prevalence of toxinogenic strains 52.2% (36/69), containing Panton-Valentine leucocidin- and EDIN-encoding genes, was identified among the S. aureus isolates in community and hospital settings. This study provides the first characterization of S. aureus clones and their genetic characteristics in Burkina Faso. Altogether, it highlights the low prevalence of antimicrobial resistance, high diversity of methicillin-sensitive S. aureus clones and high frequency of toxinogenic S. aureus strains. PMID:27679613

  20. High Nasal Carriage Rate of Staphylococcus aureus Containing Panton-Valentine leukocidin- and EDIN-Encoding Genes in Community and Hospital Settings in Burkina Faso.

    Science.gov (United States)

    Ouedraogo, Abdoul-Salam; Dunyach-Remy, Catherine; Kissou, Aimée; Sanou, Soufiane; Poda, Armel; Kyelem, Carole G; Solassol, Jérôme; Bañuls, Anne-Laure; Van De Perre, Philippe; Ouédraogo, Rasmata; Jean-Pierre, Hélène; Lavigne, Jean-Philippe; Godreuil, Sylvain

    2016-01-01

    The objectives of the present study were to investigate the rate of S.aureus nasal carriage and molecular characteristics in hospital and community settings in Bobo Dioulasso, Burkina Faso. Nasal samples (n = 219) were collected from 116 healthy volunteers and 103 hospitalized patients in July and August 2014. Samples were first screened using CHROMagar Staph aureus chromogenic agar plates, and S. aureus strains were identified by mass spectrometry. Antibiotic susceptibility was tested using the disk diffusion method on Müller-Hinton agar. All S. aureus isolates were genotyped using DNA microarray. Overall, the rate of S. aureus nasal carriage was 32.9% (72/219) with 29% in healthy volunteers and 37% in hospital patients. Among the S. aureus isolates, only four methicillin-resistant S. aureus (MRSA) strains were identified and all in hospital patients (3.9%). The 72 S. aureus isolates from nasal samples belonged to 16 different clonal complexes, particularly to CC 152-MSSA (22 clones) and CC1-MSSA (nine clones). Two clones were significantly associated with community settings: CC1-MSSA and CC45-MSSA. The MRSA strains belonged to the ST88-MRSA-IV or the CC8-MRSA-V complex. A very high prevalence of toxinogenic strains 52.2% (36/69), containing Panton-Valentine leucocidin- and EDIN-encoding genes, was identified among the S. aureus isolates in community and hospital settings. This study provides the first characterization of S. aureus clones and their genetic characteristics in Burkina Faso. Altogether, it highlights the low prevalence of antimicrobial resistance, high diversity of methicillin-sensitive S. aureus clones and high frequency of toxinogenic S. aureus strains.

  1. Staphylococcus aureus clonal dynamics and virulence factors in children with atopic dermatitis

    DEFF Research Database (Denmark)

    Lomholt, Hans; Andersen, Klaus Ejner; Kilian, Mogens

    2005-01-01

    A prospective cohort study was undertaken to determine the clonal dynamics of Staphylococcus aureus colonization and infection during 1 y in children with atopic dermatitis, and to correlate specific clones, accessory gene regulator (agr) groups, and production of virulence factors with eczema...... activity. Eleven children were examined every 6 wk with swaps taken from active eczema, anterior nose, axillae and perineum, and scoring of eczema activity by severity scoring of atopic dermatitis (SCORAD). Individual S. aureus clonal types were identified and examined for production of superantigens......, toxins, and were assigned to agr groups. S. aureus colonization patterns ranged from rare colonization over transient colonization to persistent colonization by a single clone or a dynamic exchange of up to five clones. Production of no single virulence factor including superantigens and toxins...

  2. Staphylococcus aureus clonal dynamics and virulence factors in children with atopic dermatitis

    DEFF Research Database (Denmark)

    Lomholt, Hans Bredsted; Andersen, KE; Kilian, Mogens

    2005-01-01

    SCORAD value. In 11 of 12 cases with two different clones co-colonizing a child the clones belonged to the same agr group. In conclusion, this limited group of children with atopic dermatitis showed highly variable colonization patterns of S. aureus, and communication between strains by use of agr......A prospective cohort study was undertaken to determine the clonal dynamics of Staphylococcus aureus colonization and infection during 1 y in children with atopic dermatitis, and to correlate specific clones, accessory gene regulator (agr) groups, and production of virulence factors with eczema...... activity. Eleven children were examined every 6 wk with swaps taken from active eczema, anterior nose, axillae and perineum, and scoring of eczema activity by severity scoring of atopic dermatitis (SCORAD). Individual S. aureus clonal types were identified and examined for production of superantigens...

  3. Differential Analysis of the Nasal Microbiome of Pig Carriers or Non-Carriers of Staphylococcus aureus

    DEFF Research Database (Denmark)

    Espinosa-Gongora, Carmen; Larsen, Niels; Schonning, Kristian;

    2016-01-01

    Staphylococcus aureus is presently regarded as an emerging zoonotic agent due to the spread of specific methicillin-resistant S. aureus (MRSA) clones in pig farms. Studying the microbiota can be useful for the identification of bacteria that antagonize such opportunistic veterinary and zoonotic...... pathogen in animal carriers. The aim of this study was to determine whether the nasal microbiome of pig S. aureus carriers differs from that of non-carriers. The V3-V5 region of the 16S rRNA gene was sequenced from nasal swabs of 44 S. aureus carriers and 56 non-carriers using the 454 GS FLX titanium...... system. Carriers and non-carriers were selected on the basis of quantitative longitudinal data on S. aureus carriage in 600 pigs sampled at 20 Danish herds included in two previous studies in Denmark. Raw sequences were analysed with the BION meta package and the resulting abundance matrix was analysed...

  4. Statement on Human Cloning

    Science.gov (United States)

    ... for the Advancement of Science Statement on Human Cloning Tweet The American Association for the Advancement of ... for this statement on human cloning. Ban Reproductive Cloning AAAS endorses a legally enforceable ban on efforts ...

  5. Staphylococcus aureus and surgical site infections: benefits of screening and decolonization before surgery.

    Science.gov (United States)

    Humphreys, H; Becker, K; Dohmen, P M; Petrosillo, N; Spencer, M; van Rijen, M; Wechsler-Fördös, A; Pujol, M; Dubouix, A; Garau, J

    2016-11-01

    Surgical site infections (SSIs) are among the most common healthcare-associated infections, and contribute significantly to patient morbidity and healthcare costs. Staphylococcus aureus is the most common microbial cause. The epidemiology of S. aureus is changing with the dissemination of newer clones and the emergence of mupirocin resistance. The prevention and control of SSIs is multi-modal, and this article reviews the evidence on the value of screening for nasal carriage of S. aureus and subsequent decolonization of positive patients pre-operatively. Pre-operative screening, using culture- or molecular-based methods, and subsequent decolonization of patients who are positive for meticillin-susceptible S. aureus and meticillin-resistant S. aureus (MRSA) reduces SSIs and hospital stay. This applies especially to major clean surgery, such as cardiothoracic and orthopaedic, involving the insertion of implanted devices. However, it requires a multi-disciplinary approach coupled with patient education. Universal decolonization pre-operatively without screening for S. aureus may compromise the capacity to monitor for the emergence of new clones of S. aureus, contribute to mupirocin resistance, and prevent the adjustment of surgical prophylaxis for MRSA (i.e. replacement of a beta-lactam agent with a glycopeptide or alternative).

  6. Staphylococcus aureus bacteremia.

    Science.gov (United States)

    Jensen, Allan Garlik

    2003-11-01

    Staphylococcus aureus bacteremia (SAB) is still associated with a high mortality, and knowledge on risk factors and the clinical and the therapeutic aspects of SAB is still limited. This thesis focuses on the clinical aspects of SAB and its metastatic infections. In a study of all patients with bacteremia in Copenhagen County October 1992 through April 1993 (study I) we emphasized previous findings, that S. aureus is one of the most frequent pathogens in bacteremia, and in a case control study also in Copenhagen County 1994-95 (study II) we demonstrated, that not only an inserted central venous catheter and nasal S. aureus carriage but also hyponatremia and anemia are important risk factors for hospital-acquired SAB (study II). Studies on the treatment of SAB have pointed out, that the eradication of a primary is important, but there are only limited clinical studies dealing with antibiotic treatment. By logistic regression analysis, we were able to demonstrate that focus eradication is essential, but also that treatment with dicloxacillin 1 g x 4 or 2 g x 3 are superior to 1 g x 3 (studie III), indicating that the time for serum concentration above the Minimal Inhibitory Concentration (MIC) for the bacteria plays a role in the outcome of SAB treatment. S. aureus osteomyelitis secondary to SAB is frequently observed. No other countries, however, have a centralized registration, which make it possible to evaluate a large number of these patients. Since 1960, The Staphylococcal Laboratory, Statens Serum Institut in Copenhagen, has registrated selected clinical informations from nearly all patients with positive blood cultures of S. aureus. Based on this registration, we were able to show an increased number of S. aureus osteomyelitis among older patients and a decreased number of S. aureus osteomyelitis of femur and tibia among younger infants in the period 1980-90 (study IV). By reviewing the records of a large number of patients with vertebral S. aureus

  7. Cross-talk between Staphylococcus aureus and other staphylococcal species via the agr quorum sensing system

    Directory of Open Access Journals (Sweden)

    Jaime Canovas de la Nuez

    2016-11-01

    Full Text Available Staphylococci are associated with both humans and animals. While most are non-pathogenic colonizers, Staphylococcus aureus is an opportunistic pathogen capable of causing severe infections. S. aureus virulence is controlled by the agr quorum sensing system responding to secreted auto-inducing peptides (AIPs sensed by AgrC, a two component histidine kinase. agr loci are found also in other staphylococcal species and for S. epidermidis, the encoded AIP represses expression of agr regulated virulence genes in S. aureus. In this study we aimed to determine how other staphylococci affect S. aureus agr, and if such interaction may point to new anti-virulence candidates to target S. aureus infections. Here we show that culture supernatants of 37 out of 52 staphylococcal isolates representing 17 different species inhibit S. aureus agr. The dog pathogen, S. schleiferi, expressed the most potent inhibitory activity and was active against all four agr classes found in S. aureus. By employing a S. aureus strain encoding a constitutively active AIP receptor we show that the activity is mediated via agr. Subsequent cloning and heterologous expression of the S. schleiferi AIP in S. aureus demonstrated that this molecule was likely responsible for the inhibitory activity, and further proof was provided when pure synthetic S. schleiferi AIP was able to completely abolish agr induction of an S. aureus reporter strain. To assess impact on S. aureus virulence, we co-inoculated S. aureus and S. schleiferi in vivo in the Galleria Mellonella wax moth larva, and found that expression of key S. aureus virulence factors was abrogated. Our data show that the S. aureus agr locus is highly responsive to other staphylococcal species suggesting that agr is an inter-species communication system. Based on these results we speculate that interactions between S. aureus and other colonizing staphylococci will significantly influence the ability of S. aureus to cause infection, and

  8. The Clone Factory

    Science.gov (United States)

    Stoddard, Beryl

    2005-01-01

    Have humans been cloned? Is it possible? Immediate interest is sparked when students are asked these questions. In response to their curiosity, the clone factory activity was developed to help them understand the process of cloning. In this activity, students reenact the cloning process, in a very simplified simulation. After completing the…

  9. Cloning of observables

    OpenAIRE

    Ferraro, Alessandro; Galbiati, Matteo; Paris, Matteo G. A.

    2005-01-01

    We introduce the concept of cloning for classes of observables and classify cloning machines for qubit systems according to the number of parameters needed to describe the class under investigation. A no-cloning theorem for observables is derived and the connections between cloning of observables and joint measurements of noncommuting observables are elucidated. Relationships with cloning of states and non-demolition measurements are also analyzed.

  10. Characterization of Staphylococcus aureus from Humans and a Comparison with İsolates of Animal Origin, in North Dakota, United States.

    Science.gov (United States)

    Velasco, Valeria; Buyukcangaz, Esra; Sherwood, Julie S; Stepan, Ryan M; Koslofsky, Ryan J; Logue, Catherine M

    2015-01-01

    Different clones of methicillin-susceptible (MSSA) and methicillin-resistant (MRSA) Staphylococcus aureus have been found in humans as well as in animals and retail meat. However, more information about the genetic characteristics and similarities between strains is needed. The aim of this study was to identify and characterize Staphylococcus aureus from humans, and to compare their characteristics with isolates of animal origin. A total of 550 nasal swabs were taken from healthy humans, and S. aureus was isolated and identified. Positive S. aureus isolates were subjected to molecular typing and susceptibility testing. In addition, 108 MRSA isolates recovered from clinical patients in the state of North Dakota and 133 S. aureus isolates from animals and meat previously analyzed were included. The nasal carriage of S. aureus in healthy people was 7.6% and, in general, clones were genetically diverse. None of the S. aureus strains obtained from healthy people were mecA- or PVL-positive. A total of 105 (97.2%) MRSA isolates from clinical cases harbored the mecA gene and 11 (10.2%) isolated from blood stream infections harbored the PVL gene. The most common resistance profile among S. aureus from healthy people was penicillin, and from clinical cases were erythromycin-penicillin-ciprofloxacin. The rate of multidrug resistance (MDR) was 70% in humans. Most of the S. aureus harboring mecA and PVL genes were identified as ST5 and ST8, and exhibited MDR. However, S. aureus isolates of animal origin used for comparison exhibited a lower rate of MDR. The most common resistance profiles in isolates of animal origin were penicillin-tetracycline and penicillin-tetracycline-erythromycin, in animals and raw meat, respectively. The ST5 was also found in animals and meat, with ST9 and ST398 being the major clones. The genetic similarity between clones from humans and meat suggests the risk of spread of S. aureus in the food chain.

  11. Molecular epidemiology of Staphylococcus aureus in post-earthquake northern Haiti

    Directory of Open Access Journals (Sweden)

    Marnie E. Rosenthal

    2014-12-01

    Conclusions: We found a low S. aureus carriage rate with complete vancomycin susceptibility and high tetracycline resistance, which has important public health implications with regard to treatment. Additionally, the finding of PVL-positive MSSA isolates, including the expansion of a previously described limited ‘divergent’ clone, ST152, warrants further evaluation.

  12. Transmissibility of livestock-associated methicillin-resistant Staphylococcus aureus (ST398) in Dutch hospitals

    NARCIS (Netherlands)

    Wassenberg, M. W. M.; Bootsma, M. C. J.; Troelstra, A.; Kluytmans, J. A. J. W.; Bonten, M. J. M.

    2011-01-01

    P>We quantified nosocomial transmission rates of sequence type (ST) 398 methicillin-resistant Staphylococcus aureus (MRSA) (an emerging livestock-associated MRSA clone) and non-ST398 MRSA isolates in patients hospitalized without infection control measures in 51 Dutch hospitals. Identification of 17

  13. Evolutionary dynamics of methicillin-resistant Staphylococcus aureus within a healthcare system

    NARCIS (Netherlands)

    Hsu, Li-Yang; Harris, Simon R.; Chlebowicz, Monika A.; Lindsay, Jodi A.; Koh, Tse-Hsien; Krishnan, Prabha; Tan, Thean-Yen; Hon, Pei-Yun; Grubb, Warren B.; Bentley, Stephen D.; Parkhill, Julian; Peacock, Sharon J.; Holden, Matthew T. G.

    2015-01-01

    Background: In the past decade, several countries have seen gradual replacement of endemic multi-resistant healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) with clones that are more susceptible to antibiotic treatment. One example is Singapore, where MRSA ST239, the dominant

  14. Cloning,prokaryotic expression,purification and identification of the transpeptidase domain of penicillin binding protein 2a of methicillin-resistant Staphylococcus aureus%耐甲氧西林金黄色葡萄球菌青霉素结合蛋白2a转肽酶区的克隆、表达及纯化鉴定

    Institute of Scientific and Technical Information of China (English)

    马洪玉; 兰小鹏; 陈敏

    2016-01-01

    目的:构建编码耐甲氧西林金黄色葡萄球菌(MRSA)青霉素结合蛋白2a (PBP2a)转肽酶区基因片段的原核表达载体,并表达、纯化及鉴定蛋白。方法从临床标本中分离鉴定M RSA ,设计针对编码 PBP2a转肽酶区基因片段的引物,采用聚合酶链式反应(PCR)扩增目的基因片段,克隆至pET28a(+)载体,双酶切鉴定并测序,转化大肠杆菌BL21(DE3)plysS株;用0.7 mmol/L异丙基硫代‐β‐D‐半乳糖苷(IPTG)诱导表达后,利用Ni亲和层析技术纯化目的蛋白;蛋白免疫印迹法(WB)鉴定重组蛋白。结果重组表达载体经BamHⅠ、EcoRⅠ酶切,产物在预期大小处出现条带,测序结果显示有两个碱基突变,无移码突变。所表达的PBP2a蛋白经十二烷基硫酸钠‐聚丙烯酰胺凝胶电泳(SDS‐PAGE)和WB鉴定,在相对分子质量38×103处可见一新生蛋白条带。结论成功构建了PBP2a转肽酶区原核表达载体,并获得了高效表达,制备了高纯度的目的蛋白。%Objective To construct the prokaryontic expression vector of the gene fragment which encodes the transpeptidase domain of penicillin binding protein 2a(PBP2a) of methicillin‐resistant Staphylococcus aureus(MRSA) ,and to express ,purify and i‐dentify the objective protein .Methods Strains of MRSA were isolated and identified from clinical samples ,according to the se‐quence of mecA gene recorded in GenBank ,the primers of mecA fragment which encoded the transpeptidase domain of PBP2a was designed .The gene fragment from MRSA was amplified by using polymerase chain reaction(PCR) and cloned into pET28a(+ ) plasmid .After being identified by enzyme digestion and sequencing ,the recombinant plasmid was transformed into the strain of Escherichia coli BL21(DE3)plysS .The expression of transpeptidase domain of PBP2a was induced by 0 .7 mmol/L IPTG ,the ex‐pressed products were purified by using Ni afinity

  15. Improved lux reporters for use in Staphylococcus aureus.

    Science.gov (United States)

    Mesak, Lili Rosana; Yim, Grace; Davies, Julian

    2009-05-01

    The use of luxABCDE (lux) offers certain advantages over other reporters, such as: lacZ and xylE. It is real time and its signal generation is produced without the requirement for any additional substrates. In some bacteria such as Staphylococcus spp, light production by luciferase is restricted because of a limited availability of endogenous substrates such as fatty acid aldehyde. We describe the construction of promoterless-lux cloning vectors, pGYlux and pAmilux. S. aureus carrying B. subtilis xyl/tetO promoter fused to the lux genes of pGYlux gave up to a 2.5-fold enhancement of luminescence over S. aureus carrying the xyl/tetO promoter fused to lux genes of the previously published parent vector pAL2. Furthermore, pAmilux showed a 6-fold enhancement of lux expression when compared to pGYlux in S. aureus. This was achieved by cloning the constitutive ami promoter upstream of the luxCDE genes to increase endogenous fatty acid aldehyde production while maintaining its reporter functionality by fusing promoters to the luxAB genes.

  16. New epidemiology of Staphylococcus aureus infection in Asia.

    Science.gov (United States)

    Chen, C-J; Huang, Y-C

    2014-07-01

    Not only is Asia the most populous region in the world, but inappropriate therapy, including self-medication with over-the-counter antimicrobial agents, is a common response to infectious diseases. The high antibiotic selective pressure among the overcrowded inhabitants creates an environment that is suitable for the rapid development and efficient spread of numerous multidrug-resistant pathogens. Indeed, Asia is among the regions with the highest prevalence rates of healthcare-associated methicillin-resistant Staphylococcus aureus (HA-MRSA) and community-associated methicillin-resistant S. aureus (CA-MRSA) in the world. Most hospitals in Asia are endemic for multidrug-resistant methicillin-resistant S. aureus (MRSA), with an estimated proportion from 28% (in Hong Kong and Indonesia) to >70% (in Korea) among all clinical S. aureus isolates in the early 2010s. Isolates with reduced susceptibility or a high level of resistance to glycopeptides have also been increasingly identified in the past few years. In contrast, the proportion of MRSA among community-associated S. aureus infections in Asian countries varies markedly, from 35%. Two pandemic HA-MRSA clones, namely multilocus sequence type (ST) 239 and ST5, are disseminated internationally in Asia, whereas the molecular epidemiology of CA-MRSA in Asia is characterized by clonal heterogeneity, similar to that in Europe. In this review, the epidemiology of S. aureus in both healthcare facilities and communities in Asia is addressed, with an emphasis on the prevalence, clonal structure and antibiotic resistant profiles of the MRSA strains. The novel MRSA strains from livestock animals have been considered to constitute a public health threat in western countries. The emerging livestock-associated MRSA strains in Asia are also included in this review.

  17. Draft Genome Sequences of Staphylococcus aureus AMRF1 (ST22) and AMRF2 (ST672), Ocular Methicillin-Resistant Isolates

    KAUST Repository

    Velusamy, Nithya

    2014-03-20

    Sequence type 22 (ST22) and ST672 are the two major emerging clones of community-acquired methicillin-resistant Staphylococcus aureus in India. ST672 strains were found to cause severe ocular infections. We report the draft genome sequences of two emerging strains of methicillin-resistant S. aureus, AMRF1 (ST22) and AMRF2 (ST672), isolated from patients with ocular infections.

  18. Human Staphylococcus aureus lineages among Zoological Park residents in Greece

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    E. Drougka

    2015-10-01

    Full Text Available Staphylococcus aureus is a part of the microbiota flora in many animal species. The clonal spread of S. aureus among animals and personnel in a Zoological Park was investigated. Samples were collected from colonized and infected sites among 32 mammals, 11 birds and eight humans. The genes mecA, mecC, lukF/lukS-PV (encoding Panton-Valentine leukocidin, PVL and tst (toxic shock syndrome toxin-1 were investigated by PCR. Clones were defined by Multilocus Sequence Typing (MLST, spa type and Pulsed-Field Gel Electrophoresis (PFGE. Seven S. aureus isolates were recovered from four animals and one from an employee. All were mecA, mecC and tst–negative, whereas, one carried the PVL genes and was isolated from an infected Squirrel monkey. Clonal analysis revealed the occurrence of seven STs, eight PFGE and five spa types including ones of human origin. Even though a variety of genotypes were identified among S. aureus strains colonizing zoo park residents, our results indicate that colonization with human lineages has indeed occurred.

  19. Ethical issues in cloning.

    Science.gov (United States)

    Satris, S

    2000-01-01

    There is great public concern with the ethics of human cloning. This paper briefly examines some of what I identify as pseudo-problems or myths associated with cloning, and some of the more substantial ethical concerns.

  20. Methicillin-Resistant Staphylococcus aureus in the Community in Luanda, Angola: Blurred Boundaries with the Hospital Setting.

    Science.gov (United States)

    Conceição, Teresa; Coelho, Céline; Santos Silva, Isabel; de Lencastre, Hermínia; Aires-de-Sousa, Marta

    2016-01-01

    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.

  1. Linezolid resistant Staphylococcus aureus

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    Pavani Gandham

    2014-08-01

    Full Text Available Linezolid is the only antibiotic available as an oral formulation for resistant staphylococcal infections. It is effective in skin and soft tissue infections, nosocomial pneumonias including VAP, infective endocarditis and MRSA meningitis. It is also effective in the eradication of both nasal and throat colonization of MRSA. Its high bioavailability and post antibiotic effect, ease of switching to oral therapy during its use and the fact that it can be used in patients of all ages, also in patients with liver disease and poor kidney function and its increased effectiveness over glycopeptides makes this drug a precious drug in the treatment of resistant staphylococcal infections. Linezolid resistance in staphylococcus is defined as a linezolid MIC of and #8805;8 mg/L. Reported Linezolid resistance in India and elsewhere is 2-20%. There is clonal dissemination of Linezolid Resistant Staphylococcus aureus (LRSA within or across health care settings which demands continuous surveillance to determine the emergent risk of resistance strains and to establish guidelines for appropriate use. Clinical laboratories should confirm any LRSA preferably by a second method, prior to using linezolid for serious infections. Effective surveillance, more judicious use of this antibiotic, avoiding linezolid usage for empiric therapy in hospital acquired staphylococcus infections, optimization of the pharmacological parameters of the antibiotics in specific clinical situation, decreasing bacterial load by timely surgical debridement or drainage of collections, use of combination therapies would prevent the emergence of resistance to linezolid in staphylococcus aureus. [Int J Res Med Sci 2014; 2(4.000: 1253-1256

  2. Líquen aureus "algesiogênico" "Algesiogenic" lichen aureus

    Directory of Open Access Journals (Sweden)

    Roberto Rheingantz da Cunha Filho

    2006-03-01

    Full Text Available Descreve-se caso de líquen aureus em paciente do sexo feminino, com 23 anos de idade que apresentava há dois anos lesão dolorosa, purpúrica, acastanhada tendendo por semelhante a cor de ferrugem e de aspecto liquenóide no antebraço. O exame anatomopatológico revelou denso infiltrado linfo-histiocitário na derme superior papilar, com extravasamento de hemácias. O líquen aureus é relativamente raro, sendo ainda mais raro o sintoma de dor.A case is described of lichen aureus in a 23 year old female with a 2-year history of painful, purpuric, rust-coloured to tan, lichenous lesion on forearm. A biopsy specimen demonstrated a dense lymphohistiocytic infiltrate in the upper dermis, with extravasation of red cells. The "algesiogenic" lichen aureus is a very rare dermatosis.

  3. Staphylococcus aureus: methicillin-susceptible S. aureus to methicillin-resistant S. aureus and vancomycin-resistant S. aureus.

    Science.gov (United States)

    Rehm, Susan J; Tice, Alan

    2010-09-15

    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.

  4. Quick and clean cloning.

    Science.gov (United States)

    Thieme, Frank; Marillonnet, Sylvestre

    2014-01-01

    Identification of unknown sequences that flank known sequences of interest requires PCR amplification of DNA fragments that contain the junction between the known and unknown flanking sequences. Since amplified products often contain a mixture of specific and nonspecific products, the quick and clean (QC) cloning procedure was developed to clone specific products only. QC cloning is a ligation-independent cloning procedure that relies on the exonuclease activity of T4 DNA polymerase to generate single-stranded extensions at the ends of the vector and insert. A specific feature of QC cloning is the use of vectors that contain a sequence called catching sequence that allows cloning specific products only. QC cloning is performed by a one-pot incubation of insert and vector in the presence of T4 DNA polymerase at room temperature for 10 min followed by direct transformation of the incubation mix in chemo-competent Escherichia coli cells.

  5. Population structure of Staphylococcus aureus from remote African Babongo Pygmies.

    Directory of Open Access Journals (Sweden)

    Frieder Schaumburg

    Full Text Available BACKGROUND: Pandemic community-acquired methicillin-resistant Staphylococcus aureus isolates (CA-MRSA predominantly encode the Panton-Valentine leukocidin (PVL, which can be associated with severe infections. Reports from non-indigenous Sub-Saharan African populations revealed a high prevalence of PVL-positive isolates. The objective of our study was to investigate the S. aureus carriage among a remote indigenous African population and to determine the molecular characteristics of the isolates, particularly those that were PVL-positive. METHODOLOGY/PRINCIPAL FINDINGS: Nasal S. aureus carriage and risk factors of colonization were systematically assessed in remote Gabonese Babongo Pygmies. Susceptibility to antibiotics, possession of toxin-encoding genes (i.e., PVL, enterotoxins, and exfoliative toxins, S. aureus protein A (spa types and multi-locus sequence types (MLST were determined for each isolate. The carriage rate was 33%. No MRSA was detected, 61.8% of the isolates were susceptible to penicillin. Genes encoding PVL (55.9%, enterotoxin B (20.6%, exfoliative toxin D (11.7% and the epidermal cell differentiation inhibitor B (11.7% were highly prevalent. Thirteen spa types were detected and were associated with 10 STs predominated by ST15, ST30, ST72, ST80, and ST88. CONCLUSIONS: The high prevalence of PVL-positive isolates among Babongo Pygmies demands our attention as PVL can be associated with necrotinzing infection and may increase the risk of severe infections in remote Pygmy populations. Many S. aureus isolates from Babongo Pygmies and pandemic CA-MRSA-clones have a common genetic background. Surveillance is needed to control the development of resistance to antibiotic drugs and to assess the impact of the high prevalence of PVL in indigenous populations.

  6. Role of the ESAT-6 secretion system in virulence of the emerging community-associated Staphylococcus aureus lineage ST398

    OpenAIRE

    Yanan Wang; Mo Hu; Qian Liu; Juanxiu Qin; Yingxin Dai; Lei He; Tianming Li; Bing Zheng; Fan Zhou; Kaiwen Yu; Jingyuan Fang; Xiaoyun Liu; Michael Otto; Min Li

    2016-01-01

    Novel Staphylococcus aureus clones continue to emerge that cause infections in otherwise healthy people. One example is the sequence type (ST) 398 lineage, which we show here is increasing in importance as a significant cause of community-associated (CA) human infections in China. We have a profound lack of understanding about what determines the considerable virulence potential of such newly emerging clones. Information about the contribution to virulence of the more recently discovered ESAT...

  7. Clonal profile, virulence and resistance of Staphylococcus aureus isolated from sheep milk.

    Science.gov (United States)

    Martins, Katheryne Benini; Faccioli-Martins, Patricia Yoshida; Riboli, Danilo Flávio Moraes; Pereira, Valéria Cataneli; Fernandes, Simone; Oliveira, Aline A; Dantas, Ariane; Zafalon, Luiz Francisco; da Cunha, Maria de Lourdes Ribeiro de Souza

    2015-06-01

    The objective of this study was to characterize the clonal profile, virulence factors and antimicrobial resistance, particularly oxacillin resistance, of Staphylococcus aureus isolated from sheep milk. Milk samples were collected from all teats for the California Mastitis Test (CMT), somatic cell count, identification of S. aureus, investigation in these strains of genes encoding toxins (sea, seb, sec, sed, tst), biofilm (icaA, icaC, icaD, bap), leukocidin (luk-PV) oxacillin resistance by mecA gene detection and susceptibility testing (12 antibiotics). Messenger RNA expression was evaluated by RT-PCR in isolates carrying toxin and biofilm genes. Biofilm formation was also evaluated phenotypically by adherence to polystyrene plates. The clonal profile of S. aureus was investigated by pulsed-field gel electrophoresis. A total of 473 milk samples were collected from 242 animals on three farms and 20 S. aureus strains were isolated and none carried the mecA gene. The two sec gene-positive isolates and the isolates carrying the tst and luk-PV genes were positive by RT-PCR. Staphylococcus aureus isolated from the three flocks studied showed high susceptibility to the drugs tested and none was biofilm producer, indicating that biofilm formation was not a virulence factor causing infection by these strains. The typing of 17 S. aureus isolates revealed the presence of a common clone on the three farms studied, and the presence and expression of the sec and tst genes in one strain of this clone suggest the possible acquisition of virulence genes by this clone, a fact that is important for animal health and food hygiene.

  8. Clonal profile, virulence and resistance of Staphylococcus aureus isolated from sheep milk

    Directory of Open Access Journals (Sweden)

    Katheryne Benini Martins

    2015-06-01

    Full Text Available The objective of this study was to characterize the clonal profile, virulence factors and antimicrobial resistance, particularly oxacillin resistance, of Staphylococcus aureus isolated from sheep milk. Milk samples were collected from all teats for the California Mastitis Test (CMT, somatic cell count, identification of S. aureus, investigation in these strains of genes encoding toxins (sea, seb, sec, sed, tst, biofilm (icaA, icaC, icaD, bap, leukocidin (luk-PV oxacillin resistance by mecA gene detection and susceptibility testing (12 antibiotics. Messenger RNA expression was evaluated by RT-PCR in isolates carrying toxin and biofilm genes. Biofilm formation was also evaluated phenotypically by adherence to polystyrene plates. The clonal profile of S. aureus was investigated by pulsed-field gel electrophoresis. A total of 473 milk samples were collected from 242 animals on three farms and 20 S. aureus strains were isolated and none carried the mecA gene. The two sec gene-positive isolates and the isolates carrying the tst and luk-PV genes were positive by RT-PCR. Staphylococcus aureus isolated from the three flocks studied showed high susceptibility to the drugs tested and none was biofilm producer, indicating that biofilm formation was not a virulence factor causing infection by these strains. The typing of 17 S. aureus isolates revealed the presence of a common clone on the three farms studied, and the presence and expression of the sec and tst genes in one strain of this clone suggest the possible acquisition of virulence genes by this clone, a fact that is important for animal health and food hygiene.

  9. Pathogenesis of Staphylococcus aureus abscesses.

    Science.gov (United States)

    Kobayashi, Scott D; Malachowa, Natalia; DeLeo, Frank R

    2015-06-01

    Staphylococcus aureus causes many types of human infections and syndromes-most notably skin and soft tissue infections. Abscesses are a frequent manifestation of S. aureus skin and soft tissue infections and are formed, in part, to contain the nidus of infection. Polymorphonuclear leukocytes (neutrophils) are the primary cellular host defense against S. aureus infections and a major component of S. aureus abscesses. These host cells contain and produce many antimicrobial agents that are effective at killing bacteria, but can also cause non-specific damage to host tissues and contribute to the formation of abscesses. By comparison, S. aureus produces several molecules that also contribute to the formation of abscesses. Such molecules include those that recruit neutrophils, cause host cell lysis, and are involved in the formation of the fibrin capsule surrounding the abscess. Herein, we review our current knowledge of the mechanisms and processes underlying the formation of S. aureus abscesses, including the involvement of polymorphonuclear leukocytes, and provide a brief overview of therapeutic approaches.

  10. Molecular epidemiology of heteroresistant vancomycin-intermediate Staphylococcus aureus in Brazil

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    Alessandro Conrado de Oliveira Silveira

    2015-10-01

    Full Text Available ABSTRACTTo determine the epidemiological and molecular characteristics of 12 Staphylococcus aureus isolates presenting heteroresistance to vancomycin in laboratories of two cities in Santa Catarina, southern Brazil. Epidemiological data, including the city of isolation, health institution, and date of isolation were considered, as well as the associated clinical specimen. For molecular characterization, we analyzed the staphylococcal cassette chromosome types, the erm gene presence, and the genomic diversity of isolates using pulsed-field gel electrophoresis. The 12 isolates of S. aureus were previously confirmed as heteroresistance to vancomycin using the population analysis profile-area under curve. Regarding genetic variability, two clones were detected: the main one (clone A composed of four isolates and the clones B, with two isolates. For clone A, two isolates presented identical band patterns and were related to the same hospital, with an interval of 57 days between their isolation. The other isolates of this clone showed no epidemiological link between them because they were isolated in different hospitals and had no temporal relationship. The other clone showed no detectable epidemiological relationship. The heteroresistance to vancomycin recovered in Santa Catarina State from 2009 to 2012 had, in general, heterogeneous genomic patterns based on pulsed-field gel electrophoresis results, which is in accordance with the fact that these isolates had little or no epidemiological relationship among them. Due to the characteristic phenotypic instability and often prolonged vancomycin therapy for selection, clonal spread is not as common as for other resistance mechanisms disseminated through horizontal gene transfer.

  11. Molecular Epidemiology of Staphylococcus aureus among Patients with Skin and Soft Tissue Infections in Two Chinese Hospitals

    Science.gov (United States)

    Gu, Fei-Fei; Chen, Ye; Dong, De-Ping; Song, Zhen; Guo, Xiao-Kui; Ni, Yu-Xing; Han, Li-Zhong

    2016-01-01

    Background: Staphylococcus aureus is one of the predominant causes of skin and soft tissue infections (SSTIs), but limited data were available regarding the characterization of S. aureus from SSTIs patients in Jiangsu Province in China. We aimed to investigate the molecular epidemiology of S. aureus among SSTIs patients in two hospitals of Jiangsu Province. Methods: Sixty-two patients with SSTIs from two Chinese hospitals in Jiangsu Province were enrolled in this study, and 62 S. aureus isolates were collected from February 2014 to January 2015. S. aureus isolates were characterized by antimicrobial susceptibility testing, toxin gene detection, and molecular typing with sequence type, Staphylococcus protein A gene type, accessory gene regulator (agr) group, and Staphylococcal cassette chromosome mec type. Results: Sixteen (25.8%) methicillin-resistant S. aureus (MRSA) isolates were detected, and there was no isolate found resistant to vancomycin, teicoplanin, sulfamethoxazole-trimethoprim, and linezolid. The sei was the toxin gene most frequently found, and no lukS/F-PV-positive isolates were detected among the SSTIs’ patients. Molecular analysis revealed that ST398 (10/62, 16.1%; 2 MRSA and 8 methicillin-susceptible S. aureus) to be the dominant clone, followed by ST5 (8/62, 12.9%) and ST7 (8/62, 12.9%). Conclusions: The livestock ST398 was the most common clone among patients with S. aureus SSTIs in Jiangsu Province, China. Surveillance and further studies on the important livestock ST398 clone in human infections are necessarily requested. PMID:27647191

  12. Molecular Epidemiology of Staphylococcus aureus among Patients with Skin and Soft Tissue Infections in Two Chinese Hospitals

    Institute of Scientific and Technical Information of China (English)

    Fei-Fei Gu; Ye Chen; De-Ping Dong; Zhen Song; Xiao-Kui Guo; Yu-Xing Ni; Li-Zhong Han

    2016-01-01

    Background:Staphylococcus aureus is one of the predominant causes of skin and soft tissue infections (SSTIs),but limited data were available regarding the characterization of S.aureus from SSTIs patients in Jiangsu Province in China.We aimed to investigate the molecular epidemiology ofS.aureus among SSTIs patients in two hospitals of Jiangsu Province.Methods:Sixty-two patients with SSTIs from two Chinese hospitals in Jiangsu Province were enrolled in this study,and 62 S.aureus isolates were collected from February 2014 to January 2015.S.aureus isolates were characterized by antimicrobial susceptibility testing,toxin gene detection,and molecular typing with sequence type,Staphylococcus protein A gene type,accessorygeneregulator(agr)group,and Staphylococcal cassette chromosome mec type.Results:Sixteen (25.8%) methicillin-resistant S.aureus (MRSA) isolates were detected,and there was no isolate found resistant to vancomycin,teicoplanin,sulfamethoxazole-trimethoprim,and linezolid.The sei was the toxin gene most frequently found,and no lukS/F-PV-positive isolates were detected among the SSTIs' patients.Molecular analysis revealed that ST398 (10/62,16.1%;2 MRSA and 8 methicillin-susceptible S.aureus) to be the dominant clone,followed by ST5 (8/62,12.9%) and ST7 (8/62,12.9%).Conclusions:The livestock ST398 was the most common clone among patients with S.aureus SSTIs in Jiangsu Province,China.Surveillance and further studies on the important livestock ST398 clone in human infections are necessarily requested.

  13. Staphylococcus aureus Bacteraemia

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    James Price

    2010-01-01

    Full Text Available Staphylococcus aureus bacteraemia (SAB is commonly complicated by metastatic infection or relapse after treatment. Objectives. The study aim was to determine the role of bacterial, host, and management factors in development of complicated SAB. Methods. A prospectively-conducted observational study gathered data on predisposition, management and outcome of 100 consecutive SAB cases. Antibiotic susceptibilities and genetic lineage of bacterial isolates were determined. Further clinical and microbiological data were gathered on two retrospective series from 1999–2000 (n=57 and 2004 (n=116. Results. In the prospective cases, 27% met our definition of complicated disease. Expressed as RR and 95% CI, complicated disease was associated with diabetes (1.58, 1.00–2.48, injecting-drug use (5.48, 0.88–33.49, community-onset of symptoms (1.4, 1.02–1.92, and symptom duration ≥48 hours prior to starting effective antibiotic therapy (2.10, 1.22–3.61. Uncomplicated disease was associated with the presence of a central line (0.69, 0.55–0.88 and prompt removal of a primary focus (0.71, 0.57–0.90. Neither methicillin resistance nor genetic lineage was associated with complicated disease, but methicillin resistance was associated with higher mortality. Conclusions. This study demonstrates that clinical rather than microbial factors are the major determinants of SAB outcome and underscores the importance of early treatment.

  14. New epidemiology of Staphylococcus aureus infections in the Middle East.

    Science.gov (United States)

    Tokajian, S

    2014-07-01

    Staphylococcus aureus is a bacterial pathogen that is distributed worldwide and represents an increasing problem, both in hospitals and in the community. Global transmission of methicillin-resistant S. aureus (MRSA) has been the subject of many studies. Determining the incidence of colonization with community-acquired MRSA in hospitalized patients and outpatients has been the aim of several studies conducted in the Middle East (western Asia). The local epidemiology within countries in this region is changing, owing to the introduction of new strains with the intercontinental exchange of several clones. Sequence type 80-MRSA-IV is one common clone detected in different countries within the region showing country-based differences, and hence more likely to form clonal lineages. MRSA is endemic in this region, and the burden and the difficulty in detecting imported strains are increasing. This is also increasing the risk of domestic and global transmission. To counter the threat associated with the high incidence of MRSA carriage and infections, systematic surveillance of both hospital and community isolates is required, along with appropriate measures designed to limit their spread. Additionally, antibiotic stewardship is needed to contain the further development of the observed resistance and to help in preserving antibiotics as precious therapeutic resources. It is critical for countries in this region to establish both national and international initiatives to develop better measurements designed to limit and control the spread of infections. Finally, more sequence-based studies are needed to better understand the pathogenicity and epidemiology of these important pathogens.

  15. Introduction of plasmid DNA into an ST398 livestock-associated methicillin-resistant Staphylococcus aureus strain

    Science.gov (United States)

    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...

  16. Phylogenetic Analysis of Staphylococcus aureus CC398 Reveals a Sub-Lineage Epidemiologically Associated with Infections in Horses

    DEFF Research Database (Denmark)

    Abdelbary, Mohamed M. H.; Wittenberg, Anne; Cuny, Christiane;

    2014-01-01

    -allelic polymorphisms, and phylogenetic analyses revealed that an epidemic sub-clone within CC398 (dubbed 'clade (C)') has spread within and between equine hospitals, where it causes nosocomial infections in horses and colonises the personnel. While clade (C) was strongly associated with S. aureus from horses...

  17. Genetic diversity of methicillin-resistant Staphylococcus aureus in a tertiary hospital in The Netherlands between 2002 and 2006

    NARCIS (Netherlands)

    Nulens, E; Stobberingh, E E; Smeets, E; van Dessel, H; Welling, M A; Sebastian, S; van Tiel, F H; Beisser, P S; Deurenberg, R H

    2009-01-01

    The aim of this study was to investigate the methicillin-resistant Staphylococcus aureus (MRSA) clones isolated in a Dutch university hospital, situated near the borders of Belgium and Germany, between 2002 and 2006. MRSA strains (n = 175) were characterized using spa and SCCmec typing. The presence

  18. Livestock-associated Staphylococcus aureus on Polish pig farms

    Science.gov (United States)

    Mroczkowska, Aneta; Żmudzki, Jacek; Marszałek, Natalia; Orczykowska-Kotyna, Monika; Komorowska, Iga; Nowak, Agnieszka; Grzesiak, Anna; Czyżewska-Dors, Ewelina; Dors, Arkadiusz; Pejsak, Zygmunt; Hryniewicz, Waleria; Wyszomirski, Tomasz; Empel, Joanna

    2017-01-01

    Background Livestock-associated Staphylococcus aureus (LA-SA) draws increasing attention due to its particular ability to colonize farm animals and be transmitted to people, which in turn leads to its spread in the environment. The aim of the study was to determine the dissemination of LA-SA on pig farms selected throughout Poland, characterize the population structure of identified S. aureus, and assess the prevalence of LA-SA carriage amongst farmers and veterinarians being in contact with pigs. Methods and findings The study was conducted on 123 pig farms (89 farrow-to-finish and 34 nucleus herds), located in 15 out of 16 provinces of Poland. Human and pig nasal swabs, as well as dust samples were analyzed. S. aureus was detected on 79 (64.2%) farms from 14 provinces. Amongst these farms LA-SA-positive farms dominated (71/79, 89.9%, 95% CI [81.0%, 95.5%]). The prevalence of LA-MRSA-positive farms was lower than LA-MSSA-positive (36.6% of LA-SA-positive farms, 95% CI [25.5%, 48.9%] vs. 74.6%, 95% CI [62.9%, 84.2%]). In total, 190 S. aureus isolates were identified: 72 (38%) MRSA and 118 (62%) methicillin-susceptible S. aureus (MSSA), of which 174 (92%) isolates were classified to three livestock-associated lineages: CC398 (73%), CC9 (13%), and CC30/ST433 (6%). All CC398 isolates belonged to the animal clade. Four LA-MRSA clones were detected: ST433-IVa(2B) clone (n = 8, 11%), described to the best of our knowledge for the first time, and three ST398 clones (n = 64, 89%) with the most prevalent being ST398-V(5C2&5)c, followed by ST398-V(5C2), and ST398-IVa(2B). Nasal carriage of LA-SA by pig farmers was estimated at 13.2% (38/283), CC398 carriage at 12.7% (36/283) and ST398-MRSA carriage at 3.2% (9/283), whereas by veterinarians at 21.1% (8/38), 18.4% (7/38) and 10.5% (4/38), respectively. Conclusions The prevalence of LA-MRSA-positive pig farms in Poland has increased considerably since 2008, when the first MRSA EU baseline survey was conducted in Europe. On

  19. Cloning-free CRISPR

    NARCIS (Netherlands)

    Arbab, Mandana; Srinivasan, Sharanya; Hashimoto, Tatsunori; Geijsen, Niels; Sherwood, Richard I

    2015-01-01

    We present self-cloning CRISPR/Cas9 (scCRISPR), a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA) or knockin homology construct for each targ

  20. A type IV modification-dependent restriction enzyme SauUSI from Staphylococcus aureus subsp. aureus USA300.

    Science.gov (United States)

    Xu, Shuang-Yong; Corvaglia, Anna R; Chan, Siu-Hong; Zheng, Yu; Linder, Patrick

    2011-07-01

    A gene encoding a putative DNA helicase from Staphylococcus aureus USA300 was cloned and expressed in Escherichia coli. The protein was purified to over 90% purity by chromatography. The purified enzyme, SauUSI, predominantly cleaves modified DNA containing 5mC and 5-hydroxymethylcytosine. Cleavage of 5mC-modified plasmids indicated that the sites S5mCNGS (S = C or G) are preferentially digested. The endonuclease activity requires the presence of adenosine triphosphate (ATP) or dATP whereas the non-hydrolyzable γ-S-ATP does not support activity. SauUSI activity was inhibited by ethylenediaminetetraacetic acid. It is most active in Mg(++) buffers. No companion methylase gene was found near the SauUSI restriction gene. The absence of a cognate methylase and cleavage of modified DNA indicate that SauUSI belongs to type IV restriction endonucleases, a group that includes EcoK McrBC and Mrr. SauUSI belongs to a family of highly similar homologs found in other sequenced S. aureus, S. epidermidis and S. carnosus genomes. More distant SauUSI orthologs can be found in over 150 sequenced bacterial/archaea genomes. Finally, we demonstrated the biological function of the type IV REase in restricting 5mC-modified plasmid DNA by transformation into clinical S. aureus strain SA564, and in restricting phage λ infection when the endonuclease is expressed in E. coli.

  1. Mild Staphylococcus aureus skin infection improves the course of subsequent endogenous S. aureus bacteremia in mice

    NARCIS (Netherlands)

    S. van den Berg (Sanne); C.P. de Vogel (Corné); A.F. van Belkum (Alex); I.A.J.M. Bakker-Woudenberg (Irma)

    2015-01-01

    textabstractStaphylococcus aureus carriers with S. aureus bacteremia may have a reduced mortality risk compared to non-carriers. A role for the immune system is suggested. Here, we study in mice the effect of mild S. aureus skin infection prior to endogenous or exogenous S. aureus bacteremia, and ev

  2. [Protein toxins of Staphylococcus aureus].

    Science.gov (United States)

    Shamsutdinov, A F; Tiurin, Iu A

    2014-01-01

    Main scientific-research studies regarding protein bacterial toxins of the most widespread bacteria that belong to Staphylococcus spp. genus and in particular the most pathogenic species for humans--Staphylococcus aureus, are analyzed. Structural and biological properties of protein toxins that have received the name of staphylococcus pyrogenic toxins (PTSAg) are presented. Data regarding genetic regulation of secretion and synthesis of these toxins and 3 main regulatory genetic systems (agr--accessory gene regulator, xpr--extracellular protein regulator, sar--staphylococcal accessory regulator) that coordinate synthesis of the most important protein toxins and enzymes for virulence of S. aureus, are presented.

  3. The sae locus of Staphylococcus aureus encodes a two-component regulatory system.

    Science.gov (United States)

    Giraudo, A T; Calzolari, A; Cataldi, A A; Bogni, C; Nagel, R

    1999-08-01

    Sae is a regulatory locus that activates the production of several exoproteins in Staphylococcus aureus. A 3.4-kb fragment of a S. aureus genomic library, screened with a probe adjacent to the transposon insertion of a sae::Tn551 mutant, was cloned into a bifunctional vector. This fragment was shown to carry the sae locus by restoration of exoprotein production in sae mutants. The sae locus was mapped to the SmaI-D fragment of the staphylococcal chromosome by pulse-field electrophoresis. Sequence analysis of the cloned fragment revealed the presence of two genes, designated saeR and saeS, encoding a response regulator and a histidine protein kinase, respectively, with high homology to other bacterial two-component regulatory systems.

  4. Molecular characterization and antimicrobial susceptibility of nasal Staphylococcus aureus isolates from a Chinese medical college campus.

    Directory of Open Access Journals (Sweden)

    Jimei Du

    Full Text Available Staphylococcus aureus colonization and infection occur more commonly among persons living or working in crowded conditions, but characterization of S. aureus colonization within medical communities in China is lacking. A total of 144 (15.4%, 144/935 S. aureus isolates, including 28 (3.0%, 28/935 MRSA isolates, were recovered from the nares of 935 healthy human volunteers residing on a Chinese medical college campus. All S. aureus isolates were susceptible to vancomycin, quinupristin/dalfopristin and linezolid but the majority were resistant to penicillin (96.5%, ampicillin/sulbactam (83.3% and trimethoprim/sulfamethoxazole (93.1%. 82%, (23/28 of the MRSA isolates and 66% (77/116 of the MSSA isolates were resistant to multiple antibiotics, and 3 MRSA isolates were resistant to mupirocin--an agent commonly used for nasal decolonization. 16 different sequence types (STs, as well as SCCmec genes II, III, IVd, and V, were represented among MRSA isolates. We also identified, for the first time, two novel STs (ST1778 and ST1779 and 5 novel spa types for MRSA. MRSA isolates were distributed in different sporadic clones, and ST59-MRSA-VId- t437 was found within 3 MRSA isolates. Moreover, one isolate with multidrug resistance belonging to ST398-MRSA-V- t571 associated with animal infections was identified, and 3 isolates distributed in three different clones harbored PVL genes. Collectively, these data indicate a high prevalence of nasal MRSA carriage and molecular heterogeneity of S. aureus isolates among persons residing on a Chinese medical college campus. Identification of epidemic MRSA clones associated with community infection supports the need for more effective infection control measures to reduce nasal carriage and prevent dissemination of MRSA to hospitalized patients and health care workers in this community.

  5. Prevention of Healthcare Associated Staphylococcus aureus Infections

    NARCIS (Netherlands)

    L.G.M. Bode (Lonneke)

    2014-01-01

    markdownabstract__Abstract__ S. aureus colonizes the skin and mucosae of a proportion of the human population. Carriers of S. aureus are at increased risk of developing infections with this pathogen. The aim of this thesis was to add to the prevention of healthcare associated S. aureus infections.

  6. FabH Mutations Confer Resistance to FabF-Directed Antibiotics in Staphylococcus aureus

    OpenAIRE

    Parsons, Joshua B.; Yao, Jiangwei; Frank, Matthew W.; Rock, Charles O.

    2014-01-01

    Delineating the mechanisms for genetically acquired antibiotic resistance is a robust approach to target validation and anticipates the evolution of clinical drug resistance. This study defines a spectrum of mutations in fabH that render Staphylococcus aureus resistant to multiple natural products known to inhibit the elongation condensing enzyme (FabF) of bacterial type II fatty acid synthesis. Twenty independently isolated clones resistant to platensimycin, platencin, or thiolactomycin were...

  7. Dermatology case: segmental lichen aureus

    OpenAIRE

    Fernandes, I.; S. Carvalho; Machado, S.; Alves,R.; Selores, M.

    2012-01-01

    ABSTRACT The authors describe a clinical case of a six-year-old boy with history of a segmental brownish maculopapular skin eruption on his left thoracic and lumbar wall, since the last four months. Based on clinical and histological findings he was diagnosed with segmental lichen aureus.

  8. Prevalence and Molecular Epidemiology of Staphylococcus aureus among Residents of Seven Nursing Homes in Shanghai.

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    Ji Zhang

    Full Text Available Residents in nursing homes (NHs always represent potential reservoirs for Staphylococcus aureus and methicillin-resistant S. aureus (MRSA. To our knowledge, there is no epidemiological information up till now that describes the prevalence and molecular characteristics of S. aureus in nursing home residents in Shanghai, China.Four hundred and ninety-one unique residents from 7 NHs were enrolled in this study. Specimens were collected among these residents including 491 nasal swabs, 487 axillary swabs and 119 skin swabs. S. aureus isolated and identified from the swabs was characterized according to antimicrobial susceptibility profiling, toxin gene prevalence, and multilocus sequence typing (MLST, spa and SCCmec typing.Among the 491 residents screened, S. aureus was isolated in 109 residents from 90 nasal swabs (90/491, 18.3%, 29 axillary swabs (29/487, 6.0%, and 22 skin swabs (22/119, 18.5%. Sixty-eight MRSA isolates were detected in 52 residents from 41 nasal carriers, 15 axillary carriers and 12 skin carriers. The overall prevalence rate of S. aureus and MRSA colonization was 22.2% and 10.6% respectively. Ten residents presented S. aureus in all three sample types and 12 residents presented S. aureus in two of the three sample types collected. Molecular analysis revealed CC1 (29.1% to be the dominant clone in this study, followed by CC398 (19.9%, CC188 (13.5% and CC5 (12.8%. The most common spa type was t127 (22.0%, followed by t14383 (12.8% and t002 (10.6%.A high prevalence of S. aureus and MRSA colonization was revealed in nursing home residents in Shanghai. CC1 was the most common clonal complex and t127 was the most common spa type among NH residents. The data provides an important baseline for future surveillance of S. aureus in NHs in Shanghai and other highly urbanized regions in China. Implementation of infection control strategies must be given high priority in NHs to fight such high prevalence of both MRSA and methicillin

  9. Unified Approach to Universal Cloning and Phase-Covariant Cloning

    OpenAIRE

    Hu, Jia-Zhong; Yu, Zong-Wen; Wang, Xiang-Bin

    2008-01-01

    We analyze the problem of approximate quantum cloning when the quantum state is between two latitudes on the Bloch's sphere. We present an analytical formula for the optimized 1-to-2 cloning. The formula unifies the universal quantum cloning (UQCM) and the phase covariant quantum cloning.

  10. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.; Folkers, G.E.

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning (EF

  11. Do Managers Clone Themselves?

    Science.gov (United States)

    Baron, Alma S.

    1981-01-01

    A recent questionnaire survey provides statistics on male managers' views of female managers. The author recommends that male managers break out of their cloning behavior and that the goal ought to be a plurality in management. (Author/WD)

  12. Main: Clone Detail [KOME

    Lifescience Database Archive (English)

    Full Text Available Clone Detail Mapping Pseudomolecule data detail Detail information Mapping to the TIGR japonica Pseudomolecu...les kome_mapping_pseudomolecule_data_detail.zip kome_mapping_pseudomolecule_data_detail ...

  13. BIOETHICS AND HUMAN CLONING

    Directory of Open Access Journals (Sweden)

    Željko Kaluđerović

    2011-12-01

    Full Text Available In this paper the authors analyze the process of negotiating and beginning of the United Nations Declaration on Human Cloning as well as the paragraphs of the very Declaration. The negotiation was originally conceived as a clear bioethical debate that should have led to a general agreement to ban human cloning. However, more often it had been discussed about human rights, cultural, civil and religious differences between people and about priorities in case of eventual conflicts between different value systems. In the end, a non-binding Declaration on Human Cloning had been adopted, full of numerous compromises and ambiguous formulations, that relativized the original intention of proposer states. According to authors, it would have been better if bioethical discussion and eventual regulations on cloning mentioned in the following text had been left over to certain professional bodies, and only after the public had been fully informed about it should relevant supranational organizations have taken that into consideration.

  14. MRSA in Austria--an overview.

    Science.gov (United States)

    Krziwanek, K; Luger, C; Sammer, B; Stumvoll, S; Stammler, M; Sagel, U; Witte, W; Mittermayer, H

    2008-03-01

    The aim of this study was to provide an overview of predominant and sporadic methicillin-resistant Staphylococcus aureus (MRSA) strains in large regions of Austria, and to compare the results with those from other European countries. In total, 1439 MRSA isolates, collected routinely between January 1996 and June 2006 from five Austrian federal provinces, were investigated. The isolates were confirmed as MRSA using mecA/femA multiplex PCR assays. Genes encoding Panton-Valentine leukocidin (PVL), which are characteristic of community-acquired MRSA, were also detected by PCR. Subtyping was performed using SmaI macrorestriction digestion of genomic DNA, followed by pulsed-field gel electrophoresis (PFGE) and cluster analysis. Isolates that could not be assigned to clusters were further analysed by spa typing and/or multilocus sequence typing. The predominant clones detected in Austria were ST228 (southern German epidemic clone), ST5 (Rhine-Hessen MRSA), the ST8 Austrian clone and CC8/ST8. Whereas the frequencies of lineages corresponding to ST247, ST45 and ST22 remained comparably low, an increase in the frequency of lineages corresponding to ST5 and to ST228 was recorded. Overall, 20 different MRSA types and 321 subtypes were recognised according to PFGE analysis. The prevalence of different strains varied considerably in the different Austrian regions. When compared to other European countries, the situation in Austria was most similar to that found in Germany.

  15. Phosphorylation of Staphylococcus aureus Protein-Tyrosine Kinase Affects the Function of Glucokinase and Biofilm Formation

    Science.gov (United States)

    Vasu, Dudipeta; Kumar, Pasupuleti Santhosh; Prasad, Uppu Venkateswara; Swarupa, Vimjam; Yeswanth, Sthanikam; Srikanth, Lokanathan; Sunitha, Manne Mudhu; Choudhary, Abhijith; Krishna Sarma, Potukuchi Venkata Gurunadha

    2017-01-01

    Background: When Staphylococcus aureus is grown in the presence of high concentration of external glucose, this sugar is phosphorylated by glucokinase (glkA) to form glucose-6-phosphate. This product subsequently enters into anabolic phase, which favors biofilm formation. The presence of ROK (repressor protein, open reading frame, sugar kinase) motif, phosphate-1 and -2 sites, and tyrosine kinase sites in glkA of S. aureus indicates that phosphorylation must regulate the glkA activity. The aim of the present study was to identify the effect of phosphorylation on the function of S. aureus glkA and biofilm formation. Methods: Pure glkA and protein-tyrosine kinase (BYK) of S. aureus ATCC 12600 were obtained by fractionating the cytosolic fractions of glkA1 and BYK-1 expressing recombinant clones through nickel metal chelate column. The pure glkA was used as a substrate for BYK, and the phosphorylation of glkA was confirmed by treating with reagent A and resolving in SDS-PAGE, as well as staining with reagent A. The kinetic parameters of glkA and phosphorylated glkA were determined spectrophotometrically, and in silico tools were used for validation. S. aureus was grown in brain heart infusion broth, which was supplemented with glucose, and then biofilm units were calculated. Results: Fourfold elevated glkA activity was observed upon the phosphorylation by BYK. Protein-protein docking analysis revealed that glkA structure docked close to the adenosine triphosphate-binding site of BYK structure corroborating the kinetic results. Further, S. aureus grown in the presence of elevated glucose concentration exhibited an increase in the rate of biofilm formation. Conclusion: The elevated function of glkA is an essential requirement for increased biofilm units in S. aureus, a key pathogenic factor that helps its survival and the progress of infection. PMID:27695030

  16. Comparative Whole-Genome Mapping To Determine Staphylococcus aureus Genome Size, Virulence Motifs, and Clonality

    Science.gov (United States)

    Pantrang, Madhulatha; Stahl, Buffy; Briska, Adam M.; Stemper, Mary E.; Wagner, Trevor K.; Zentz, Emily B.; Callister, Steven M.; Lovrich, Steven D.; Henkhaus, John K.; Dykes, Colin W.

    2012-01-01

    Despite being a clonal pathogen, Staphylococcus aureus continues to acquire virulence and antibiotic-resistant genes located on mobile genetic elements such as genomic islands, prophages, pathogenicity islands, and the staphylococcal chromosomal cassette mec (SCCmec) by horizontal gene transfer from other staphylococci. The potential virulence of a S. aureus strain is often determined by comparing its pulsed-field gel electrophoresis (PFGE) or multilocus sequence typing profiles to that of known epidemic or virulent clones and by PCR of the toxin genes. Whole-genome mapping (formerly optical mapping), which is a high-resolution ordered restriction mapping of a bacterial genome, is a relatively new genomic tool that allows comparative analysis across entire bacterial genomes to identify regions of genomic similarities and dissimilarities, including small and large insertions and deletions. We explored whether whole-genome maps (WGMs) of methicillin-resistant S. aureus (MRSA) could be used to predict the presence of methicillin resistance, SCCmec type, and Panton-Valentine leukocidin (PVL)-producing genes on an S. aureus genome. We determined the WGMs of 47 diverse clinical isolates of S. aureus, including well-characterized reference MRSA strains, and annotated the signature restriction pattern in SCCmec types, arginine catabolic mobile element (ACME), and PVL-carrying prophage, PhiSa2 or PhiSa2-like regions on the genome. WGMs of these isolates accurately characterized them as MRSA or methicillin-sensitive S. aureus based on the presence or absence of the SCCmec motif, ACME and the unique signature pattern for the prophage insertion that harbored the PVL genes. Susceptibility to methicillin resistance and the presence of mecA, SCCmec types, and PVL genes were confirmed by PCR. A WGM clustering approach was further able to discriminate isolates within the same PFGE clonal group. These results showed that WGMs could be used not only to genotype S. aureus but also to

  17. Comparison of genotypes and enterotoxin genes between Staphylococcus aureus isolates from blood and nasal colonizers in a Korean hospital.

    Science.gov (United States)

    Peck, Kyong Ran; Baek, Jin Yang; Song, Jae-Hoon; Ko, Kwan Soo

    2009-08-01

    In this study, we investigated the genetic background of 70 Staphylococcus aureus isolates (36 methicillin-resistant S. aureus [MRSA] and 34 methicillin-susceptible S. aureus [MSSA]) obtained from blood at a Korean tertiary-care hospital, using spa typing, multilocus sequence typing, and SCCmec typing. In addition, the prevalence of enterotoxin (sea, seb, sec, sed, see, seg, seh, sei, and sek), tst, and pvl genes among the samples was assessed via polymerase chain reaction, and the results were compared with those of 95 isolates of S. aureus obtained from nasal swabs. All MRSA isolates from blood, except one, belonged to three major clones: sequence type (ST)5-MRSA-II, ST72-MRSA-II (or IVA), and ST239-MRSA-III, among which ST5-MRSA-II was the predominant clone. The prevalence of enterotoxin genes in the S. aureus isolates obtained from blood differed significantly from those from the nasal swabs for the sea, seb, sec, and seh gene. In particular, the seb and sec genes were detected exclusively in the MRSA isolates of ST5 or spa-CC002, thereby suggesting the co-adaptation of virulence genes with the genetic background and their contribution to biological fitness.

  18. Genomic information on multidrug-resistant livestock-associated methicillin-resistant Staphylococcus aureus ST398 isolated from a Brazilian patient with cystic fibrosis

    Science.gov (United States)

    Lima, Danielle F; Cohen, Renata WF; Rocha, Géssica A; Albano, Rodolpho M; Marques, Elizabeth A; Leão, Robson S

    2017-01-01

    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

  19. Preliminary treatment of bovine mastitis caused by Staphylococcus aureus, with trx-SA1, recombinant endolysin of S. aureus bacteriophage IME-SA1.

    Science.gov (United States)

    Fan, Jindai; Zeng, Zhiliang; Mai, Kaijie; Yang, Yu; Feng, Jiaqi; Bai, Yang; Sun, Baoli; Xie, Qingmei; Tong, Yigang; Ma, Jingyun

    2016-08-15

    Methicillin-resistant Staphylococcus aureus (MRSA) has become a great threat to human and animal health and there is an urgent need to develop novel antibacterial agents to control this pathogen. The objective of this study was to obtain an active recombinant endolysin from the novel bacteriophage (IME-SA1), and conduct an efficacy trial of its effectiveness against bovine mastitis. We isolated a phage that was virulent and specific for S. aureus with an optimal multiplicity of infection of 0.01. Electron microscopy revealed that IME-SA1 was a member of the family Myoviridae, with an isometric head (98nm) and a long contractile tail (200nm). Experimental lysis experiments indicated the phage had an incubation period of 20min with a burst size of 80. When host bacteria were in early exponential growth stages, a multiplicity of infection of 0.01 resulted in a complete bacterial lysis after 9h. The endolysin gene (804bp) was cloned into the pET-32a bacterial expression vector and recombinant endolysin Trx-SA1 was successfully obtained with molecular size of about 47kDa. Preliminary results of therapeutic trials in cow udders showed that Trx-SA1 could effectively control mild clinical mastitis caused by S. aureus. The endolysin Trx-SA1 might be an alternative treatment strategy for infections caused by S. aureus, including MRSA.

  20. Evasion of Neutrophil Killing by Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Will A. McGuinness

    2016-03-01

    Full Text Available Staphylococcus aureus causes many types of infections, ranging from self-resolving skin infections to severe or fatal pneumonia. Human innate immune cells, called polymorphonuclear leukocytes (PMNs or neutrophils, are essential for defense against S. aureus infections. Neutrophils are the most prominent cell type of the innate immune system and are capable of producing non-specific antimicrobial molecules that are effective at eliminating bacteria. Although significant progress has been made over the past few decades, our knowledge of S. aureus-host innate immune system interactions is incomplete. Most notably, S. aureus has the capacity to produce numerous molecules that are directed to protect the bacterium from neutrophils. Here we review in brief the role played by neutrophils in defense against S. aureus infection, and correspondingly, highlight selected S. aureus molecules that target key neutrophil functions.

  1. Cloning of an agr homologue of Staphylococcus saprophyticus.

    Science.gov (United States)

    Sakinc, Türkan; Kulczak, Pawel; Henne, Karsten; Gatermann, Sören G

    2004-08-01

    An agr homologue of Staphylococcus saprophyticus was identified, cloned and sequenced. The gene locus shows homologies to other staphylococcal agr systems, especially to those of S. epidermidis and S. lugdunensis. A putative RNAIII was identified and found to be differentially expressed during the growth phases. In contrast to the RNAIII molecules of S. epidermidis and S. aureus it does not contain an open reading frame that codes for a protein with homologies to the delta-toxin. Using PCR, the agr was found to be present in clinical isolates of S. saprophyticus.

  2. Staphylococcus aureus complex from animals and humans in three remote African regions.

    Science.gov (United States)

    Schaumburg, Frieder; Pauly, Maude; Anoh, Etile; Mossoun, Arsene; Wiersma, Lidewij; Schubert, Grit; Flammen, Arnaud; Alabi, Abraham S; Muyembe-Tamfum, Jean-Jacques; Grobusch, Martin P; Karhemere, Stomy; Akoua-Koffi, Chantal; Couacy-Hymann, Emmanuel; Kremsner, Peter G; Mellmann, Alexander; Becker, Karsten; Leendertz, Fabian H; Peters, Georg

    2015-04-01

    Staphylococcus schweitzeri has been recently considered to be a highly divergent Staphylococcus aureus clade and usually colonises nonhuman primates and bats in sub-Saharan Africa. Its transmissibility to humans remains unclear. We therefore investigated the transmission of S. aureus and S. schweitzeri among humans, domestic animals, and wildlife in three remote African regions. A cross-sectional study on nasal and pharyngeal colonisation in humans (n = 1288) and animals (n = 698) was performed in Côte d'Ivoire, Gabon, and Democratic Republic of Congo (DR Congo). Isolates were subjected to spa typing and multilocus sequence typing. Antimicrobial susceptibility and selected virulence factors were tested. S. schweitzeri was found in monkeys from all study sites but no transmission to humans was evident, despite frequent contact of humans with wildlife. In contrast, human-associated S. aureus sequence types (ST1, ST6, ST15) were detected in domestic animals and nonhuman primates, pointing toward a human-to-monkey transmission in the wild. The proportion of methicillin-resistant S. aureus (MRSA) among all S. aureus was 0% (Gabon), 1.7% (DR Congo), and 5.3% (Côte d'Ivoire). The majority of MRSA isolates belonged to the African clone ST88. In conclusion, we did not find any evidence for a transmission of S. schweitzeri from animals to humans. However, such a transmission might remain possible due to the close phylogenetic relation of humans and nonhuman primates. The ST88-MRSA clone was widespread in Côte d'Ivoire but not in Gabon and DR Congo.

  3. Postoperative Staphylococcus aureus infections in Medicare beneficiaries.

    Directory of Open Access Journals (Sweden)

    Moaven Razavi

    Full Text Available Staphylococcus aureus (S. aureus infections are important because of their increasing frequency, resistance to antibiotics, and high associated rates of disabilities and deaths. We examined the incidence and correlates of S. aureus infections following 219,958 major surgical procedures in a 5% random sample of fee-for-service Medicare beneficiaries from 2004-2007. Of these surgical patients, 0.3% had S. aureus infections during the hospitalizations when index surgical procedures were performed; and 1.7% and 2.3%, respectively, were hospitalized with infections within 60 days or 180 days following admissions for index surgeries. S. aureus infections occurred within 180 days in 1.9% of patients following coronary artery bypass graft surgery, 2.3% following hip surgery, and 5.9% following gastric or esophageal surgery. Of patients first hospitalized with any major infection reported during the first 180 days after index surgery, 15% of infections were due to S. aureus, 18% to other documented organisms, and no specific organism was reported on claim forms in 67%. Patient-level predictors of S. aureus infections included transfer from skilled nursing facilities or chronic hospitals and comorbid conditions (e.g., diabetes, congestive heart failure, chronic obstructive pulmonary disease, and chronic renal disease. In a logarithmic regression, elective index admissions with S. aureus infection stayed 130% longer than comparable patients without that infection. Within 180 days of the index surgery, 23.9% of patients with S. aureus infection and 10.6% of patients without this infection had died. In a multivariate logistic regression of death within 180 days of admission for the index surgery with adjustment for demographics, co-morbidities, and other risks, S. aureus was associated with a 42% excess risk of death. Due to incomplete documentation of organisms in Medicare claims, these statistics may underestimate the magnitude of S. aureus infection

  4. Placentation in cloned cattle

    DEFF Research Database (Denmark)

    Miglino, M A; Pereira, F T V; Visintin, J A

    2007-01-01

    To elucidate the morphological differences between placentas from normal and cloned cattle pregnancies reaching term, the umbilical cord, placentomes and interplacentomal region of the fetal membranes were examined macroscopically as well as by light and scanning electron microscopy. In pregnancies...... than one primary villus, as opposed to a single villus in non-cloned placentae. Scanning electron microscopy of blood vessel casts revealed that there was also more than one stem artery per villous tree and that the ramification of the vessels failed to form dense complexes of capillary loops...

  5. Molecular epidemiology of hospital-onset methicillin-resistant Staphylococcus aureus infections in Southern Chile.

    Science.gov (United States)

    Medina, G; Egea, A L; Otth, C; Otth, L; Fernández, H; Bocco, J L; Wilson, M; Sola, C

    2013-12-01

    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.

  6. CHROMOSOMAL MAPPING IN STRAINS OF STAPHYLOCOCCUS AUREUS,

    Science.gov (United States)

    STAPHYLOCOCCUS AUREUS , CHROMOSOMES), (*CHROMOSOMES, MAPPING), NITROSO COMPOUNDS, GUANIDINES, GENETICS, MUTATIONS, DRUGS, TOLERANCES(PHYSIOLOGY), TEST METHODS, DEOXYRIBONUCLEIC ACIDS, INHIBITION, RESISTANCE(BIOLOGY).

  7. Mastite com lesões sistêmicas por Staphylococus aureus subesp. aureus em coelhos Mastitis with systemic lesions due to Staphylococus aureus subesp. aureus in rabbits

    OpenAIRE

    Sandra Davi Traverso; Leonardo da Cunha; Joaquim César Teixeira Fernandes; Alexandre Paulino Loretti; Adriana Rhoden; Elsio Wunder Jr.; David Driemeier

    2003-01-01

    Em uma criação composta por 1800 coelhos, 33% das matrizes apresentaram mastite e lesões cutâneas crostosas e purulentas. Estes animais apresentavam-se entre 10 a- 12 meses de idade e em segunda parição. Quinze coelhos afetados foram sacrificados e necropsiados. Na necropsia, além das lesões cutâneas haviam microabscessos em diversos órgãos. Das amostras coletadas isolou-se Staphylococcus aureus subesp. aureus. S. aureus subesp. aureus também foi isolado de "swab" nasal coletado do tratador e...

  8. Cloning and occurrence of czrC, a gene conferring cadmium and zinc resistance in MRSA CC398 Isolates

    DEFF Research Database (Denmark)

    Cavaco, Lina; Hasman, Henrik; Stegger, Marc

    2010-01-01

    determinant, czrC, by PCR. The cloning of czrC confirmed that the zinc chloride and cadmium acetate MICs for isogenic constructs carrying this gene were increased compared to those for S. aureus RN4220. No difference in susceptibility to sodium arsenate, copper sulfate, or silver nitrate was observed. Seventy...

  9. Mild Staphylococcus aureus Skin Infection Improves the Course of Subsequent Endogenous S. aureus Bacteremia in Mice.

    Directory of Open Access Journals (Sweden)

    Sanne van den Berg

    Full Text Available Staphylococcus aureus carriers with S. aureus bacteremia may have a reduced mortality risk compared to non-carriers. A role for the immune system is suggested. Here, we study in mice the effect of mild S. aureus skin infection prior to endogenous or exogenous S. aureus bacteremia, and evaluate protection in relation to anti-staphylococcal antibody levels. Skin infections once or twice by a clinical S. aureus isolate (isolate P or S. aureus strain 8325-4 were induced in mice free of S. aureus and anti-staphylococcal antibodies. Five weeks later, immunoglobulin G (IgG levels in blood against 25 S. aureus antigens were determined, and LD50 or LD100 bacteremia caused by S. aureus isolate P was induced. S. aureus skin infections led to elevated levels of anti-staphylococcal IgG in blood. One skin infection improved the course of subsequent severe endogenous bacteremia only. A second skin infection further improved animal survival rate, which was associated with increased pre-bacteremia IgG levels against Efb, IsaA, LukD, LukE, Nuc, PrsA and WTA. In conclusion, S. aureus isolate P skin infection in mice reduces the severity of subsequent endogenous S. aureus bacteremia only. Although cellular immune effects cannot be rules out, anti-staphylococcal IgG against specified antigens may contribute to this effect.

  10. Expression of Four Methionine Sulfoxide Reductases in Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Kuldeep Singh

    2012-01-01

    Full Text Available Staphylococcus aureus possesses three MsrA enzymes (MsrA1, MsrA2, MsrA3 that reduce the S-epimer of methionine sulfoxide (MetO and an MsrB enzyme that reduces R-MetO. The four msr genes are expressed from three different promoters. The msrA1/msrB genes are coexpressed. To determine the expression pattern of msr genes, three independent reporter strains were constructed where msr promoter was cloned in front of a promoterless lacZ and the resulting construct was integrated in the chromosome. Using these strains, it was determined that the msrA1/B expression is significantly higher in S. aureus compared to msrA2 or msrA3. Expression of msrA1/B was highest during stationary phase growth, but the expression of msrA2 and msrA3 was highest during the early to midexponential growth phase. Expression of msrA1/B was induced by oxacillin and the expression of msrA3 was upregulated by salt. Expression of msrA2 remained unchanged under all tested conditions.

  11. Why clone flies? Using cloned Drosophila to monitor epigenetic defects.

    Science.gov (United States)

    Haigh, Andrew J; Lloyd, Vett K

    2007-01-01

    Since the birth of the first cloned sheep in 1996, advances in nuclear transplantation have led to both the creation of genetically tailored stem cells and the generation of a number of cloned organisms. The list of cloned animals reared to adulthood currently includes the frog, sheep, mouse, cow, goat, pig, rabbit, cat, zebrafish, mule, horse, rat and dog. The addition of Drosophila to this elite bestiary of cloned animals has prompted the question - why clone flies? Organisms generated by nuclear transplantation suffer from a high rate of associated defects, and many of these defects appear to be related to aberrant genomic imprinting. Imprinted gene expression also appears to be compromised in Drosophila clones. Proper imprinted gene regulation relies on a suite of highly conserved chromatin-modifying genes first identified in Drosophila. Thus, Drosophila can potentially be used to study epigenetic dysfunction in cloned animals and to screen for genetic and epigenetic conditions that promote the production of healthy clones.

  12. Genomic analysis of ST88 community-acquired methicillin resistant Staphylococcus aureus in Ghana

    Science.gov (United States)

    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

    2017-01-01

    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

  13. Characterisation of Staphylococcus aureus bacteraemia at Tygerberg hospital

    NARCIS (Netherlands)

    Orth, H.; Dreyer, Z.S.; Makgotlho, E.; Oosthuysen, W.; Sinha, B.; Wasserman, E.

    2013-01-01

    To elucidate the local epidemiology of Staphylococcus aureus bacteraemia, we characterised blood culture isolates using molecular methods and prospectively collected clinical data to determine the occurrence of community-acquired, methicillinresistant S. aureus (MRSA). Consecutive S. aureus blood cu

  14. Levels of alpha-toxin correlate with distinct phenotypic response profiles of blood mononuclear cells and with agr background of community-associated Staphylococcus aureus isolates

    NARCIS (Netherlands)

    S.M. Shambat (Srikanth Mairpady); A. Haggar (Axana); F. Vandenesch (Francois); G. Lina (Gérard); W.J.B. van Wamel (Willem); G. Arakere (Gayathri); M. Svensson (Mattias); A. Norrby-Teglund (Anna)

    2014-01-01

    textabstractEpidemiological studies of Staphylococcus aureus have shown a relation between certain clones and the presence of specific virulence genes, but how this translates into virulence-associated functional responses is not fully elucidated. Here we addressed this issue by analyses of communit

  15. The Cloning of America.

    Science.gov (United States)

    Dobson, Judith E.; Dobson, Russell L.

    1981-01-01

    Proposes that the U.S. school system purports to prize human variability, but many educators are engaged in activities that seek to homogenize students. Describes these activities, including diagnosis, labeling, ability grouping, and positive reinforcement. Presents suggestions for counselors to combat sources of cloning and self-validation. (RC)

  16. Clip, connect, clone

    DEFF Research Database (Denmark)

    Fujima, Jun; Lunzer, Aran; Hornbæk, Kasper

    2010-01-01

    using three mechanisms: clipping of input and result elements from existing applications to form cells on a spreadsheet; connecting these cells using formulas, thus enabling result transfer between applications; and cloning cells so that multiple requests can be handled side by side. We demonstrate...

  17. Asian Yellow Goat Cloned

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    @@ It was released on August 24,2005 by Prof. CHEN Dayuan (Da-Yuan Chen) from the CAS Institute of Zoology that the first success in cloning the Asian Yellow Goat by nuclear transfer had recently been achieved in east China's Shandong Province.

  18. Surgimiento y diseminación de Staphylococcus aureus meticilinorresistente Staphylococcus aureus methicillin-resistant: emergence and dissemination

    Directory of Open Access Journals (Sweden)

    Maria Elena Velázquez-Meza

    2005-10-01

    Full Text Available Las infecciones nosocomiales ocasionadas por cepas de Staphylococcus aureus meticilinorresistentes (SAMR son un problema de salud importante en todo el mundo. Este microorganismo produce una gran variedad de infecciones incluyendo osteomielitis, endocarditis invasora, artritis séptica y septicemia. La multirresistencia es un factor que influye en la persistencia de los SAMR dentro del ámbito hospitalario. La introducción de técnicas de tipificación molecular dentro de las investigaciones epidemiológicas ha provisto nuevas herramientas para conocer el origen y las vías de diseminación de este microorganismo. Una de las conclusiones importantes que han surgido de este tipo de estudios es que un número pequeño de clonas son las responsables de las infecciones estafilocócicas en todo el mundo.Nosocomial infections due to methicillin-resistant Staphylococcus aureus (MRSA is an important health problem worldwide. This microorganism causes a variety of clinical infections, including osteomyelitis, invasive endocarditis, septic arthritis and septicemia. Antimicrobial resistance is a factor that influences the persistence of MRSA in the hospital environment. The introduction of molecular typing techniques in epidemiological investigations has provided new tools for identifying the microorganism's origin and routes of dissemination. One of the most important conclusions that have resulted from these types of studies is that a small number of clones are responsible for most of the staphylococcal infections throughout the world.

  19. Triclosan promotes Staphylococcus aureus nasal colonization.

    Science.gov (United States)

    Syed, Adnan K; Ghosh, Sudeshna; Love, Nancy G; Boles, Blaise R

    2014-04-08

    The biocide triclosan is used in many personal care products, including toothpastes, soaps, clothing, and medical equipment. Consequently, it is present as a contaminant in the environment and has been detected in some human fluids, including serum, urine, and milk. Staphylococcus aureus is an opportunistic pathogen that colonizes the noses and throats of approximately 30% of the population. Colonization with S. aureus is known to be a risk factor for several types of infection. Here we demonstrate that triclosan is commonly found in the nasal secretions of healthy adults and the presence of triclosan trends positively with nasal colonization by S. aureus. We demonstrate that triclosan can promote the binding of S. aureus to host proteins such as collagen, fibronectin, and keratin, as well as inanimate surfaces such as plastic and glass. Lastly, triclosan-exposed rats are more susceptible to nasal colonization with S. aureus. These data reveal a novel factor that influences the ability of S. aureus to bind surfaces and alters S. aureus nasal colonization. IMPORTANCE Triclosan has been used as a biocide for over 40 years, but the broader effects that it has on the human microbiome have not been investigated. We demonstrate that triclosan is present in nasal secretions of a large portion of a test population and its presence correlates with Staphylococcus aureus nasal colonization. Triclosan also promotes the binding of S. aureus to human proteins and increases the susceptibility of rats to nasal colonization by S. aureus. These findings are significant because S. aureus colonization is a known risk factor for the development of several types of infections. Our data demonstrate the unintended consequences of unregulated triclosan use and contribute to the growing body of research demonstrating inadvertent effects of triclosan on the environment and human health.

  20. Adhesive polypeptides of Staphylococcus aureus identified using a novel secretion library technique in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Holm Liisa

    2011-05-01

    Full Text Available Abstract Background Bacterial adhesive proteins, called adhesins, are frequently the decisive factor in initiation of a bacterial infection. Characterization of such molecules is crucial for the understanding of bacterial pathogenesis, design of vaccines and development of antibacterial drugs. Because adhesins are frequently difficult to express, their characterization has often been hampered. Alternative expression methods developed for the analysis of adhesins, e.g. surface display techniques, suffer from various drawbacks and reports on high-level extracellular secretion of heterologous proteins in Gram-negative bacteria are scarce. These expression techniques are currently a field of active research. The purpose of the current study was to construct a convenient, new technique for identification of unknown bacterial adhesive polypeptides directly from the growth medium of the Escherichia coli host and to identify novel proteinaceous adhesins of the model organism Staphylococcus aureus. Results Randomly fragmented chromosomal DNA of S. aureus was cloned into a unique restriction site of our expression vector, which facilitates secretion of foreign FLAG-tagged polypeptides into the growth medium of E. coli ΔfliCΔfliD, to generate a library of 1663 clones expressing FLAG-tagged polypeptides. Sequence and bioinformatics analyses showed that in our example, the library covered approximately 32% of the S. aureus proteome. Polypeptides from the growth medium of the library clones were screened for binding to a selection of S. aureus target molecules and adhesive fragments of known staphylococcal adhesins (e.g coagulase and fibronectin-binding protein A as well as polypeptides of novel function (e.g. a universal stress protein and phosphoribosylamino-imidazole carboxylase ATPase subunit were detected. The results were further validated using purified His-tagged recombinant proteins of the corresponding fragments in enzyme-linked immunoassay and

  1. The First Human Cloned Embryo.

    Science.gov (United States)

    Cibelli, Jose B.; Lanza, Robert P.; West, Michael D.; Ezzell, Carol

    2002-01-01

    Describes a process known as parthenogenesis which produces cloned, early-stage embryos and human embryos generated only from eggs. Speculates that this technology puts therapeutic cloning within reach. (DDR)

  2. Animal Cloning and Food Safety

    Science.gov (United States)

    ... For Consumers Home For Consumers Consumer Updates Animal Cloning and Food Safety Share Tweet Linkedin Pin it ... evaluate the issue. back to top FDA Studies Cloning For more than five years, CVM scientists studied ...

  3. Staphylococcus aureus outbreak in the intensive care unit of the largest public hospital in Quito, Ecuador.

    Science.gov (United States)

    Cardenas, Paul A; Alarcón, Marta; Narvaez, Inés; Salazar, Ramiro; Falconí, Guillermo; Espinel, Mauricio; Trueba, Gabriel

    2013-06-01

    Staphylococcus aureus is a frequent cause of nosocomial pneumonia and bacteremia worldwide. Classical and molecular epidemiology approaches were used to study a S. aureus outbreak in the intensive care unit (ICU) of one of the largest public hospitals in Quito. Staphylococcus aureus isolates from 17 patients and 19 potential carriers from the staff were collected from March 2007 to February 2008 and analyzed by pulsed-field gel electrophoresis (PFGE) to determine their clonal relationships. During this period the hospital reported 16 cases of hospital-acquired staphylococcal pneumonia and an apparent outbreak occurred from June to September 2007. DNA from these isolates formed six different PFGE patterns: four clonal groups, and two groups of clonally related isolates. Molecular typing failed to identify any staphylococcal reservoir among staff members. The current study suggested that a staphylococcal outbreak that occurred in the summer of 2007 was caused by different bacterial clones, although some clones were shared by two patients. Historical analysis of the staphylococcal infections in the ICU showed a higher incidence during the summer months, which coincided with the programmed personnel shift. This observation suggests that outbreaks might be produced by the introduction of improperly trained personnel.

  4. Probabilistic Cloning and Quantum Computation

    Institute of Scientific and Technical Information of China (English)

    GAO Ting; YAN Feng-Li; WANG Zhi-Xi

    2004-01-01

    @@ We discuss the usefulness of quantum cloning and present examples of quantum computation tasks for which the cloning offers an advantage which cannot be matched by any approach that does not resort to quantum cloning.In these quantum computations, we need to distribute quantum information contained in the states about which we have some partial information. To perform quantum computations, we use a state-dependent probabilistic quantum cloning procedure to distribute quantum information in the middle of a quantum computation.

  5. A privileged intraphagocyte niche is responsible for disseminated infection of Staphylococcus aureus in a zebrafish model.

    Science.gov (United States)

    Prajsnar, Tomasz K; Hamilton, Ruth; Garcia-Lara, Jorge; McVicker, Gareth; Williams, Alexander; Boots, Michael; Foster, Simon J; Renshaw, Stephen A

    2012-10-01

    The innate immune system is the primary defence against the versatile pathogen, Staphylococcus aureus. How this organism is able to avoid immune killing and cause infections is poorly understood. Using an established larval zebrafish infection model, we have shown that overwhelming infection is due to subversion of phagocytes by staphylococci, allowing bacteria to evade killing and found foci of disease. Larval zebrafish coinfected with two S. aureus strains carrying different fluorescent reporter gene fusions (but otherwise isogenic) had bacterial lesions, at the time of host death, containing predominantly one strain. Quantitative data using two marked strains revealed that the strain ratios, during overwhelming infection, were often skewed towards the extremes, with one strain predominating. Infection with passaged bacterial clones revealed the phenomenon not to bedue to adventitious mutations acquired by the pathogen. After infection of the host, all bacteria are internalized by phagocytes and the skewing of population ratios is absolutely dependent on the presence of phagocytes. Mathematical modelling of pathogen population dynamics revealed the data patterns are consistent with the hypothesis that a small number of infected phagocytes serve as an intracellular reservoir for S. aureus, which upon release leads to disseminated infection. Strategies to specifically alter neutrophil/macrophage numbers were used to map the potential subpopulation of phagocytes acting as a pathogen reservoir, revealing neutrophils as the likely 'niche'. Subsequently in a murine sepsis model, S. aureus abscesses in kidneys were also found to be predominantly clonal, therefore likely founded by an individual cell, suggesting a potential mechanism analogous to the zebrafish model with few protected niches. These findings add credence to the argument that S. aureus control regimes should recognize both the intracellular as well as extracellular facets of the S. aureus life

  6. Recent emergence of Staphylococcus aureus clonal complex 398 in human blood cultures.

    Directory of Open Access Journals (Sweden)

    Erwin Verkade

    Full Text Available BACKGROUND: Recently, a clone of MRSA with clonal complex 398 (CC398 has emerged that is related to an extensive reservoir in animals, especially pigs and veal calves. It has been reported previously that methicillin-susceptible variants of CC398 circulate among humans at low frequency, and these have been isolated in a few cases of bloodstream infections (BSI. The purpose of this study was to determine the prevalence of S. aureus CC398 in blood cultures taken from patients in a geographic area with a high density of pigs. METHODOLOGY/PRINCIPAL FINDINGS: In total, 612 consecutive episodes of S. aureus BSI diagnosed before and during the emergence of CC398 were included. Three strains (2 MSSA and 1 MRSA that were isolated from bacteremic patients between 2010-2011 were positive in a CC398 specific PCR. There was a marked increase in prevalence of S. aureus CC398 BSI isolated between 2010-2011 compared to the combined collections that were isolated between 1996-1998 and 2002-2005 (3/157, 1.9% vs. 0/455, 0.0%; p = 0.017. CONCLUSIONS/SIGNIFICANCE: In conclusion, in an area with a relative high density of pigs, S. aureus CC398 was found as a cause of BSI in humans only recently. This indicates that S. aureus CC398 is able to cause invasive infections in humans and that the prevalence is rising. Careful monitoring of the evolution and epidemiology of S. aureus CC398 in animals and humans is therefore important.

  7. Nasal Carriage of Staphylococcus aureus among Children in the Ashanti Region of Ghana

    Science.gov (United States)

    Hogan, Benedikt; Azuure, Clinton; Krumkamp, Ralf; Dekker, Denise; Gajdiss, Mike; Brunke, Melanie; Sarpong, Nimako; Owusu-Dabo, Ellis; May, Jürgen

    2017-01-01

    Background Nasal carriage with Staphylococcus aureus is a common risk factor for invasive infections, indicating the necessity to monitor prevalent strains, particularly in the vulnerable paediatric population. This surveillance study aims to identify carriage rates, subtypes, antimicrobial susceptibilities and virulence markers of nasal S. aureus isolates collected from children living in the Ashanti region of Ghana. Methods Nasal swabs were obtained from children < 15 years of age on admission to the Agogo Presbyterian Hospital between April 2014 and January 2015. S. aureus isolates were characterized by their antimicrobial susceptibility, the presence of genes encoding for Panton-Valentine leukocidin (PVL) and toxic shock syndrome toxin-1 (TSST-1) and further differentiated by spa-typing and multi-locus-sequence-typing. Results Out of 544 children 120 (22.1%) were colonized with S. aureus, with highest carriage rates during the rainy seasons (27.2%; p = 0.007), in females aged 6–8 years (43.7%) and males aged 8–10 years (35.2%). The 123 isolates belonged to 35 different spa-types and 19 sequence types (ST) with the three most prevalent spa-types being t355 (n = 25), t84 (n = 18), t939 (n = 13), corresponding to ST152, ST15 and ST45. Two (2%) isolates were methicillin-resistant S. aureus (MRSA), classified as t1096 (ST152) and t4454 (ST45), and 16 (13%) were resistant to three or more different antimicrobial classes. PVL and TSST-1 were detected in 71 (58%) and 17 (14%) isolates respectively. Conclusion S. aureus carriage among Ghanaian children seems to depend on age, sex and seasonality. While MRSA rates are low, the high prevalence of PVL is of serious concern as these strains might serve not only as a source for severe invasive infections but may also transfer genes, leading to highly virulent MRSA clones. PMID:28107412

  8. Rise and subsequent decline of community-associated methicillin resistant Staphylococcus aureus ST30-IVc in Copenhagen, Denmark through an effective search and destroy policy

    DEFF Research Database (Denmark)

    Bartels, M D; Kristoffersen, K; Boye, K;

    2010-01-01

    The number of patients with community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has increased rapidly in Copenhagen, Denmark since 2003. Patients with the typical Panton-Valentine leukocidin-positive CA-MRSA clone ST30-IVc were contacted with the aim of treating MRSA carrie....... Treatment of MRSA carriers was difficult and required many resources, but the clone was eventually successfully eliminated. The import of ST30-IVc to Denmark will continue, but the spread of the clone in Denmark can be kept to a minimum by direct intervention in the affected families....

  9. Colonização microbiana precoce de pacientes identificados por triagem neonatal para fibrose cística, com ênfase em Staphylococcus aureus Early microbial colonization of cystic fibrosis patients identified by neonatal screening, with emphasis on Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Helena A. P. H. M. Souza

    2006-10-01

    Full Text Available OBJETIVOS: Avaliar prospectivamente a colonização bacteriana de pacientes com fibrose cística identificados por triagem neonatal. Avaliar a suscetibilidade a antimicrobianos e caracterizar molecularmente as cepas de Staphylococcus aureus isoladas da orofaringe dos pacientes no período do estudo. MÉTODOS: Foram estudados 25 pacientes com fibrose cística, identificados por tripsina imunorreativa e com diagnóstico confirmado por duas ou mais provas de suor, atendidos regularmente no ambulatório de fibrose cística do Hospital de Clínicas da Universidade Federal do Paraná. Foram coletadas amostras de orofaringe com swab e cultivadas por métodos rotineiros; as colônias bacterianas foram identificadas fenotipicamente e testadas quanto à suscetibilidade a antimicrobianos. Os isolados de S. aureus foram submetidos a tipagem molecular por eletroforese em campo pulsado. RESULTADOS: De um total de 234 amostras de orofaringe, S. aureus foi isolado em maior número (76% dos pacientes, 42% das amostras, seguido de Pseudomonas aeruginosa (36% dos pacientes, 16% das amostras e Haemophilus spp. (76% dos pacientes; 19% das amostras. Dos 19 pacientes colonizados com S. aureus, foram obtidos 73 isolados, 18 oxacilina-resistentes (24,6%, isolados de dois pacientes, com perfis eletroforéticos idênticos ao do clone brasileiro. Os demais isolados oxacilina-sensíveis distribuíram-se entre 18 perfis eletroforéticos distintos. CONCLUSÃO: Observou-se uma maior prevalência de S. aureus, com isolamento mais precoce em relação aos outros patógenos pesquisados. Os isolados multissensíveis distribuíram-se em clones distintos, caracterizando a não transmissibilidade entre as cepas comunitárias. Os S. aureus resistentes a oxacilina isolados apresentaram perfis eletroforéticos idênticos, provavelmente adquiridos no ambiente hospitalar. P. aeruginosa foi pouco freqüente na população estudada.OBJECTIVES: To assess bacterial colonization prospectively

  10. Chromones from an ascomycete,Chaetomium aureus

    Institute of Scientific and Technical Information of China (English)

    Li Mei Li; Qiang Zou; Guo You Li

    2010-01-01

    A novel chromone,named chaetoaurin (1),along with six known chromone derivatives (2-7),was isolated from the ethyl acetate extract of a solid-state fermented culture of Chaetomium aureus.Their structures were elucidated by extensive spectral analysis.All of these compounds were reported from C.aureus for the first time.

  11. Immunogenicity of toxins during Staphylococcus aureus infection

    NARCIS (Netherlands)

    N.J. Verkaik (Nelianne); O. Dauwalder (Olivier); K. Antri (Kenza); I. Boubekri (Ilhem); C.P. de Vogel (Corné); C. Badiou (Cédric); M. Bes (Michèle); F. Vandenesch (François); M. Tazir (Mohammed); H. Hooijkaas (Herbert); H.A. Verbrugh (Henri); A.F. van Belkum (Alex); J. Etienne (Jerome); G. Lina (Gérard); N. Ramdani-Bouguessa (Nadjia); W.J.B. van Wamel (Willem)

    2010-01-01

    textabstractAB - BACKGROUND: Toxins are important Staphylococcus aureus virulence factors, but little is known about their immunogenicity during infection. Here, additional insight is generated. METHODS: Serum samples from 206 S. aureus-infected patients and 201 hospital-admitted control subjects we

  12. Staphylococcus aureus and hand eczema severity

    DEFF Research Database (Denmark)

    Haslund, P; Bangsgaard, N; Jarløv, J O

    2009-01-01

    BACKGROUND: The role of bacterial infections in hand eczema (HE) remains to be assessed. OBJECTIVES: To determine the prevalence of Staphylococcus aureus in patients with HE compared with controls, and to relate presence of S. aureus, subtypes and toxin production to severity of HE. METHODS......: Bacterial swabs were taken at three different visits from the hand and nose in 50 patients with HE and 50 controls. Staphylococcus aureus was subtyped by spa typing and assigned to clonal complexes (CCs), and isolates were tested for exotoxin-producing S. aureus strains. The Hand Eczema Severity Index...... was used for severity assessment. RESULTS: Staphylococcus aureus was found on the hands in 24 patients with HE and four controls (P hands...

  13. Subpopulations of Staphylococcus aureus clonal complex 121 are associated with distinct clinical entities.

    Directory of Open Access Journals (Sweden)

    Kevin Kurt

    Full Text Available We investigated the population structure of Staphylococcus aureus clonal complex CC121 by mutation discovery at 115 genetic housekeeping loci from each of 154 isolates, sampled on five continents between 1953 and 2009. In addition, we pyro-sequenced the genomes from ten representative isolates. The genome-wide SNPs that were ascertained revealed the evolutionary history of CC121, indicating at least six major clades (A to F within the clonal complex and dating its most recent common ancestor to the pre-antibiotic era. The toxin gene complement of CC121 isolates was correlated with their SNP-based phylogeny. Moreover, we found a highly significant association of clinical phenotypes with phylogenetic affiliations, which is unusual for S. aureus. All isolates evidently sampled from superficial infections (including staphylococcal scalded skin syndrome, bullous impetigo, exfoliative dermatitis, conjunctivitis clustered in clade F, which included the European epidemic fusidic-acid resistant impetigo clone (EEFIC. In comparison, isolates from deep-seated infections (abscess, furuncle, pyomyositis, necrotizing pneumonia were disseminated in several clades, but not in clade F. Our results demonstrate that phylogenetic lineages with distinct clinical properties exist within an S. aureus clonal complex, and that SNPs serve as powerful discriminatory markers, able to identify these lineages. All CC121 genomes harboured a 41-kilobase prophage that was dissimilar to S. aureus phages sequenced previously. Community-associated MRSA and MSSA from Cambodia were extremely closely related, suggesting this MRSA arose in the region.

  14. Parallel Epidemics of Community-Associated Methicillin-Resistant Staphylococcus aureus USA300 Infection in North and South America

    OpenAIRE

    Paul J Planet; Diaz, Lorena; Kolokotronis, Sergios-Orestis; Narechania, Apurva; Reyes, Jinnethe; Xing, Galen; Rincon, Sandra; Smith, Hannah; Panesso, Diana; Ryan, Chanelle; Smith, Dylan P.; Guzman, Manuel De; Zurita, Jeannete; Sebra, Robert; Deikus, Gintaras

    2015-01-01

    Background. The community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) epidemic in the United States is attributed to the spread of the USA300 clone. An epidemic of CA-MRSA closely related to USA300 has occurred in northern South America (USA300 Latin-American variant, USA300-LV). Using phylogenomic analysis, we aimed to understand the relationships between these 2 epidemics.

  15. Entering the Clone Age

    Institute of Scientific and Technical Information of China (English)

    1995-01-01

    Suppose you make your parents so happy,they decide to have another baby just like you.It might be flattering,but how would you feel about having a little brother or sister who is also your twin? A laboratory experiment conducted last fall suggests it may someday be possible.For the first time ever,scientists made exact copies, or clones, of a human embryo.

  16. Ethical issues in livestock cloning.

    Science.gov (United States)

    Thompson, P B

    1999-01-01

    Although cloning may eventually become an important technology for livestock production, four ethical issues must be addressed before the practice becomes widespread. First, researchers must establish that the procedure is not detrimental to the health or well-being of affected animals. Second, animal research institutions should evaluate the net social benefits to livestock producers by weighing the benefits to producers against the opportunity cost of research capacity lost to biomedical projects. Third, scientists should consider the indirect effects of cloning research on the larger ethical issues surrounding human cloning. Finally, the market structure for products of cloned animals should protect individual choice, and should recognize that many individuals find the prospect of cloning (or consuming cloned animals) repugnant. Analysis of these four issues is complicated by spurious arguments alleging that cloning will have a negative impact on environment and genetic diversity.

  17. Cloning-free CRISPR

    Directory of Open Access Journals (Sweden)

    Mandana Arbab

    2015-11-01

    Full Text Available We present self-cloning CRISPR/Cas9 (scCRISPR, a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA or knockin homology construct for each target locus. We introduce a self-cleaving palindromic sgRNA plasmid and a short double-stranded DNA sequence encoding the desired locus-specific sgRNA into target cells, allowing them to produce a locus-specific sgRNA plasmid through homologous recombination. scCRISPR enables efficient generation of gene knockouts (∼88% mutation rate at approximately one-sixth the cost of plasmid-based sgRNA construction with only 2 hr of preparation for each targeted site. Additionally, we demonstrate efficient site-specific knockin of GFP transgenes without any plasmid cloning or genome-integrated selection cassette in mouse and human embryonic stem cells (2%–4% knockin rate through PCR-based addition of short homology arms. scCRISPR substantially lowers the bar on mouse and human transgenesis.

  18. Sequential cloning of chromosomes

    Science.gov (United States)

    Lacks, S.A.

    1995-07-18

    A method for sequential cloning of chromosomal DNA of a target organism is disclosed. A first DNA segment homologous to the chromosomal DNA to be sequentially cloned is isolated. The first segment has a first restriction enzyme site on either side. A first vector product is formed by ligating the homologous segment into a suitably designed vector. The first vector product is circularly integrated into the target organism`s chromosomal DNA. The resulting integrated chromosomal DNA segment includes the homologous DNA segment at either end of the integrated vector segment. The integrated chromosomal DNA is cleaved with a second restriction enzyme and ligated to form a vector-containing plasmid, which is replicated in a host organism. The replicated plasmid is then cleaved with the first restriction enzyme. Next, a DNA segment containing the vector and a segment of DNA homologous to a distal portion of the previously isolated DNA segment is isolated. This segment is then ligated to form a plasmid which is replicated within a suitable host. This plasmid is then circularly integrated into the target chromosomal DNA. The chromosomal DNA containing the circularly integrated vector is treated with a third, retrorestriction (class IIS) enzyme. The cleaved DNA is ligated to give a plasmid that is used to transform a host permissive for replication of its vector. The sequential cloning process continues by repeated cycles of circular integration and excision. The excision is carried out alternately with the second and third enzymes. 9 figs.

  19. Secure the Clones

    CERN Document Server

    Jensen, Thomas; Pichardie, David

    2012-01-01

    Exchanging mutable data objects with untrusted code is a delicate matter because of the risk of creating a data space that is accessible by an attacker. Consequently, secure programming guidelines for Java stress the importance of using defensive copying before accepting or handing out references to an internal mutable object. However, implementation of a copy method (like clone()) is entirely left to the programmer. It may not provide a sufficiently deep copy of an object and is subject to overriding by a malicious sub-class. Currently no language-based mechanism supports secure object cloning. This paper proposes a type-based annotation system for defining modular copy policies for class-based object-oriented programs. A copy policy specifies the maximally allowed sharing between an object and its clone. We present a static enforcement mechanism that will guarantee that all classes fulfil their copy policy, even in the presence of overriding of copy methods, and establish the semantic correctness of the ove...

  20. Molecular characterization of Staphylococcus aureus isolates causing skin and soft tissue infections in patients from Malakand, Pakistan.

    Science.gov (United States)

    Madzgalla, S; Syed, M A; Khan, M A; Rehman, S S; Müller, E; Reissig, A; Ehricht, R; Monecke, S

    2016-09-01

    Comparatively few studies have been published describing Staphylococcus aureus/MRSA epidemiology in Central Asia including Pakistan. Here, we report the genotyping of Staphylococcus aureus strains (that include both methicillin-susceptible and methicillin-resistant Staphylococcus aureus) from community- and hospital-acquired skin and soft-tissue infections in a tertiary care hospital in the Malakand district of the Khyber Pakhtunkhwa Province of Pakistan. Forty-five isolates of Staphylococcus aureus were characterized by microarray hybridization. Twenty isolates (44 %) were MRSA, whereas 22 (49 %) were PVL-positive. Fourteen isolates (31 %) harboured both mecA and PVL genes. The dominant clones were CC121-MSSA (n = 15, 33 %) and the PVL-positive "Bengal Bay Clone" (ST772-MRSA-V; n = 13, 29 %). The PVL-positive CC8-MRSA-IV strain "USA300" was found once. The pandemic ST239-MRSA-III strain was absent, although it has previously been observed in Pakistan. These observations require a re-assessment of schemes for initial antibiotic therapy to cover MRSA and they emphasise the need for a rapid and non-molecular test for PVL.

  1. Role of the ESAT-6 secretion system in virulence of the emerging community-associated Staphylococcus aureus lineage ST398.

    Science.gov (United States)

    Wang, Yanan; Hu, Mo; Liu, Qian; Qin, Juanxiu; Dai, Yingxin; He, Lei; Li, Tianming; Zheng, Bing; Zhou, Fan; Yu, Kaiwen; Fang, Jingyuan; Liu, Xiaoyun; Otto, Michael; Li, Min

    2016-04-26

    Novel Staphylococcus aureus clones continue to emerge that cause infections in otherwise healthy people. One example is the sequence type (ST) 398 lineage, which we show here is increasing in importance as a significant cause of community-associated (CA) human infections in China. We have a profound lack of understanding about what determines the considerable virulence potential of such newly emerging clones. Information about the contribution to virulence of the more recently discovered ESAT-6 secretion system (ESS) has remained particularly scarce. The Chinese ST398 isolates exhibited significantly increased expression of ESS genes as compared to predominant hospital-associated clones, which we found is likely due to increased expression of the accessory gene regulator (Agr) system and control of ESS by Agr. Importantly, deletion of essB in ST398 resulted in significantly reduced resistance to neutrophil killing and decreased virulence in murine skin and blood infection models. Our results demonstrate a key function of ESS in promoting virulence and mechanisms of resistance to innate host defense in an important emerging CA-S. aureus lineage. They suggest that ESS has a so far underestimated role in promoting aggressive virulence and epidemiological success of S. aureus.

  2. Exfoliative Toxins of Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Michal Bukowski

    2010-05-01

    Full Text Available Staphylococcus aureus is an important pathogen of humans and livestock. It causes a diverse array of diseases, ranging from relatively harmless localized skin infections to life-threatening systemic conditions. Among multiple virulence factors, staphylococci secrete several exotoxins directly associated with particular disease symptoms. These include toxic shock syndrome toxin 1 (TSST-1, enterotoxins, and exfoliative toxins (ETs. The latter are particularly interesting as the sole agents responsible for staphylococcal scalded skin syndrome (SSSS, a disease predominantly affecting infants and characterized by the loss of superficial skin layers, dehydration, and secondary infections. The molecular basis of the clinical symptoms of SSSS is well understood. ETs are serine proteases with high substrate specificity, which selectively recognize and hydrolyze desmosomal proteins in the skin. The fascinating road leading to the discovery of ETs as the agents responsible for SSSS and the characterization of the molecular mechanism of their action, including recent advances in the field, are reviewed in this article.

  3. Clonal expansion during Staphylococcus aureus infection dynamics reveals the effect of antibiotic intervention.

    Directory of Open Access Journals (Sweden)

    Gareth McVicker

    2014-02-01

    Full Text Available To slow the inexorable rise of antibiotic resistance we must understand how drugs impact on pathogenesis and influence the selection of resistant clones. Staphylococcus aureus is an important human pathogen with populations of antibiotic-resistant bacteria in hospitals and the community. Host phagocytes play a crucial role in controlling S. aureus infection, which can lead to a population "bottleneck" whereby clonal expansion of a small fraction of the initial inoculum founds a systemic infection. Such population dynamics may have important consequences on the effect of antibiotic intervention. Low doses of antibiotics have been shown to affect in vitro growth and the generation of resistant mutants over the long term, however whether this has any in vivo relevance is unknown. In this work, the population dynamics of S. aureus pathogenesis were studied in vivo using antibiotic-resistant strains constructed in an isogenic background, coupled with systemic models of infection in both the mouse and zebrafish embryo. Murine experiments revealed unexpected and complex bacterial population kinetics arising from clonal expansion during infection in particular organs. We subsequently elucidated the effect of antibiotic intervention within the host using mixed inocula of resistant and sensitive bacteria. Sub-curative tetracycline doses support the preferential expansion of resistant microorganisms, importantly unrelated to effects on growth rate or de novo resistance acquisition. This novel phenomenon is generic, occurring with methicillin-resistant S. aureus (MRSA in the presence of β-lactams and with the unrelated human pathogen Pseudomonas aeruginosa. The selection of resistant clones at low antibiotic levels can result in a rapid increase in their prevalence under conditions that would previously not be thought to favor them. Our results have key implications for the design of effective treatment regimes to limit the spread of antimicrobial

  4. Molecular characterization of methicillin resistant Staphylococcus aureus isolated from hospitals environments and patients in Northern Palestine

    Directory of Open Access Journals (Sweden)

    Ghaleb Adwan

    2015-09-01

    Full Text Available BACKGROUND: Staphylococcus aureus (S. aureus is considered one of the most common pathogen to humans. Infections caused by this mocroorganism can be acquired through both hospital and community settings. This study was carried out to investigate molecular characterization of MRSA strains isolated from the patients and their environment in two hospitals (Rafidia hospital and Thabet hospital inNorthern Palestine, and to determine the clonal identity between these strains and their possible contribution to nosocomial infections.METHODS: Two hundred sixty five swabbed samples were collected from these hospitals, S. aureus was isolated,  antibiotic resistant genes were Panton–Valentin leukocidin (PVL gene were detected and SCCmec and spA were typed by PCR and/or sequencing.RESULTS: The prevalence of MRSA among S. aureus isolates was 29% and 8.2% in Rafidia hospital and Thabet hospital, respectively. All strains resistant to oxacilllin disk were carried mecA gene. Majority of strains (84.6% carried SCCmec type II (n = 11, type IVa and non-typeable were also detected. In addition, PVL was detected in 2 (14.3% clinical strains. ERIC PCR patterns revealed that 2 strains recovered from patient bed and nasal swab isolated from Thabet Hospital were nontypeable, spA typing showed that they belonged to type t386 and have identical DNA sequences. Other 2 clinical isolates were spa typed, one belonged to clone t044, while the other is new clone not exist in database.CONCLUSIONS: Results may give evidence that environmental contamination possibly contributing to nosocomial infections.

  5. 21 CFR 866.3700 - Staphylococcus aureus serological reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Staphylococcus aureus serological reagents. 866... Staphylococcus aureus serological reagents. (a) Identification. Staphylococcus aureus serological reagents are... epidemiological information on these diseases. Certain strains of Staphylococcus aureus produce an...

  6. Ethical issues in animal cloning.

    Science.gov (United States)

    Fiester, Autumn

    2005-01-01

    The issue of human reproductive cloning has recently received a great deal attention in public discourse. Bioethicists, policy makers, and the media have been quick to identify the key ethical issues involved in human reproductive cloning and to argue, almost unanimously, for an international ban on such attempts. Meanwhile, scientists have proceeded with extensive research agendas in the cloning of animals. Despite this research, there has been little public discussion of the ethical issues raised by animal cloning projects. Polling data show that the public is decidedly against the cloning of animals. To understand the public's reaction and fill the void of reasoned debate about the issue, we need to review the possible objections to animal cloning and assess the merits of the anti-animal cloning stance. Some objections to animal cloning (e.g., the impact of cloning on the population of unwanted animals) can be easily addressed, while others (e.g., the health of cloned animals) require more serious attention by the public and policy makers.

  7. To clone or not to clone--whither the law?

    Science.gov (United States)

    Lupton, M L

    1999-01-01

    The cloning of Dolly the lamb from adult cells by scientists at the Roslin Laboratories near Edinburgh in February 1997 has startled the world because it now opens the way to clone adult human beings. The reaction to Ian Wilmut's breakthrough has been instant and largely negative. Bills were rushed into both the US Senate and House of Representatives aimed at banning the cloning of human beings. Human cloning is premature at this stage, but there are many positive spin-offs of cloning in the field of genetic engineering, such as the production of human proteins such as blood clotting factors which aid in healing wounds. Progress by means of cloning can also be made into devising a cure for Parkinson's Disease amongst others. No lesser ethicist than John C. Fletcher of the University of Virginia foresees circumstances in which human cloning is acceptable e.g. to enable a couple to replace a dying child, to enable a couple, one of whom is infertile, to clone a child from either partner. Extensive regulation of cloning by the law is inevitable but, in doing so, the legislation should be careful not to outlaw research in this area which could be beneficial to mankind.

  8. Prevalence of Staphylococcus Aureus Colonizing the Health Care Personnel of a Hospital in the City of Cali

    Directory of Open Access Journals (Sweden)

    Leidy Catherine Arteaga Delgado

    2016-01-01

    Full Text Available Objecitve: The aim of this study was to determine the prevalence of isolates of Staphylococcus aureus in workers at a hospital in Cali. Materials and methods: A descriptive study was conducted with samples of nasal swabs, skin smears to 30 health care workers. Phenotypic characterization of isolates was based on susceptibility antibiogram testing and pcr amplification of the identified mecA and agr genes. The origin of isolates was established by analysis of agr gene, identifying the agr groups. Results: Eleven (26.7 % workers were colonized with S. aureus. The frequency of S. aureus was higher in health care personnel who were in the operating room (20 %; OR = 2.077; P> 0.05. Four antibiotypes were identified, this feature is compatible with community clones that have proven to be highly diverse with a large capacity to spread in the community. 36.4 % of the isolates were resistant to cefotaxime and/or oxacillin, suggestive mrsa phenotype in these isolated the mecA gene was identified. Agr i was found primarily among isolates metillicin-sensitive S. aureus (mssa, compatible with community origin, and mrsa isolates belong to agr ii, with hospital waste. Conclusion: The prevalence of S. aureus resistant to antibiotics that colonize health care personnel was demonstrated, mainly in those working in the operating room. Regu­lar monitoring of personnel should be regularly conducted to prevent the spread of pathogens.

  9. Lessons learned from cloning dogs.

    Science.gov (United States)

    Kim, M J; Oh, H J; Kim, G A; Park, J E; Park, E J; Jang, G; Ra, J C; Kang, S K; Lee, B C

    2012-08-01

    The aim of this article is to review dog cloning research and to suggest its applications based on a discussion about the normality of cloned dogs. Somatic cell nuclear transfer was successfully used for production of viable cloned puppies despite limited understanding of in vitro dog embryo production. Cloned dogs have similar growth characteristics to those born from natural fertilization, with no evidence of serious adverse effects. The offspring of cloned dogs also have similar growth performance and health to those of naturally bred puppies. Therefore, cloning in domestic dogs can be applied as an assisted reproductive technique to conserve endangered species, to treat sterile canids or aged dogs, to improve reproductive performance of valuable individuals and to generate disease model animals.

  10. Mastite com lesões sistêmicas por Staphylococus aureus subesp. aureus em coelhos

    OpenAIRE

    Traverso Sandra Davi; Cunha Leonardo da; Fernandes Joaquim César Teixeira; Loretti Alexandre Paulino; Rhoden Adriana; Wunder Jr. Elsio; Driemeier David

    2003-01-01

    Em uma criação composta por 1800 coelhos, 33% das matrizes apresentaram mastite e lesões cutâneas crostosas e purulentas. Estes animais apresentavam-se entre 10 a- 12 meses de idade e em segunda parição. Quinze coelhos afetados foram sacrificados e necropsiados. Na necropsia, além das lesões cutâneas haviam microabscessos em diversos órgãos. Das amostras coletadas isolou-se Staphylococcus aureus subesp. aureus. S. aureus subesp. aureus também foi isolado de "swab" nasal coletado do tratador e...

  11. Molecular Cloning of Adenosinediphosphoribosyl Transferase.

    Science.gov (United States)

    1987-09-08

    ACCESSION NO.D,. 03261102F 2312 A~5 11. TITLE (include Securqt Classification) 0 Molecular Cloning of Adenosinediphosphoribosyl Transferase 12. PERSONAL...I’:- AFOSR.Tlt. 8 7 - 0 9 8,2 0IL * pi AFOSR- 85 -0377 PROGRESS REPORT Molecular Cloning of Adenosinediphosphoribosyl Transferase 5." Period of...Pharmacology and the Cardiovascular Research Institute September 8, 1987 .’, 5.’- "’S ". -f, AFOSR - 85 -0377 PROGRESS REPORT Molecular Cloning of

  12. Characterization of inosine monophosphate dehydrogenase from Staphylococcus aureus ATCC12600 and its involvement in biofilm formation

    Directory of Open Access Journals (Sweden)

    S. Yeswanth

    2013-10-01

    Full Text Available Background: In Staphylococcus aureus purine metabolism plays a crucial role in the formation of biofilm which is a key pathogenic factor. The present study is aimed in the characterization of inosine monophosphate dehydrogenase (IMPDH from Staphylococcus aureus ATCC 12600. Methods: IMPDH gene was amplified using primers designed from IMPDH gene sequence of S. aureus reported in the database. Then polymerase chain reaction (PCR product was cloned in the Sma I site of M13mp18 and expressed in Escherichia coli JM109. The recombinant IMPDH (rIMPDH was overexpressed with 1 mM isopropyl beta-D-1- thiogalactopyranoside (IPTG; Michaelis constant (Km, maximum enzyme velocity (Vmax and catalytic constant (Kcat of expressed IMPDH were determined. Results: The enzyme kinetics of IMPDH grown under aerobic conditions showed a Km of 43.71±1.56 µM, Vmax of 0.247±0.84/µM/mg/min and Kcat of 2.74±0.015/min while in anaerobic conditions the kinetics showed Km of 42.81±3.154/ µM, Vmax of 0.378±0.036 µM/mg/min and Kcat of 4.78±0.021 /min, indicating elevated levels of IMPDH activity under anaerobic conditions. Three-folds increased activity in the presence of 1 mM adenosine triphosphate (ATP correlated with biofilm formation. The kinetics of pure rIMPDH were close to the native IMPDH of S. aureus ATCC12600 and the enzyme showed single band in sodium dodecyl sulphate polyacrylamide gel electrophoresis with a molecular weight of 53 KDa. Conclusions: Elevated activity of IMPDH was observed in S. aureus grown under anaerobic conditions and this was correlated with the biofilm formation indicating the linkage between purine metabolism and pathogenesis.

  13. Antimicrobial Resistance and Molecular Characteristics of Nasal Staphylococcus aureus Isolates From Newly Admitted Inpatients.

    Science.gov (United States)

    Chen, Xu; Sun, Kangde; Dong, Danfeng; Luo, Qingqiong; Peng, Yibing; Chen, Fuxiang

    2016-05-01

    Staphylococcus aureus, or methicillin-resistant S. aureus (MRSA), is a significant pathogen in both nosocomial and community infections. Community-associated MRSA (CA-MRSA) strains tend to be multi-drug resistant and to invade hospital settings. This study aimed to assess the antimicrobial resistance and molecular characteristicsof nasal S. aureus among newlyadmitted inpatients.In the present study, 66 S. aureus isolates, including 10 healthcare-associated MRSA (HA-MRSA), 8 CA-MRSA, and 48 methicillin-sensitive S. aureus (MSSA) strains, were found in the nasal cavities of 62 patients by screening 292 newlyadmitted patients. Antimicrobial resistance and molecular characteristics of these isolates, including spa-type, sequence type (ST) and SCCmec type, were investigated. All isolates were sensitive to linezolid, teicoplanin, and quinupristin/dalfopristin, but high levels of resistance to penicillin and erythromycin were detected. According to D-test and erm gene detection results, the cMLS(B) and iMLS(B) phenotypes were detected in 24 and 16 isolates, respectively. All 10 HA-MRSA strains displayed the cMLS(B) phenotypemediated by ermA or ermA/ermC, while the cMLS(B) CA-MRSA and MSSA strains carried the ermB gene. Molecular characterization revealedall 10 HA-MRSA strains were derived from the ST239-SCCmec III clone, and four out of eight CA-MRSA strains were t437-ST59-SCCmec V. The results suggest that patients play an indispensable role in transmitting epidemic CA-MRSA and HA-MRSA strains.

  14. Etudes structurales du ribosome de Staphylococcus aureus

    OpenAIRE

    Khusainov, Iskander

    2015-01-01

    The ribosome is a large cellular machinery that performs the protein synthesis in every living cell. Therefore, the ribosome is one of the major targets of naturally produced antibiotics, which can kill bacterial cells by blocking protein synthesis. However, some bacteria are resistant to these antibiotics due to small modifications of their ribosomes. Among them, Staphylococcus aureus (S. aureus) is a severe pathogen that causes numerous infections in humans. The crystal structures of comple...

  15. Carotenoid Formation by Staphylococcus aureus

    Science.gov (United States)

    Hammond, Ray K.; White, David C.

    1970-01-01

    The carotenoid pigments of Staphylococcus aureus U-71 were identified as phytoene; ζ-carotene; δ-carotene; phytofluenol; a phytofluenol-like carotenoid, rubixanthin; and three rubixanthin-like carotenoids after extraction, saponification, chromatographic separation, and determination of their absorption spectra. There was no evidence of carotenoid esters or glycoside ethers in the extract before saponification. During the aerobic growth cycle the total carotenoids increased from 45 to 1,000 nmoles per g (dry weight), with the greatest increases in the polar, hydroxylated carotenoids. During the anaerobic growth cycle, the total carotenoids increased from 20 nmoles per g (dry weight) to 80 nmoles per g (dry weight), and only traces of the polar carotenoids were formed. Light had no effect on carotenoid synthesis. About 0.14% of the mevalonate-2-14C added to the culture was incorporated into the carotenoids during each bacterial doubling. The total carotenoids did not lose radioactivity when grown in the absence of 14C for 2.5 bacterial doublings. The total carotenoids did not lose radioactivity when grown in the absence of 14C for 2.5 bacterial doublings. The incorporation and turnover of 14C indicated the carotenes were sequentially desaturated and hydroxylated to form the polar carotenoids. PMID:5423369

  16. CATO: The Clone Alignment Tool.

    Directory of Open Access Journals (Sweden)

    Peter V Henstock

    Full Text Available High-throughput cloning efforts produce large numbers of sequences that need to be aligned, edited, compared with reference sequences, and organized as files and selected clones. Different pieces of software are typically required to perform each of these tasks. We have designed a single piece of software, CATO, the Clone Alignment Tool, that allows a user to align, evaluate, edit, and select clone sequences based on comparisons to reference sequences. The input and output are designed to be compatible with standard data formats, and thus suitable for integration into a clone processing pipeline. CATO provides both sequence alignment and visualizations to facilitate the analysis of cloning experiments. The alignment algorithm matches each of the relevant candidate sequences against each reference sequence. The visualization portion displays three levels of matching: 1 a top-level summary of the top candidate sequences aligned to each reference sequence, 2 a focused alignment view with the nucleotides of matched sequences displayed against one reference sequence, and 3 a pair-wise alignment of a single reference and candidate sequence pair. Users can select the minimum matching criteria for valid clones, edit or swap reference sequences, and export the results to a summary file as part of the high-throughput cloning workflow.

  17. CATO: The Clone Alignment Tool.

    Science.gov (United States)

    Henstock, Peter V; LaPan, Peter

    2016-01-01

    High-throughput cloning efforts produce large numbers of sequences that need to be aligned, edited, compared with reference sequences, and organized as files and selected clones. Different pieces of software are typically required to perform each of these tasks. We have designed a single piece of software, CATO, the Clone Alignment Tool, that allows a user to align, evaluate, edit, and select clone sequences based on comparisons to reference sequences. The input and output are designed to be compatible with standard data formats, and thus suitable for integration into a clone processing pipeline. CATO provides both sequence alignment and visualizations to facilitate the analysis of cloning experiments. The alignment algorithm matches each of the relevant candidate sequences against each reference sequence. The visualization portion displays three levels of matching: 1) a top-level summary of the top candidate sequences aligned to each reference sequence, 2) a focused alignment view with the nucleotides of matched sequences displayed against one reference sequence, and 3) a pair-wise alignment of a single reference and candidate sequence pair. Users can select the minimum matching criteria for valid clones, edit or swap reference sequences, and export the results to a summary file as part of the high-throughput cloning workflow.

  18. [The discrete horror of cloning].

    Science.gov (United States)

    Guibourg, Ricardo A

    2009-01-01

    The author raises the topic of cloning after the decision of the Argentine government, which concerned for the "dignity of the human person", passed a decree of need and urgency, No. 200/97 (Annex), prohibiting cloning experiments with human beings. Therefore, considering that the topic is so terribly urgent and necessary, the author feels it is timely to consider it.

  19. Quantum probabilistically cloning and computation

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    In this article we make a review on the usefulness of probabilistically cloning and present examples of quantum computation tasks for which quantum cloning offers an advantage which cannot be matched by any approach that does not resort to it.In these quantum computations,one needs to distribute quantum information contained in states about which we have some partial information.To perform quantum computations,one uses state-dependent probabilistic quantum cloning procedure to distribute quantum information in the middle of a quantum computation.And we discuss the achievable efficiencies and the efficient quantum logic network for probabilistic cloning the quantum states used in implementing quantum computation tasks for which cloning provides enhancement in performance.

  20. Clone Networks, Clone Extensions and Biregularizations of Varieties of Algebras

    Institute of Scientific and Technical Information of China (English)

    J. Plonka

    2001-01-01

    We consider algebras of type τ- without nullary operations. An identity ψ≈ψ of type τ is clone compatible if ψ and ψ are the same variable or the sets of fundamental operation symbols in ψ and ψ are non-empty and identical. For a variety V, we denote by Vc the variety defined by all clone compatible identities from Id(V). In this paper, we give a construction of algebras called a clone network. Under some assumptions, we describe algebras from Vc by means of this construction. We find some properties of Vc and applications.

  1. Clonal diversity of Staphylococcus aureus originating from the small ruminants goats and sheep.

    Science.gov (United States)

    Porrero, M Concepción; Hasman, Henrik; Vela, Ana I; Fernández-Garayzábal, Jose F; Domínguez, Lucas; Aarestrup, Frank M

    2012-04-23

    Staphylococcus aureus is an important pathogen in humans and many animal species. The prevalence of different clonal types in animal species remains largely unknown. We analyzed 267 S. aureus from intramammary infections in goats (47) and sheep (220) by spa typing, multi-locus sequence typing (MLST) and antimicrobial susceptibility. The most frequent spa types in goats were t337 (N=9), t759 (N=6) and t1534 (N=5). Sheep isolates mainly belonged to spa types t1534 (N=72), t2678 (N=29) and t3576 (N=20). Eighteen novel spa-types were observed; two from goat strains, 13 from sheep and three in both species. The majority of the goat strains grouped in MLST CC133 (N=10) and ST522 (N=10), followed by CC9 (N=9), while the majority of the sheep strains were of ST522 (N=108) followed by CC133 (N=86) and CC130 (N=11). Nine new MLST types were detected; three in goat and sheep isolates (ST1739, ST1758 and ST1780), two identified in goats only (ST1740 and ST2061) and four in sheep only (ST1742, ST1743, ST1781 and ST2011). Strains showed resistance below 20% against penicillin and tetracycline; a strong association between CC-types and penicillin resistance was observed. No resistance was detected to cefoxitin, quinupristin-dalfopristin, rifampicin and vancomycin. This study suggests that ST522 is the most common S. aureus clone associated with small ruminants followed by CC133.

  2. In vitro characterization of representative clinical South African Staphylococcus aureus isolates from various clonal lineages

    Directory of Open Access Journals (Sweden)

    W.F. Oosthuysen

    2014-07-01

    Full Text Available Data concerning the virulence and pathogenesis of South African strains of Staphylococcus aureus are limited. We investigated host–pathogen interactions of randomly selected clinical S. aureus isolates representing various clones. We characterized the ability of isolates to adhere to fibronectin, fibrinogen, collagens IV and VI, to invade host cells and to induce cell death in vitro. We analysed the possible association of these results with characteristics such as methicillin resistance, Panton–Valentine leucocidin (PVL positivity and clonality. The S. aureus isolates displayed diversity in their abilities to adhere to various human ligands. All isolates were highly invasive except for ST121. PVL-negative isolates were significantly more invasive than the PVL-positive isolates (p 0.004. Isolates of CC5, CC30 and CC121 were non-cytotoxic, whereas isolates of CC22, CC8, CC15, CC45 and CC88 were very cytotoxic. No statistical association was identified between cell death and methicillin resistance, bacterial PVL status, clonality or patient HIV status. The vast majority of isolates were invasive and induced significant cell death. PVL-negative isolates were more invasive than PVL-positive isolates, while methicillin-resistant isolates were not found to be more invasive or cytotoxic than methicillin-susceptible isolates.

  3. A common variant of staphylococcal cassette chromosome mec type IVa in isolates from Copenhagen, Denmark, is not detected by the BD GeneOhm methicillin-resistant Staphylococcus aureus assay

    DEFF Research Database (Denmark)

    Bartels, Mette Damkjaer; Boye, Kit; Rohde, Susanne Mie

    2009-01-01

    -susceptible Staphylococcus aureus isolates were included as negative controls. Forty-four MRSA isolates were undetectable; of these, 95% harbored SCCmec type IVa, and these included the most-common clone in Copenhagen, spa t024-sequence type 8-IVa. The false-negative MRSA isolates were tested with new primers (analyte...

  4. Rapid bench identification of methicillin-sensitive and methicillin-resistant Staphylococcus aureus: A multicenter comparative evaluation of Alere PBP2a Culture Colony Test (Alere) Versus Slidex MRSA detection (bioMérieux).

    Science.gov (United States)

    Tasse, Jason; Dupieux, Céline; Caillon, Jocelyne; Lanotte, Philippe; Lamy, Brigitte; Aissa, Nejla; Bemer, Pascale; Mereghetti, Laurent; Michon, Anne-Laure; Lozniewski, Alain; Bes, Michèle; Trouillet-Assant, Sophie; Laurent, Frédéric

    2016-08-01

    Using 30 clinical isolates of Staphylococcus aureus representative of the most prevalent clones circulating in France, the performance of the Alere™ PBP2a Culture Colony Test (CCT) and the Slidex(®) MRSA detection kit (SMD) were compared in 5 different labs. CCT demonstrated better performance and was easier to conduct in routine.

  5. Limitations on Cloning in Classical Mechanics

    OpenAIRE

    Fenyes, Aaron

    2010-01-01

    In this paper, we show that a result precisely analogous to the traditional quantum no-cloning theorem holds in classical mechanics. This classical no-cloning theorem does not prohibit classical cloning, we argue, because it is based on a too-restrictive definition of cloning. Using a less popular, more inclusive definition of cloning, we give examples of classical cloning processes. We also prove that a cloning machine must be at least as complicated as the object it is supposed to clone.

  6. A single copy integration vector that integrates at an engineered site on the Staphylococcus aureus chromosome

    Directory of Open Access Journals (Sweden)

    Lei Mei G

    2012-01-01

    Full Text Available Abstract Background Single-copy integration vectors based upon the site-specific recombination systems of bacteriophage are invaluable tools in the study of bacterial pathogenesis. The utility of such vectors is often limited, however, by the fact that integration often results in the inactivation of bacterial genes or has undesirable effects on gene transcription. The aim of this study is to develop an integration vector that does not have a detectable effect on gene transcription upon integration. Findings We have developed a single-copy integration system that enables the cloning vector to integrate at a specific engineered site, within an untranscribed intergenic region, in the chromosome of Staphylococcus aureus. This system is based on the lysogenic phage L54a site-specific recombination system in which the L54a phage (attP and chromosome (attB attachment sites, which share an 18-bp identical core sequence, were modified with identical mutations. The integration vector, pLL102, was constructed to contain the modified L54a attP site (attP2 that was altered at 5 nucleotide positions within the core sequence. In the recipient strain, the similarly modified attB site (attB2 was inserted in an intergenic region devoid of detectable transcription read-through. Integration of the vector, which is unable to replicate in S. aureus extrachromosomally, was achieved by providing the L54a integrase gene in a plasmid in the recipient. We showed that pLL102 integrated specifically at the engineered site rather than at the native L54a attB site and that integration did not have a significant effect on transcription of genes immediately upstream or downstream of the integration site. Conclusions In this work, we describe an E. coli-S. aureus shuttle vector that can be used to introduce any cloned gene into the S. aureus chromosome at a select site without affecting gene expression. The vector should be useful for genetic manipulation of S. aureus and for

  7. The Active Component of Aspirin, Salicylic Acid, Promotes Staphylococcus aureus Biofilm Formation in a PIA-dependent Manner.

    Science.gov (United States)

    Dotto, Cristian; Lombarte Serrat, Andrea; Cattelan, Natalia; Barbagelata, María S; Yantorno, Osvaldo M; Sordelli, Daniel O; Ehling-Schulz, Monika; Grunert, Tom; Buzzola, Fernanda R

    2017-01-01

    Aspirin has provided clear benefits to human health. But salicylic acid (SAL) -the main aspirin biometabolite- exerts several effects on eukaryote and prokaryote cells. SAL can affect, for instance, the expression of Staphylococcus aureus virulence factors. SAL can also form complexes with iron cations and it has been shown that different iron chelating molecules diminished the formation of S. aureus biofilm. The aim of this study was to elucidate whether the iron content limitation caused by SAL can modify the S. aureus metabolism and/or metabolic regulators thus changing the expression of the main polysaccharides involved in biofilm formation. The exposure of biofilm to 2 mM SAL induced a 27% reduction in the intracellular free Fe(2+) concentration compared with the controls. In addition, SAL depleted 23% of the available free Fe(2+) cation in culture media. These moderate iron-limited conditions promoted an intensification of biofilms formed by strain Newman and by S. aureus clinical isolates related to the USA300 and USA100 clones. The slight decrease in iron bioavailability generated by SAL was enough to induce the increase of PIA expression in biofilms formed by methicillin-resistant as well as methicillin-sensitive S. aureus strains. S. aureus did not produce capsular polysaccharide (CP) when it was forming biofilms under any of the experimental conditions tested. Furthermore, SAL diminished aconitase activity and stimulated the lactic fermentation pathway in bacteria forming biofilms. The polysaccharide composition of S. aureus biofilms was examined and FTIR spectroscopic analysis revealed a clear impact of SAL in a codY-dependent manner. Moreover, SAL negatively affected codY transcription in mature biofilms thus relieving the CodY repression of the ica operon. Treatment of mice with SAL induced a significant increase of S aureus colonization. It is suggested that the elevated PIA expression induced by SAL might be responsible for the high nasal

  8. The Active Component of Aspirin, Salicylic Acid, Promotes Staphylococcus aureus Biofilm Formation in a PIA-dependent Manner

    Science.gov (United States)

    Dotto, Cristian; Lombarte Serrat, Andrea; Cattelan, Natalia; Barbagelata, María S.; Yantorno, Osvaldo M.; Sordelli, Daniel O.; Ehling-Schulz, Monika; Grunert, Tom; Buzzola, Fernanda R.

    2017-01-01

    Aspirin has provided clear benefits to human health. But salicylic acid (SAL) -the main aspirin biometabolite- exerts several effects on eukaryote and prokaryote cells. SAL can affect, for instance, the expression of Staphylococcus aureus virulence factors. SAL can also form complexes with iron cations and it has been shown that different iron chelating molecules diminished the formation of S. aureus biofilm. The aim of this study was to elucidate whether the iron content limitation caused by SAL can modify the S. aureus metabolism and/or metabolic regulators thus changing the expression of the main polysaccharides involved in biofilm formation. The exposure of biofilm to 2 mM SAL induced a 27% reduction in the intracellular free Fe2+ concentration compared with the controls. In addition, SAL depleted 23% of the available free Fe2+ cation in culture media. These moderate iron-limited conditions promoted an intensification of biofilms formed by strain Newman and by S. aureus clinical isolates related to the USA300 and USA100 clones. The slight decrease in iron bioavailability generated by SAL was enough to induce the increase of PIA expression in biofilms formed by methicillin-resistant as well as methicillin-sensitive S. aureus strains. S. aureus did not produce capsular polysaccharide (CP) when it was forming biofilms under any of the experimental conditions tested. Furthermore, SAL diminished aconitase activity and stimulated the lactic fermentation pathway in bacteria forming biofilms. The polysaccharide composition of S. aureus biofilms was examined and FTIR spectroscopic analysis revealed a clear impact of SAL in a codY-dependent manner. Moreover, SAL negatively affected codY transcription in mature biofilms thus relieving the CodY repression of the ica operon. Treatment of mice with SAL induced a significant increase of S aureus colonization. It is suggested that the elevated PIA expression induced by SAL might be responsible for the high nasal colonization

  9. Molecular typing and phenotype characterization of methicillin-resistant Staphylococcus aureus isolates from blood in Taiwan.

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    Wei-Yao Wang

    Full Text Available BACKGROUND: Staphylococcus aureus causes a variety of severe infections such as bacteremia and sepsis. At present, 60-80% of S. aureus isolates from Taiwan are methicillin resistant (MRSA. It has been shown that certain MRSA clones circulate worldwide. The goals of this study were to identify MRSA clones in Taiwan and to correlate the molecular types of isolates with their phenotypes. METHODS: A total of 157 MRSA isolates from bacteremic patients were collected from nine medical centers. They were typed based on polymorphisms in agr, SCCmec, MLST, spa, and dru. Phenotypes characterized included Panton-Valentine leucocidin (pvl, inducible macrolide-lincosamide-streptogramin B resistance (MLSBi, vancomycin (VA and daptomycin (DAP minimal inhibitory concentrations (MIC, and superantigenic toxin gene profiles. Difference between two consecutive samples was determined by Mann-Whitney-U test, and difference between two categorical variables was determined by Fisher's exact test. RESULTS: Four major MRSA clone complexes CC1, CC5, CC8, and CC59 were found, including 4 CC1, 9 CC5, 111 CC8, and 28 CC59 isolates. These clones had the following molecular types: CC1: SCCmecIV and ST573; CC5: SCCmecII and ST5; CC8: SCCmecIII, ST239, and ST241, and CC59: SCCmecIV, SCCmecV(T, ST59, and ST338. The toxin gene profiles of these clones were CC1: sec-seg-(sei-sell-selm-(seln-selo; CC5: sec-seg-sei-sell-selm-(seln-selp-tst1; CC8: sea-selk-selq, and CC59: seb-selk-selq. Most isolates with SCCmecV(T, ST59, spat437, and dru11 types were pvl(+ (13 isolates, while multidrug resistance (≥4 antimicrobials were associated with SCCmecIII, ST239, spa t037, agrI, and dru14 (119 isolates (p<0.001. One hundred and twenty four isolates with the following molecular types had higher VA MIC: SCCmecII and SCCmecIII; ST5, ST239, and ST241; spa t002, t037, and t421; dru4, dru10, dru12, dru13, and dru14 (p<0.05. No particular molecular types were found to be associated with MLSBi

  10. Seamless Ligation Cloning Extract (SLiCE) cloning method.

    Science.gov (United States)

    Zhang, Yongwei; Werling, Uwe; Edelmann, Winfried

    2014-01-01

    SLiCE (Seamless Ligation Cloning Extract) is a novel cloning method that utilizes easy to generate bacterial cell extracts to assemble multiple DNA fragments into recombinant DNA molecules in a single in vitro recombination reaction. SLiCE overcomes the sequence limitations of traditional cloning methods, facilitates seamless cloning by recombining short end homologies (15-52 bp) with or without flanking heterologous sequences and provides an effective strategy for directional subcloning of DNA fragments from bacterial artificial chromosomes or other sources. SLiCE is highly cost-effective and demonstrates the versatility as a number of standard laboratory bacterial strains can serve as sources for SLiCE extract. We established a DH10B-derived E. coli strain expressing an optimized λ prophage Red recombination system, termed PPY, which facilitates SLiCE with very high efficiencies.

  11. Cloning: revisiting an old debate.

    Science.gov (United States)

    Verhey, Allen D

    1994-09-01

    The debate about cloning that took place 25 years ago, although directed toward a different sort of cloning, elucidates fundamental issues currently at stake in reproductive technologies and research. Paul Ramsey and Joseph Fletcher were participants in this early debate. The differences between Ramsey and Fletcher about the meaning and sufficiency of freedom, the understanding and weighing of good and evil, the connection between embodiment and personhood, the relationship of humans with nature, and the meaning of parenthood suggest both a broader agenda for the debate about cloning and a cautious move forward in the development of embryo-splitting.

  12. Staphylococcus aureus survival in human blood.

    Science.gov (United States)

    Malachowa, Natalia; DeLeo, Frank R

    2011-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is abundant in hospitals and in the United States is a leading cause of mortality due to infectious agents. Community-associated MRSA (CA-MRSA) strains such as USA300, which typically cause disease outside of healthcare settings, are also prevalent in the United States. Although most CA-MRSA infections affect skin and soft tissue, the pathogen can enter the bloodstream and ultimately cause severe disease. In a recent paper, we used USA300-specific microarrays to generate a comprehensive view of the molecules that facilitate S. aureus immune evasion and survival in human blood. Notably, genes encoding proteins involved in iron-uptake and utilization and gamma-hemolysin (hlgABC) are highly up-regulated by USA300 during culture in human blood. Here we discuss the potential implication of these findings and the possible role of gamma-hemolysin in the success of S. aureus as a human pathogen.

  13. Geographic distribution of Staphylococcus aureus causing invasive infections in Europe: a molecular-epidemiological analysis.

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    Hajo Grundmann

    2010-01-01

    Full Text Available BACKGROUND: Staphylococcus aureus is one of the most important human pathogens and methicillin-resistant variants (MRSAs are a major cause of hospital and community-acquired infection. We aimed to map the geographic distribution of the dominant clones that cause invasive infections in Europe. METHODS AND FINDINGS: In each country, staphylococcal reference laboratories secured the participation of a sufficient number of hospital laboratories to achieve national geo-demographic representation. Participating laboratories collected successive methicillin-susceptible (MSSA and MRSA isolates from patients with invasive S. aureus infection using an agreed protocol. All isolates were sent to the respective national reference laboratories and characterised by quality-controlled sequence typing of the variable region of the staphylococcal spa gene (spa typing, and data were uploaded to a central database. Relevant genetic and phenotypic information was assembled for interactive interrogation by a purpose-built Web-based mapping application. Between September 2006 and February 2007, 357 laboratories serving 450 hospitals in 26 countries collected 2,890 MSSA and MRSA isolates from patients with invasive S. aureus infection. A wide geographical distribution of spa types was found with some prevalent in all European countries. MSSA were more diverse than MRSA. Genetic diversity of MRSA differed considerably between countries with dominant MRSA spa types forming distinctive geographical clusters. We provide evidence that a network approach consisting of decentralised typing and visualisation of aggregated data using an interactive mapping tool can provide important information on the dynamics of MRSA populations such as early signalling of emerging strains, cross border spread, and importation by travel. CONCLUSIONS: In contrast to MSSA, MRSA spa types have a predominantly regional distribution in Europe. This finding is indicative of the selection and spread

  14. Methicillin-resistant Staphylococcus aureus associated with animals and its relevance to human health

    Directory of Open Access Journals (Sweden)

    Annalisa ePantosti

    2012-04-01

    Full Text Available Staphylococcus aureus is a typical human pathogen. Some animal S. aureus lineages have derived from human strains following profound genetic adaptation determining a change in host specificity. Due to the close relationship of animals with the environmental microbioma and resistoma, animal staphylococcal strains also represent a source of resistance determinants. Methicillin-resistant S. aureus (MRSA emerged fifty years ago as a nosocomial pathogen but in the last decade it has also become a frequent cause of infections in the community. The recent finding that MRSA frequently colonizes animals, especially livestock, has been a reason for concern, as it has revealed an expanded reservoir of MRSA. While MRSA strains recovered from companion animals are generally similar to human nosocomial MRSA, MRSA strains recovered from food animals appear to be specific animal-adapted clones. Since 2005, MRSA belonging to ST398 was recognized as a colonizer of pigs and human subjects professionally exposed to pig farming. The pig MRSA was also found to colonize other species of farmed animals, including horses, cattle and poultry and was therefore designated livestock-associated (LA-MRSA. LA-MRSA ST398 can cause infections in humans in contact with animals, and can infect hospitalized people, although at the moment this occurrence is relatively rare. Other animal-adapted MRSA clones have been detected in livestock, such as ST1 and ST9. Recently, ST130 MRSA isolated from bovine mastitis has been found to carry a novel mecA gene that eludes detection by conventional PCR tests. Similar ST130 strains have been isolated from human infections in UK, Denmark and Germany at low frequency. It is plausible that the increased attention to animal MRSA will reveal other strains with peculiar characteristics that can pose a risk to human health.

  15. Methylotroph cloning vehicle

    Science.gov (United States)

    Hanson, Richard S.; Allen, Larry N.

    1989-04-25

    A cloning vehicle comprising: a replication determinant effective for replicating the vehicle in a non-C.sub.1 -utilizing host and in a C.sub.1 -utilizing host; DNA effective to allow the vehicle to be mobilized from the non-C.sub.1 -utilizing host to the C.sub.1 -utilizing host; DNA providing resistance to two antibiotics to which the wild-type C.sub.1 -utilizing host is susceptible, each of the antibiotic resistance markers having a recognition site for a restriction endonuclease; a cos site; and a means for preventing replication in the C.sub.1 -utilizing host. The vehicle is used for complementation mapping as follows. DNA comprising a gene from the C.sub.1 -utilizing organism is inserted at the restriction nuclease recognition site, inactivating the antibiotic resistance marker at that site. The vehicle can then be used to form a cosmid structure to infect the non-C.sub.1 -utilizing (e.g., E. coli) host, and then conjugated with a selected C.sub.1 -utilizing mutant. Resistance to the other antibiotic by the mutant is a marker of the conjugation. Other phenotypical changes in the mutant, e.g., loss of an auxotrophic trait, is attributed to the C.sub.1 gene. The vector is also used to inactivate genes whose protein products catalyze side reactions that divert compounds from a biosynthetic pathway to a desired product, thereby producing an organism that makes the desired product in higher yields.

  16. Propionibacterium acnes biofilm - A sanctuary for Staphylococcus aureus?

    Science.gov (United States)

    Tyner, Harmony; Patel, Robin

    2016-08-01

    The purpose of this study was to measure the effect of combined culture of Propionibacterium acnes and Staphylococcus aureus on biofilm formation under different oxygen concentrations. We measured planktonic growth and biofilm formation of P. acnes and S. aureus alone and together under aerobic and anaerobic conditions. Both P. acnes and S. aureus grew under anaerobic conditions. When grown under anaerobic conditions, P. acnes with or without S. aureus formed a denser biomass biofilm than did S. aureus alone. Viable S. aureus was recovered from a16-day old combined P. acnes and S. aureus biofilm, but not a monomicrobial S. aureus biofilm.

  17. Mastite com lesões sistêmicas por Staphylococus aureus subesp. aureus em coelhos Mastitis with systemic lesions due to Staphylococus aureus subesp. aureus in rabbits

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    Sandra Davi Traverso

    2003-04-01

    Full Text Available Em uma criação composta por 1800 coelhos, 33% das matrizes apresentaram mastite e lesões cutâneas crostosas e purulentas. Estes animais apresentavam-se entre 10 a- 12 meses de idade e em segunda parição. Quinze coelhos afetados foram sacrificados e necropsiados. Na necropsia, além das lesões cutâneas haviam microabscessos em diversos órgãos. Das amostras coletadas isolou-se Staphylococcus aureus subesp. aureus. S. aureus subesp. aureus também foi isolado de "swab" nasal coletado do tratador encarregado de fazer o diagnóstico de gestação nas coelhas. Histologicamente, havia formação de múltiplos abscessos, presença de bactérias gram positivas em vasos sangüíneos e linfáticos, além de êmbolos bacterianos nos tecidos. Nas mamas, observou-se tecido glandular normal associado a abscessos multifocais delimitados.At a commercial rabbitry which was composed of 1800 New Zealand White rabbits, 30% of the does had presented mastitis and purulent cutaneal lesions. The age of the animals ranged from 10 to 12 months and were at the second parturition. At necropsy, microabscesses were observed in several organs. Bacteriological samples collected from affected animals resulted Staphylococcus aureus subesp. aureus.. Additionally, the same agent has been isolated from a nasal swab collected from the person responsible for the pregnancy diagnosis. Histologically, there were multiple abscesses, gram positive bacteria within blood and lymphatic vessels, and bacterial emboli scattered in the tissues. In the mammas, normal glandular tissue associated with multifocal abscesses were observed.

  18. Epidemiology of Staphylococcus aureus during space flight

    Science.gov (United States)

    Pierson, D. L.; Chidambaram, M.; Heath, J. D.; Mallary, L.; Mishra, S. K.; Sharma, B.; Weinstock, G. M.

    1996-01-01

    Staphylococcus aureus was isolated over 2 years from Space Shuttle mission crewmembers to determine dissemination and retention of bacteria. Samples before and after each mission were from nasal, throat, urine, and feces and from air and surface sampling of the Space Shuttle. DNA fingerprinting of samples by digestion of DNA with SmaI restriction endonuclease followed by pulsed-field gel electrophoresis showed S. aureus from each crewmember had a unique fingerprint and usually only one strain was carried by an individual. There was only one instance of transfer between crewmembers. Strains from interior surfaces after flight matched those of crewmembers, suggesting microbial fingerprinting may have forensic application.

  19. Massive dissemination of methicillin resistant Staphylococcus aureus in bloodstream infections in a high MRSA prevalence country: establishment and diversification of EMRSA-15.

    Science.gov (United States)

    Faria, Nuno A; Miragaia, Maria; de Lencastre, Hermínia

    2013-12-01

    Portugal is the European country with the highest prevalence of methicillin resistant Staphylococcus aureus (MRSA), in which EMRSA-15 (ST22-IVh) has been the dominant clone since soon after its introduction in Portuguese hospitals in 2001. In this study, we intend to not only, assess the evolution of the invasive MRSA in Portuguese hospitals, but also to evaluate the invasive methicillin susceptible S. aureus (MSSA) population and the relationship between both populations. In the current study, two major MRSA clones were identified: EMRSA-15 that has been dominant for more than 10 years and accounts for 75% of the MRSA isolates, and ST105-II, a clone related with the New York/Japan clone (ST5-II). In contrast, among MSSA, several clonal backgrounds were identified. Despite of the massive predominance of EMRSA-15 in the last decade, an increase in spa diversity has been observed in the last few years, which suggests a recent and local diversification of this clone. Interestingly, MRSA and MSSA populations with related clonal backgrounds appear to have increased as a result of the dissemination of MRSA to the community environment.

  20. A Clone of Your Own.

    Science.gov (United States)

    Bilodeau, Kirsten

    1997-01-01

    Describes an activity used at the Washington Park Arboretum that helps students understand cloning through plant propagation. Students also learn how to make a pot from recycled newspapers and how to make soil that is appropriate for the plants. (DDR)

  1. Human Cloning: Let's Discuss It.

    Science.gov (United States)

    Taras, Loretta; Stavroulakis, Anthea M.; Ortiz, Mary T.

    1999-01-01

    Describes experiences with holding discussions on cloning at a variety of levels in undergraduate biology courses. Discusses teaching methods used and student reactions to the discussions. Contains 12 references. (WRM)

  2. Human cloning and 'posthuman' society.

    Science.gov (United States)

    Blackford, Russell

    2005-01-01

    Since early 1997, when the creation of Dolly the sheep by somatic cell nuclear transfer was announced in Nature, numerous government reports, essays, articles and books have considered the ethical problems and policy issues surrounding human reproductive cloning. In this article, I consider what response a modern liberal society should give to the prospect of human cloning, if it became safe and practical. Some opponents of human cloning have argued that permitting it would place us on a slippery slope to a repugnant future society, comparable to that portrayed in Aldous Huxley's novel, Brave New World. I conclude that, leaving aside concerns about safety, none of the psychological or social considerations discussed in this article provides an adequate policy justification for invoking the state's coercive powers to prevent human cloning.

  3. Cloning Sequencing and Structural Manipulation of the Enterotoxin D and E Genes from Staphylococcus aureus

    Science.gov (United States)

    1990-07-01

    time. Further characterization of the plasmid was carried out by restriction mapping of pIB485 was performed. pIB485 DNA was digested with EcoRI...the interruption of the gene by insertion of the phage DNA. To characterize this unique regulation of gene expression, we sequenced the lipase gene...in a solution of 0.1% carboxymethylcellulose (added to stabilize the emulsion) by sonication for 7 - 10 minutes at 50w. This suspension was used to

  4. The molecular evolution of methicillin-resistant Staphylococcus aureus

    NARCIS (Netherlands)

    Deurenberg, R H; Vink, C; Kalenic, S; Friedrich, A W; Bruggeman, C A; Stobberingh, E E

    2007-01-01

    Staphylococcus aureus is a potentially pathogenic bacterium that causes a broad spectrum of diseases. S. aureus can adapt rapidly to the selective pressure of antibiotics, and this has resulted in the emergence and spread of methicillin-resistant S. aureus (MRSA). Resistance to methicillin and other

  5. 9 CFR 113.115 - Staphylococcus Aureus Bacterin-Toxoid.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Staphylococcus Aureus Bacterin-Toxoid... REQUIREMENTS Inactivated Bacterial Products § 113.115 Staphylococcus Aureus Bacterin-Toxoid. Staphylococcus... Staphylococcus aureus which has been inactivated and is nontoxic. Each serial of biological product...

  6. A pig model of acute Staphylococcus aureus induced pyemia

    DEFF Research Database (Denmark)

    Nielsen, O. L.; Iburg, T.; Aalbæk, B.;

    2009-01-01

    Background: Sepsis caused by Staphylococcus aureus constitutes an important cause of morbidity and mortality in humans, and the incidence of this disease-entity is increasing. In this paper we describe the initial microbial dynamics and lesions in pigs experimentally infected with S. aureus....... aureus isolated from man and an extension of the timeframe aiming at inducing sepsis, severe sepsis and septic shock....

  7. Sequence diversity in the A domain of Staphylococcus aureus fibronectin-binding protein A

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    Speziale Pietro

    2008-05-01

    Full Text Available Abstract Background Fibronectin-binding protein A (FnBPA mediates adhesion of Staphylococcus aureus to fibronectin, fibrinogen and elastin. We previously reported that S. aureus strain P1 encodes an FnBPA protein where the fibrinogen/elastin-binding domain (A domain is substantially divergent in amino acid sequence from the archetypal FnBPA of S. aureus NCTC8325, and that these variations created differences in antigenicity. In this study strains from multilocus sequence types (MLST that spanned the genetic diversity of S.aureus were examined to determine the extent of FnBPA A domain variation within the S. aureus population and its effect on ligand binding and immuno-crossreactivity. Results Seven different isotype forms (I – VII of the FnBPA A domain were identified which were between 66 to 76% identical in amino acid sequence in any pair-wise alignment. The fnbA allelic variants in strains of different multilocus sequence type were identified by DNA hybridization using probes specific for sequences encoding the highly divergent N3 sub-domain of different isotypes. Several isotypes were not restricted to specific clones or clonal complexes but were more widely distributed. It is highly likely that certain fnbA genes have been transferred horizontally. Residues lining the putative ligand-binding trench were conserved, which is consistent with the ability of each A domain isotype to bind immobilized fibrinogen and elastin by the dock-latch-lock mechanism. Variant amino acid residues were mapped on a three-dimensional model of the FnBPA A domain and were predicted to be surface-exposed. Polyclonal antibodies raised against the recombinant isotype I A domain bound that protein with a 4 – 7 fold higher apparent affinity compared to the A domains of isotypes II – VII, while some monoclonal antibodies generated against the isotype I A domain showed reduced or no binding to the other isotypes. Conclusion The FnBPA A domain occurs in at least 7

  8. Identification and Characterization of Staphylococcus aureus Strains with an Incomplete Hemolytic Phenotype

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    Haifang Zhang

    2016-11-01

    Staphylococcus aureus. We also found that IL-2, IL-6 and IL-17A secreted in the supernatant of SIHP infected macrophages increased significantly compared to tst negative control strains infected ones. MLST analysis showed that all of SIHP strains were classified into ST5 clone. To our knowledge, this study firstly showed that SIHP strains are

  9. Artificial cloning of domestic animals.

    Science.gov (United States)

    Keefer, Carol L

    2015-07-21

    Domestic animals can be cloned using techniques such as embryo splitting and nuclear transfer to produce genetically identical individuals. Although embryo splitting is limited to the production of only a few identical individuals, nuclear transfer of donor nuclei into recipient oocytes, whose own nuclear DNA has been removed, can result in large numbers of identical individuals. Moreover, clones can be produced using donor cells from sterile animals, such as steers and geldings, and, unlike their genetic source, these clones are fertile. In reality, due to low efficiencies and the high costs of cloning domestic species, only a limited number of identical individuals are generally produced, and these clones are primarily used as breed stock. In addition to providing a means of rescuing and propagating valuable genetics, somatic cell nuclear transfer (SCNT) research has contributed knowledge that has led to the direct reprogramming of cells (e.g., to induce pluripotent stem cells) and a better understanding of epigenetic regulation during embryonic development. In this review, I provide a broad overview of the historical development of cloning in domestic animals, of its application to the propagation of livestock and transgenic animal production, and of its scientific promise for advancing basic research.

  10. Methicillin/Oxacillin-resistant Staphylococcus aureus as a hospital and public health threat in Brazil

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    Karinne Spirandelli Carvalho

    2010-02-01

    Full Text Available Methicillin-resistant Staphylococcus aureus is an established nosocomial pathogen (HA-MRSA, hospital acquired MRSA, but has recently begun to appear in the community (CA-MRSA, community acquired MRSA. The cause of resistance to methicillin and all other β-lactam antibiotics is the mecA gene, which is situated on a mobile genetic element, the Staphylococcal Cassette Chromosome mec (SCCmec. Seven major variants of SCCmec, type I to VII are distinguished. HA-MRSA disseminated worldwide and causes the majority of S. aureus nosocomial infections with a limited number of clones disseminated including the Brazilian Epidemic Clone (BEC, ST239-MRSA-III. CA-MRSA isolates are susceptible to non-β-lactam antibiotics, usually isolated from healthy individuals which do not possess any unknown risk factors for MRSA infection and are associated with a larger clonal diversity compared with HA-MRSA. However, during recent years distinction between HA-MRSA and CA-MRSA is beginning to fade. Actually, knowledge about MRSA disseminating clones is required to implement any strategies to control the transmission of MRSA either within hospitals or in community. For this reason, rapid identification of strains is an important issue. The rate of HA-MRSA can be reduced substantially through the implementation of interventions strategies, even in settings where MRSA is endemic as in most Brazilian hospitals. However, these policies could be quite complicated in the light of an increasing CA-MRSA prevalence in healthcare facilities, considering that distinction between HA-MRSA and CA-MRSA has started to disappear.

  11. Structured Review of Code Clone Literature

    NARCIS (Netherlands)

    Hordijk, Wiebe; Ponisio, María Laura; Wieringa, Roel

    2008-01-01

    This report presents the results of a structured review of code clone literature. The aim of the review is to assemble a conceptual model of clone-related concepts which helps us to reason about clones. This conceptual model unifies clone concepts from a wide range of literature, so that findings ab

  12. Characterization of community acquired Staphylococcus aureus associated with skin and soft tissue infection in Beijing: high prevalence of PVL+ ST398.

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    Chunjiang Zhao

    Full Text Available Adult community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA and methicillin-susceptible S. aureus (CA-MSSA skin and soft tissue infection (SSTI in China is not well described. A prospective cohort of adults with SSTI was established between January 2009 and August 2010 at 4 hospitals in Beijing. Susceptibility testing and molecular typing, including multilocus sequence typing, spa, agr typing, and toxin detection were assessed for all S. aureus isolates. Overall, 501 SSTI patients were enrolled. Cutaneous abscess (40.7% was the most common infection, followed by impetigo (6.8% and cellulitis (4.8%. S. aureus accounted for 32.7% (164/501 of SSTIs. Five isolates (5/164, 3.0% were CA-MRSA. The most dominant ST in CA-MSSA was ST398 (17.6%. The prevalence of Panton-Valentine Leukocidin (pvl gene was 41.5% (66/159 in MSSA. Female, younger patients and infections requiring incision or drainage were more commonly associated with pvl-positive S. aureus (P<0.03; sec gene was more often identified in CC5 (P<0.03; seh gene was more prevalent in CC1 (P = 0.001. Importantly, ST59 isolates showed more resistance to erythromycin, clindamycin and tetracycline, and needed more surgical intervention. In conclusion, CA-MRSA infections were rare among adult SSTI patients in Beijing. Six major MSSA clones were identified and associated with unique antimicrobial susceptibility, toxin profiles, and agr types. A high prevalence of livestock ST398 clone (17.1% of all S. aureus infections was found with no apparent association to animal contact.

  13. Associated risk factors and pulsed field gel electrophoresis of nasal isolates of Staphylococcus aureus from medical students in a tertiary hospital in Lagos, Nigeria

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    Solayide A. Adesida

    2007-02-01

    Full Text Available Staphylococcus aureus infections are growing problems worldwide with important implications in hospitals. The organism is normally present in the nasal vestibule of about 35% of apparently healthy individuals and its carriage varies between different ethnic and age groups. Staphylococcal nasal carriage among health workers is particularly important to establish new clones and track origin of infections during outbreak situations. To determine the carriage rate and compare the pulsed field gel patterns of the strains, nasal swabs were collected from 185 medical students in a teaching hospital in Lagos, Nigeria. Isolates of S. aureus were tested for heamolysin production, methicillin sensitivity and Pulsed Field Gel Electrophoresis (PFGE was performed. The results showed S.aureus nasal carrier rate of 14% with significant rate among males compared to females. All the isolates produced heamolysin. Antibiotic susceptibility pattern revealed that majority of the isolates was susceptible. Five strains (19% harboured resistant determinants to penicillin and tetracycline. None of the strains was resistant to methicillin. 44% of the isolates typed by PFGE had type B, the most predominant pulsotype. PFGE A clone exhibited a single resistance phenotype suggesting a strong clonal relationship that could punctual an outbreak in the hospital. The results speculate that nasal carriage among medical personnel could be a function of various risk factors. Personal hygiene and behaviour may however be the means to reducing colonization and spread of S.aureus in our hospitals.

  14. Molecular characterization of Staphylococcus aureus isolates from skin and soft tissue infections samples and healthy carriers in the Central Slovenia region.

    Science.gov (United States)

    Svent-Kucina, Natasa; Pirs, Mateja; Kofol, Romina; Blagus, Rok; Smrke, Dragica Maja; Bilban, Marjan; Seme, Katja

    2016-04-01

    Staphylococcus aureus is among the most important human pathogens. It is associated with different infections and is a major cause of skin and soft tissue infections (SSTIs). The aim of our study was to compare S. aureus isolates associated with SSTIs with isolates obtained from healthy carriers in the Central Slovenia region in terms of antimicrobial susceptibility, genetic diversity by clonal complex (CC)/sequence type, spa type, and by toxin gene profiling. In total, 274 S. aureus isolates were collected prospectively by culturing wound samples from 461 SSTI patients and nasal samples from 451 healthy carriers. We have demonstrated high heterogeneity in terms of CCs and spa type in both groups of isolates. The main clone among SSTI strains was Panton-Valentine leukocidin gene (pvl) positive CC121, whereas the main clone among carrier strains was CC45 carrying a large range of toxin genes. The main spa type in both groups was t091. Pvl was more frequently present in SSTI strains (31.2% SSTI vs 3.6% carrier strains) and staphylococcal enterotoxin C was more frequently present in carrier strains (1.6% SSTI vs 17.0% carrier strains). We have also demonstrated that methicillin-resistant S. aureus was a rare cause (2.8%) of SSTIs in our region.

  15. Staphylococcus aureus entrance into the dairy chain: Tracking S. aureus from dairy cow to cheese

    Directory of Open Access Journals (Sweden)

    Judith Kümmel

    2016-10-01

    Full Text Available Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle. Due to its zoonotic potential, control of S. aureus is not only of great economic importance in the dairy industry but also a significant public health concern. The aim of this study was to decipher the potential of bovine udder associated S. aureus as reservoir for S. aureus contamination in dairy production and processing. From 18 farms, delivering their milk to an alpine dairy plant for the production of smeared semi-hard and hard cheese. 1176 quarter milk (QM samples of all cows in lactation (n = 294 and representative samples form bulk tank milk (BTM of all farms were surveyed for coagulase positive (CPS and coagulase negative Staphylococci (CNS. Furthermore, samples from different steps of the cheese manufacturing process were tested for CPS and CNS. As revealed by chemometric-assisted FTIR spectroscopy and molecular subtyping (spa typing and multi locus sequence typing, dairy cattle represent indeed an important, yet underreported, entrance point of S. aureus into the dairy chain. Our data clearly show that certain S. aureus subtypes are present in primary production as well as in the cheese processing at the dairy plant. However, although a considerable diversity of S. aureus subtypes was observed in QM and BTM at the farms, only certain S. aureus subtypes were able to enter and persist in the cheese manufacturing at the dairy plant and could be isolated from cheese until day fourteen of ripening. Farm strains belonging to the FTIR cluster B1 and B3, which show genetic characteristics (t2953, ST8, enterotoxin profile: sea/sed/sej of the recently described S. aureus genotype B, most successfully contaminated the cheese production at the dairy plant. Thus our study fosters the hypothesis that genotype B S. aureus represent a specific challenge in control of S. aureus in the dairy chain that requires effective clearance strategies and hygienic

  16. Staphylococcus aureus Entrance into the Dairy Chain: Tracking S. aureus from Dairy Cow to Cheese

    Science.gov (United States)

    Kümmel, Judith; Stessl, Beatrix; Gonano, Monika; Walcher, Georg; Bereuter, Othmar; Fricker, Martina; Grunert, Tom; Wagner, Martin; Ehling-Schulz, Monika

    2016-01-01

    Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle. Due to its zoonotic potential, control of S. aureus is not only of great economic importance in the dairy industry but also a significant public health concern. The aim of this study was to decipher the potential of bovine udder associated S. aureus as reservoir for S. aureus contamination in dairy production and processing. From 18 farms, delivering their milk to an alpine dairy plant for the production of smeared semi-hard and hard cheese. one thousand hundred seventy six one thousand hundred seventy six quarter milk (QM) samples of all cows in lactation (n = 294) and representative samples form bulk tank milk (BTM) of all farms were surveyed for coagulase positive (CPS) and coagulase negative Staphylococci (CNS). Furthermore, samples from different steps of the cheese manufacturing process were tested for CPS and CNS. As revealed by chemometric-assisted FTIR spectroscopy and molecular subtyping (spa typing and multi locus sequence typing), dairy cattle represent indeed an important, yet underreported, entrance point of S. aureus into the dairy chain. Our data clearly show that certain S. aureus subtypes are present in primary production as well as in the cheese processing at the dairy plant. However, although a considerable diversity of S. aureus subtypes was observed in QM and BTM at the farms, only certain S. aureus subtypes were able to enter and persist in the cheese manufacturing at the dairy plant and could be isolated from cheese until day 14 of ripening. Farm strains belonging to the FTIR cluster B1 and B3, which show genetic characteristics (t2953, ST8, enterotoxin profile: sea/sed/sej) of the recently described S. aureus genotype B, most successfully contaminated the cheese production at the dairy plant. Thus, our study fosters the hypothesis that genotype B S. aureus represent a specific challenge in control of S. aureus in the dairy chain that requires

  17. Meningitis causada por staphylococcus aureus catalasa negativa

    OpenAIRE

    Álvarez Moreno, Carlos Arturo; Arroyo A., Claudia Patricia; Rodríguez, Elizabeth; Martínez R., Luz Marina; Quevedo S., Ruth

    2011-01-01

    En un paciente con cáncer se aisló del liquido cefaloraquideo y ascitico un coco gram positivo coagulasa positivo. El germen aislado mostró las características típicas de un Staphylococcus aureus, a excepción de la actividad de la catalasa, la cual no pudo ser encontrada.

  18. Meticillin-resistant Staphylococcus aureus (MRSA)

    DEFF Research Database (Denmark)

    Stefani, Stefania; Chung, Doo Ryeon; Lindsay, Jodi A;

    2012-01-01

    This article reviews recent findings on the global epidemiology of healthcare-acquired/associated (HA), community-acquired/associated (CA) and livestock-associated (LA) meticillin-resistant Staphylococcus aureus (MRSA) and aims to reach a consensus regarding the harmonisation of typing methods...

  19. Methicillin-resistant Staphylococcus aureus transmission

    DEFF Research Database (Denmark)

    Andersen, Leif Percival; Nielsen, Xiaohui

    2015-01-01

    INTRODUCTION: Even though methicillin-resistant Staphylococcus aureus (MRSA) is a common cause of nosocomial infections, it may often be difficult to evaluate the exact route of transmission. METHODS: In this study, we describe four cases of nosocomial transmission of MRSA in a hospital with a low...

  20. Profiling the surfacome of Staphylococcus aureus

    NARCIS (Netherlands)

    Dreisbach, Annette; Hempel, Kristina; Buist, Girbe; Hecker, Michael; Becher, Doerte; van Dijl, Jan Maarten

    2010-01-01

    Staphylococcus aureus is a widespread opportunistic pathogen that can cause a wide variety of life-threatening diseases. Especially for the colonization of human tissues and the development of invasiveness, surface-exposed proteins are of major importance. In the present studies, we optimized a prot

  1. Staphylococcus aureus resistente a la meticilina (SARM)

    Centers for Disease Control (CDC) Podcasts

    2007-10-22

    Datos importantes sobre las infecciones por SARM en Estados Unidos, en las escuelas y los entornos médicos. (Title: Methicillin-resistant Staphylococcus aureus (MRSA)Created: 10/2007).  Created: 10/22/2007 by National Center for Preparedness, Detection, and Control of Infectious Diseases.   Date Released: 11/9/2007.

  2. Caracterización molecular de Staphylococcus aureus de origen nosocomial por Electroforesis de Enzimas Multilocus (MLEE

    Directory of Open Access Journals (Sweden)

    Suárez Zulma

    2002-12-01

    Full Text Available Se caracterizaron 41 aislamientos se Staphylococcus aureus tomados a pacientes y personal de la unidad de cuidados intensivos (UCI del Hospital San Juan de Dios, en el periodo comprendido entre septiembre de 1998 y septiembre de 1999 por medio del método de caracterización de Electroforesis de Enzimas Multilocus(MLEE. Se establecieron las relaciones genéticas entre los individuos, así como influencia del perfil de resistencia, el biotipo y el periodo de recolección. La evidencia experimental sugirió que la estructura de la población analizada encaja en el modelo clonal descrito previamente para las poblaciones de Staphylococcus aureus, coincidiendo con la prevalencia de un pequeño número de clones durante el periodo de recolección. 

  3. Evolutionary Dynamics of Pandemic Methicillin-Sensitive Staphylococcus aureus ST398 and Its International Spread via Routes of Human Migration

    Science.gov (United States)

    McAdam, Paul R.; Sullivan, Sean B.; Knox, Justin R.; Khiabanian, Hossein; Rabadan, Raul; Davies, Peter R.; Fitzgerald, J. Ross; Lowy, Franklin D.

    2017-01-01

    ABSTRACT Methicillin-susceptible Staphylococcus aureus (MSSA) accounts for the majority of S. aureus infections globally, and yet surprisingly little is known about its clonal evolution. We applied comparative whole-genome sequencing (WGS) analyses to epidemiologically and geographically diverse ST398-MSSA, a pandemic lineage affecting both humans and livestock. Bayesian phylogenetic analysis predicted divergence of human-associated ST398-MSSA ~40 years ago. Isolates from Midwestern pigs and veterinarians differed substantially from those in New York City (NYC). Pig ST398 strains contained a large region of recombination representing imports from multiple sequence types (STs). Phylogeographic analyses supported the spread of ST398-MSSA along local cultural and migratory links between parts of the Caribbean, North America, and France, respectively. Applying pairwise single-nucleotide polymorphism (SNP) distances as a measure of genetic relatedness between isolates, we observed that ST398 not only clustered in households but also frequently extended across local social networks. Isolates collected from environmental surfaces reflected the full diversity of colonizing individuals, highlighting their potentially critical role as reservoirs for transmission and diversification. Strikingly, we observed high within-host SNP variability compared to our previous studies on the dominant methicillin-resistant Staphylococcus aureus (MRSA) clone USA300. Our data indicate that the dynamics of colonization, persistence, and transmission differ substantially between USA300-MRSA and ST398-MSSA. Taken together, our study reveals local and international routes of transmission for a major MSSA clone, indicating key impacts of recombination and mutation on genetic diversification and highlighting important ecological differences from epidemic USA300. Our study demonstrates extensive local and international routes of transmission for a major MSSA clone despite the lack of substantial

  4. [Mystery and problems of cloning].

    Science.gov (United States)

    Nikitin, V A

    2010-01-01

    The attention of investigators is attracted to the fact that, in spite of great efforts in mammalian cloning, advances that have been made in this area of research are not great, and cloned animals have developmental pathologies often incompatible with life and/or reproduction ability. It is yet not clear what technical or biological factors underlie this, and how they are connected or interact with each other, which is more realistic strategically. There is a great number of articles dealing with the influence of cloning with the nuclear transfer on genetic and epigenetic reprogramming of donor cells. At the same time we can see the practical absence of analytical investigations concerning the technology of cloning as such, its weak points, and possible sources of cellular trauma in the course of microsurgery of nuclear transfer or twinning. This article discusses step by step several nuclear transfer techniques and the methods of dividing early preimplanted embryos for twinning with the aim to reveal possible sources of cell damage during micromanipulation that may have negative influence on the development of cloned organisms. Several new author's technologies based on the study of cell biophysical characteristics are described, which allow one to avoid cellular trauma during manipulation and minimize the possibility of cell damage at any rate.

  5. [Cloning and law in Hungary].

    Science.gov (United States)

    Julesz, Máté

    2015-03-01

    Reproductive human cloning is prohibited in Hungary, as in many other countries. Therapeutic human cloning is not prohibited, just like in many other countries. Stem cell therapy is also allowed. Article III, paragraph (3) of the Hungarian basic law (constitution) strictly forbids total human cloning. Article 1 of the Additional Protocol to the Oviedo Convention, on the Prohibition of Cloning Human Beings (1998) stipulates that any intervention seeking to create a human being genetically identical to another human being, whether living or dead, is prohibited. In Hungary, according to Article 174 of the Criminal Code, total human cloning constitutes a crime. Article 180, paragraph (3) of the Hungarian Act on Health declares that embryos shall not be brought about for research purposes; research shall be conducted only on embryos brought about for reproductive purposes when this is authorized by the persons entitled to decide upon its disposal, or when the embryo is damaged. Article 180, paragraph (5) of the Hungarian Act on Health stipulates that multiple individuals who genetically conform to one another shall not be brought about. According to Article 181, paragraph (1) of the Hungarian Act on Health, an embryo used for research shall be kept alive for not longer than 14 days, not counting the time it was frozen for storage and the time period of research.

  6. Staphylococcus aureus Shifts toward Commensalism in Response to Corynebacterium Species.

    Science.gov (United States)

    Ramsey, Matthew M; Freire, Marcelo O; Gabrilska, Rebecca A; Rumbaugh, Kendra P; Lemon, Katherine P

    2016-01-01

    Staphylococcus aureus-human interactions result in a continuum of outcomes from commensalism to pathogenesis. S. aureus is a clinically important pathogen that asymptomatically colonizes ~25% of humans as a member of the nostril and skin microbiota, where it resides with other bacteria including commensal Corynebacterium species. Commensal Corynebacterium spp. are also positively correlated with S. aureus in chronic polymicrobial diabetic foot infections, distinct from acute monomicrobial S. aureus infections. Recent work by our lab and others indicates that microbe-microbe interactions between S. aureus and human skin/nasal commensals, including Corynebacterium species, affect S. aureus behavior and fitness. Thus, we hypothesized that S. aureus interactions with Corynebacterium spp. diminish S. aureus virulence. We tested this by assaying for changes in S. aureus gene expression during in vitro mono- versus coculture with Corynebacterium striatum, a common skin and nasal commensal. We observed a broad shift in S. aureus gene transcription during in vitro growth with C. striatum, including increased transcription of genes known to exhibit increased expression during human nasal colonization and decreased transcription of virulence genes. S. aureus uses several regulatory pathways to transition between commensal and pathogenic states. One of these, the quorum signal accessory gene regulator (agr) system, was strongly inhibited in response to Corynebacterium spp. Phenotypically, S. aureus exposed to C. striatum exhibited increased adhesion to epithelial cells, reflecting a commensal state, and decreased hemolysin activity, reflecting an attenuation of virulence. Consistent with this, S. aureus displayed diminished fitness in experimental in vivo coinfection with C. striatum when compared to monoinfection. These data support a model in which S. aureus shifts from virulence toward a commensal state when exposed to commensal Corynebacterium species.

  7. Towards Clone Detection in UML Domain Models

    DEFF Research Database (Denmark)

    Störrle, Harald

    2010-01-01

    Code clones - that is, duplicate fragments of code - have been studied for a long time. There is strong evidence that code clones are a major source of software faults. Anecdotal evidence suggests that this phenomenon is not restricted to code, but occurs in models in a very similar way. So...... it is likely that model clones are as detrimental to model quality as they are to code quality. However, programming language code and visual models also have significant differences so that notions and algorithms developed in the code clone arena cannot be transferred directly to model clones. In this article......, we discuss how model clones arise by analyzing several practical scenarios. We propose a formal definition of models and clones, that allows us to specify a generic clone detection algorithm. Through a thorough analysis of the detail structure of sample UML domain models, recommendations for clone...

  8. The topsy-turvy cloning law.

    Science.gov (United States)

    Brassington, Iain; Oultram, Stuart

    2011-03-01

    In debates about human cloning, a distinction is frequently drawn between therapeutic and reproductive uses of the technology. Naturally enough, this distinction influences the way that the law is framed. The general consensus is that therapeutic cloning is less morally problematic than reproductive cloning--one can hold this position while holding that both are morally unacceptable--and the law frequently leaves the way open for some cloning for the sake of research into new therapeutic techniques while banning it for reproductive purposes. We claim that the position adopted by the law has things the wrong way around: if we accept a moral distinction between therapeutic and reproductive cloning, there are actually more reasons to be morally worried about therapeutic cloning than about reproductive cloning. If cloning is the proper object of legal scrutiny, then, we ought to make sure that we are scrutinising the right kind of clone.

  9. Public perceptions of animal cloning

    DEFF Research Database (Denmark)

    Jelsøe, Erling; Vincentsen, Ulla; Andersen, Ida-Elisabeth

    What was from the outset meant to be a survey testing predefined categories of ethical positions related to new biotechnologies with animal cloning as an example was subsequently developed into a process of broader involvement of groups of citizens in the issue. The survey was conducted at meetings...... in four different cities in Denmark. The participants were introduced to animal cloning and after that they filled out the questionnaire. Finally, the issue was discussed in focus groups. The process as a whole was run in a dialogue oriented way. Through the information they received in combination...... with reflecting on the survey questions the participants were well prepared for discussions in the focus groups. This approach made it possible, on the one hand to get a measure of the citizen's perceptions of the ethical aspects of animal cloning, but also to go deeper into their own thoughts of the issue...

  10. Animal cloning: advances and prospects

    Directory of Open Access Journals (Sweden)

    Chuaire Lilian

    2004-05-01

    Full Text Available Few recent advances have revolutionized the developmental biology as the animal cloning has. Since the birth of Dolly, the sheep, in 1996, which was the first derived clone of a mature animal, a new scientific era began. It has been characterized by growing demystification that differentiated cells are unalterable entities in its nuclear organization and chromatin structure, and by a better understanding of the mechanisms that regulate the development. Throughout this paper, we will review some of the achievements and limitations of the techniques used, both in therapeutic and in the reproductive cloning, as well as the perspectives that its application allows to glimpse within a close future. At the same time, we will point out some considerations regarding the ethical debate that surrounds such a controversial issue.

  11. Drug resistance and genetic characteristics of clinical isolates of staphylococci in Myanmar: high prevalence of PVL among methicillin-susceptible Staphylococcus aureus belonging to various sequence types

    Directory of Open Access Journals (Sweden)

    M.S. Aung

    2016-03-01

    Full Text Available Prevalence, drug resistance and genetic characteristics were analysed for a total of 128 clinical isolates of staphylococci obtained from a tertiary hospital in Myanmar. The dominant species were S. aureus (39% and S. haemolyticus (35%, followed by S. epidermidis (6% and S. saprophyticus (5%. The majority of S. haemolyticus isolates (71.1% harboured mecA, showing high resistance rates to ampicillin, cephalosporins, erythromycin and levofloxacin, while methicillin-resistant S. aureus (MRSA was only 8% (four isolates among S. aureus with type IV SCCmec. Panton-Valentine leukocidin (PVL genes were detected in 20 isolates of S. aureus (40%, among which only one isolate was MRSA belonging to sequence type (ST 88/agr-III/coa-IIIa, and the other 19 methicillin-susceptible S. aureus (MSSA isolates were classified into six STs (ST88, ST121, ST1153, ST1155, ST1930, ST3206. An ST1153 MSSA isolate with PVL was revealed to belong to a novel coa type, XIIIa. ST121 S. aureus was the most common in the PVL-positive MSSA (47%, 9/19, harbouring genes of bone sialoprotein and variant of elastin binding protein as a distinctive feature. Although PVL-positive MSSA was susceptible to most of the antimicrobial agents examined, ST1930 isolates were resistant to erythromycin and levofloxacin. ST59 PVL-negative MRSA and MSSA had more resistance genes than other MRSA and PVL-positive MSSA, showing resistance to more antimicrobial agents. This study indicated higher prevalence of mecA associated with multiple drug resistance in S. haemolyticus than in S. aureus, and dissemination of PVL genes to multiple clones of MSSA, with ST121 being dominant, among hospital isolates in Myanmar.

  12. Toward an understanding of the evolution of Staphylococcus aureus strain USA300 during colonization in community households.

    Science.gov (United States)

    Uhlemann, Anne-Catrin; Kennedy, Adam D; Martens, Craig; Porcella, Stephen F; Deleo, Frank R; Lowy, Franklin D

    2012-01-01

    Staphylococcus aureus is a frequent cause of serious infections and also a human commensal. The emergence of community-associated methicillin-resistant S. aureus led to a dramatic increase in skin and soft tissue infections worldwide. This epidemic has been driven by a limited number of clones, such as USA300 in the United States. To better understand the extent of USA300 evolution and diversification within communities, we performed comparative whole-genome sequencing of three clinical and five colonizing USA300 isolates collected longitudinally from three unrelated households over a 15-month period. Phylogenetic analysis that incorporated additional geographically diverse USA300 isolates indicated that all but one likely arose from a common recent ancestor. Although limited genetic adaptation occurred over the study period, the greatest genetic heterogeneity occurred between isolates from different households and within one heavily colonized household. This diversity allowed for a more accurate tracking of interpersonal USA300 transmission. Sequencing of persisting USA300 isolates revealed mutations in genes involved in major aspects of S. aureus function: adhesion, cell wall biosynthesis, virulence, and carbohydrate metabolism. Genetic variations also included accumulation of multiple polymorphisms within select genes of two multigene operons, suggestive of small genome rearrangements rather than de novo single point mutations. Such rearrangements have been underappreciated in S. aureus and may represent novel means of strain variation. Subtle genetic changes may contribute to USA300 fitness and persistence. Elucidation of small genome rearrangements reveals a potentially new and intriguing mechanism of directed S. aureus genome diversification in environmental niches and during pathogen-host interactions.

  13. Foods from black market at EU border as a neglected route of potential methicillin-resistant Staphylococcus aureus transmission.

    Science.gov (United States)

    Oniciuc, Elena-Alexandra; Ariza-Miguel, Jaime; Bolocan, Andrei-Sorin; Diez-Valcarce, Marta; Rovira, Jordi; Hernández, Marta; Fernández-Natal, Isabel; Nicolau, Anca Ioana; Rodríguez-Lázaro, David

    2015-09-16

    The illegal entrance of foods to EU through black markets at the EU borders can constitute a neglected route of dissemination of foodborne pathogens, and in particular of methicillin-resistant Staphylococcus aureus (MRSA). In this study, we have assessed the presence of MRSA in foods sold in a black market at an EU border (the southeast part of Romania, on the border with Republic of Moldavia). We performed a search for MRSA among 200 food samples collected from 2012 to 2013. All S. aureus were studied by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing. MRSA isolates were further characterized by multilocus sequence typing (MLST) and SCCmec typing, and tested for the presence of Panton-Valentine leukocidin (PVL) virulence factors. Overall, 32 S. aureus isolates were recovered from 16 food samples (8%). One isolate detected in a pork lard sample was MRSA (0.5%). PFGE with the restriction enzyme SmaI revealed 12 genotypes among the 32 S. aureus isolates. The MRSA isolate belonged to sequence type 398, harbored SCCmec type V, tested negative for the presence of the PVL genes and was resistant to ciprofloxacin, tetracycline and cefazolin, besides all β-lactams. Among 31 methicillin-sensitive S. aureus (MSSA), 29% were resistant to penicillin, 9.7% to tetracycline and 3.2% to ciprofloxacin. In conclusion, in this study we report the presence of livestock-associated MRSA in foods sold in a black market at an EU border: ST398-MRSA-V. These results confirm the potential role of food in the dissemination of MRSA lineages among population, and the potential role of illegally introduced food to EU in the prevalence and evolution of MRSA clones in the community.

  14. Staphylococcus aureus shifts towards commensalism in response to Corynebacterium species

    Directory of Open Access Journals (Sweden)

    Matthew M Ramsey

    2016-08-01

    Full Text Available Staphylococcus aureus–human interactions result in a continuum of outcomes from commensalism to pathogenesis. S. aureus is a clinically important pathogen that asymptomatically colonizes ~25% of humans as a member of the nostril and skin microbiota, where it resides with other bacteria including commensal Corynebacterium species. Commensal Corynebacterium spp. are also positively correlated with S. aureus in chronic polymicrobial diabetic foot infections, distinct from acute monomicrobial S. aureus infections. Recent work by our lab and others indicates that microbe-microbe interactions between S. aureus and human skin/nasal commensals, including Corynebacterium species, affect S. aureus behavior and fitness. Thus, we hypothesized that S. aureus interactions with Corynebacterium spp. diminish S. aureus virulence. We tested this by assaying for changes in S. aureus gene expression during in vitro mono- versus coculture with Corynebacterium striatum, a common skin and nasal commensal. We observed a broad shift in S. aureus gene transcription during in vitro growth with C. striatum, including increased transcription of genes known to exhibit increased expression during human nasal colonization and decreased transcription of virulence genes. S. aureus uses several regulatory pathways to transition between commensal and pathogenic states. One of these, the quorum signal accessory gene regulator (agr system, was strongly inhibited in response to Corynebacterium spp. Phenotypically, S. aureus exposed to C. striatum exhibited increased adhesion to epithelial cells, reflecting a commensal state, and decreased hemolysin activity, reflecting an attenuation of virulence. Consistent with this, S. aureus displayed diminished fitness in experimental in vivo coinfection with C. striatum when compared to monoinfection. These data support a model in which S. aureus shifts from virulence towards a commensal state when exposed to commensal Corynebacterium species.

  15. Aspartate inhibits Staphylococcus aureus biofilm formation.

    Science.gov (United States)

    Yang, Hang; Wang, Mengyue; Yu, Junping; Wei, Hongping

    2015-04-01

    Biofilm formation renders Staphylococcus aureus highly resistant to conventional antibiotics and host defenses. Four D-amino acids (D-Leu, D-Met, D-Trp and D-Tyr) have been reported to be able to inhibit biofilm formation and disassemble established S. aureus biofilms. We report here for the first time that both D- and L-isoforms of aspartate (Asp) inhibited S. aureus biofilm formation on tissue culture plates. Similar biofilm inhibition effects were also observed against other staphylococcal strains, including S. saprophyticus, S. equorum, S. chromogenes and S. haemolyticus. It was found that Asp at high concentrations (>10 mM) inhibited the growth of planktonic N315 cells, but at subinhibitory concentrations decreased the cellular metabolic activity without influencing cell growth. The decreased cellular metabolic activity might be the reason for the production of less protein and DNA in the matrix of the biofilms formed in the presence of Asp. However, varied inhibition efficacies of Asp were observed for biofilms formed by clinical staphylococcal isolates. There might be mechanisms other than decreasing the metabolic activity, e.g. the biofilm phenotypes, affecting biofilm formation in the presence of Asp.

  16. Toxin-Antitoxin Systems of Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Christopher F. Schuster

    2016-05-01

    Full Text Available Toxin-antitoxin (TA systems are small genetic elements found in the majority of prokaryotes. They encode toxin proteins that interfere with vital cellular functions and are counteracted by antitoxins. Dependent on the chemical nature of the antitoxins (protein or RNA and how they control the activity of the toxin, TA systems are currently divided into six different types. Genes comprising the TA types I, II and III have been identified in Staphylococcus aureus. MazF, the toxin of the mazEF locus is a sequence-specific RNase that cleaves a number of transcripts, including those encoding pathogenicity factors. Two yefM-yoeB paralogs represent two independent, but auto-regulated TA systems that give rise to ribosome-dependent RNases. In addition, omega/epsilon/zeta constitutes a tripartite TA system that supposedly plays a role in the stabilization of resistance factors. The SprA1/SprA1AS and SprF1/SprG1 systems are post-transcriptionally regulated by RNA antitoxins and encode small membrane damaging proteins. TA systems controlled by interaction between toxin protein and antitoxin RNA have been identified in S. aureus in silico, but not yet experimentally proven. A closer inspection of possible links between TA systems and S. aureus pathophysiology will reveal, if these genetic loci may represent druggable targets. The modification of a staphylococcal TA toxin to a cyclopeptide antibiotic highlights the potential of TA systems as rather untapped sources of drug discovery.

  17. Toxin-Antitoxin Systems of Staphylococcus aureus.

    Science.gov (United States)

    Schuster, Christopher F; Bertram, Ralph

    2016-05-05

    Toxin-antitoxin (TA) systems are small genetic elements found in the majority of prokaryotes. They encode toxin proteins that interfere with vital cellular functions and are counteracted by antitoxins. Dependent on the chemical nature of the antitoxins (protein or RNA) and how they control the activity of the toxin, TA systems are currently divided into six different types. Genes comprising the TA types I, II and III have been identified in Staphylococcus aureus. MazF, the toxin of the mazEF locus is a sequence-specific RNase that cleaves a number of transcripts, including those encoding pathogenicity factors. Two yefM-yoeB paralogs represent two independent, but auto-regulated TA systems that give rise to ribosome-dependent RNases. In addition, omega/epsilon/zeta constitutes a tripartite TA system that supposedly plays a role in the stabilization of resistance factors. The SprA1/SprA1AS and SprF1/SprG1 systems are post-transcriptionally regulated by RNA antitoxins and encode small membrane damaging proteins. TA systems controlled by interaction between toxin protein and antitoxin RNA have been identified in S. aureus in silico, but not yet experimentally proven. A closer inspection of possible links between TA systems and S. aureus pathophysiology will reveal, if these genetic loci may represent druggable targets. The modification of a staphylococcal TA toxin to a cyclopeptide antibiotic highlights the potential of TA systems as rather untapped sources of drug discovery.

  18. NVC-422 inactivates Staphylococcus aureus toxins.

    Science.gov (United States)

    Jekle, Andreas; Yoon, Jungjoo; Zuck, Meghan; Najafi, Ramin; Wang, Lu; Shiau, Timothy; Francavilla, Charles; Rani, Suriani Abdul; Eitzinger, Christian; Nagl, Markus; Anderson, Mark; Debabov, Dmitri

    2013-02-01

    Bacterial pathogens have specific virulence factors (e.g., toxins) that contribute significantly to the virulence and infectivity of microorganisms within the human hosts. Virulence factors are molecules expressed by pathogens that enable colonization, immunoevasion, and immunosuppression, obtaining nutrients from the host or gaining entry into host cells. They can cause pathogenesis by inhibiting or stimulating certain host functions. For example, in systemic Staphylococcus aureus infections, virulence factors such as toxic shock syndrome toxin 1 (TSST-1), staphylococcal enterotoxin A (SEA), and staphylococcal enterotoxin B (SEB) cause sepsis or toxic shock by uncontrolled stimulation of T lymphocytes and by triggering a cytokine storm. In vitro, these superantigens stimulate the proliferation of human peripheral blood mononuclear cells (PBMC) and the release of many cytokines. NVC-422 (N,N-dichloro-2,2-dimethyltaurine) is a broad-spectrum, fast-acting topical anti-infective agent against microbial pathogens, including antibiotic-resistant microbes. Using mass spectrometry, we demonstrate here that NVC-422 oxidizes methionine residues of TSST-1, SEA, SEB, and exfoliative toxin A (ETA). Exposure of virulence factors to 0.1% NVC-422 for 1 h prevented TSST-1-, SEA-, SEB-, and ETA-induced cell proliferation and cytokine release. Moreover, NVC-422 also delayed and reduced the protein A- and clumping factor-associated agglutination of S. aureus cultures. These results show that, in addition to its well-described direct microbicidal activity, NVC-422 can inactivate S. aureus virulence factors through rapid oxidation of methionines.

  19. Quantum cloning machines and the applications

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Heng, E-mail: hfan@iphy.ac.cn [Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190 (China); Collaborative Innovation Center of Quantum Matter, Beijing 100190 (China); Wang, Yi-Nan; Jing, Li [School of Physics, Peking University, Beijing 100871 (China); Yue, Jie-Dong [Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing 100190 (China); Shi, Han-Duo; Zhang, Yong-Liang; Mu, Liang-Zhu [School of Physics, Peking University, Beijing 100871 (China)

    2014-11-20

    No-cloning theorem is fundamental for quantum mechanics and for quantum information science that states an unknown quantum state cannot be cloned perfectly. However, we can try to clone a quantum state approximately with the optimal fidelity, or instead, we can try to clone it perfectly with the largest probability. Thus various quantum cloning machines have been designed for different quantum information protocols. Specifically, quantum cloning machines can be designed to analyze the security of quantum key distribution protocols such as BB84 protocol, six-state protocol, B92 protocol and their generalizations. Some well-known quantum cloning machines include universal quantum cloning machine, phase-covariant cloning machine, the asymmetric quantum cloning machine and the probabilistic quantum cloning machine. In the past years, much progress has been made in studying quantum cloning machines and their applications and implementations, both theoretically and experimentally. In this review, we will give a complete description of those important developments about quantum cloning and some related topics. On the other hand, this review is self-consistent, and in particular, we try to present some detailed formulations so that further study can be taken based on those results.

  20. EasyClone-MarkerFree

    DEFF Research Database (Denmark)

    Fabre, Mathew Malcolm Jessop; Jakociunas, Tadas; Stovicek, Vratislav

    2016-01-01

    Clone-MarkerFree. The integration of linearized expression cassettes into defined genomic loci is facilitated by CRISPR/Cas9. Cas9 is recruited to the chromosomal location by specific guide RNAs (gRNAs) expressed from a set of gRNA helper vectors. Using our genome engineering vector suite, single and triple insertions are obtained...

  1. Clone Poems and the Microcomputer.

    Science.gov (United States)

    Irizarry, Estelle

    1989-01-01

    Describes how students can use the computer to study and create clone poems (altering original Spanish-language poems by substituting words and expressions), and how students can gain a deeper appreciation of the original poem's poetic structure and semantics. (CB)

  2. Graph rewriting with polarized cloning

    CERN Document Server

    Duval, Dominique; Prost, Frédéric

    2009-01-01

    We tackle the problem of graph transformation with a particular focus on node cloning. We propose a graph rewriting framework where nodes can be cloned zero, one or more times. A node can be cloned together with all its incident edges, with only the outgoing edges, with only the incoming edges or without any of the incident edges. We thus subsume previous works such as the sesqui-pushout, the heterogeneous pushout and the adaptive star grammars approaches. A rule is defined as a span $\\spa{\\grpol{L}}{l}{\\grpol{K}}{r}{R}$ where the right-hand side $R$ is a multigraph, the left-hand side $\\grpol{L}$ and the interface $\\grpol{K}$ are polarized multigraphs. A polarized multigraph is a multigraph endowed with some cloning annotations on nodes and edges. We introduce the notion of polarized multigraphs and define a rewriting step as pushback followed by a pushout in the same way as in the sesqui-pushout approach.

  3. Operads, clones, and distributive laws

    CERN Document Server

    Curien, Pierre-Louis

    2012-01-01

    We show how non-symmetric operads (or multicategories), symmetric operads, and clones, arise from three suitable monads on Cat, each extending to a (pseudo-)monad on the bicategory of categories and profunctors. We also explain how other previous categorical analyses of operads (via Day's tensor products, or via analytical functors) fit with the profunctor approach.

  4. Mammalian cloning: possibilities and threats.

    Science.gov (United States)

    Mitalipov, S M; Wolf, D P

    2000-10-01

    The cloning of mammals originated with the production of limited numbers of genetically identical offspring by blastomere separation or embryo splitting. In the past few years, remarkable progress has been reported in cloning by nuclear transfer (NT) with donor nuclei recovered from embryonic, fetal or adult cells. Factors that contribute to the successful reprogramming of the transferred nucleus and the normal term development of the newly reconstructed embryo include the cell cycle stage of both the donor nucleus and recipient cytoplast, the timing of fusion and cytoplast activation, and the source of donor nuclei. The possibility of producing live offspring by somatic cell NT carries potential applications in animal husbandry, biotechnology, transgenic and pharmaceutical production, biomedical research, and the preservation of endangered species. However, the low efficiencies of cloning by NT coupled with high embryonic, fetal and neonatal losses may restrict immediate commercial applications in agriculture. These limitations notwithstanding, the greatest benefits and practical implications of this new technology could be in transplantation medicine and therapeutic cloning.

  5. A 12-year survey of methicillin-resistant Staphylococcus aureus infections in Greece: ST80-IV epidemic?

    Science.gov (United States)

    Drougka, E; Foka, A; Liakopoulos, A; Doudoulakakis, A; Jelastopulu, E; Chini, V; Spiliopoulou, A; Levidiotou, S; Panagea, T; Vogiatzi, A; Lebessi, E; Petinaki, E; Spiliopoulou, I

    2014-11-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of both healthcare-associated MRSA (HA-MRSA) and community-associated MRSA (CA-MRSA) infections. Severe MRSA infections have been associated with the virulence factor Panton-Valentine leukocidin (PVL). The aim of this study was to investigate susceptibility patterns, the presence of toxin genes, including that encoding PVL, and clonality among MRSA isolates collected from patients in Greece over a 12-year period. MRSA isolates were collected from January 2001 to December 2012 from six different hospitals. Antibiotic susceptibility was determined with the disk diffusion method and the Etest. The presence of the toxic shock syndrome toxin-1 gene (tst), the enterotoxin gene cluster (egc) and the PVL gene was tested with PCR. The genotypic characteristics of the strains were analysed by SCCmec and agr typing, and clonality was determined with pulsed-field gel electrophoresis and multilocus sequence typing. An increasing rate of MRSA among S. aureus infections was detected up to 2008. The majority of PVL-positive MRSA isolates belonged to a single clone, sequence type (ST)80-IV, which was disseminated both in the community and in hospitals, especially during the warmest months of the year. Carriage of tst was associated with ST30-IV, whereas egc was distributed in different clones. CA-MRSA isolates were recovered mainly from skin and soft tissue infections, whereas HA-MRSA isolates were associated with surgical and wound infections. During the period 2001-2012, ST80-IV predominated in the community and infiltrated the hospital settings in Greece, successfully replacing other PVL-positive clones. The predominance of ST239-III in HA-MRSA infections was constant, whereas new clones have also emerged. Polyclonality was statistically significantly higher among CA-MRSA isolates and isolates from adult patients.

  6. SAMMD: Staphylococcus aureus Microarray Meta-Database

    Directory of Open Access Journals (Sweden)

    Elasri Mohamed O

    2007-10-01

    Full Text Available Abstract Background Staphylococcus aureus is an important human pathogen, causing a wide variety of diseases ranging from superficial skin infections to severe life threatening infections. S. aureus is one of the leading causes of nosocomial infections. Its ability to resist multiple antibiotics poses a growing public health problem. In order to understand the mechanism of pathogenesis of S. aureus, several global expression profiles have been developed. These transcriptional profiles included regulatory mutants of S. aureus and growth of wild type under different growth conditions. The abundance of these profiles has generated a large amount of data without a uniform annotation system to comprehensively examine them. We report the development of the Staphylococcus aureus Microarray meta-database (SAMMD which includes data from all the published transcriptional profiles. SAMMD is a web-accessible database that helps users to perform a variety of analysis against and within the existing transcriptional profiles. Description SAMMD is a relational database that uses MySQL as the back end and PHP/JavaScript/DHTML as the front end. The database is normalized and consists of five tables, which holds information about gene annotations, regulated gene lists, experimental details, references, and other details. SAMMD data is collected from the peer-reviewed published articles. Data extraction and conversion was done using perl scripts while data entry was done through phpMyAdmin tool. The database is accessible via a web interface that contains several features such as a simple search by ORF ID, gene name, gene product name, advanced search using gene lists, comparing among datasets, browsing, downloading, statistics, and help. The database is licensed under General Public License (GPL. Conclusion SAMMD is hosted and available at http://www.bioinformatics.org/sammd/. Currently there are over 9500 entries for regulated genes, from 67 microarray

  7. Human reproductive cloning: a conflict of liberties.

    Science.gov (United States)

    Havstad, Joyce C

    2010-02-01

    Proponents of human reproductive cloning do not dispute that cloning may lead to violations of clones' right to self-determination, or that these violations could cause psychological harms. But they proceed with their endorsement of human reproductive cloning by dismissing these psychological harms, mainly in two ways. The first tactic is to point out that to commit the genetic fallacy is indeed a mistake; the second is to invoke Parfit's non-identity problem. The argument of this paper is that neither approach succeeds in removing our moral responsibility to consider and to prevent psychological harms to cloned individuals. In fact, the same commitment to personal liberty that generates the right to reproduce by means of cloning also creates the need to limit that right appropriately. Discussion of human reproductive cloning ought to involve a careful and balanced consideration of both the relevant aspects of personal liberty - the parents' right to reproductive freedom and the cloned child's right to self-determination.

  8. Bacillithiol: a key protective thiol in Staphylococcus aureus.

    Science.gov (United States)

    Perera, Varahenage R; Newton, Gerald L; Pogliano, Kit

    2015-01-01

    Bacillithiol is a low-molecular-weight thiol analogous to glutathione and is found in several Firmicutes, including Staphylococcus aureus. Since its discovery in 2009, bacillithiol has been a topic of interest because it has been found to contribute to resistance during oxidative stress and detoxification of electrophiles, such as the antibiotic fosfomycin, in S. aureus. The rapid increase in resistance of methicillin-resistant Staphylococcus aureus (MRSA) to available therapeutic agents is a great health concern, and many research efforts are focused on identifying new drugs and targets to combat this organism. This review describes the discovery of bacillithiol, studies that have elucidated the physiological roles of this molecule in S. aureus and other Bacilli, and the contribution of bacillithiol to S. aureus fitness during pathogenesis. Additionally, the bacillithiol biosynthesis pathway is evaluated as a novel drug target that can be utilized in combination with existing therapies to treat S. aureus infections.

  9. [Infections caused by multi-resistant Gram-positive bacteria (Staphylococcus aureus and Enterococcus spp.)].

    Science.gov (United States)

    Cantón, Rafael; Ruiz-Garbajosa, Patricia

    2013-10-01

    Methicillin -resistant Staphylocccus aureus (MRSA) and multirresistant entorococci are still problematic in nosocomial infections and new challenges have emerged for their containment. MRSA has increased the multiresistant profile; it has been described vancomycin and linezolid resistant isolates and isolates with decreased daptomycin susceptibility. Moreover, new clones (ST398) have emerged, initially associated with piggeries, and new mec variants (mecC) with livestock origin that escape to the detection with current molecular methods based on mecA gene have been detected. In enterococci, linzeolid resistant isolates and isolates with deceased susceptibility to daptomycin have been described. Moreover, ampicillin resistant Enterococcus faecium due to β-lactamase production has been recently found in Europe. Control of MRSA isolates and multiresistant enteroccocci should combined antibiotic stewardship strategies and epidemiological measures, including detection of colonized patients in order to reduce colonization pressure and their transmission.

  10. Economical Phase-Covariant Cloning of Qudits

    CERN Document Server

    Buscemi, F; Macchiavello, C; Buscemi, Francesco; Ariano, Giacomo Mauro D'; Macchiavello, Chiara

    2004-01-01

    We derive the optimal $N\\to M$ phase-covariant quantum cloning for equatorial states in dimension $d$ with $M=kd+N$, $k$ integer. The cloning maps are optimal for both global and single-qudit fidelity. The map is achieved by an ``economical'' cloning machine, which works without ancilla. The connection between optimal phase-covariant cloning and optimal multi-phase estimation is finally established.

  11. Phagocytosis and Killing of Staphylococcus aureus by Human Neutrophils

    OpenAIRE

    Lu, Thea; Porter, Adeline R.; Kennedy, Adam D.; Kobayashi, Scott D.; Frank R DeLeo

    2014-01-01

    Neutrophils are essential for host defense against Staphylococcus aureus infections. Although significant progress has been made, our understanding of neutrophil interactions with S. aureus remains incomplete. To provide a more comprehensive view of this process, we investigated phagocytosis and killing of S. aureus by human neutrophils using varied assay conditions in vitro. A greater percentage of bacteria were internalized by adherent neutrophils compared to those in suspension, and unexpe...

  12. Dealing with clones in the tracking

    CERN Document Server

    Rodrigues, E

    2006-01-01

    The note describes the way clone tracks are found and eliminated in the LHCb tracking. Both the "clone killer" algorithm and the related "clone finder" tool are presented. The performance of the algorithm as it is used at present in Brunel is also discussed.

  13. A single-copy galK promoter cloning vector suitable for cloning strong promoters

    DEFF Research Database (Denmark)

    Dandanell, Gert; Court, Donald L.; Hammer, Karin

    1986-01-01

    We report the construction of lambda galK promoter cloning vectors for cloning and characterization of strong promoters. This phage, which contains a unique HindIII cloning site, was applied to the cloning and analysis of transcription initiations of the regulatory region of the deo-operon of...

  14. Resistance in Staphylococcus Aureus: The Never-Ending Story

    Directory of Open Access Journals (Sweden)

    Orlović Jovan

    2016-09-01

    Full Text Available Combating Staphylococcus aureus (S. aureus infections using antibacterial drugs is actually an ongoing effort to overcome resistance mechanism of this microorganism. In this paper, we discussed (1 the mechanisms of resistance to some of the most commonly used antimicrobial agents in the treatment of S. aureus: methicillin, vancomicyn and quinolones. In addition, (2 efflux pump mechanisms involved in maintaining homeostasis in the presence of compounds that inhibit S. aureus growth and reproduction, as well as mechanisms of resistance to a number of antibiotics, have been reviewed.

  15. A porcine model of haematogenous brain infectionwith staphylococcus aureus

    DEFF Research Database (Denmark)

    Astrup, Lærke Boye; Agerholm, Jørgen Steen; Nielsen, Ole Lerberg;

    2012-01-01

    A PORCINE MODEL OF HAEMATOGENOUS BRAIN INFECTION WITH STAPHYLOCOCCUS AUREUS Astrup Lærke1, Agerholm Jørgen1, Nielsen Ole1, Jensen Henrik1, Leifsson Páll1, Iburg Tine2. 1: Faculty of Health and Medical Sciences, University of Copenhagen, Denmark boye@life.ku.dk 2: National Veterinary Institute......, Uppsala, Sweden Introduction Staphylococcus aureus (S.aureus) is a common cause of sepsis and brain abscesses in man and a frequent cause of porcine pyaemia. Here we present a porcine model of haematogenous S. aureus-induced brain infection. Materials and Methods Four pigs had two intravenous catheters...

  16. Phagocytosis and killing of Staphylococcus aureus by human neutrophils.

    Science.gov (United States)

    Lu, Thea; Porter, Adeline R; Kennedy, Adam D; Kobayashi, Scott D; DeLeo, Frank R

    2014-01-01

    Neutrophils are essential for host defense against Staphylococcus aureus infections. Although significant progress has been made, our understanding of neutrophil interactions with S. aureus remains incomplete. To provide a more comprehensive view of this process, we investigated phagocytosis and killing of S. aureus by human neutrophils using varied assay conditions in vitro. A greater percentage of bacteria were internalized by adherent neutrophils compared to those in suspension, and, unexpectedly, uptake of S. aureus by adherent neutrophils occurred efficiently in the absence of opsonins. An antibody specific for S. aureus promoted uptake of unopsonized bacteria in suspension, but had little or no capacity to enhance phagocytosis of S. aureus opsonized with normal human serum or by adherent neutrophils. Collectively, these results indicate that assay conditions can have a significant influence on the phagocytosis and killing of S. aureus by neutrophils. More importantly, the results suggest a vaccine approach directed to enhance opsonophagocytosis alone is not sufficient to promote increased killing of S. aureus by human neutrophils. With the emergence and reemergence of antibiotic-resistant microorganisms, establishing parameters that are optimal for studying neutrophil-S. aureus interactions will pave the way towards developing immune-directed strategies for anti-staphylococcal therapies.

  17. Characterization of a Bacteriophage-Derived Murein Peptidase for Elimination of Antibiotic-Resistant Staphylococcus aureus.

    Science.gov (United States)

    Keary, Ruth; Sanz-Gaitero, Marta; van Raaij, Mark J; O'Mahony, Jim; Fenton, Mark; McAuliffe, Olivia; Hill, Colin; Ross, R Paul; Coffey, Aidan

    2016-01-01

    Staphylococcus aureus is a major cause of infection in humans and animals, causing a wide variety of diseases, from local inflammations to fatal sepsis. The bacterium is commonly multi-drug resistant and thus many front-line antibiotics have been rendered ineffective for treating such infections. Research on murein/peptidoglycan hydrolases, derived from bacterial viruses (bacteriophages), has demonstrated that such proteins are attractive candidates for development as novel antibacterial agents for combatting Gram-positive pathogens. Here we review the research produced to-date on the bacteriophage-derived CHAPK murein peptidase. Initially, we sequenced and annotated the genome of anti-staphylococcal bacteriophage K and cloned the gene for the bacteriophage endolysin, a murein hydrolase which plays a role in cell killing during the bacteriophage life cycle. An highly active domain of the enzyme, a cysteine, histidine-dependent amido hydrolase/peptidase (CHAPK), was cloned, overexpressed in E. coli and purified. This CHAPK enzyme was demonstrated to rapidly lyse several strains of methicillin resistant S. aureus and both disrupted and prevented the formation of a staphylococcal biofilm. The staphylolytic activity of the peptidase was demonstrated in vivo using a mouse model, without adverse effects on the animals. The crystal structure of the enzyme was elucidated, revealing a calcium ion close to the active site. Site-directed mutagenesis indicated that this calcium ion is involved in the catalytic mechanism of the enzyme. The crystal structure of this enzyme is a valuable source of information for efficient engineering of this and similar CHAP-domain-containing proteins. Overall, the data collected to date on CHAPK has demonstrated its strong potential as a novel therapeutic candidate for treatment of staphylococcal infections and has provided us with insight into the fundamental enzymatic mechanisms of CHAP domain-containing peptidoglycan hydrolases.

  18. Staphylococcus epidermidis ΔSortase A strain elicits protective immunity against Staphylococcus aureus infection.

    Science.gov (United States)

    Tan, Chao; Wang, Jun; Hu, Yifang; Wang, Peng; Zou, Lili

    2017-01-01

    Staphylococcus aureus and Staphylococcus epidermidis are two of the most significant opportunistic human pathogens, causing medical implant and nosocomial infections worldwide. These bacteria contain surface proteins that play crucial roles in multiple biological processes. It has become apparent that they have evolved a number of unique mechanisms by which they can immobilise proteins on their surface. Notably, a conserved cell membrane-anchored enzyme, sortase A (SrtA), can catalyse the covalent attachment of precursor bacterial cell wall-attached proteins to peptidoglycan. Considering its indispensable role in anchoring substrates to the cell wall and its effects on virulence, SrtA has attracted great attention. In this study, a 549-bp gene was cloned from a pathogenic S. epidermidis strain, YC-1, which shared high identity with srtA from other Staphylococcus spp. A mutant strain, YC-1ΔsrtA, was then constructed by allelic exchange mutagenesis. The direct survival rate assay suggested that YC-1ΔsrtA had a lower survival capacity in healthy mice blood compare with the wild-type strain, indicating that the deletion of srtA affects the virulence and infectious capacity of S. epidermidis YC-1. YC-1ΔsrtA was then administered via intraperitoneal injection and it provided a relative percent survival value of 72.7 % in mice against S. aureus TC-1 challenge. These findings demonstrate the possbility that YC-1ΔsrtA might be used as a live attenuated vaccine to produce cross-protection against S. aureus.

  19. MRSA and multidrug-resistant Staphylococcus aureus in U.S. retail meats, 2010-2011.

    Science.gov (United States)

    Ge, Beilei; Mukherjee, Sampa; Hsu, Chih-Hao; Davis, Johnnie A; Tran, Thu Thuy T; Yang, Qianru; Abbott, Jason W; Ayers, Sherry L; Young, Shenia R; Crarey, Emily T; Womack, Niketta A; Zhao, Shaohua; McDermott, Patrick F

    2017-04-01

    Methicillin-resistant Staphylococcus aureus (MRSA) has been detected in retail meats, although large-scale studies are scarce. We conducted a one-year survey in 2010-2011 within the framework of the National Antimicrobial Resistance Monitoring System. Among 3520 retail meats collected from eight U.S. states, 982 (27.9%) contained S. aureus and 66 (1.9%) were positive for MRSA. Approximately 10.4% (107/1032) of S. aureus isolates, including 37.2% (29/78) of MRSA, were multidrug-resistant (MDRSA). Turkey had the highest MRSA prevalence (3.5%), followed by pork (1.9%), beef (1.7%), and chicken (0.3%). Whole-genome sequencing was performed for all 66 non-redundant MRSA. Among five multilocus sequence types identified, ST8 (72.7%) and ST5 (22.7%) were most common and livestock-associated MRSA ST398 was assigned to one pork isolate. Eleven spa types were represented, predominately t008 (43.9%) and t2031 (22.7%). All four types of meats harbored t008, whereas t2031 was recovered from turkey only. The majority of MRSA (84.8%) possessed SCCmec IV and 62.1% harbored Panton-Valentine leukocidin. Pulsed-field gel electrophoresis showed that all ST8 MRSA belonged to the predominant human epidemic clone USA300, and others included USA100 and USA200. We conclude that a diverse MRSA population was present in U.S. retail meats, albeit at low prevalence.

  20. SarA positively controls bap-dependent biofilm formation in Staphylococcus aureus.

    Science.gov (United States)

    Trotonda, María Pilar; Manna, Adhar C; Cheung, Ambrose L; Lasa, Iñigo; Penadés, José R

    2005-08-01

    The biofilm-associated protein Bap is a staphylococcal surface protein involved in biofilm formation. We investigated the influence of the global regulatory locus sarA on bap expression and Bap-dependent biofilm formation in three unrelated Staphylococcus aureus strains. The results showed that Bap-dependent biofilm formation was diminished in the sarA mutants by an agr-independent mechanism. Complementation studies using a sarA clone confirmed that the defect in biofilm formation was due to the sarA mutation. As expected, the diminished capacity to form biofilms in the sarA mutants correlated with the decreased presence of Bap in the bacterial surface. Using transcriptional fusion and Northern analysis data, we demonstrated that the sarA gene product acts as an activator of bap expression. Finally, the bap promoter was characterized and the transcriptional start point was mapped by the rapid amplification of cDNA ends technique. As expected, we showed that purified SarA protein binds specifically to the bap promoter, as determined by gel shift and DNase I footprinting assays. Based on the previous studies of others as well as our work demonstrating the role for SarA in icaADBC and bap expression, we propose that SarA is an essential regulator controlling biofilm formation in S. aureus.

  1. First description of PVL-positive methicillin-resistant Staphylococcus aureus (MRSA) in wild boar meat.

    Science.gov (United States)

    Kraushaar, Britta; Fetsch, Alexandra

    2014-09-01

    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.

  2. Isolation, molecular characteristics and disinfection of methicillin-resistant Staphylococcus aureus from ICU units in Brazil.

    Science.gov (United States)

    Campos, Guilherme B; Souza, Simone G; Lob O, Tassia N; Da Silva, Danilo C C; Sousa, Daniel S; Oliveira, Pollianna S; Santos, Verena M; Amorim, Aline T; Farias, S Vio T; Cruz, Mariluze P; Yatsuda, Regiane; Marques, Lucas M

    2012-04-01

    The aim of the present study was to isolate S. aureus strains resistant to antibiotics, characterize the genotype profiles of resistance staphylococci, and evaluate the efficacy of antiseptic agents and disinfectants used in two public hospitals of Vitoria da Conquista, Bahia, Brazil. Clinical samples were obtained from ICU environments and equipment surfaces in two public hospitals in Vitoria da Conquista. Broth cultures were plated onto mannitol salt agar, and antimicrobial susceptibility testing was performed by the broth microdilution method according to CLSI. MRSA strains were submitted to PCR for detecting the mecA gene. PCR products were purified and sequenced for SCCmec type identification. Moreover, the strains were tested for efficacy of different disinfectant solutions. S. aureus were isolated from 31 and 67 sites in each hospital, respectively. Among the isolates from hospital 1, 07 (22.6%) were resistant to oxacillin while 28 (41.8%) were resistant in hospital 2. Thirty-one were positive for the mecA gene. All isolates showed SCCmec type III genotype characteristics of the Brazilian epidemic clone. In disinfectant tests, sodium hypochlorite (0.5, 1.0 and 2.0%), 2% chlorhexidine gluconate, quaternary ammonium, peracetic acid and formaldehyde were effective against the isolates tested. The strains showed higher resistance to vinegar (4% acetic acid), alcohol and glutaraldehyde. The findings of this study should assist in reducing the occurrence of nosocomial infections and therefore the morbidity, mortality and socio-economic burden caused by prolonged hospitalization.

  3. Clone DB: an integrated NCBI resource for clone-associated data.

    Science.gov (United States)

    Schneider, Valerie A; Chen, Hsiu-Chuan; Clausen, Cliff; Meric, Peter A; Zhou, Zhigang; Bouk, Nathan; Husain, Nora; Maglott, Donna R; Church, Deanna M

    2013-01-01

    The National Center for Biotechnology Information (NCBI) Clone DB (http://www.ncbi.nlm.nih.gov/clone/) is an integrated resource providing information about and facilitating access to clones, which serve as valuable research reagents in many fields, including genome sequencing and variation analysis. Clone DB represents an expansion and replacement of the former NCBI Clone Registry and has records for genomic and cell-based libraries and clones representing more than 100 different eukaryotic taxa. Records provide details of library construction, associated sequences, map positions and information about resource distribution. Clone DB is indexed in the NCBI Entrez system and can be queried by fields that include organism, clone name, gene name and sequence identifier. Whenever possible, genomic clones are mapped to reference assemblies and their map positions provided in clone records. Clones mapping to specific genomic regions can also be searched for using the NCBI Clone Finder tool, which accepts queries based on sequence coordinates or features such as gene or transcript names. Clone DB makes reports of library, clone and placement data on its FTP site available for download. With Clone DB, users now have available to them a centralized resource that provides them with the tools they will need to make use of these important research reagents.

  4. New shuttle vector-based expression system to generate polyhistidine-tagged fusion proteins in Staphylococcus aureus and Escherichia coli.

    Science.gov (United States)

    Schwendener, Sybille; Perreten, Vincent

    2015-05-01

    Four Staphylococcus aureus-Escherichia coli shuttle vectors were constructed for gene expression and production of tagged fusion proteins. Vectors pBUS1-HC and pTSSCm have no promoter upstream of the multiple cloning site (MCS), and this allows study of genes under the control of their native promoters, and pBUS1-Pcap-HC and pTSSCm-Pcap contain the strong constitutive promoter of S. aureus type 1 capsule gene 1A (Pcap) upstream of a novel MCS harboring codons for the peptide tag Arg-Gly-Ser-hexa-His (rgs-his6). All plasmids contained the backbone derived from pBUS1, including the E. coli origin ColE1, five copies of terminator rrnB T1, and tetracycline resistance marker tet(L) for S. aureus and E. coli. The minimum pAMα1 replicon from pBUS1 was improved through either complementation with the single-strand origin oriL from pUB110 (pBUS1-HC and pBUS1-Pcap-HC) or substitution with a pT181-family replicon (pTSSCm and pTSSCm-Pcap). The new constructs displayed increased plasmid yield and segregational stability in S. aureus. Furthermore, pBUS1-Pcap-HC and pTSSCm-Pcap offer the potential to generate C-terminal RGS-His6 translational fusions of cloned genes using simple molecular manipulation. BcgI-induced DNA excision followed by religation converts the TGA stop codon of the MCS into a TGC codon and links the rgs-his6 codons to the 3' end of the target gene. The generation of the rgs-his6 codon-fusion, gene expression, and protein purification were demonstrated in both S. aureus and E. coli using the macrolide-lincosamide-streptogramin B resistance gene erm(44) inserted downstream of Pcap. The new His tag expression system represents a helpful tool for the direct analysis of target gene function in staphylococcal cells.

  5. Staphylococcus aureus: resistance pattern and risk factors

    Directory of Open Access Journals (Sweden)

    Mohammad Naghavi-Behzad

    2015-03-01

    Full Text Available Introduction: Methicillin resistant Staphylococcus aureus (MRSA has emerged as a nosocomial pathogen of major worldwide importance and is an increasingly frequent cause of community-acquired infections. In this study, different risk factors and MRSA resistance pattern were investigated. Methods: In a 24 months period, all of the patients who were confined to bed in the surgery ward were included in the study. Then they were assessed to find out as if they had MRSA infection when hospitalized and once when they were discharged. Almost 48 h after admission, when patients were discharged, social and medical histories were acquired. Acquired samples were examined. Results: During the present study of 475 patients, 108 patients (22.8% had S. aureus. About frequency of antibiotic resistance among collected S. aureus colonies, erythromycin resistance, was the most frequent antibiotic resistance, also resistance to vancomycin was 0.4% that was the least. Only hospitalization duration had statistically significant correlation with antibiotic resistance, also resistance to erythromycin had statistically significant relation with history of surgery and alcohol consumption. Of all 34 MRSA species, 22 (64.7% samples were resistant to erythromycin, 17 (50.0% resistant to cefoxitin, 5 (14.7% resistant to mupirocin, 1 (2.9% resistant to vancomycin and 1 (2.9% resistant to linezolid. Conclusion: The results of the current study show that among hospitalized patients, there is resistance against methicillin. Since based on results of the study there is resistance against oxacillin and erythromycin in most cases, administering appropriate antibiotics have an important role in minimizing the resistance burden among bacterial species.

  6. Curcumin Reverse Methicillin Resistance in Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Su-Hyun Mun

    2014-11-01

    Full Text Available Curcumin, a natural polyphenolic flavonoid extracted from the rhizome of Curcuma longa L., was shown to possess superior potency to resensitize methicillin-resistant Staphylococcus aureus (MRSA to antibiotics. Previous studies have shown the synergistic activity of curcumin with β-lactam and quinolone antibiotics. Further, to understand the anti-MRSA mechanism of curcumin, we investigated the potentiated effect of curcumin by its interaction in diverse conditions. The mechanism of anti-MRSA action of curcumin was analyzed by the viability assay in the presence of detergents, ATPase inhibitors and peptidoglycan (PGN from S. aureus, and the PBP2a protein level was analyzed by western blotting. The morphological changes in the curcumin-treated MRSA strains were investigated by transmission electron microscopy (TEM. We analyzed increased susceptibility to MRSA isolates in the presence of curcumin. The optical densities at 600 nm (OD600 of the suspensions treated with the combinations of curcumin with triton X-100 and Tris were reduced to 63% and 59%, respectively, compared to curcumin without treatment. N,N'-dicyclohexylcarbodiimide (DCCD and sodium azide (NaN3 were reduced to 94% and 55%, respectively. When peptidoglycan (PGN from S. aureus was combined with curcumin, PGN (0–125 μg/mL gradually blocked the antibacterial activity of curcumin (125 μg/mL; however, at a concentration of 125 µg/mL PGN, it did not completely block curcumin. Curcumin has a significant effect on the protein level of PBP2a. The TEM images of MRSA showed damage of the cell wall, disruption of the cytoplasmic contents, broken cell membrane and cell lysis after the treatment of curcumin. These data indicate a remarkable antibacterial effect of curcumin, with membrane permeability enhancers and ATPase inhibitors, and curcumin did not directly bind to PGN on the cell wall. Further, the antimicrobial action of curcumin involved in the PBP2a-mediated resistance mechanism was

  7. Staphylococcus aureus reservoirs during traditional Austrian raw milk cheese production.

    Science.gov (United States)

    Walcher, Georg; Gonano, Monika; Kümmel, Judith; Barker, Gary C; Lebl, Karin; Bereuter, Othmar; Ehling-Schulz, Monika; Wagner, Martin; Stessl, Beatrix

    2014-11-01

    Sampling approaches following the dairy chain, including microbiological hygiene status of critical processing steps and physicochemical parameters, contribute to our understanding of how Staphylococcus aureus contamination risks can be minimised. Such a sampling approach was adopted in this study, together with rapid culture-independent quantification of Staph. aureus to supplement standard microbiological methods. A regional cheese production chain, involving 18 farms, was sampled on two separate occasions. Overall, 51·4% of bulk milk samples were found to be Staph. aureus positive, most of them (34·3%) at the limit of culture-based detection. Staph. aureus positive samples >100 cfu/ml were recorded in 17·1% of bulk milk samples collected mainly during the sampling in November. A higher number of Staph. aureus positive bulk milk samples (94·3%) were detected after applying the culture-independent approach. A concentration effect of Staph. aureus was observed during curd processing. Staph. aureus were not consistently detectable with cultural methods during the late ripening phase, but >100 Staph. aureus cell equivalents (CE)/ml or g were quantifiable by the culture-independent approach until the end of ripening. Enterotoxin gene PCR and pulsed-field gel electrophoresis (PFGE) typing provided evidence that livestock adapted strains of Staph. aureus mostly dominate the post processing level and substantiates the belief that animal hygiene plays a pivotal role in minimising the risk of Staph. aureus associated contamination in cheese making. Therefore, the actual data strongly support the need for additional sampling activities and recording of physicochemical parameters during semi-hard cheese-making and cheese ripening, to estimate the risk of Staph. aureus contamination before consumption.

  8. El envejecimiento de los clones

    OpenAIRE

    Trippi, Victorio S.

    2007-01-01

    El envejecimiento de los clones se observa en plantas que muestran crecimiento definido por un determinismo genético, cuando se multiplican con tejidos que evolucionan hacia el crecimiento reproductivo. Las plantas fuertemente influenciadas por el ambiente, pueden mostrar fenómenos de senescencia cuando la condición de ambiente determina el crecimiento reproductivo. Los cambios asociados con la edad resultan de alteraciones del citoplasma como un tipo de diferenciación cel...

  9. Rice's Salt Tolerance Gene Cloned

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    @@ In cooperation with US colleagues, CAS researchers have made significant progress in their studies into functional genes for key agronomic traits by cloning SKC1, a salt-tolerant functional gene of rice and making clear its biological functions and mechanisms. This pioneering work,which was reported in the Oct. issue of Nature Genetics (37:1141-1146), is believed to hold promise to increase the output of the crop plant in this country.

  10. Conotoxins Are Purified and Cloned

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    @@ A group of CAS scientists have succeeded in purifying many conotoxins and cloning more than 100 new genes from six species of cone snails living in waters off the coast of the South China Sea, paving the way for the development of new drugs to relieve neuropathic pains. The work has been honored with a first prize from the 2005 Awards for S&T Progress in Shanghai.

  11. Methicillin-resistant Staphylococcus aureus laryngitis.

    Science.gov (United States)

    Liakos, Tracey; Kaye, Keith; Rubin, Adam D

    2010-09-01

    Infections due to methicillin-resistant Staphylococcus aureus (MRSA) have become more prevalent, in part because of the emergence and spread of community-acquired MRSA. This trend is particularly concerning because of the significant rates of morbidity and mortality associated with MRSA infections, and because MRSA strains are often resistant to many classes of antibiotics. Reports of infections of the head and neck, including wound infections, cellulitis, sinusitis, otitis media, and otitis externa, are well documented. However, to our knowledge, there have been no reports of bacterial laryngitis due to MRSA. We report the first published case of bacterial laryngitis caused by MRSA.

  12. Resistencia antimicrobiana de cepas de Staphylococcus aureus, Costa Rica Antimicrobial resistance of Staphylococcus aureus, Costa Rica

    Directory of Open Access Journals (Sweden)

    Víctor Hugo Alvarado

    2011-12-01

    Full Text Available Objetivo: Determinar y comparar los perfiles de resistencia de cepas de S. aureus aisladas de quesos, producidos en la Zona Sur de Costa Rica y de un centro hospitalario de la misma región. Materiales y Métodos: Se analizaron 35 muestras de queso fresco, adquiridas durante los meses de setiembre y octubre del 2010 en la zona de San Vito de Coto Brus. A cada muestra se le realizaron recuentos de coliformes totales, coliformes fecales y Staphylococcus aureus. Adicionalmente se analizó presencia/ausencia de Listeria monocytogenes en 25 gramos del producto. A las cepas identificadas como S. aureus se les realizó la prueba de sensibilidad a los antibióticos mediante el sistema automatizado Vitek y la interpretación de los datos se realizó siguiendo las pautas del Clinical and Laboratory Standards Institute antimicrobial susceptibility testing 2011. Adicional a esto se recolectaron datos acerca de la sensibilidad de las cepas de S. aureus aisladas e identificadas en el Hospital de San Vito de Coto Brus en el mismo período. Resultados: El promedio obtenido para el recuento de coliformes totales fue de 9,7 X 10(6 UFC/g, para coliformes fecales de 6,7 X 10(5 y para S. aureus de 2,8 X 10(5 UFC/g, obteniéndose un 83 % de muestras positivas por esta bacteria. En cuanto a la resistencia antimicrobiana, se obtuvieron porcentajes de resistencia mayores en las cepas de origen clínico. Se encontró también que 23 de las cepas (96% provenientes de muestras clínicas, presentaban resistencia a más de un antibiótico, mientras que siete de las obtenidas a partir de queso (27% presentaban esta característica. Con respecto a los betalactamicos (ampicilina, oxacilina y penicilina se observó la existencia de una diferencia estadísticamente significativa (pObjective: determined and compared the resistance patters of S. aureus strains isolated from cheese produced in the southern zone of Costa Rica and from clinical samples isolated at the hospital center

  13. Cloning expeditions: risky but rewarding.

    Science.gov (United States)

    Lodish, Harvey

    2013-12-01

    In the 1980s, a good part of my laboratory was using the then-new recombinant DNA techniques to clone and characterize many important cell surface membrane proteins: GLUT1 (the red cell glucose transporter) and then GLUT2 and GLUT4, the red cell anion exchange protein (Band 3), asialoglycoprotein receptor subunits, sucrase-isomaltase, the erythropoietin receptor, and two of the subunits of the transforming growth factor β (TGF-β) receptor. These cloned genes opened many new fields of basic research, including membrane insertion and trafficking of transmembrane proteins, signal transduction by many members of the cytokine and TGF-β families of receptors, and the cellular physiology of glucose and anion transport. They also led to many insights into the molecular biology of several cancers, hematopoietic disorders, and diabetes. This work was done by an exceptional group of postdocs and students who took exceptionally large risks in developing and using novel cloning technologies. Unsurprisingly, all have gone on to become leaders in the fields of molecular cell biology and molecular medicine.

  14. Staphylococcus aureus bacteriuria as a prognosticator for outcome of Staphylococcus aureus bacteremia: a case-control study

    Directory of Open Access Journals (Sweden)

    Weinstein Robert A

    2010-07-01

    Full Text Available Abstract Background When Staphylococcus aureus is isolated in urine, it is thought to usually represent hematogenous spread. Because such spread might have special clinical significance, we evaluated predictors and outcomes of S. aureus bacteriuria among patients with S. aureus bacteremia. Methods A case-control study was performed at John H. Stroger Jr. Hospital of Cook County among adult inpatients during January 2002-December 2006. Cases and controls had positive and negative urine cultures, respectively, for S. aureus, within 72 hours of positive blood culture for S. aureus. Controls were sampled randomly in a 1:4 ratio. Univariate and multivariable logistic regression analyses were done. Results Overall, 59% of patients were African-American, 12% died, 56% of infections had community-onset infections, and 58% were infected with methicillin-susceptible S. aureus (MSSA. Among 61 cases and 247 controls, predictors of S. aureus bacteriuria on multivariate analysis were urological surgery (OR = 3.4, p = 0.06 and genitourinary infection (OR = 9.2, p = 0.002. Among patients who died, there were significantly more patients with bacteriuria than among patients who survived (39% vs. 17%; p = 0.002. In multiple Cox regression analysis, death risks in bacteremic patients were bacteriuria (hazard ratio 2.9, CI 1.4-5.9, p = 0.004, bladder catheter use (2.0, 1.0-4.0, p = 0.06, and Charlson score (1.1, 1.1-1.3, p = 0.02. Neither length of stay nor methicillin-resistant Staphylococcus aureus (MRSA infection was a predictor of S. aureus bacteriuria or death. Conclusions Among patients with S. aureus bacteremia, those with S. aureus bacteriuria had 3-fold higher mortality than those without bacteriuria, even after adjustment for comorbidities. Bacteriuria may identify patients with more severe bacteremia, who are at risk of worse outcomes.

  15. Staphylococcus aureus and the ecology of the nasal microbiome

    DEFF Research Database (Denmark)

    Liu, Cindy M; Price, Lance B; Hungate, Bruce A

    2015-01-01

    The human microbiome can play a key role in host susceptibility to pathogens, including in the nasal cavity, a site favored by Staphylococcus aureus. However, what determines our resident nasal microbiota-the host or the environment-and can interactions among nasal bacteria determine S. aureus...

  16. Duplex Identification of Staphylococcus aureus by Aptamer and Gold Nanoparticles.

    Science.gov (United States)

    Chang, Tianjun; Wang, Libo; Zhao, Kexu; Ge, Yu; He, Meng; Li, Gang

    2016-06-01

    Staphylococcus aureus is the top common pathogen causing infections and food poisoning. Identification of S. aureus is crucial for the disease diagnosis and regulation of food hygiene. Herein, we report an aptamer-AuNPs based method for duplex identification of S. aureus. Using AuNPs as an indicator, SA23, an aptamer against S. aureus, can well identify its target from Escherichia coli, Listeria monocytogenes and Pseudomonas aeruginosa. Furthermore, we find citrate-coated AuNPs can strongly bind to S. aureus, but not bind to Salmonella enterica and Proteus mirabilis, which leads to different color changes in salt solution. This colorimetric response is capable of distinguishing S. aureus from S. enteritidis and P. mirabilis. Thus, using the aptasensor and AuNPs together, S. aureus can be accurately identified from the common pathogens. This duplex identification system is a promising platform for simple visual identification of S. aureus. Additionally, in the aptasensing process, bacteria are incubated with aptamers and then be removed before the aptamers adding to AuNPs, which may avoid the interactions between bacteria and AuNPs. This strategy can be potentially applied in principle to detect other cells by AuNPs-based aptasensors.

  17. The changing epidemiology of Staphylococcus aureus bloodstream infection

    DEFF Research Database (Denmark)

    Laupland, K B; Lyytikäinen, O; Søgaard, M;

    2012-01-01

    Clin Microbiol Infect ABSTRACT: Although the epidemiology of Staphylococcus aureus bloodstream infection (BSI) has been changing, international comparisons are lacking. We sought to determine the incidence of S. aureus BSI and assess trends over time and by region. Population-based surveillance...

  18. Daya Hambat Ekstrak Aloe Vera terhadap pertumbuhan Staphylococcus Aureus

    OpenAIRE

    Rahmat, drg.Sp,Pros

    2011-01-01

    Dari hasil penelitian , maka dapat disimpulkan bahwa ekstrak Aloe Vera dapat menghambat pertumbuhan bakteri Stafhylococcus aureus, dan kadar hambat minimal ekstrak Aloe Vera adalah pada konsentrasi 25%. Tujuan Penelitan Ini adalah untuk mengetahui efektifitas ekstrak Aloe vera dalam menghambat pertumbuhan bakteri Stafhylococcus aureus dan daya hambat menimal, (DHM) terhadap pertumbuhan bakteri tersebut. Metode yang digunakan adalah pertumbuhan ekstrak Aloe vera, penegnceran ekstrak , pemur...

  19. Intracellular Staphylococcus aureus : live-in and let die

    NARCIS (Netherlands)

    Fraunholz, Martin; Sinha, Bhanu

    2012-01-01

    Staphylococcus aureus uses a plethora of virulence factors to accommodate a diversity of niches in its human host. Aside from the classical manifestations of S. aureus-induced diseases, the pathogen also invades and survives within mammalian host cells. The survival strategies of the pathogen are as

  20. New method for early detection of two random amplified polymorphic DNA (RAPD groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil

    Directory of Open Access Journals (Sweden)

    Dicezar Gonçalves

    2010-04-01

    Full Text Available The aim of this work was to develop a fast and accurate molecular approach to allow early detection of two RAPD groups of S. aureus causing bovine mastitis. Seventy five S. aureus isolates from infected animals were characterized by RAPD. Genomic fragments isolated from the unique bands present in either group were cloned and sequenced. Based on the DNA sequences, specific primers were designed to allow for the simultaneous detection of either group by multiplex PCR of S. aureus DNA isolated from clinical and subclinical bovine mastitis. Results showed that these proposed primers set could be used to detect various clinical and subclinical S. aureus isolates as well as the detection of the microorganism in bulk milk. Their use as a specific method for effective and early diagnostic tool for S. aureus infection in dairy herds is suggested.Esta pesquisa objetivou o desenvolvimento de técnica rápida e eficiente para diagnosticar precocemente diferentes linhagens de S. aureus causadoras de mastite bovina. Como resultados da metodologia empregada, foram isoladas duas linhagens destas bactérias que causam diferentes tipos de mastite bovina. Os fragmentos de DNA genômico caracterizando ambas as linhagens, por meio de RAPD foram inseridos em vetor plasmidial pGEM e clonados por meio de clones T10 F1 de Escherichia coli. As seqüências obtidas permitiram desenhar iniciadores específicos para o reconhecimento de ambas as linhagens, os quais foram testados com amostras de S. aureus e com outras linhagens próximas. O diagnóstico por meios moleculares, pode ser realizado diretamente de amostras coletadas de rebanhos leiteiros assim como dos equipamentos de ordenha. A significância deste estudo consiste em um rápido e acurado método para localizar animais infectados, representando importante ferramenta no manejo do rebanho, na redução de custos com tratamentos e, rápida recuperação de rebanhos infectados.

  1. Survival of Staphylococcus aureus on fomites.

    Science.gov (United States)

    Cuesta, Alicia; Nastri, Natalia; Bernat, Maria; Brusca, Maria; Turcot, Liliana; Nastri, Maria; Rosa, Alcira C

    2008-01-01

    The aim of this study was to evaluate duration of survival of Staphylococcus aureus on contaminated standardized fomites, such as sterilization paper (SP) and polyester previously sterilized in a steam autoclave, and to determine the potential inhibitory effects of the substrates (fabrics used to manufacture garments and special wrapping paper used in the dental setting) using the bacteriostasis test. The test was performed on two types of sterile standardized samples (T1 and T2). Sterility of the samples was validated following the protocol in use at the Department of Microbiology, after which the samples were inoculated with 50 microl of a calibrated suspension of Staphylococcus aureus (reference strain ATCC 25923) in the exponential growth phase, in a final concentration of 10(7) cfu/ml and 10(6) cfu/ml). The samples were incubated at 27 degrees C and survival and concentration of microorganisms attached to the surface of the substrates was determined at the following experimental time points: immediately post-contamination, and 3 hours, 24 hours, 3 days, and 7 days post-contamination. Recovery was determined and expressed as a percentage; the bacteriostasis test was performed and showed negative results. Our results suggest that the quantity of recovered microorganisms varies according to the type of substrate and that there is a relation between survival and incubation time of the inoculated substrate serving as an artificial niche.

  2. Antimicrobial drug resistance ofStaphylococcus aureus in dairy products

    Institute of Scientific and Technical Information of China (English)

    Sasidharan S; Prema B; Yoga Latha L

    2011-01-01

    Objective:To evaluate the prevalence of multidrug resistantStaphylococcus aureus(S. aureus) in dairy products.Methods:Isolation and identification ofS. aureus were performed in3 dairy-based food products. The isolates were tested for their susceptibility to5 different common antimicrobial drugs.Results:Of50 samples examined,5 (10%) were contaminated with S. aureus. Subsequently, the5 isolates were subjected to antimicrobial resistance pattern using five antibiotic discs (methicillin, vancomycin, kanamycin, chloramphenicol and tetracycline). Sample 29 showed resistance to methicillin and vancomycin. Sample18 showed intermediate response to tetracycline. The other samples were susceptible to all the antibiotics tested.Conclusions:The results provide preliminary data on sources of food contamination which may act as vehicles for the transmission of antimicrobial-resistantStaphylococcus.Therefore, it enables us to develop preventive strategies to avoid the emergence of new strains of resistantS. aureus.

  3. Quantitation of Staphylococcus aureus in seawater using CHROMagar SA.

    Science.gov (United States)

    Tice, Alan D; Pombo, David; Hui, Jennifer; Kurano, Michelle; Bankowski, Matthew J; Seifried, Steven E

    2010-01-01

    A microbiological algorithm has been developed to analyze beach water samples for the determination of viable colony forming units (CFU) of Staphylococcus aureus (S. aureus). Membrane filtration enumeration of S. aureus from recreational beach waters using the chromogenic media CHROMagar SA alone yields a positive predictive value (PPV) of 70%. Presumptive CHROMagar SA colonies were confirmed as S. aureus by 24-hour tube coagulase test. Combined, these two tests yield a PPV of 100%. This algorithm enables accurate quantitation of S. aureus in seawater in 72 hours and could support risk-prediction processes for recreational waters. A more rapid protocol, utilizing a 4-hour tube coagulase confirmatory test, enables a 48-hour turnaround time with a modest false negative rate of less than 10%.

  4. Specificity for human hemoglobin enhances Staphylococcus aureus infection

    Science.gov (United States)

    Pishchany, Gleb; McCoy, Amanda L.; Torres, Victor J.; Krause, Jens C.; Crowe, James E.; Fabry, Mary E.; Skaar, Eric P.

    2010-01-01

    SUMMARY Iron is required for bacterial proliferation and Staphylococcus aureus steals this metal from host hemoglobin during invasive infections. This process involves hemoglobin binding to the cell wall of S. aureus, heme extraction, passage through the cell envelope, and degradation to release free iron. Herein we demonstrate an enhanced ability of S. aureus to bind hemoglobin derived from humans as compared to other mammals. Increased specificity for human hemoglobin (hHb) translates into an improved ability to acquire iron and is entirely dependent on the staphylococcal hemoglobin receptor IsdB. This feature affects host-pathogen interaction as demonstrated by the increased susceptibility of hHb expressing mice to systemic staphylococcal infection. Interestingly, enhanced utilization of human hemoglobin is not a uniform property of all bacterial pathogens. These results suggest a step in the evolution of S. aureus to better colonize the human host and establish hHb expressing mice as a model of S. aureus pathogenesis. PMID:21147468

  5. Genetic diversity of community-associated methicillin-resistant Staphylococcus aureus isolated from Tenerife Island, Spain.

    Science.gov (United States)

    Rivero-Pérez, Belinda; Alcoba-Flórez, Julia; Méndez-Álvarez, Sebastián

    2012-04-01

    With the recent detection of MRSA (methicillin-resistant Staphylococcus aureus) infections in patients lacking health care-related risk factors, the term community-acquired MRSA (CA-MRSA) has been become widely recognised. Many cases of CA-MRSA spreading to the community have been described worldwide. The aim of this study was to determine the features of CA-MRSA isolates from Tenerife Island. Toward this end, one hundred MRSA isolates were collected from eight different health regions, and their molecular features were investigated. This study revealed a wide variety of MRSA clones, including an emergent ST: ST1434 (CC8) and two new spa types, t7575 (ST125) and t7678 (ST22). The PVL genes were found in only five isolates belonging to unrelated lineages, ST8, ST30 and ST22, which could indicate at least three independent introductions of PVL(+) strains to Tenerife. Moreover, we detected that hospital MRSA clones, like EMRSA-15 and EMRSA-16, had spread to the community and are now circulating in both environments. Therefore, in our study, the CDC's rules were not specific enough to distinguish CA-MRSA from HA-MRSA. Thus, we think that the current epidemiological information is not enough to discriminate between both MRSAs, and it is necessary for prevention guidelines to include the routine determination of at least the genetic background, the antimicrobial susceptibility profile, and the PVL genes of each MRSA isolate.

  6. Animal and human Staphylococcus aureus associated clonal lineages and high rate of Staphylococcus pseudintermedius novel lineages in Spanish kennel dogs: predominance of S. aureus ST398.

    Science.gov (United States)

    Gómez-Sanz, Elena; Torres, Carmen; Benito, Daniel; Lozano, Carmen; Zarazaga, Myriam

    2013-10-25

    Methicillin-susceptible Staphylococcus aureus (MSSA) and Staphylococcus pseudintermedius (MSSP) are gaining interest to track the evolution of emerging methicillin-resistant strains in animals and humans. We focused on the characterization of the methicillin-susceptible coagulase-positive staphylococci (MSCoPS) recovered from nasal samples of 98 healthy kennel-dogs. Isolates were typed by spa, agr, MLST and SmaI/ApaI-PFGE. Antimicrobial resistance and virulence profiles were investigated. Presence of the human-associated Immune-Evasion-Cluster (IEC) genes was analyzed in MSSA. Twenty-four MSSA, 16 MSSP and one MS Staphylococcus schleiferi subspecies coagulans were obtained. Thirteen spa-types and 12 sequence-types (STs) were detected among MSSA, with ST398 predominance (7/24, 29.2%). MSSA isolates were enclosed within 6 clonal complexes (no. of isolates): CC5 (8), CC398 (7), CC88 (4), CC45 (2), CC133 (1), and CC22 (1), and one singleton. High clonal diversity was observed among MSSP, and 14 STs (10 of them new) were detected. Twelve (50%) MSSA and 12 (75%) MSSP isolates showed resistance to at least one of the tested antimicrobials, with low MSSA penicillin resistance (5 isolates) and high MSSP tetracycline resistance (9 isolates). MSSA isolates ST398, ST133, ST1 and ST2329[new] were susceptible to all antimicrobials and were the only ones lacking the scn, chp and/or sak IEC genes. High diversity of enterotoxin genes was detected among non-ST398/ST133 MSSA isolates. MSSP showed a more homogeneous virulence genes profile. Our results give evidence that dogs can be S. aureus carriers of not only typical human associated lineages but also lineages commonly detected among other animal species. Continue surveillance on CoPS in dogs is required to unveil their role in the dissemination of clones adapted to other animal species.

  7. Cloning

    Science.gov (United States)

    ... sheep that have been genetically modified to produce milk that contains a human protein essential for blood clotting. The hope is that someday this protein can be purified from the milk and given to humans whose blood does not ...

  8. Cloning

    Institute of Scientific and Technical Information of China (English)

    彭荣华

    2002-01-01

    As we come near to the 21st century, it is clear than ever that science and technology are changing the way we live and work. The breakthroughs1 in bioengineering2 science are helping to uncover the mysteries of life, holding out new hope for life-saving cures to some of our greatly terrible diseases.

  9. Efektivitas Ekstrak Daun Jambu Biji Buah Putih Terhadap Pertumbuhan Staphylococcus aureus Dari Abses Dan Staphylococcus aureus (ATCC® 29213™)

    OpenAIRE

    Sinurat, Jojor

    2016-01-01

    Daun jambu biji mengandung senyawa aktif seperti tanin, triterpenoid, flavonoid, saponin yang mempunyai efek antibakteri. Mekanisme tanin sebagai antibakteri dengan mengkerutkan dinding sel dan membran sel, inaktivasi enzim, inaktivasi fungsi materi genetik bakteri. Flavonoid merusak sel bakteri, denaturasi protein, inaktivasi enzim dan menyebabkan lisis. Triterpenoid dan saponin menghambat pertumbuhan Staphylococcus aureus dengan cara merusak struktur membran sel. Staphylococcus aureus adala...

  10. Co-colonization and clonal diversity of methicillin-sensitive and methicillin-resistant Staphylococcus aureus in sows.

    Science.gov (United States)

    Fetsch, Alexandra; Roesler, Uwe; Kraushaar, Britta; Friese, Anika

    2016-03-15

    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.

  11. Petrifilm rapid S. aureus Count Plate method for rapid enumeration of Staphylococcus aureus in selected foods: collaborative study.

    Science.gov (United States)

    Silbernagel, K M; Lindberg, K G

    2001-01-01

    A rehydratable dry-film plating method for Staphylococcus aureus in foods, the 3M Petrifilm Rapid S. aureus Count Plate method, was compared with AOAC Official Method 975.55 (Staphylococcus aureus in Foods). Nine foods-instant nonfat dried milk, dry seasoned vegetable coating, frozen hash browns, frozen cooked chicken patty, frozen ground raw pork, shredded cheddar cheese, fresh green beans, pasta filled with beef and cheese, and egg custard-were analyzed for S. aureus by 13 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample and 3 levels of inoculated test sample, each in duplicate. The mean log counts for the methods were comparable for pasta filled with beef and cheese; frozen hash browns; cooked chicken patty; egg custard; frozen ground raw pork; and instant nonfat dried milk. The repeatability and reproducibility variances of the Petrifilm Rapid S. aureus Count Plate method were similar to those of the standard method.

  12. Prevalence and characteristics of community carriage of methicillin-resistant Staphylococcus aureus in Malta.

    Science.gov (United States)

    Scerri, Jeanesse; Monecke, Stefan; Borg, Michael A

    2013-09-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen worldwide. Malta is one of the countries with the highest MRSA prevalence in Europe, as identified from hospital blood cultures [1]. However, community prevalence of MRSA has never previously been investigated. This study aimed at establishing the prevalence of community MRSA nasal colonization in Maltese individuals and identifying the clonal characteristics of the detected isolates. Nasal swabs were collected from 329 healthy individuals who were also asked to complete a brief questionnaire about risk factors commonly associated with MRSA carriage and infection. The swabs were transported and enriched in a nutrient broth supplemented with NaCl. The presence of MRSA was then determined by culturing on MRSA Select chromogenic agar and then confirming by several assays, including catalase, coagulase and PBP2a agglutination tests. The isolates were assayed for antibiotic susceptibilities and typed by microarray analysis to determine the clonal characteristics of each strain. The prevalence of MRSA nasal colonization in the healthy Maltese population was found to be 8.81% (95% confidence interval [CI], 5.75-11.87%), much higher than that found in other studies carried out in several countries. No statistical association was found between MRSA carriage and demographics or risk factors; however, this was hindered by the small sample size. Almost all the isolates were fusidic-acid resistant. The majority were found to belong to a local endemic clone (CC5) which seems to be replacing the previously prevalent European clone UK-EMRSA-15 in the country. A new clone (CC50-MRSA-V) was also characterized. The presence of such a significant community reservoir of MRSA increases the burdens already faced by the local healthcare system to control the MRSA epidemic. Colonization of MRSA in otherwise healthy individuals may represent a risk for endogenous infection and transmission to hospitalized

  13. Cloning cattle: the methods in the madness.

    Science.gov (United States)

    Oback, Björn; Wells, David N

    2007-01-01

    Somatic cell nuclear transfer (SCNT) is much more widely and efficiently practiced in cattle than in any other species, making this arguably the most important mammal cloned to date. While the initial objective behind cattle cloning was commercially driven--in particular to multiply genetically superior animals with desired phenotypic traits and to produce genetically modified animals-researchers have now started to use bovine SCNT as a tool to address diverse questions in developmental and cell biology. In this paper, we review current cattle cloning methodologies and their potential technical or biological pitfalls at any step of the procedure. In doing so, we focus on one methodological parameter, namely donor cell selection. We emphasize the impact of epigenetic and genetic differences between embryonic, germ, and somatic donor cell types on cloning efficiency. Lastly, we discuss adult phenotypes and fitness of cloned cattle and their offspring and illustrate some of the more imminent commercial cattle cloning applications.

  14. Towards Clone Detection in UML Domain Models

    DEFF Research Database (Denmark)

    Störrle, Harald

    2013-01-01

    Code clones (i.e., duplicate fragments of code) have been studied for long, and there is strong evidence that they are a major source of software faults. Anecdotal evidence suggests that this phenomenon occurs similarly in models, suggesting that model clones are as detrimental to model quality...... as they are to code quality. However, programming language code and visual models have significant differences that make it difficult to directly transfer notions and algorithms developed in the code clone arena to model clones. In this article, we develop and propose a definition of the notion of “model clone” based...... on the thorough analysis of practical scenarios. We propose a formal definition of model clones, specify a clone detection algorithm for UML domain models, and implement it prototypically. We investigate different similarity heuristics to be used in the algorithm, and report the performance of our approach. While...

  15. Effective and efficient model clone detection

    DEFF Research Database (Denmark)

    Störrle, Harald

    2015-01-01

    Code clones are a major source of software defects. Thus, it is likely that model clones (i.e., duplicate fragments of models) have a significant negative impact on model quality, and thus, on any software created based on those models, irrespective of whether the software is generated fully...... automatically (“MDD-style”) or hand-crafted following the blueprint defined by the model (“MBSD-style”). Unfortunately, however, model clones are much less well studied than code clones. In this paper, we present a clone detection algorithm for UML domain models. Our approach covers a much greater variety...... of model types than existing approaches while providing high clone detection rates at high speed....

  16. Nuclear transfer technology in mammalian cloning.

    Science.gov (United States)

    Wolf, D P; Mitalipov, S; Norgren, R B

    2001-01-01

    The past several years have witnessed remarkable progress in mammalian cloning using nuclear transfer (NT). Until 1997 and the announcement of the successful cloning of sheep from adult mammary gland or fetal fibroblast cells, our working assumption was that cloning by NT could only be accomplished with relatively undifferentiated embryonic cells. Indeed, live offspring were first produced by NT over 15 years ago from totipotent, embryonic blastomeres derived from early cleavage-stage embryos. However, once begun, the progression to somatic cell cloning or NT employing differentiated cells as the source of donor nuclei was meteoric, initially involving differentiated embryonic cell cultures in sheep in 1996 and quickly thereafter, fetal or adult somatic cells in sheep, cow, mouse, goat, and pig. Several recent reviews provide a background for and discussion of these successes. Here we will focus on the potential uses of reproductive cloning along with recent activities in the field and a discussion concerning current interests in human reproductive and therapeutic cloning.

  17. Impact of target site distribution for Type I restriction enzymes on the evolution of methicillin-resistant Staphylococcus aureus (MRSA) populations.

    Science.gov (United States)

    Roberts, Gareth A; Houston, Patrick J; White, John H; Chen, Kai; Stephanou, Augoustinos S; Cooper, Laurie P; Dryden, David T F; Lindsay, Jodi A

    2013-08-01

    A limited number of Methicillin-resistant Staphylococcus aureus (MRSA) clones are responsible for MRSA infections worldwide, and those of different lineages carry unique Type I restriction-modification (RM) variants. We have identified the specific DNA sequence targets for the dominant MRSA lineages CC1, CC5, CC8 and ST239. We experimentally demonstrate that this RM system is sufficient to block horizontal gene transfer between clinically important MRSA, confirming the bioinformatic evidence that each lineage is evolving independently. Target sites are distributed randomly in S. aureus genomes, except in a set of large conjugative plasmids encoding resistance genes that show evidence of spreading between two successful MRSA lineages. This analysis of the identification and distribution of target sites explains evolutionary patterns in a pathogenic bacterium. We show that a lack of specific target sites enables plasmids to evade the Type I RM system thereby contributing to the evolution of increasingly resistant community and hospital MRSA.

  18. Occurrence of enterotoxin genes and macrorestriction analysis of Staphylococcus aureus isolated from bovine mastitis and bulk-tank milk samples in Italy. An epidemiological study

    Directory of Open Access Journals (Sweden)

    Roberto Rosmini

    2010-01-01

    Full Text Available The goal of the study was to genotypically compare S. aureus isolates from mastitis milk and raw milk to identify therelation between strains and to assess the enterotoxigenicity of the isolates. Eighty-three Staphylococcus aureus isolatesrecovered from cows and bulk tank milk of five farms in northern Italy were compared genotypically. The genes for theenterotoxins A, D, G and I, but not for B, C, E and H and the toxic shock syndrome toxin 1 (TSST-1, were detected byPCR amplification. Macrorestriction analysis with the restrictions enzyme SmaI revealed 14 pulsed-field gel electrophoresispatterns. These were in part different from each other only in a few fragments and thus displayed a closeclonal relation. The results of the present investigation showed that identical or closely related clones seemed to beresponsible for the cases of bovine mastitis in the farms investigated and partly responsible for contamination of bulktank milk.

  19. The molecular changing mechanism of Ampicillin-Sulbactam resistant Staphylococcus aureus towards Methicillin resistant Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Mieke Hemiawati Satari

    2005-12-01

    Full Text Available The aim of this study was to determine the molecular changing of S.aureus, which is resistant to Ampicillin-Sulbactam and then become resistant to Methicillin as a result of improper dosage. The study was conducted by isolating Ampicillin-Sulbactam resistant and Methicillin Resistant S.aureus (MRSA, afterwards an amplification process was performed by PCR (Polymerase Chain Reaction. to isolate the betalactamase enzyme regulator and PBP 2a genes. The result of this research showed that there were a deletion of few amino acids from the regulator gene, and a suspicion that the DNA sequence had been substituted from PBP 2 gene into PBP 2a (gen mec. This process had formed MRSA.

  20. Optimal quantum cloning via stimulated emission

    CERN Document Server

    Simon, C; Zeilinger, Anton; Simon, Christoph; Weihs, Gregor; Zeilinger, Anton

    2000-01-01

    We show that optimal universal quantum cloning can be realized via stimulated emission. Universality of the cloning procedure is achieved by choosing systems that have appropriate symmetries. We first discuss a scheme based on stimulated emission in certain three-level-systems, e.g. atoms in a cavity. Then we present a way of realizing optimal universal cloning based on stimulated parametric down-conversion. This scheme also implements the optimal universal NOT operation.

  1. Quantum cloning with an optical fiber amplifier

    CERN Document Server

    Fasel, S; Ribordy, G; Scarani, V; Zbinden, H; Fasel, Sylvain; Gisin, Nicolas; Ribordy, Gregoire; Scarani, Valerio; Zbinden, Hugo

    2002-01-01

    It has been shown theoretically that a light amplifier working on the physical principle of stimulated emission should achieve optimal quantum cloning of the polarization state of light. We demonstrate close-to-optimal universal quantum cloning of polarization in a standard fiber amplifier for telecom wavelengths. For cloning $1\\to 2$ we find a fidelity of 0.82, the optimal value being ${5/6}=0.83$.

  2. Experimental Quantum Cloning of Single Photons

    CERN Document Server

    Lamas-Linares, A; Howell, J C; Bouwmeester, D; Lamas-Linares, Antia; Simon, Christoph; Howell, John C.; Bouwmeester, Dik

    2002-01-01

    Although perfect copying of unknown quantum systems is forbidden by the laws of quantum mechanics, approximate cloning is possible. A natural way of realizing quantum cloning of photons is by stimulated emission. In this context the fundamental quantum limit to the quality of the clones is imposed by the unavoidable presence of spontaneous emission. In our experiment a single input photon stimulates the emission of additional photons from a source based on parametric down-conversion. This leads to the production of quantum clones with near optimal fidelity. We also demonstrate universality of the copying procedure by showing that the same fidelity is achieved for arbitrary input states.

  3. Metabolomic phenotyping of a cloned pig model

    Directory of Open Access Journals (Sweden)

    Callesen Henrik

    2011-08-01

    Full Text Available Abstract Background Pigs are widely used as models for human physiological changes in intervention studies, because of the close resemblance between human and porcine physiology and the high degree of experimental control when using an animal model. Cloned animals have, in principle, identical genotypes and possibly also phenotypes and this offer an extra level of experimental control which could possibly make them a desirable tool for intervention studies. Therefore, in the present study, we address how phenotype and phenotypic variation is affected by cloning, through comparison of cloned pigs and normal outbred pigs. Results The metabolic phenotype of cloned pigs (n = 5 was for the first time elucidated by nuclear magnetic resonance (NMR-based metabolomic analysis of multiple bio-fluids including plasma, bile and urine. The metabolic phenotype of the cloned pigs was compared with normal outbred pigs (n = 6 by multivariate data analysis, which revealed differences in the metabolic phenotypes. Plasma lactate was higher for cloned vs control pigs, while multiple metabolites were altered in the bile. However a lower inter-individual variability for cloned pigs compared with control pigs could not be established. Conclusions From the present study we conclude that cloned and normal outbred pigs are phenotypically different. However, it cannot be concluded that the use of cloned animals will reduce the inter-individual variation in intervention studies, though this is based on a limited number of animals.

  4. Quantum cloning disturbed by thermal Davies environment

    Science.gov (United States)

    Dajka, Jerzy; Łuczka, Jerzy

    2016-06-01

    A network of quantum gates designed to implement universal quantum cloning machine is studied. We analyze how thermal environment coupled to auxiliary qubits, `blank paper' and `toner' required at the preparation stage of copying, modifies an output fidelity of the cloner. Thermal environment is described in terms of the Markovian Davies theory. We show that such a cloning machine is not universal any more but its output is independent of at least a part of parameters of the environment. As a case study, we consider cloning of states in a six-state cryptography's protocol. We also briefly discuss cloning of arbitrary input states.

  5. Species-specific challenges in dog cloning.

    Science.gov (United States)

    Kim, G A; Oh, H J; Park, J E; Kim, M J; Park, E J; Jo, Y K; Jang, G; Kim, M K; Kim, H J; Lee, B C

    2012-12-01

    Somatic cell nuclear transfer (SCNT) is now an established procedure used in cloning of several species. SCNT in dogs involves multiple steps including the removal of the nuclear material, injection of a donor cell, fusion, activation of the reconstructed oocytes and finally transfer to a synchronized female recipient. There are therefore many factors that contribute to cloning efficiency. By performing a retrospective analysis of 2005-2012 published papers regarding dog cloning, we define the optimum procedure and summarize the specific feature for dog cloning.

  6. Salicylic acid enhances Staphylococcus aureus extracellular adhesin protein expression.

    Science.gov (United States)

    Alvarez, Lucía P; Barbagelata, María S; Cheung, Ambrose L; Sordelli, Daniel O; Buzzola, Fernanda R

    2011-11-01

    One of the virulence factors required by Staphylococcus aureus at the early stages of infection is Eap, a secreted adhesin that binds many host proteins and is upregulated by the two-component regulatory system saeRS. The S. aureus Newman strain harbors a mutation in saeS that is thought to be responsible for the high level of Eap expression in this strain. This study was designed to ascertain whether salicylic acid (SAL) affects the expression of Eap and the internalization of S. aureus into epithelial cells. The strain Newman treated with SAL exhibited increased levels of eap transcription and protein expression. Furthermore, SAL treatment increased the eap promoter activity. SAL treatment enhanced Eap expression in the Newman and in other S. aureus strains that do not carry the mutation in saeS. Internalization of S. aureus eap and sae mutants into the MAC-T epithelial cells was significantly decreased compared with the wild-type counterparts. In conclusion, we demonstrated that a low concentration of SAL increased S. aureus Eap expression possibly due to enhancement of sae. SAL may create the conditions for S. aureus persistence in the host, not only by decreasing the capsular polysaccharide expression as shown before, but also by enhancing Eap expression.

  7. Neutrophil-mediated phagocytosis of Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Jos A.G. Van Strijp

    2014-09-01

    Full Text Available For invading staphylococci, phagocytosis an killing bij human neutrophils is the biggest threat. Neutrophils are the only cells that can effectively kill staphylococci by engulfment and subsequent bombardment with proteases, amidases, antimicrobial peptides and proteins in concert with reactive oxygen species that are generated during the metabolic burst.Both complement and antibodies are crucial for effective uptake and neutrophil activation. S. aureus is not an innocent bystander in this process. It actively secretes several proteins to impair every single step in this process from receptor modulation, to complement inhibition to neutrophil lysis to protease, antimicrobial peptide inhibition and resistance to reactive oxygen species. For the design of future novel antimicrobial strategies: therapeutic antibodies, vaccines, novel antibiotics, all this should be taken into account. Still the best way to treat diseases is to help to enhance the natural defence mechanism that are already in place.

  8. Cloning and sequence analysis of genes encoding Staphylococcus hyicus exfoliative toxin types A, B, C, and D

    DEFF Research Database (Denmark)

    Ahrens, Peter; Andresen, Lars Ole

    2004-01-01

    Exfoliative toxins produced by certain strains of Staphylococcus hyicus mediate exudative epidermitis in pigs. In this study the genes coding for four different exfoliative toxin from S. hyicus (ExhA, ExhB, ExhC, and ExhD) were cloned and sequenced. The coding sequence of the four toxin genes...... ranged from 816 to 834 bp. The amino acid sequences of these four toxins were homologous to the earlier described exfoliative toxins SHETB from S. hyicus and ETA, ETB, and ETD from Staphylococcus aureus. The homology between the S. hyicus toxins was at the same level as the homology to the exfoliative...

  9. Structural and functional characterization of Staphylococcus aureus dihydrodipicolinate synthase.

    Science.gov (United States)

    Girish, Tavarekere S; Sharma, Eshita; Gopal, B

    2008-08-20

    Lysine biosynthesis is crucial for cell-wall formation in bacteria. Enzymes involved in lysine biosynthesis are thus potential targets for anti-microbial therapeutics. Dihydrodipicolinate synthase (DHDPS) catalyzes the first step of this pathway. Unlike its homologues, Staphylococcus aureus DHDPS is a dimer both in solution and in the crystal and is not feedback inhibited by lysine. The crystal structure of S. aureus DHDPS in the free and substrate bound forms provides a structural rationale for its catalytic mechanism. The structure also reveals unique conformational features of the S. aureus enzyme that could be crucial for the design of specific non-competitive inhibitors.

  10. The Effect of Essential Oils on Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Seda Ozdikmenli

    2014-05-01

    Full Text Available Diseases caused by Staphylococcus aureus are widespread through the world in spite of developing technology. S. aureus is an important pathogen causing food intoxications besides hospital infections by its antibiotic resistant strains. Nowadays, there has been worldwide increasing concern on usage of natural products to control microorganisms. One of these natural products is essential oils. They are produced from plants especially from spices and composed of many components and volatiles. This review summarizes informative literature on essential oils and their mode of antimicrobial action. In addition, current knowledge on in vitro researches on antibacterial activity of essential oils and food applications to control S. aureus has been discussed.

  11. ES cells derived from cloned embryos in monkey - a jump toward human therapeutic cloning

    Institute of Scientific and Technical Information of China (English)

    Xiangzhong Yang; Sadie L Smith

    2007-01-01

    @@ Therapeutic cloning refers to the derivation of embryonic stem cells (ntESC) from embryos derived from somatic cell nuclear transfer (SCNT) also known as cloning. Cloning involves transplanting a differentiated cell into an oocyte that has had its nucleus (DNA) removed.

  12. Cloning non-transformed sheep B cells.

    Science.gov (United States)

    Griebel, P J; Beskorwayne, T; Godson, D L; Popowych, Y; Hein, W

    2000-04-03

    The capacity to clone B cells and establish permanent B cell lines has greatly facilitated a wide variety of studies characterising the growth, differentiation, and gene expression of murine and human B cells. Similar investigations of B cell biology for other species have been severely restricted by an inability to culture or clone B cells. This is the first report of a method to clone non-transformed sheep B cells using a culture system based on murine CD154 and a combination of human gamma chain-common cytokines. Sheep Peyer's patch B cells were cultured for 120 days and then cloned by limiting dilution culture. The parental B cell culture contained both surface immunoglobulin (sIg)M(+) and sIgG1(+) B cells and both types of B cell were cloned. Clonality was confirmed by PCR analysis of Ig heavy chain (HC) and light chain (LC) expression and DNA sequencing of HC V genes. There was agreement between the PCR and flow cytometric analyses of HC isotype expression on the B cell clones but the available monoclonal antibodies specific for sheep lambda and kappa LC did not react with all clones. Soluble Ig was detected in the culture supernatant of sIgG1(+) clones but not sIgM(+) clones. The B cell clones remained dependent upon CD154 and gamma chain-common cytokine co-stimulation for sustained growth and maintained stable Ig expression. The cloning of non-transformed sheep B cells should provide a valuable tool for studying sheep B cell biology, establishing Ig HC- and LC-specific monoclonal antibodies, analysing the B cell Ig repertoire, and may be used to produce sheep monoclonal antibodies.

  13. Staphylococcus aureus intestinal colonization is associated with increased frequency of S. aureus on skin of hospitalized patients

    Directory of Open Access Journals (Sweden)

    Donskey Curtis J

    2007-09-01

    Full Text Available Abstract Background Intestinal colonization by Staphylococcus aureus among hospitalized patients has been associated with increased risk of staphylococcal infection and could potentially contribute to transmission. We hypothesized that S. aureus intestinal colonization is associated with increased frequency of S. aureus on patients' skin and nearby environmental surfaces. Methods Selected inpatients were cultured weekly for S. aureus from stool, nares, skin (groin and axilla, and environmental surfaces (bed rail and bedside table. Investigator's hands were cultured after contacting the patients' skin and the environmental surfaces. Results Of 71 subjects, 32 (45.1% had negative nares and stool cultures, 23 (32.4% had positive nares and stool cultures, 13 (18.3% were nares carriers only, and 3 (4.2% were stool carriers only. Of the 39 patients with S. aureus carriage, 30 (76.9% had methicillin-resistant isolates. In comparison to nares colonization only, nares and intestinal colonization was associated with increased frequency of positive skin cultures (41% versus 77%; p = 0.001 and trends toward increased environmental contamination (45% versus 62%; p = 0.188 and acquisition on investigator's hands (36% versus 60%; p = 0.057. Patients with negative nares and stool cultures had low frequency of S. aureus on skin and the environment (4.8% and 11.3%, respectively. Conclusion We found that hospitalized patients with S. aureus nares and/or stool carriage frequently had S. aureus on their skin and on nearby environmental surfaces. S. aureus intestinal colonization was associated with increased frequency of positive skin cultures, which could potentially facilitate staphylococcal infections and nosocomial transmission.

  14. Staphylococcus aureus in former Portuguese colonies from Africa and the Far East: missing data to help fill the world map.

    Science.gov (United States)

    Conceição, T; Coelho, C; Silva, I Santos; de Lencastre, H; Aires-de-Sousa, M

    2015-09-01

    The aim of the present study was to determine the prevalence and risk factors for methicillin-resistant Staphylococcus aureus (MRSA) nasal carriage among patients and healthcare workers in Angola (ANG), São Tomé and Príncipe (STP), Cape Verde (CV) and East Timor (ET), and to characterize the antimicrobial susceptibility, virulence content and population structure of all S. aureus. Despite the importance of MRSA as a major human pathogen, data from these former Portuguese colonies in Africa and Asia are scarce. A total of 2065 nasal swabs recovered between 2010-14 were included in the study. Antimicrobial susceptibility testing and molecular characterization of S. aureus showed: (i) a very high MRSA prevalence in ANG (61.6%), moderate in STP (25.5%), low in CV (5.6%) and null in ET; (ii) a high prevalence of Panton-Valentine leukocidin in STP (36.8%), ET (29.2%) and CV (28.3%) contrasting with ANG (7.9%); (iii) ST5-SCCmecIVa, ST8-IV/V and ST5-VI were the major MRSA clones in ANG (65.2%), STP (44.8%) and CV (50%), respectively; (iv) a high resistance to trimethoprim-sulfamethoxazole in ANG (66.5%) and STP (50.9%), to rifampin in ANG (77.3%), and to tetracycline in STP (26.3%) and ET (20.8%); (v) three major methicillin-susceptible S. aureus clones (ST15, ST508, ST152) were present in all four countries. Age <18 years (OR 2.03, 95% CI 1.24-3.31), previous surgery (OR 2.45, 95% CI 1.24-4.83), no smoking (OR 4.04, 95% CI 1.05-15.50), and longer hospitalization (OR 2.53, 95% CI 1.49-4.28) were risk factors for MRSA carriage. This study provided the first comprehensive overview on MRSA in former Portuguese colonies in Africa and Asia, missing data in the world map.

  15. Retrospective study of necrotizing fasciitis and characterization of its associated Methicillin-resistant Staphylococcus aureus in Taiwan

    Directory of Open Access Journals (Sweden)

    Changchien Chih-Hsuan

    2011-10-01

    Full Text Available Abstract Background Methicillin-resistant Staphylococcus aureus (MRSA has emerged as a prevalent pathogen of necrotizing fasciitis (NF in Taiwan. A four-year NF cases and clinical and genetic differences between hospital acquired (HA- and community-acquired (CA-MRSA infection and isolates were investigated. Methods A retrospective study of 247 NF cases in 2004-2008 and antimicrobial susceptibilities, staphylococcal chromosomal cassette mec (SCCmec types, pulsed field gel electrophoresis (PFGE patterns, virulence factors, and multilocus sequence typing (MLST of 16 NF-associated MRSA in 2008 were also evaluated. Results In 247 cases, 42 microbial species were identified. S. aureus was the major prevalent pathogen and MRSA accounted for 19.8% of NF cases. Most patients had many coexisting medical conditions, including diabetes mellitus, followed by hypertension, chronic azotemia and chronic hepatic disease in order of decreasing prevalence. Patients with MRSA infection tended to have more severe clinical outcomes in terms of amputation rate (p S. aureus or non-S. aureus infection. NF patients infected by HA-MRSA had a significantly higher amputation rate, comorbidity, C-reactive protein level, and involvement of lower extremity than those infected by CA-MRSA. In addition to over 90% of MRSA resistant to erythromycin and clindamycin, HA-MRSA was more resistant than CA-MRSA to trimethoprim-sulfamethoxazole (45.8% vs. 4%. ST59/pulsotype C/SCCmec IV and ST239/pulsotype A/SCCmec III isolates were the most prevalent CA- and HA-MRSA, respectively in 16 isolates obtained in 2008. In contrast to the gene for γ-hemolysin found in all MRSA, the gene for Panton-Valentine leukocidin was only identified in ST59 MRSA isolates. Other three virulence factors TSST-1, ETA, and ETB were occasionally identified in MRSA isolates tested. Conclusion NF patients with MRSA infection, especially HA-MRSA infection, had more severe clinical outcomes than those infected by

  16. Challenges in regulating farm animal cloning

    DEFF Research Database (Denmark)

    Gunning, Jennifer; Hartlev, Mette; Gamborg, Christian

    Report from the project Cloning in Public: A specific support action within the 6th framework programme, priority 5: Food quality and safety......Report from the project Cloning in Public: A specific support action within the 6th framework programme, priority 5: Food quality and safety...

  17. Progress in interspecies cloning of mammals

    Institute of Scientific and Technical Information of China (English)

    WEN Duancheng; BI Chunming; CHEN Dayuan

    2004-01-01

    Interspecies mammalian cloning can be achieved by application of two key techniques, i.e.the technique of interspecies nuclear transfer and the technique of interspecies pregnancy.The general principles, problems and possible solutions, as well as the recent advances of interspecies mammalian cloning have been summarized in this review.

  18. Cloning of endangered mammalian species: any progress?

    Science.gov (United States)

    Loi, Pasqualino; Galli, Cesare; Ptak, Grazyna

    2007-05-01

    Attempts through somatic cell nuclear transfer to expand wild populations that have shrunk to critical numbers is a logical extension of the successful cloning of mammals. However, although the first mammal was cloned 10 years ago, nuclear reprogramming remains phenomenological, with abnormal gene expression and epigenetic deregulation being associated with the cloning process. In addition, although cloning of wild animals using host oocytes from different species has been successful, little is known about the implication of partial or total mitochondrial DNA heteroplasmy in cloned embryos, fetuses and offspring. Finally, there is a need for suitable foster mothers for inter-intra specific cloned embryos. Considering these issues, the limited success achieved in cloning endangered animals is not surprising. However, optimism comes from the rapid gain in the understanding of the molecular clues underlying nuclear reprogramming. If it is possible to achieve a controlled reversal of the differentiated state of a cell then it is probable that other issues that impair the cloning of endangered animals, such as the inter-intra species oocyte or womb donor, will be overcome in the medium term.

  19. Probabilistic cloning with supplementary information

    CERN Document Server

    Azuma, K; Koashi, M; Imoto, N; Azuma, Koji; Shimamura, Junichi; Koashi, Masato; Imoto, Nobuyuki

    2005-01-01

    We consider probabilistic cloning of a state chosen from a mutually nonorthogonal set of pure states, with the help of a party holding supplementary information in the form of pure states. When the number of states is two, we show that the best efficiency of producing m copies is always achieved by a two-step protocol in which the helping party first attempts to produce m-1 copies from the supplementary state, and if it fails, then the original state is used to produce m copies. On the other hand, when the number of states exceeds two, the best efficiency is not always achieved by such a protocol. We give examples in which the best efficiency is not achieved even if we allow any amount of one-way classical communication from the helping party.

  20. Finding Code Clones for Refactoring with Clone Metrics : A Case Study of Open Source Software

    OpenAIRE

    Choi, Eunjong; Yoshida, Norihiro; IshioTakashi; Inoue, Katsuro; Sano, Tateki

    2011-01-01

    A code clone is a code fragment that has identical or similar code fragments to it in the source code. Code clone has been regarded as one of the factors that makes software maintenance more difficult. Therefore, to refactor code clones into one method is promising way to reduce maintenance cost in the future. In our previous study, we proposed a method to extract code clones for refactoring using clone metrics. We had conducted an empirical study on Java application developed by NEC Corporat...

  1. Cloned Sheep May Age Prematurely

    Institute of Scientific and Technical Information of China (English)

    Joseph; B.Verrengia; 孙颖

    1999-01-01

    1996年的头条科技新闻之一是:多利羊被克隆成功。世人曾为消息雀跃,以为克隆技术马上可以造福人类了,而且科幻作家也开始忙碌起来。而今,当多利羊过3岁生日时,人们却伤感地发现: In Dolly’s case,she is 3,but her genetic material is aging at the rate of the6-year-old sheep from which she was cloned. 这就是所谓aging prematurely。这则消息给人们带来的忧虑有两条。一是:被克隆的动物的预期寿命比人们想象的要短;二是:人们是否能够有效利用克隆的人体细胞去治疗疾病。目前,科学家们的担心还是集中于后者。本书收入的另一篇有关克隆的文章(It’s A Boy!Scientists Clone First Male Mammal)和本篇构成了强烈的对照,可谓一喜一忧。然而,无论喜忧,人类在克隆技术方面正在以坚实的步伐向前迈进。

  2. [Recovery of Staphylococcus aureus after acid injury in milk products].

    Science.gov (United States)

    Assis, E M; De Carvalho, E P; Asquieri, E R; Robbs, P G

    1994-01-01

    The growth behavior of Staphylococcus aureus in fresh Cheese (Minas and Muzzarella) during their shelf-life was studied. The possible injury of this microorganism caused by the increasing acidity was also investigated. Raw milk was inoculated with 10(6) cells/ml (S. aureus FRIA-100) and the cheese production was performed according to normal procedures. Minas and muzzarella cheese were stored at 7 degrees C for 40 and 60 days, respectively. At 2-3 days intervals, the following analysis were performed: acidity, pH, S. aureus counting using agar Baird Parker by the traditional methods and by the method recommended by the American Public Health Association to evaluate the reparation of injured cells. We had a secure indication of the presence of injured S. aureus when acidity was in the range of 0.7 to 0.8% expressed in lactic acid and when the cycle was 1.3 log higher than the traditional one.

  3. Improving Diagnosis and Treatment of Staphylococcus aureus Infections : Experimental Studies

    NARCIS (Netherlands)

    S. van den Berg (Sanne)

    2015-01-01

    markdownabstract__Abstract__ Staphylococcus aureus is an opportunistic pathogen that causes a variety of infections, ranging from mild skin infections like furuncles and impetigo, to severe, lifethreatening infections including endocarditis, osteomyelitis and pneumonia. Invasive infections are freq

  4. Environmental Staphylococcus aureus contamination in a Tunisian hospital.

    Science.gov (United States)

    Gharsa, Haythem; Dziri, Raoudha; Klibi, Naouel; Chairat, Sarra; Lozano, Carmen; Torres, Carmen; Bellaaj, Ridha; Slama, Karim Ben

    2016-12-01

    One hundred hospital environment samples were obtained in 2012 in a Tunisian hospital and tested for Staphylococcus aureus recovery. Antimicrobial resistance profile and virulence gene content were determined. Multilocus-sequence-typing (MLST), spa-typing, agr-typing and SmaI-pulsed-field gel electrophoresis (PFGE) were performed. Two methicillin-resistant S. aureus (MRSA) isolates typed as: ST247-t052-SCCmecI-agrI were recovered from the intensive care unit (ICU). Ten samples contained methicillin-susceptible S. aureus (MSSA) and these samples were collected in different services, highlighting the presence of the tst gene encoding the toxic shock syndrome toxin as well as the lukED, hla, hlb, hld and hlgv virulence genes in some of the isolates. In conclusion, we have shown that the hospital environment could be a reservoir contributing to dissemination of virulent S. aureus and MRSA.

  5. METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS STRAINS IN FOOD AND ANIMAL

    Directory of Open Access Journals (Sweden)

    A. Traversa

    2010-03-01

    Full Text Available Some authors reported the possibility of a methicillin-resistant Staphylococcus aureus (MRSA human infections from meat and dairy products and methicillin-resistant Staphylococcus intermedius isolation in animals. The aim of this study is to investigate the methicillin-resistance in S. aureus strains and in S. intermedius strains (food and wild animals. 236 S.aureus strains from food, 36 S.aureus strains and 1 S. intermedius strain from wild animals were analyzed. 2 (0.74% MRSA strains from bovine milk were phenotipically resistant to cefoxitin, grew on chromogenic medium (MRSA Brilliance Oxoid and were mecA positive. All MRSA strains had the spa-type t899. All mecA positive strains showed at least resistance to eight of the antibiotics tested but none to glicopeptides. Both MRSA strains were enterotoxigenic.

  6. Rapid, Culture-Free Detection of Staphylococcus aureus Bacteremia

    Science.gov (United States)

    Burghardt, Elliot L.; Flenker, Katie S.; Clark, Karen C.; Miguel, Jeff; Ince, Dilek; Winokur, Patricia; Ford, Bradley; McNamara, James O.

    2016-01-01

    S. aureus bacteremia (SAB) is a common condition with high rates of morbidity and mortality. Current methods used to diagnose SAB take at least a day, and often longer. Patients with suspected bacteremia must therefore be empirically treated, often unnecessarily, while assay results are pending. In this proof-of-concept study, we describe an inexpensive assay that detects SAB via the detection of micrococcal nuclease (an enzyme secreted by S. aureus) in patient plasma samples in less than three hours. In total, 17 patient plasma samples from culture-confirmed S. aureus bacteremic individuals were tested. 16 of these yielded greater nuclease assay signals than samples from uninfected controls or individuals with non-S. aureus bacteremia. These results suggest that a nuclease-detecting assay may enable the rapid and inexpensive diagnosis of SAB, which is expected to substantially reduce the mortality and morbidity that result from this condition. PMID:27305148

  7. [Human cloning in Muslim and Arab law].

    Science.gov (United States)

    Aldeeb Abu-Sahlieh, Sami A

    2009-01-01

    Cloning is a modern medical procedure that Muslim religious authorities treat en resorting to the general principles established by classical Muslim law based on the Koran and the Sunnah of Muhhamad as the messenger of God. In this regard, human beings are not capable of deciding what is or what is not lawful without resorting to divine norms. Cloning clashes with several principles. Firstly, the principle of the respect for life in relation to surpernumeraries, but Muslim authors are not in unanimous agreement on the determination of the moment at which life begins. Secondly, is the respect of progeny: cloning could only take place between a married couple. But even if these two principles are respected, cloning poses two major problems: the diversity of species expounded by the Koran and the Sunnah and a lack of interest. Which explains the quasi-unanimous opposition of Muslim writings regarding cloning.

  8. Identification of gene clusters associated with host adaptation and antibiotic resistance in Chinese Staphylococcus aureus isolates by microarray-based comparative genomics.

    Directory of Open Access Journals (Sweden)

    Henan Li

    Full Text Available A comparative genomic microarray comprising 2,457 genes from two whole genomes of S. aureus was employed for the comparative genome hybridization analysis of 50 strains of divergent clonal lineages, including methicillin-resistant S. aureus (MRSA, methicillin-susceptible S. aureus (MSSA, and swine strains in China. Large-scale validation was confirmed via polymerase chain reaction in 160 representative clinical strains. All of the 50 strains were clustered into seven different complexes by phylogenetic tree analysis. Thirteen gene clusters were specific to different S. aureus clones. Ten gene clusters, including seven known (vSa3, vSa4, vSaα, vSaβ, Tn5801, and phage ϕSa3 and three novel (C8, C9, and C10 gene clusters, were specific to human MRSA. Notably, two global regulators, sarH2 and sarH3, at cluster C9 were specific to human MRSA, and plasmid pUB110 at cluster C10 was specific to swine MRSA. Three clusters known to be part of SCCmec, vSa4 or Tn5801, and vSaα as well as one novel gene cluster C12 with homology with Tn554 of S. epidermidis were identified as MRSA-specific gene clusters. The replacement of ST239-spa t037 with ST239-spa t030 in Beijing may be a result of its acquisition of vSa4, phage ϕSa1, and ϕSa3. In summary, thirteen critical gene clusters were identified to be contributors to the evolution of host specificity and antibiotic resistance in Chinese S. aureus.

  9. Long-term mortality after Staphylococcus aureus spondylodiscitis

    DEFF Research Database (Denmark)

    Aagaard, Theis; Larsen, Anders R; Roed-Petersen, Casper;

    2014-01-01

    Patients diagnosed with Staphylococcus aureus spondylodiscitis have increased long-term mortality compared with the background population mainly due to infectious, endocrine, cardiovascular, gastrointestinal and alcohol and drug abuse-related diseases.......Patients diagnosed with Staphylococcus aureus spondylodiscitis have increased long-term mortality compared with the background population mainly due to infectious, endocrine, cardiovascular, gastrointestinal and alcohol and drug abuse-related diseases....

  10. Tea Tree Oil-Induced Transcriptional Alterations in Staphylococcus aureus

    OpenAIRE

    Cuaron, Jesus A.; Dulal, Santosh; Song, Yang; Singh, Atul K; Montelongo, Cesar E.; Yu, Wanqin; Nagarajan, Vijayaraj; Jayaswal, Radheshyam K.; Wilkinson, Brian J; Gustafson, John E.

    2012-01-01

    Tea tree oil (TTO) is a steam distillate of Melaleuca alternifolia that demonstrates broad-spectrum antibacterial activity. This study was designed to document how TTO challenge influences the Staphylococcus aureus transcriptome. Overall, bioinformatic analyses (S. aureus microarray meta-database) revealed that both ethanol and TTO induce related transcriptional alterations. TTO challenge led to the down-regulation of genes involved with energy-intensive transcription and translation, and alt...

  11. Phenotype Switching Is a Natural Consequence of Staphylococcus aureus Replication

    OpenAIRE

    Edwards, Andrew M.

    2012-01-01

    The pathogen Staphylococcus aureus undergoes phenotype switching in vivo from its normal colony phenotype (NCP) to a slow-growing, antibiotic-resistant small-colony-variant (SCV) phenotype that is associated with persistence in host cells and tissues. However, it is not clear whether phenotype switching is the result of a constitutive process that is selected for under certain conditions or is triggered by particular environmental stimuli. Examination of cultures of diverse S. aureus strains ...

  12. Staphylococcus aureus in the community: colonization versus infection.

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    Maureen Miller

    Full Text Available BACKGROUND: Antibiotic-resistant Staphylococcus aureus infections have increased dramatically in the community, yet S. aureus nasal colonization has remained stable. The objectives of this study were to determine if S. aureus colonization is a useful proxy measure to study disease transmission and infection in community settings, and to identify potential community reservoirs. METHODOLOGY/PRINCIPAL FINDINGS: Randomly selected households in Northern Manhattan, completed a structured social network questionnaire and provided nasal swabs that were typed by pulsed field gel electrophoresis to identify S. aureus colonizing strains. The main outcome measures were: 1 colonization with S. aureus; and 2 recent serious skin infection. Risk factor analyses were conducted at both the individual and the household levels; logistic regression models identified independent risks for household colonization and infection. RESULTS: 321 surveyed households contained 914 members. The S. aureus prevalence was 25% and MRSA was 0.4%. More than 40% of households were colonized. Recent antibiotic use was the only significant correlate for household colonization (p = .002. Seventy-eight (24% households reported serious skin infection. In contrast with colonization, five of the six risk factors that increased the risk of skin infection in the household at the univariate level remained independently significant in multivariable analysis: international travel, sports participation, surgery, antibiotic use and towel sharing. S. aureus colonization was not significantly associated with serious skin infection in any analysis. Among multiperson households with more than one person colonized, 50% carried the same strain. CONCLUSIONS/SIGNIFICANCE: The lack of association between S. aureus nasal colonization and serious skin infection underscores the need to explore alternative venues or body sites that may be crucial to transmission. Moreover, the magnitude of colonization and

  13. Staphylococcus aureus nasal carriage in hemodialysis centers of Fez, Morocco.

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    Idrissa Diawara

    2014-06-01

    Full Text Available Staphylococcus aureus (S. aureus nasal carriage may be responsible for some serious infections in hemodialyzed patients. The main target of this study was to estimate the prevalence of S. aureus nasal carriage in hemodialysis outpatients and medical staff in hemodialysis centers specifically in Fez region. The second target is to identify the risks of colonization, resistance pattern of isolates and their virulence toxin genes.Nasal swab specimens were obtained from 143 hemodialyzed outpatients and 32 medical staff from January to June 2012. Each participant completed a short questionnaire. Nasal carriage of S. aureus was demographically related (age, gender, hemodialysis duration, comorbidity (diabetes, malignancy and exposure to health care (dialysis staff, hospitalization. PCR (Polymerase Chain Reaction were used on all the isolates in the research of twelve staphylococcal enterotoxins genes. Also the PCR was used to investigate on the three factors epidermal cell differentiation inhibitors; three exfoliatin toxins; two leukotoxins; the toxic shock syndrome toxin-1 and the hemolysin beta genes.Nasal screening revealed 38.16%, 50% and 18.75% S. aureus carries in chronic, acute hemodialysis patients and medical staff, respectively. Only young participants were likely to be S. aureus carries (p = 0.002. But there were no gender differences between the isolate carriers and non-carriers or some comorbidity factors such as viral hepatitis B and C, HIV (Human Immunodeficiency Virus infections, diabetes, chronic smoking, recent hospitalization or antibiotic therapy. Out of all isolates, only one (1.61% was methicillin-resistant and Twenty-one (33.87% had at least two virulence toxin genes.Knowledge and monitoring of antibiotic resistance profile and virulence of S. aureus carriage are essential in the treatment of infections generated by this pathogen, as well as in the control of clonal dissemination and prevent the spread of S. aureus resistance.

  14. Prevalence of Staphylococcus aureus in Shrimps in Tehran during 2013

    OpenAIRE

    Mohammad Mehdi Soltan Dallal; Abbas Rahimi Foroushani; Sara Sharifi –Yazdi; Mohammad Kazem Sharifi -Yazdi; Noushin Arfatahery

    2016-01-01

    Background During fishing and transport, preservation and quality of fish products are importantas well as storage to prevent the growth of pathogenic and toxin producing bacteria.Staphylococcus aureus is one of the most common causes of sea food-borne diseases worldwidedue to contamination of food by preformed enterotoxins. The aim of this study was to compare theprevalence and contamination of S. aureus in marine and farmed shrimps in Tehran fishery center.Methods: A total of 300 samples, i...

  15. IDENTIFIKASI MIKROORGANISE STAPHYLOCOCCUS AUREUS PADA PENDERITA ANGULAR CHEILITIS

    OpenAIRE

    MINARTI, NURHAERATUL

    2012-01-01

    2011 Pada suatu penelitian tentang Angular cheilitis ditemukan Staphylococcus aureus hampir dua kali dari candida albicans. Oleh karena itu tujuan penelitian ini adalah untuk mengidentifikasi pengaruh Staphylococcus aureus pada penyakit Angular cheilitis. Sampel penelitian adalah 30 pasien yang datang ke Rumah Sakit Gigi dan Mulut Halimah Daeng Sikati Kandea Bagian Penyakit Gigi dan Mulut dalam periode waktu bulan Oktober-November 2011. Apusan pada permukaan lesi angular cheilitis dima...

  16. AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach.

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    Hannes M Beyer

    Full Text Available Assembly cloning is increasingly replacing conventional restriction enzyme and DNA-ligase-dependent cloning methods for reasons of efficiency and performance. Here, we describe AQUA (advanced quick assembly, a simple and versatile seamless assembly cloning approach. We demonstrate the applicability and versatility of AQUA Cloning in selected proof-of-principle applications including targeted insertion-, deletion- and site-directed point-mutagenesis, and combinatorial cloning. Furthermore, we show the one pot de novo assembly of multiple DNA fragments into a single circular plasmid encoding a complex light- and chemically-regulated Boolean A NIMPLY B logic operation. AQUA Cloning harnesses intrinsic in vivo processing of linear DNA fragments with short regions of homology of 16 to 32 bp mediated by Escherichia coli. It does not require any kits, enzymes or preparations of reagents and is the simplest assembly cloning protocol to date.

  17. Rise and subsequent decline of community-associated methicillin resistant Staphylococcus aureus ST30-IVc in Copenhagen, Denmark through an effective search and destroy policy

    DEFF Research Database (Denmark)

    Bartels, M D; Kristoffersen, K; Boye, K;

    2010-01-01

    , evaluating the effect of MRSA eradication therapy (ET), and finding links among patients. Twenty-three index patients infected with the ST30-IVc clone from November 2003 to September 2005 were contacted and transmission chains were studied. The majority of ST30-IVc patients had a connection......The number of patients with community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has increased rapidly in Copenhagen, Denmark since 2003. Patients with the typical Panton-Valentine leukocidin-positive CA-MRSA clone ST30-IVc were contacted with the aim of treating MRSA carriers...... to the Philippines. Household members were screened for MRSA and all members of families with MRSA carriers were offered treatment of the carrier state and were followed for 1 year. MRSA carriers were found in seven of 16 households and transmission occurred among close contacts and in kindergartens. Five days of ET...

  18. Molecular characterisation of meticillin-resistant Staphylococcus aureus isolates from two ceftobiprole Phase 3 complicated skin and skin-structure infection clinical trials.

    Science.gov (United States)

    Davies, Todd A; Shang, Wenchi; Amsler, Karen M; Bajaksouzian, Saralee; Jacobs, Michael R; Bush, Karen

    2009-08-01

    Meticillin-resistant Staphylococcus aureus (MRSA) isolates from two worldwide ceftobiprole Phase 3 clinical trials for the treatment of complicated skin and skin-structure infections were characterised by clonality, staphylococcal cassette chromosome mec (SCCmec) type and the presence of Panton-Valentine leukocidin (PVL). PVL was predominantly found in US isolates (196/231 vs. 13/110 non-US isolates). SCCmec type IV was the most common (253/329) owing to the predominance of clone USA300 in isolates from the USA (197/226). In Europe, SCCmec type III was the most prevalent (30/74). Ceftobiprole minimum inhibitory concentrations (MICs) ranged from 0.25 microg/mL to 4 microg/mL, with MICs clone type.

  19. Detoxification of toxins by bacillithiol in Staphylococcus aureus.

    Science.gov (United States)

    Newton, Gerald L; Fahey, Robert C; Rawat, Mamta

    2012-04-01

    Bacillithiol (BSH), an α-anomeric glycoside of l-cysteinyl-d-glucosaminyl-l-malate, is a major low-molecular-mass thiol found in bacteria such as Bacillus sp., Staphylococcus aureus and Deinococcus radiodurans. Like other low-molecular-mass thiols such as glutathione and mycothiol, BSH is likely to be involved in protection against environmental toxins including thiol-reactive antibiotics. We report here a BSH-dependent detoxification mechanism in S. aureus. When S. aureus Newman strain was treated with monobromobimane and monochlorobimane, the cellular BSH was converted to the fluorescent S-conjugate BS-bimane. A bacillithiol conjugate amidase activity acted upon the BS-bimane to produce Cys-bimane, which was then acetylated by an N-acetyltransferase to generate N-acetyl-Cys-bimane, a mercapturic acid. An S. aureus mutant lacking BSH did not produce mercapturic acid when treated with monobromobimane and monochlorobimane, confirming the involvement of bacillithiol. Furthermore, treatment of S. aureus Newman with rifamycin, the parent compound of the first-line anti-tuberculosis drug, rifampicin, indicated that this thiol-reactive antibiotic is also detoxified in a BSH-dependent manner, since mercapturic acids of rifamycin were observed in the culture medium. These data indicate that toxins and thiol-reactive antibiotics are detoxified to less potent mercapturic acids in a BSH-dependent manner and then exported out of the cell in S. aureus.

  20. Determining of antibiotic resistance profile inStaphylococcus aureus isolates

    Institute of Scientific and Technical Information of China (English)

    Hossein Motamedi; Hadis Mirzabeigi; Tahere Shirali

    2010-01-01

    Objective:To determine the pattern of antibiotic resistance amongStaphylococcus aureus (S. aureus) isolates from clinical specimens and to identify community-acquired methicillin-resistantStaphylococcus aureus(CA-MRSA)in specimens that have been collected from patients referring to one of the hospitals of Ahvaz.Methods:S. aureus isolates from a hospital in Ahvaz were screened for resistance to various antibiotics including methicillin. The susceptibility of the isolates was determined by Kirby-Bauer disc diffusion method. TheMRSA was also treated with ethidium bromide to find the origin of resistance.Results: Among the bacterial isolates, all of 11S. aureus were resistant to methicillin and cefixime,2 were resistant to ciprofloxacine,6 were resistant to tetracycline and the reminder were sensitive or intermediate to other antibiotics. The treated isolates were reminded resistant to methicillin and this suggested that the plasmid was not the origin of resistance in these isolates.Conclusions: These results showed that infection due toMRSA is widespread in Ahvaz and with respect to the spread of vancomycin resistance among MRSA and appearance of overwhelming infections. It is necessary to identify continuously the profile of antibiotic resistance amongS. aureus isolates in other regions and finding appropriate antibiotic for infection control and eradication.

  1. Genotyping of methicillin resistant staphylococcus aureus from tertiary care hospitals in Coimbatore, South India

    Directory of Open Access Journals (Sweden)

    Toms John Peedikayil Neetu

    2016-01-01

    Full Text Available Background: Globally, methicillin resistant Staphylococcus aureus (MRSA is one of the most common pathogens that causes hospital- and community-acquired infections. The use of molecular typing methods is essential for determining the origin of the isolates, their clonal relations, and also epidemiological investigations. Objective: The purpose of this study was to determine the prevalence of antibiotic-resistant MRSA investigate the accessory gene regulator (agr and staphylococcal cassette chromosome mec (SCCmec types and perform multilocus sequence typing (MLST. Furthermore, the minimum inhibitory concentration of MRSA isolates was determined for vancomycin and daptomycin. Materials and Methods: Two hundred and fifty-nine MRSA isolates were collected from Tertiary Care Hospitals in Coimbatore. Disk diffusion method was employed to assess the sensitivity of MRSA isolates to selected antibiotics and genetic analysis was performed using SCCmec, agr, and MLST typing by multiplex-polymerase chain reaction strategy. Minimal inhibitory concentration (MIC was determined using Ezy MIC (vancomycin and Biomerieux (daptomycin E-test strip. Results: Of 259 MRSA isolates, 209 (80.7% were confirmed as methicillin resistant. Antibiotic susceptibility pattern revealed that all the MRSA isolates were 100% sensitive to linezolid, rifampicin, teicoplanin, and vancomycin. MIC results showed that of 209 MRSA isolates, 10 were found to be vancomycin intermediate S. aureus and 100% of the MRSA isolates were daptomycin-susceptible. The agr group I and SCCmec Type III were the major type among MRSA isolates. In addition to these MLST typing revealed the prevalence of sequence type (ST 239 (SLV of ST8 among the MRSA isolates. Conclusion: This study confirms that ST239 (Brazilian clone of MRSA is predominant in this region which is responsible for the hospital-acquired MRSA infections. Thus, the study also suggests that vancomycin and daptomycin can still be used as an

  2. Subtle genetic changes enhance virulence of methicillin resistant and sensitive Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Hawes Alicia C

    2007-11-01

    Full Text Available Abstract Background Community acquired (CA methicillin-resistant Staphylococcus aureus (MRSA increasingly causes disease worldwide. USA300 has emerged as the predominant clone causing superficial and invasive infections in children and adults in the USA. Epidemiological studies suggest that USA300 is more virulent than other CA-MRSA. The genetic determinants that render virulence and dominance to USA300 remain unclear. Results We sequenced the genomes of two pediatric USA300 isolates: one CA-MRSA and one CA-methicillin susceptible (MSSA, isolated at Texas Children's Hospital in Houston. DNA sequencing was performed by Sanger dideoxy whole genome shotgun (WGS and 454 Life Sciences pyrosequencing strategies. The sequence of the USA300 MRSA strain was rigorously annotated. In USA300-MRSA 2658 chromosomal open reading frames were predicted and 3.1 and 27 kilobase (kb plasmids were identified. USA300-MSSA contained a 20 kb plasmid with some homology to the 27 kb plasmid found in USA300-MRSA. Two regions found in US300-MRSA were absent in USA300-MSSA. One of these carried the arginine deiminase operon that appears to have been acquired from S. epidermidis. The USA300 sequence was aligned with other sequenced S. aureus genomes and regions unique to USA300 MRSA were identified. Conclusion USA300-MRSA is highly similar to other MRSA strains based on whole genome alignments and gene content, indicating that the differences in pathogenesis are due to subtle changes rather than to large-scale acquisition of virulence factor genes. The USA300 Houston isolate differs from another sequenced USA300 strain isolate, derived from a patient in San Francisco, in plasmid content and a number of sequence polymorphisms. Such differences will provide new insights into the evolution of pathogens.

  3. Production and applications of an N-terminally-truncated recombinant beta-haemolysin from Staphylococcus aureus.

    Science.gov (United States)

    Singh, M; Singh, A; Sharma, A

    2014-07-01

    The beta-haemolysin of Staphylococcus aureus (SA-hlb) is a secreted neutral sphingomyelinase (nSMase) implicated in the pathogenesis of infection and responsible for the characteristic in vitro 'hot-cold' haemolytic ability of the bacterium. Here, we describe the production of a biologically active N-terminally-truncated recombinant SA-hlb protein for use in in vitro assays and as a research tool. Using local isolates of S. aureus, we PCR-amplified an SA-hlb DNA sequence of 891 nucleotides, 99 nucleotides shorter than the full-length molecule, before cloning and sequencing (GenBank accession no. JN580071). The pQE.TriSystem vector (Qiagen, Germany) was used to express recombinant SA-hlb (r-SA-hlb) with a C-terminal 8xHis tag in Escherichia coli JM107 cells. Both JM107 lysate and the purified r-SA-hlb possessed hot-cold lytic activity against sheep and buffalo erythrocytes, which was abolished by incubation at ≥90 °C for 30 min or exposure to dithiothreitol, and could be neutralized by bovine immune sera. Purified r-SA-hlb was also cytotoxic to buffalo mononuclear cells and was effective as a coating antigen for indirect ELISA to screen for reactive sera. Importantly, the r-SA-hlb was suitable for use as a β-toxin in the modified CAMP test. We conclude that the r-SA-hlb protein produced was functionally active and has numerous potential applications.

  4. Prevalence of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus in Retail Ready-to-Eat Foods in China.

    Science.gov (United States)

    Yang, Xiaojuan; Zhang, Jumei; Yu, Shubo; Wu, Qingping; Guo, Weipeng; Huang, Jiahui; Cai, Shuzhen

    2016-01-01

    Staphylococcus aureus, particularly methicillin-resistant S.aureus (MRSA), is a life-threatening pathogen in humans, and its presence in food is a public health concern. MRSA has been identified in foods in China, but little information is available regarding MRSA in ready-to-eat (RTE) foods. We aimed to investigate the prevalence of S. aureus and MRSA in Chinese retail RTE foods. All isolated S. aureus were tested for antimicrobial susceptibility, and MRSA isolates were further characterized by multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. Of the 550 RTE foods collected from 2011 to 2014, 69 (12.5%) were positive for S. aureus. Contamination levels were mostly in the range of 0.3-10 most probable number (MPN)/g, with five samples exceeding 10 MPN/g. Of the 69 S. aureus isolates, seven were identified as MRSA by cefoxitin disc diffusion test. Six isolates were mecA-positive, while no mecC-positive isolates were identified. In total, 75.8% (47/62) of the methicillin-susceptible S. aureus isolates and all of the MRSA isolates were resistant to three or more antibiotics. Amongst the MRSA isolates, four were identified as community-acquired strains (ST59-MRSA-IVa (n = 2), ST338-MRSA-V, ST1-MRSA-V), while one was a livestock-associated strain (ST9, harboring an unreported SCCmec type 2C2). One novel sequence type was identified (ST3239), the SCCmec gene of which could not be typed. Overall, our findings showed that Chinese retail RTE foods are likely vehicles for transmission of multidrug-resistant S. aureus and MRSA lineages. This is a serious public health risk and highlights the need to implement good hygiene practices.

  5. Control of Staphylococcus aureus pathogenicity island excision.

    Science.gov (United States)

    Mir-Sanchis, Ignacio; Martínez-Rubio, Roser; Martí, Miguel; Chen, John; Lasa, Íñigo; Novick, Richard P; Tormo-Más, María Ángeles; Penadés, José R

    2012-09-01

    Staphylococcus aureus pathogenicity islands (SaPIs) are a group of related 15-17 kb mobile genetic elements that commonly carry genes for superantigen toxins and other virulence factors. The key feature of their mobility is the induction of SaPI excision and replication by certain phages and their efficient encapsidation into specific small-headed phage-like infectious particles. Previous work demonstrated that chromosomal integration depends on the SaPI-encoded recombinase, Int. However, although involved in the process, Int alone was not sufficient to mediate efficient SaPI excision from chromosomal sites, and we expected that SaPI excision would involve an Xis function, which could be encoded by a helper phage or by the SaPI, itself. Here we report that the latter is the case. In vivo recombination assays with plasmids in Escherichia coli demonstrate that SaPI-coded Xis is absolutely required for recombination between the SaPI att(L) and att(R) sites, and that both sites, as well as their flanking SaPI sequences, are required for SaPI excision. Mutational analysis reveals that Xis is essential for efficient horizontal SaPI transfer to a recipient strain. Finally, we show that the master regulator of the SaPI life cycle, Stl, blocks expression of int and xis by binding to inverted repeats present in the promoter region, thus controlling SaPI excision.

  6. Inhibition of major integrin αV β3 reduces Staphylococcus aureus attachment to sheared human endothelial cells.

    Science.gov (United States)

    McDonnell, C J; Garciarena, C D; Watkin, R L; McHale, T M; McLoughlin, A; Claes, J; Verhamme, P; Cummins, P M; Kerrigan, S W

    2016-12-01

    Essentials Staphylococcus aureus (S. aureus) binds and impairs function of vascular endothelial cells (EC). We investigated the molecular signals triggered by S. aureus adhesion to EC. Inhibition of the EC integrin αVβ3 reduces S. aureus binding and rescues EC function. αVβ3 blockade represents an attractive target to treat S. aureus bloodborne infections.

  7. ANTISTAPHYBASE: database of antimicrobial peptides (AMPs) and essential oils (EOs) against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus.

    Science.gov (United States)

    Zouhir, Abdelmajid; Taieb, Malek; Lamine, Mohamed Ashraf; Cherif, Ammar; Jridi, Taoufik; Mahjoubi, Basma; Mbarek, Sarra; Fliss, Ismail; Nefzi, Adel; Sebei, Khaled; Ben Hamida, Jeannette

    2017-03-01

    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/ .

  8. Aberrant DNA methylation in cloned ovine embryos

    Institute of Scientific and Technical Information of China (English)

    LIU Lei; HOU Jian; LEI TingHua; BAI JiaHua; GUAN Hong; AN XiaoRong

    2008-01-01

    By using the approach of immunofluorescence staining with an antibody against 5-methylcytosine (5MeC), the present study detected the DNA methylation patterns of cloned ovine embryos. The em-bryos derived from in vitro fertilization were also examined for reference purpose. The results showed that: (1) during the pre-implantation development, cloned embryos displayed a similar demethylation profile to the fertilized embryos; that is, the methylation level decreased to the lowest at 8-cell stage, and then increased again at morulae stage. However, methylation level was obviously higher in cloned embryos than in stage-matched fertilized embryos, especially at 8-cell stage and afterwards; (2) at blastocyst stage, the methylation pattern in cloned embryos was different from that in fertilized em-bryos. In cloned blastocyst, inner cell mass (ICM) exhibited a comparable level to trophectoderm cells (TE), while in in-vitro fertilized blastocyst the methylation level of ICM was lower than that of TE, which is not consistent with that reported by other authors. These results indicate that DNA methylation is abnormally reprogrammed in cloned embryos, implying that aberrant DNA methylation reprogramming may be one of the factors causing cloned embryos developmental failure.

  9. Human embryo cloning prohibited in Hong Kong.

    Science.gov (United States)

    Liu, Athena

    2005-12-01

    Since the birth of Dolly (the cloned sheep) in 1997, debates have arisen on the ethical and legal questions of cloning-for-biomedical-research (more commonly termed "therapeutic cloning") and of reproductive cloning using human gametes. Hong Kong enacted the Human Reproductive Technology Ordinance (Cap 561) in 2000. Section 15(1)(e) of this Ordinance prohibits the "replacing of the nucleus of a cell of an embryo with a nucleus taken from any other cell," i.e., nucleus substitution. Section 15(1)(f) prohibits the cloning of any embryo. The scope of the latter, therefore, is arguably the widest, prohibiting all cloning techniques such as cell nucleus replacement, embryo splitting, parthenogenesis, and cloning using stem cell lines. Although the Human Reproductive Technology Ordinance is not yet fully operative, this article examines how these prohibitions may adversely impact on basic research and the vision of the Hong Kong scientific community. It concludes that in light of recent scientific developments, it is time to review if the law offers a coherent set of policies in this area.

  10. The molecular epidemiology of the highly virulent ST93 Australian community Staphylococcus aureus strain.

    Directory of Open Access Journals (Sweden)

    Geoffrey W Coombs

    Full Text Available In Australia the PVL-positive ST93-IV [2B], colloquially known as "Queensland CA-MRSA" has become the dominant CA-MRSA clone. First described in the early 2000s, ST93-IV [2B] is associated with skin and severe invasive infections including necrotizing pneumonia. A singleton by multilocus sequence typing (MLST eBURST analysis ST93 is distinct from other S. aureus clones. To determine if the increased prevalence of ST93-IV [2B] is due to the widespread transmission of a single strain of ST93-IV [2B] the genetic relatedness of 58 S. aureus ST93 isolated throughout Australia over an extended period were studied in detail using a variety of molecular methods including pulsed-field gel electrophoresis, spa typing, MLST, microarray DNA, SCCmec typing and dru typing. Identification of the phage harbouring the lukS-PV/lukF-PV Panton Valentine leucocidin genes, detection of allelic variations in lukS-PV/lukF-PV, and quantification of LukF-PV expression was also performed. Although ST93-IV [2B] is known to have an apparent enhanced clinical virulence, the isolates harboured few known virulence determinants. All PVL-positive isolates carried the PVL-encoding phage ΦSa2USA and the lukS-PV/lukF-PV genes had the same R variant SNP profile. The isolates produced similar expression levels of LukF-PV. Although multiple rearrangements of the spa sequence have occurred, the core genome in ST93 is very stable. The emergence of ST93-MRSA is due to independent acquisitions of different dru-defined type IV and type V SCCmec elements in several spa-defined ST93-MSSA backgrounds. Rearrangement of the spa sequence in ST93-MRSA has subsequently occurred in some of these strains. Although multiple ST93-MRSA strains were characterised, little genetic diversity was identified for most isolates, with PVL-positive ST93-IVa [2B]-t202-dt10 predominant across Australia. Whether ST93-IVa [2B] t202-dt10 arose from one PVL-positive ST93-MSSA-t202, or by independent acquisitions of

  11. Meticillin-resistant Staphylococcus aureus: spread of specific lineages among patients in different wards at a Brazilian teaching hospital.

    Science.gov (United States)

    Cavalcante, F S; Schuenck, R P; Ferreira, D C; da Costa, C R; Nouér, S A; dos Santos, K R N

    2014-02-01

    This study aimed to characterize meticillin-resistant Staphylococcus aureus (MRSA) lineages circulating in a Brazilian teaching hospital. MRSA isolates from nasal swabs were evaluated to assess antimicrobial susceptibility, staphylococcal cassette chromosome mec (SCCmec), Panton-Valentine leucocidin status, pulsed-field gel electrophoresis profile and multi-locus sequence type (MLST) analysis. Eighty-three MRSA isolates were analysed. SCCmec III (43.4%) and IV (49.4%) were predominant. ST1-IV (USA400) was more common in internal medicine (P = 0.002) whereas 'clone M' (SCCmec III) was more common in the medical and surgical intensive care unit (P = 0.004), and all isolates were ST5-IV (USA800) in dermatology (P inside the hospital and helped to establish effective control measures.

  12. The presence of heterogeneous vancomycin-lntermediate Staphylococcus aureus (heteroVISA) in a major Malaysian hospital.

    Science.gov (United States)

    Norazah, Ahmad; Law, Ngiik Ling; Kamel, Abd Ghani Mohamed; Salbiah, Nawi

    2012-06-01

    This study was conducted to detect the presence of heterogenous vancomycin-intermediate Staphylococcus aureus (heteroVISA) among MRSA isolates in a major hospital. Forty-three MRSA isolates with vancomycin MIC 2 microg/ml collected in 2009 was screened for heteroVISA using Etest Glycopeptide Resistance Detection (GRD) and confirmed by population analysis profile-area under curve method. The genetic relatedness of heteroVISA strains with other MRSA was examined by pulsed-field gel electrophoresis (PFGE) method. Two isolates were shown to be heteroVISA and derived from the same clone. This showed that heteroVISA strains were already present among our local strains since 2009 and were genetically related to other susceptible strains.

  13. The carriage of the serine-aspartate repeat protein-encoding sdr genes among Staphylococcus aureus lineages.

    Science.gov (United States)

    Liu, Huanle; Lv, Jingnan; Qi, Xiuqin; Ding, Yu; Li, Dan; Hu, Longhua; Wang, Liangxing; Yu, Fangyou

    2015-01-01

    The serine-aspartate repeat proteins (Sdr) are members of a family of surface proteins and contribute to the pathogenicity of Staphylococcus aureus. Among 288 S. aureus isolates including 158 and 130 associated with skin and soft tissue infections and bloodstream infection, respectively; 275 (95.5%) were positive for at least one of three sdr genes tested. The positivity rates for sdrC, sdrD, and sdrE among S. aureus isolates were 87.8% (253/288), 63.9% (184/288), and 68.1% (196/288), respectively. 224 (77.8%) of 288 isolates were concomitantly positive for two or three sdr genes. There was an association between carriage of sdrE and methicillin-resistant S. aureus (MRSA) isolates, while the carriage rates of sdrC and sdrD in MRSA isolates were similar to those in methicillin-sensitive S. aureus (MSSA) isolates. The prevalence of co-existence of sdrC and sdrE among MRSA isolates was significantly higher than that among MSSA isolates (p<0.05). All ST1, ST5, ST7, and ST25 isolates were positive for sdrD. While all ST121 and ST398 isolates were negative for sdrD. All ST59 and ST88 isolates were positive for sdrE. All ST1 isolates were concomitantly positive for sdrC and sdrD. Concomitant carriage of sdrC, sdrD, and sdrE was found among all ST5, 75.0% (9/12) of ST1, 69.2% (9/13) of ST6, 78.6% (11/14) of ST25, and 90.9% (20/22) of ST88 isolates. sdrD was linked to CC5, CC7 and CC88 isolates, especially CC88 isolates. There was a strong association between the presence of sdrE and CC59, CC88, and CC5 isolates. A significant correlation between concomitant carriage of sdrC, sdrD, and sdrE and CC88 isolates was found. sdrC-positive, sdrD-positive and sdrE-negative gene profile was significantly associated with CC7 clone. There was an association between sdrC-positive, sdrD-negative, and sdrE-positive gene profile and CC59 isolates. A correlation between sdrC-positive, sdrD-negative, and sdrE-negative gene profile and CC121 clone was found. More CC59 isolates carried sdr

  14. Maximum confidence measurements via probabilistic quantum cloning

    Institute of Scientific and Technical Information of China (English)

    Zhang Wen-Hai; Yu Long-Bao; Cao Zhuo-Liang; Ye Liu

    2013-01-01

    Probabilistic quantum cloning (PQC) cannot copy a set of linearly dependent quantum states.In this paper,we show that if incorrect copies are allowed to be produced,linearly dependent quantum states may also be cloned by the PQC.By exploiting this kind of PQC to clone a special set of three linearly dependent quantum states,we derive the upper bound of the maximum confidence measure of a set.An explicit transformation of the maximum confidence measure is presented.

  15. Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) at ambient freshwater beaches

    Science.gov (United States)

    Fogarty, Lisa R.; Haack, Sheridan K.; Johnson, Heather E.; Brennan, Angela K.; Isaacs, Natasha M.; Spencer, Chelsea

    2015-01-01

    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.

  16. Staphylococcus aureus small colony variants in diabetic foot infections

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    Estrella Cervantes-García

    2015-03-01

    Full Text Available Background: Staphylococcus aureus (S. aureus is one of the major pathogens causing chronic infections. The ability of S. aureus to acquire resistance to a diverse range of antimicrobial compounds results in limited treatment options, particularly in methicillin-resistant S. aureus (MRSA. A mechanism by which S. aureus develops reduced susceptibility to antimicrobials is through the formation of small colony variants (SCVs. Infections by SCVs of S. aureus are an upcoming problem due to difficulties in laboratory diagnosis and resistance to antimicrobial therapy. Methods: A prospective study was performed on 120 patients diagnosed with both type 2 diabetes mellitus and infected diabetic foot ulcers. The study was carried out from July 2012 to December 2013 in Hospital General de Mexico. The samples were cultured in blood agar, mannitol salt agar, and MacConkey agar media, and incubated at 37°C in aerobic conditions. Results: We describe the first known cases of diabetic foot infections caused by MRSA-SCVs in patients diagnosed with type 2 diabetes mellitus and infected diabetic foot ulcers. In all of our cases, the patients had not received any form of gentamicin therapy. Conclusions: The antibiotic therapy commonly used in diabetic patients with infected diabetic foot ulcers fails in the case of MRSA-SCVs because the intracellular location protects S. aureus-SCVs from the host's defenses and also helps them resist antibiotics. The cases studied in this article add to the spectrum of persistent and relapsing infections attributed to MRSA-SCVs and emphasizes that these variants may also play a relevant role in diabetic foot infections.

  17. Staphylococcus aureus small colony variants in diabetic foot infections

    Science.gov (United States)

    Cervantes-García, Estrella; García-Gonzalez, Rafael; Reyes-Torres, Angélica; Resendiz-Albor, Aldo Arturo; Salazar-Schettino, Paz María

    2015-01-01

    Background Staphylococcus aureus (S. aureus) is one of the major pathogens causing chronic infections. The ability of S. aureus to acquire resistance to a diverse range of antimicrobial compounds results in limited treatment options, particularly in methicillin-resistant S. aureus (MRSA). A mechanism by which S. aureus develops reduced susceptibility to antimicrobials is through the formation of small colony variants (SCVs). Infections by SCVs of S. aureus are an upcoming problem due to difficulties in laboratory diagnosis and resistance to antimicrobial therapy. Methods A prospective study was performed on 120 patients diagnosed with both type 2 diabetes mellitus and infected diabetic foot ulcers. The study was carried out from July 2012 to December 2013 in Hospital General de Mexico. The samples were cultured in blood agar, mannitol salt agar, and MacConkey agar media, and incubated at 37°C in aerobic conditions. Results We describe the first known cases of diabetic foot infections caused by MRSA-SCVs in patients diagnosed with type 2 diabetes mellitus and infected diabetic foot ulcers. In all of our cases, the patients had not received any form of gentamicin therapy. Conclusions The antibiotic therapy commonly used in diabetic patients with infected diabetic foot ulcers fails in the case of MRSA-SCVs because the intracellular location protects S. aureus-SCVs from the host's defenses and also helps them resist antibiotics. The cases studied in this article add to the spectrum of persistent and relapsing infections attributed to MRSA-SCVs and emphasizes that these variants may also play a relevant role in diabetic foot infections. PMID:25787018

  18. Pathogenic Staphylococcus aureus Isolates from Postoperative Wounds of Hospitalized Patients

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    Smritikana Biswas

    2010-07-01

    Full Text Available Staphylococcus sp., gram positive pyogenic bacteria located on skin, nose etc, secretes toxin that causes toxic shock syndrome, abscess, food poisoning and other infectious diseases. This study was carried out to identify and characterize the type of Staphylococcus sp. bacteria especially Staphylococcus aureus in the pus from postoperative wounds of hospitalized patients. From pus samples collected from twenty-four patients from Kharagpur Hospital, Paschim Medinipur, West Bengal, twenty-eight bacterial isolates were obtained. Among them twenty-five (89.2% were appeared with golden yellow colonies which is usually formed by Staphylococcus aureus. Twenty-three (82.14% of the bacterial isolates were Gram positive. Among them twenty isolates (86.9% were further confirmed to be Staphylococcus aureus by their ability to produce Catalase enzyme (positive in Catalase test and Coagulase enzyme (positive in Coagulase Test. Eighteen (90.00% of these Staphylococcus aureus were found to liquefy gelatin (Gelatin hydrolysis test, were able to hydrolyze urea (Urea hydrolysis test and were also l positive in Mannitol Fermentation Test. But there was no growth found of these isolates on MacConkey Agar, while sixteen isolates (80.00% of Staphylococcus aureus were resistant to penicillin (50µg/ml. Moreover eighteen (90.00% Staphylococcus aureus isolates were able to elaborate Hemolysin (Hemolysis test on Blood Agar media. Hence the bacterial isolates obtained from pus of postoperative wounds were predominantly pathogenic Staphylococcus aureus. So it can be concluded that careful treatment and postoperative measures to be taken to avoid serious health problem that may often be life threatening.

  19. A genomic portrait of the emergence, evolution, and global spread of a methicillin-resistant Staphylococcus aureus pandemic

    Science.gov (United States)

    Holden, Matthew T.G.; Hsu, Li-Yang; Kurt, Kevin; Weinert, Lucy A.; Mather, Alison E.; Harris, Simon R.; Strommenger, Birgit; Layer, Franziska; Witte, Wolfgang; de Lencastre, Herminia; Skov, Robert; Westh, Henrik; Žemličková, Helena; Coombs, Geoffrey; Kearns, Angela M.; Hill, Robert L.R.; Edgeworth, Jonathan; Gould, Ian; Gant, Vanya; Cooke, Jonathan; Edwards, Giles F.; McAdam, Paul R.; Templeton, Kate E.; McCann, Angela; Zhou, Zhemin; Castillo-Ramírez, Santiago; Feil, Edward J.; Hudson, Lyndsey O.; Enright, Mark C.; Balloux, Francois; Aanensen, David M.; Spratt, Brian G.; Fitzgerald, J. Ross; Parkhill, Julian; Achtman, Mark; Bentley, Stephen D.; Nübel, Ulrich

    2013-01-01

    The widespread use of antibiotics in association with high-density clinical care has driven the emergence of drug-resistant bacteria that are adapted to thrive in hospitalized patients. Of particular concern are globally disseminated methicillin-resistant Staphylococcus aureus (MRSA) clones that cause outbreaks and epidemics associated with health care. The most rapidly spreading and tenacious health-care-associated clone in Europe currently is EMRSA-15, which was first detected in the UK in the early 1990s and subsequently spread throughout Europe and beyond. Using phylogenomic methods to analyze the genome sequences for 193 S. aureus isolates, we were able to show that the current pandemic population of EMRSA-15 descends from a health-care-associated MRSA epidemic that spread throughout England in the 1980s, which had itself previously emerged from a primarily community-associated methicillin-sensitive population. The emergence of fluoroquinolone resistance in this EMRSA-15 subclone in the English Midlands during the mid-1980s appears to have played a key role in triggering pandemic spread, and occurred shortly after the first clinical trials of this drug. Genome-based coalescence analysis estimated that the population of this subclone over the last 20 yr has grown four times faster than its progenitor. Using comparative genomic analysis we identified the molecular genetic basis of 99.8% of the antimicrobial resistance phenotypes of the isolates, highlighting the potential of pathogen genome sequencing as a diagnostic tool. We document the genetic changes associated with adaptation to the hospital environment and with increasing drug resistance over time, and how MRSA evolution likely has been influenced by country-specific drug use regimens. PMID:23299977

  20. Molecular architecture of the regulatory Locus sae of Staphylococcus aureus and its impact on expression of virulence factors.

    Science.gov (United States)

    Steinhuber, Andrea; Goerke, Christiane; Bayer, Manfred G; Döring, Gerd; Wolz, Christiane

    2003-11-01

    We characterized the sae operon, a global regulator for virulence gene expression in Staphylococcus aureus. A Tn917 sae mutant was obtained by screening a Tn917 library of the agr mutant ISP479Mu for clones with altered hemolytic activity. Sequence analysis of the sae operon revealed two additional open reading frames (ORFs) (ORF3 and ORF4) upstream of the two-component regulatory genes saeR and saeS. Four overlapping sae-specific transcripts (T1 to T4) were detected by Northern blot analysis, and the transcriptional initiation points were mapped by primer extension analysis. The T1, T2, and T3 mRNAs are probably terminated at the same stem-loop sequence downstream of saeS. The T1 message (3.1 kb) initiates upstream of ORF4, T2 (2.4 kb) initiates upstream of ORF3, and T3 (2.0 kb) initiates in front of saeR. T4 (0.7 kb) represents a monocistronic mRNA encompassing ORF4 only. sae-specific transcripts were detectable in all of the 40 different clinical S. aureus isolates investigated. Transcript levels were at maximum during the post-exponential growth phase. The sae mutant showed a significantly reduced rate of invasion of human endothelial cells, consistent with diminished transcription and expression of fnbA. The expression of type 5 capsular polysaccharide is activated in the sae mutant of strain Newman, as shown by immunofluorescence and promoter-reporter fusion experiments. In summary, the sae operon constitutes a four-component regulator system which acts on virulence gene expression in S. aureus.

  1. Detecção de Staphylococcus aureus utilizando a técnica de REP-PCR no monitoramento da qualidade do leite de cabra Detection of Staphylococcus aureus using the REP-PCR technique to monitor goat milk quality

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    Cellyneude de Souza Olivindo

    2009-07-01

    Full Text Available Este estudo foi realizado com o objetivo de aplicar a técnica de REP-PCR no monitoramento da qualidade do leite de cabra por meio da detecção de Staphylococcus aureus em amostras de mãos de ordenhador, tetos das cabras, leite, ordenhadeira e água para estabelecimento e implantação do sistema de Análise de Perigos e Pontos Críticos de Controle (APPCC. Verificaram-se vários fingerprints de todos os isolados coletados das diferentes fontes estudadas (mãos de ordenhador, tetos das cabras, leite, ordenhadeira e água. Observaram-se comportamentos muito similares das bandas, o que indica que os isolados podem ser relatados como clones epidemiológicos. As mãos do ordenhador caracterizaram-se como ponto crítico de controle, pois se destacaram como iniciador de contaminação nas amostras Staphylococcus aureus. A técnica demonstrou ser eficiente para a análise da similaridade entre indivíduos da espécie Staphylococcus aureus e, portanto, constitui ferramenta útil para investigação de falhas no manejo e na busca de controle mais eficiente para evitar ou reduzir a disseminação de microrganismos patogênicos causadores de sérias enfermidades em humanos e animais, que muitas vezes podem ser transmitidas por produtos como o leite e seus derivados.The objective of the present study was to apply REP-PCR sequences in the monitoring goat milk quality, by detecting Staphylococcus aureus, in samples of from milker hands, goat teats, milk, milking machine and water, for the future establishment and implantation of the system of Hazard Analysis Critical Control Points (HACCP. Several fingerprints were verified of all the isolates collected from the different sources studied (milker hands, goat teats, milk, milking machine and water. Very similar band behaviors were observed that indicated that the isolates can be reported as epidemic clones. Milker´s hands were was characterized as a critical control point (CCP, because it stands out as an

  2. Characterization of a novel cell wall binding domain-containing Staphylococcus aureus endolysin LysSA97.

    Science.gov (United States)

    Chang, Yoonjee; Ryu, Sangryeol

    2017-01-01

    Endolysin from Staphylococcus aureus phage SA97 (LysSA97) was cloned and investigated. LysSA97 specifically lyse the staphylococcal strains and effectively disrupted staphylococcal biofilms. Bioinformatic analysis of LysSA97 revealed a novel putative cell wall binding domain (CBD) as well as two enzymatically active domains (EADs) containing cysteine, histidine-dependent amidohydrolases/peptidases (CHAP, PF05257) and N-acetylmuramoyl-L-alanine amidase (Amidase-3, PF01520) domains. Comparison of 98 endolysin genes of S. aureus phages deposited in GenBank showed that they can be classified into six groups based on their domain composition. Interestingly, approximately 80.61 % of the staphylococcal endolysins have a src-homology 3 (SH3, PF08460) domain as CBD, but the remaining 19.39 %, including LysSA97, has a putative C-terminal CBD with no homology to the known CBD. The fusion protein containing green fluorescent protein and the putative CBD of LysSA97 showed a specific binding spectrum against staphylococcal cells comparable to SH3 domain (PF08460), suggesting that the C-terminal domain of LysSA97 is a novel CBD of staphylococcal endolysins.

  3. Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Ito, T; Katayama, Y; Asada, K; Mori, N; Tsutsumimoto, K; Tiensasitorn, C; Hiramatsu, K

    2001-05-01

    The beta-lactam resistance gene mecA of Staphylococcus aureus is carried by a novel mobile genetic element, designated staphylococcal cassette chromosome mec (SCCmec), identified in the chromosome of a Japanese methicillin-resistant S. aureus (MRSA) strain. We now report identification of two additional types of mecA-carrying genetic elements found in the MRSA strains isolated in other countries of the world. There were substantial differences in the size and nucleotide sequences between the elements and the SCCmec. However, new elements shared the chromosomal integration site with the SCCmec. Structural analysis of the new elements revealed that they possessed all of the salient features of the SCCmec: conserved terminal inverted repeats and direct repeats at the integration junction points, conserved genetic organization around the mecA gene, and the presence of cassette chromosome recombinase (ccr) genes responsible for the movements of SCCmec. The elements, therefore, were considered to comprise the SCCmec family of staphylococcal mobile genetic elements together with the previously identified SCCmec. Among 38 epidemic MRSA strains isolated in 20 countries, 34 were shown to possess one of the three typical SCCmec elements on the chromosome. Our findings indicated that there are at least three distinct MRSA clones in the world with different types of SCCmec in their chromosome.

  4. Genome-Wide Transcriptional Profiling of the Response of Staphylococcus aureus to Cryptotanshinone

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    Haihua Feng

    2009-01-01

    Full Text Available Staphylococcus aureus (S. aureus strains with multiple antibiotic resistances are increasingly widespread, and new agents are required for the treatment of S. aureus. Cryptotanshinone (CT, a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated effective in vitro antibacterial activity against all 21 S. aureus strains tested in this experiment. Affymetrix GeneChips were utilized to determine the global transcriptional response of S. aureus ATCC 25923 to treatment with subinhibitory concentrations of CT. Transcriptome profiling indicated that the antibacterial action of CT may be associated with its action as active oxygen radical generator; S. aureus undergoes an oxygen-limiting state upon exposure to CT.

  5. Epidemiology of Emerging Methicillin-Resistant Staphylococcus aureus (MRSA) in Denmark: a Nationwide Study in a Country with Low Prevalence of MRSA Infection

    Science.gov (United States)

    Faria, Nuno A.; Oliveira, Duarte C.; Westh, Henrik; Monnet, Dominique L.; Larsen, Anders R.; Skov, Robert; de Lencastre, Hermínia

    2005-01-01

    Strict infection control measures introduced during the 1970s have kept the incidence of methicillin-resistant Staphylococcus aureus (MRSA) infections extremely low in Denmark. Nevertheless, similarly to other countries, MRSA infections began to appear in the community in the late 1990s. A nationwide surveillance program has collected and stored all MRSA isolates since 1988 and, since 1999, clinical information has been also recorded. We used this information and isolates in a detailed epidemiological and molecular analysis of the 81 MRSA infections identified in Denmark in 2001. MRSA isolates were characterized by pulsed-field gel electrophoresis (PFGE), spa typing, multilocus sequence typing, and SCCmec typing. Comparison of the 45 community-onset MRSA (CO-MRSA) infections with the 36 hospital-acquired MRSA (HA-MRSA) infections showed several striking contrasts. Most CO-MRSA were recovered from skin and soft tissue infections caused by isolates carrying the Panton-Valentine leucocidin toxin genes, and the majority (84%) of isolates belonged to a single clonal type, ST80-IV, which has been found in the community in other European countries. Clone ST80-IV could be traced in Denmark back to 1993. ST80-IV was rarely found in HA-MRSA infections, which belonged to a large number of clonal types, including some pandemic MRSA clones. The low number of HA-MRSA infections and the diversity of MRSA clones in Danish hospitals may be the result of successful infection control measures that prevent spread of clones in hospitals. The mechanism of spread of the ST80-IV clone in the Danish community is not known, and new control measures are needed to control further spread of this and other CA-MRSA clones. PMID:15815005

  6. Royana: Successful Experience in Cloning the Sheep

    OpenAIRE

    Saeed Kazemi Ashtiani; Mohammad Hossein Nasr-Esfahani; Sayyed Mortaza Hosseini; Fariba Moulavi; Mahdi Hajian; Mohsen Frouzanfar; Parvaneh Abedi; Maryam Meamar; Mojtaba Rezazadeh Valojerdi; Hamid Gourabi; Abdolhossein Shahverdi; Hossein Baharvand; Ahmad Vosough Dizaj; Hossein Imani; Poopak Eftekhari-Yazdi

    2008-01-01

    Objective: This study describes our experiences in reproductive cloning using two differentprocedures resulting in birth of the first successfully cloned sheep in Iran and theMiddle-East, nick-named "Royana".Materials and Methods: Abattoir-derived sheep oocytes were enucleated after in vitromaturation for 18-20hrs and then reconstructed by ear-derived sheep somatic cells usingtwo different procedures of renucleation (subzonary, intracytoplasmic), embryo culture (coculture,sequential medium) a...

  7. Economical phase-covariant cloning with multiclones

    Institute of Scientific and Technical Information of China (English)

    Zhang Wen-Hai; Ye Liu

    2009-01-01

    This paper presents a very simple method to derive the explicit transformations of the optimal economical to M phase-covariant cloning. The fidelity of clones reaches the theoretic bound [D'Ariano G M and Macchiavello C 2003 Phys. Rcv. A 67 042306]. The derived transformations cover the previous contributions [Delgado Y,Lamata L et al,2007 Phys. Rev. Lett. 98 150502] in which M must be odd.

  8. Endangered wolves cloned from adult somatic cells.

    Science.gov (United States)

    Kim, Min Kyu; Jang, Goo; Oh, Hyun Ju; Yuda, Fibrianto; Kim, Hye Jin; Hwang, Woo Suk; Hossein, Mohammad Shamim; Kim, Joung Joo; Shin, Nam Shik; Kang, Sung Keun; Lee, Byeong Chun

    2007-01-01

    Over the world, canine species, including the gray wolf, have been gradually endangered or extinct. Many efforts have been made to recover and conserve these canids. The aim of this study was to produce the endangered gray wolf with somatic cell nuclear transfer (SCNT) for conservation. Adult ear fibroblasts from a female gray wolf (Canis lupus) were isolated and cultured in vitro as donor cells. Because of limitations in obtaining gray wolf matured oocytes, in vivo matured canine oocytes obtained by flushing the oviducts from the isthmus to the infundibulum were used. After removing the cumulus cells, the oocyte was enucleated, microinjected, fused with a donor cell, and activated. The reconstructed cloned wolf embryos were transferred into the oviducts of the naturally synchronized surrogate mothers. Two pregnancies were detected by ultrasonography at 23 days of gestation in recipient dogs. In each surrogate dog, two fetal sacs were confirmed by early pregnancy diagnosis at 23 days, but only two cloned wolves were delivered. The first cloned wolf was delivered by cesarean section on October 18, 2005, 60 days after embryo transfer. The second cloned wolf was delivered on October 26, 2005, at 61 days postembryo transfer. Microsatellite analysis was performed with genomic DNA from the donor wolf, the two cloned wolves, and the two surrogate female recipients to confirm the genetic identity of the cloned wolves. Analysis of 19 microsatellite loci confirmed that the cloned wolves were genetically identical to the donor wolf. In conclusion, we demonstrated live birth of two cloned gray wolves by nuclear transfer of wolf somatic cells into enucleated canine oocyte, indicating that SCNT is a practical approach for conserving endangered canids.

  9. Prevalence of Salmonella and Staphylococcus aureus in chorizo and longaniza

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    Refugio Torres-Vitela

    2011-12-01

    Full Text Available Epidemiological research in developed and developing countries, had found meat products as the principal cause for foodbourne diseases. In addition, Salmonella and Staphyococcus aureus are well known pathogens for their mayor impact in public health. The objective for the present study consisted on determinate the sanitary quality from chorizo and longaniza samples from several butcheries in Guadalajara, Jalisco, Mexico. Samples of chorizo (50 and longaniza (50 were obtained from different points in Guadalajara metropolis. Presence of Salmonella and recounts for S. aureus were tested in 25 g samples. Procedure was followed according Mexican NOM 145-SSA1-1995 methods. In chorizo, 18 samples were positive to Salmonella. The count of S. aureus showed a mean of 24,600 UFC/g. On the other hand, 24 samples of longaniza were positive to Salmonella spp. In this case, the mean of S. aureus was 7,800 UFC/g. The serotypes of Salmonella spp were: Derby (30%, Adelaile (17%, Azteca (15%, Infantis (15%, Muenster(10% y Anatum (13 %. The high positivity of Salmonella spp. and S. aureus is a potential hazard to consumers.

  10. Antibacterial Action of Curcumin against Staphylococcus aureus: A Brief Review

    Science.gov (United States)

    Liew, Kitson; Ali, Syed A.; Khoo, Alan Soo-Beng; Peh, Suat-Cheng

    2016-01-01

    Curcumin, the major constituent of Curcuma longa L. (Zingiberaceae family) or turmeric, commonly used for cooking in Asian cuisine, is known to possess a broad range of pharmacological properties at relatively nontoxic doses. Curcumin is found to be effective against Staphylococcus aureus (S. aureus). As demonstrated by in vitro experiment, curcumin exerts even more potent effects when used in combination with various other antibacterial agents. Hence, curcumin which is a natural product derived from plant is believed to have profound medicinal benefits and could be potentially developed into a naturally derived antibiotic in the future. However, there are several noteworthy challenges in the development of curcumin as a medicine. S. aureus infections, particularly those caused by the multidrug-resistant strains, have emerged as a global health issue and urgent action is needed. This review focuses on the antibacterial activities of curcumin against both methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA). We also attempt to highlight the potential challenges in the effort of developing curcumin into a therapeutic antibacterial agent. PMID:27956904

  11. Antibacterial Action of Curcumin against Staphylococcus aureus: A Brief Review

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    Sin-Yeang Teow

    2016-01-01

    Full Text Available Curcumin, the major constituent of Curcuma longa L. (Zingiberaceae family or turmeric, commonly used for cooking in Asian cuisine, is known to possess a broad range of pharmacological properties at relatively nontoxic doses. Curcumin is found to be effective against Staphylococcus aureus (S. aureus. As demonstrated by in vitro experiment, curcumin exerts even more potent effects when used in combination with various other antibacterial agents. Hence, curcumin which is a natural product derived from plant is believed to have profound medicinal benefits and could be potentially developed into a naturally derived antibiotic in the future. However, there are several noteworthy challenges in the development of curcumin as a medicine. S. aureus infections, particularly those caused by the multidrug-resistant strains, have emerged as a global health issue and urgent action is needed. This review focuses on the antibacterial activities of curcumin against both methicillin-sensitive S. aureus (MSSA and methicillin-resistant S. aureus (MRSA. We also attempt to highlight the potential challenges in the effort of developing curcumin into a therapeutic antibacterial agent.

  12. Superantigen Profiling of Staphylococcus aureus Infective Endocarditis Isolates

    Science.gov (United States)

    Chung, Jin-Won; Karau, Melissa J.; Greenwood-Quaintance, Kerryl E.; Ballard, Alessandro D.; Tilahun, Ashenafi; Khaleghi, Shahryar Rostamkolaei; David, Chella S.; Patel, Robin; Rajagopalan, Govindarajan

    2014-01-01

    The frequency of superantigen production among Staphylococcus aureus isolates associated with endocarditis is not well defined. We tested 154 S. aureus isolates from definite infective endocarditis cases for the presence of staphylococcal enterotoxins A-E, H and TSST-1 by PCR, ELISA and using an HLA-DR3 transgenic mouse splenocyte proliferation assay. Sixty-three isolates (50.8%) tested positive for at least one superantigen gene, with 21 (16.9%) testing positive for more than two. tst (28.6%) was most common, followed by seb (27%), sea (22.2%), sed (20.6%), see (17.5%), and sec (11.1%). Of 41 methicillin-resistant S. aureus, 21 had superantigen genes, with sed being more frequently detected in this group compared to methicillin-susceptible S. aureus (P<0.05). Superantigen genes were not associated with mortality (P=0.81). 75% of PCR-positive isolates induced robust splenocyte proliferation. Overall, more than half of S. aureus isolates causing endocarditis carry superantigen genes of which most are functional. PMID:24745820

  13. Staphylococcus aureus isolated from tonsillectomized adult patients with recurrent tonsillitis.

    Science.gov (United States)

    Katkowska, Marta; Garbacz, Katarzyna; Stromkowski, Józef

    2017-01-01

    The aim of this study was to analyze the prevalence and antibiotic resistance of Staphylococcus aureus strains from 118 tonsillectomized adults due to recurrent tonsillitis (RT). The study included strains isolated from the tonsillar surface prior to tonsillectomy, recovered from the tonsillar core at the time of surgery, and from the posterior throat 2-4 weeks after the procedure. Susceptibility of isolates to 19 antibiotics was tested in line with the Clinical and Laboratory Standards Institute recommendations. Irrespective of the stage, the most commonly isolated bacteria were gram-positive cocci, and among them S. aureus. The tonsillar core was the most common site of S. aureus isolation (30.5%), followed by the tonsillar surface (10.8%) and the posterior pharynx (5.9%). This difference turned out to be statistically significant (p aureus seems to be the most common pathogen isolated from patients tonsillectomized due to RT. Staphylococcal isolates associated with RT are present mostly within the tonsillar core and susceptible to most antibiotics. They are typically isolated from patients between 21 and 30 years of age. Tonsillectomy results in less frequent isolation of S. aureus strains.

  14. Prevalence of Staphylococcus aureus in Shrimps in Tehran during 2013

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    Mohammad Mehdi Soltan Dallal

    2015-12-01

    Full Text Available Background During fishing and transport, preservation and quality of fish products are importantas well as storage to prevent the growth of pathogenic and toxin producing bacteria.Staphylococcus aureus is one of the most common causes of sea food-borne diseases worldwidedue to contamination of food by preformed enterotoxins. The aim of this study was to compare theprevalence and contamination of S. aureus in marine and farmed shrimps in Tehran fishery center.Methods: A total of 300 samples, including 150 marine, 150 farmed shrimps were selected duringSeptember 2013 to December 2013. Isolation and identification of S. aureus from isolated sampleswere carried out according to conventional methods, and antibiotic susceptibility test wasperformed by modified Kirby-Bauer disc diffusion methodResults: The results of this study showed that 30% of marine and 20% off armed shrimps werecontaminated with S. aureus. The highest resistance was observed with penicillin and ampicillin,whereas 100% were sensitive to vancomycin, clindamycin, ciprofloxacin, and rifampin.Conclusions: Due to relatively high contamination of shrimp by S. aureus more attention shouldbe given during processing and manufacturing.

  15. Prevalence of Staphylococcus aureus in Shrimps in Tehran during 2013

    Directory of Open Access Journals (Sweden)

    Mohammad Mehdi Soltan Dallal

    2016-02-01

    Full Text Available Background During fishing and transport, preservation and quality of fish products are importantas well as storage to prevent the growth of pathogenic and toxin producing bacteria.Staphylococcus aureus is one of the most common causes of sea food-borne diseases worldwidedue to contamination of food by preformed enterotoxins. The aim of this study was to compare theprevalence and contamination of S. aureus in marine and farmed shrimps in Tehran fishery center.Methods: A total of 300 samples, including 150 marine, 150 farmed shrimps were selected duringSeptember 2013 to December 2014. Isolation and identification of S. aureus from isolated sampleswere carried out according to conventional methods, and antibiotic susceptibility test wasperformed by modified Kirby-Bauer disc diffusion method.Results: The results of this study showed that 30% of marine and 20% off armed shrimps werecontaminated with S. aureus. The highest resistance was observed with penicillin and ampicillin,whereas 100% were sensitive to vancomycin, clindamycin, ciprofloxacin, and rifampin.Conclusions: Due to relatively high contamination of shrimp by S. aureus more attention shouldbe given during processing and manufacturing.

  16. A systematic review of animal models for Staphylococcus aureus osteomyelitis

    Directory of Open Access Journals (Sweden)

    W Reizner

    2014-03-01

    Full Text Available Staphylococcus aureus (S. aureus osteomyelitis is a significant complication for orthopaedic patients undergoing surgery, particularly with fracture fixation and arthroplasty. Given the difficulty in studying S. aureus infections in human subjects, animal models serve an integral role in exploring the pathogenesis of osteomyelitis, and aid in determining the efficacy of prophylactic and therapeutic treatments. Animal models should mimic the clinical scenarios seen in patients as closely as possible to permit the experimental results to be translated to the corresponding clinical care. To help understand existing animal models of S. aureus, we conducted a systematic search of PubMed and Ovid MEDLINE to identify in vivo animal experiments that have investigated the management of S. aureus osteomyelitis in the context of fractures and metallic implants. In this review, experimental studies are categorised by animal species and are further classified by the setting of the infection. Study methods are summarised and the relevant advantages and disadvantages of each species and model are discussed. While no ideal animal model exists, the understanding of a model’s strengths and limitations should assist clinicians and researchers to appropriately select an animal model to translate the conclusions to the clinical setting.

  17. Detection of Staphylococcus Aureus Enterotoxin Genes A-E

    Directory of Open Access Journals (Sweden)

    Dadgar, T. (PhD

    2014-06-01

    Full Text Available Background and Objective: The main cause of spreading staphylococcal infections among patients is the healthy carriers working in hospitals. With the secretion of different sorts of toxins such as entrotoxin, this bacteria can provide the conditions for attacking on the host. The main objective of this study is identification of the characteristics and differences in the Staphylococcus aureus isolated from healthy carriers and from the patients on the basis of enterotoxin genes (sea-see. Material and Methods: One hundred and twenty of the patients and 80 of healthy carriers worked in health centers of Gorgan, north of Iran, were investigated for S. aureus isolate. The isolates were evaluated by PCR for Enterotoxin Genes A-E (SEA to SEE. Results: Enterotoxin genes (SEA to SEE was found in 87.5% of the total isolates and the most frequent one was enterotoxin gene sea (N= 124. The prevalence of these isolates in healthy carriers was significantly higher than those of the patients. Conclusion: Based on the results, the high percentage of S. aureus isolated from clinical samples contains enterotoxin genes. Therefore, Human as the source and carrier of S. aureus is paramount importance, which is due to significant relationship between being toxigenic strains and the source of isolation. Key words: Staphylococcus Aureus; Enterotoxin; Patient; Carrier

  18. Prevalence and antibiotic susceptibility of Staphylococcus aureus from bovine mastitis

    Directory of Open Access Journals (Sweden)

    C. G. Unakal and B. B. Kaliwal

    2010-04-01

    Full Text Available The primary objective of this study was to determine the prevalence and antimicrobial susceptibility of mastitic Staphylococcus aureus in dairy cows. Milk samples for microbiological culture were collected from dairy herds. A total of 105 samples were screened and 68 confirmed Staphylococcus aureus were obtained. The a, ß and non haemolytic activity revealed 20.58%, 75% and 4.41% respectively in 68 isolated strains of Staphylococcus aureus. Resistance of Staphylococcus aureus against 10 antimicrobial agents was tested using the disc diffusion method. The highest 86.76% isolates were resistant to penicillin followed by ampicillin 70.50%, amoxicillin 63.23%, gentamycin 47.05%, amikacin 30.80%, erythromycin 27.94%, Ciprofloxacin 26.47%, methicillin 23.52%, cefotaxime 20.58% and the lowest resistant was shown in ceftriaxone 19.11%. The study revealed that the increase in prevalence and antibiotic resistance pattern of the Staphylococcus aureus isolated from bovine mastitis. [Vet. World 2010; 3(2.000: 65-67

  19. A modified version of the digestion-ligation cloning method for more efficient molecular cloning.

    Science.gov (United States)

    Gao, Song; Li, Yanling; Zhang, Jiannan; Chen, Hongman; Ren, Daming; Zhang, Lijun; An, Yingfeng

    2014-05-15

    Here we describe a modified version of the digestion-ligation approach for efficient molecular cloning. In comparison with the original method, the modified method has the additional steps of gel purification and a second ligation after the first ligation of the linearized vector and DNA insert. During this process, the efficiency and reproducibility could be significantly improved for both stick-end cloning and blunt-end cloning. As an improvement of the very important molecular cloning technique, this method may find a wide range of applications in bioscience and biotechnology.

  20. Do Clones Dream of Love? Images of Clones in Popular Culture

    Directory of Open Access Journals (Sweden)

    Dragana Antonijević

    2016-02-01

    Full Text Available Fantasies about clones, cyborgs and androids have become part and parcel of the mythology of modern times – the mythologies of the biotechnological era in which the achievements of genetic engineering have inflamed fears of possible abuse of scientific knowledge and the consequences of such abuse. The paper considers the phenomenon of reproductive cloning of human beings as it is represented in popular culture, especially film as it is one of the most important sources of representations and constructions of ideas about clones. After the introductory consideration of this phenomenon in scientific, ethical and media debates which are imbued with rejection of reproductive cloning, I have analyzed the different uses of the clone motif in selected movies. I have examined the structure and content of the genre formula of "social melodrama" which is present in films about clones, and have analyzed the mythical patterns pertaining to the topic of cloning, such as the myth of immortality, the myth of twins, the myth of the uniqueness of human kind etc. Ultimately, the nature and origins of the fear of clones and disgust that clones cause have been examined, and it has been shown that they mostly boil down to the fear of the dehumanization of human beings, the fear of the loss of difference and the transgression of biological, sociocultural and metaphysical boundaries.

  1. Identification and characterization of methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus pettenkoferi from a small animal clinic.

    Science.gov (United States)

    Weiss, Sonja; Kadlec, Kristina; Fessler, Andrea T; Schwarz, Stefan

    2013-12-27

    The aim of this study was to isolate and characterize methicillin-resistant staphylococci (MRS) in a small animal clinic and to investigate their distribution and possible transmission. Swabs (n=72) were taken from hospitalized pets, the environment and employees of a small animal clinic and screened for the presence of MRS. The staphylococcal species was confirmed biochemically or by 16S rDNA sequencing. Susceptibility to antimicrobial agents was tested by broth dilution. The presence of mecA and other resistance genes was confirmed by PCR. Molecular typing of the isolates followed standard procedures. In total, 34 MRS belonging to the four species Staphylococcus aureus (n=5), Staphylococcus epidermidis (n=21), Staphylococcus haemolyticus (n=6) or Staphylococcus pettenkoferi (n=2) were isolated. All isolates were multidrug-resistant with resistance to at least three classes of antimicrobial agents. Among the five methicillin-resistant S. aureus (MRSA) isolates, four belonged to the clonal complex CC398; two of them were isolated from cats, the remaining two from pet cages. Overall, the MRS isolates differed in their characteristics, except for one S. epidermidis clone (n=9) isolated from hospitalized cats without clinical staphylococcal infections, pet cages, the clinic environment as well as from a healthy employee. This MRSE clone was resistant to 10 classes of antimicrobial agents, including aminocyclitols, β-lactams, fluoroquinolones, lincosamides, macrolides, phenicols, pleuromutilins, sulfonamides, tetracyclines and trimethoprim. These findings suggest a possible transmission of specific MRS isolates between animal patients, employees and the clinic environment.

  2. Antibacterial Activity of Defensin PaDef from Avocado Fruit (Persea americana var. drymifolia Expressed in Endothelial Cells against Escherichia coli and Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Jaquelina Julia Guzmán-Rodríguez

    2013-01-01

    Full Text Available Antimicrobial therapy is a useful tool to control infectious diseases in general and rising antibiotic resistant microorganisms in particular. Alternative strategies are desirable, and antimicrobial peptides (AMP represent attractive control agents. Mexican avocado (Persea americana var. drymifolia is used in traditional medicine; however, the AMP production has not been reported in this plant. We obtained a cDNA library from avocado fruit and clone PaDef was identified, which has a cDNA (249 bp encoding a protein (78 aa homologous with plant defensins (>80%. We expressed the defensin PaDef cDNA (pBME3 in the bovine endothelial cell line BVE-E6E7. Polyclonal and clonal populations were obtained and their activity was evaluated against Escherichia coli, Staphylococcus aureus, and Candida albicans. E. coli viability was inhibited with 100 μg/mL of total protein from clones (>55%. Also, S. aureus viability was inhibited from 50 μg/mL total protein (27–38% but was more evident at 100 μg/mL (52–65%. This inhibition was higher than the effect showed by polyclonal population (~23%. Finally, we did not detect activity against C. albicans. These results are the first report that shows antimicrobial activity of a defensin produced by avocado and suggest that this AMP could be used in the control of pathogens.

  3. Antibacterial Activity of Defensin PaDef from Avocado Fruit (Persea americana var. drymifolia) Expressed in Endothelial Cells against Escherichia coli and Staphylococcus aureus

    Science.gov (United States)

    Guzmán-Rodríguez, Jaquelina Julia; López-Gómez, Rodolfo; Suárez-Rodríguez, Luis M.; Salgado-Garciglia, Rafael; Rodríguez-Zapata, Luis C.; Ochoa-Zarzosa, Alejandra; López-Meza, Joel E.

    2013-01-01

    Antimicrobial therapy is a useful tool to control infectious diseases in general and rising antibiotic resistant microorganisms in particular. Alternative strategies are desirable, and antimicrobial peptides (AMP) represent attractive control agents. Mexican avocado (Persea americana var. drymifolia) is used in traditional medicine; however, the AMP production has not been reported in this plant. We obtained a cDNA library from avocado fruit and clone PaDef was identified, which has a cDNA (249 bp) encoding a protein (78 aa) homologous with plant defensins (>80%). We expressed the defensin PaDef cDNA (pBME3) in the bovine endothelial cell line BVE-E6E7. Polyclonal and clonal populations were obtained and their activity was evaluated against Escherichia coli, Staphylococcus aureus, and Candida albicans. E. coli viability was inhibited with 100 μg/mL of total protein from clones (>55%). Also, S. aureus viability was inhibited from 50 μg/mL total protein (27–38%) but was more evident at 100 μg/mL (52–65%). This inhibition was higher than the effect showed by polyclonal population (~23%). Finally, we did not detect activity against C. albicans. These results are the first report that shows antimicrobial activity of a defensin produced by avocado and suggest that this AMP could be used in the control of pathogens. PMID:24319695

  4. Antibacterial activity of defensin PaDef from avocado fruit (Persea americana var. drymifolia) expressed in endothelial cells against Escherichia coli and Staphylococcus aureus.

    Science.gov (United States)

    Guzmán-Rodríguez, Jaquelina Julia; López-Gómez, Rodolfo; Suárez-Rodríguez, Luis M; Salgado-Garciglia, Rafael; Rodríguez-Zapata, Luis C; Ochoa-Zarzosa, Alejandra; López-Meza, Joel E

    2013-01-01

    Antimicrobial therapy is a useful tool to control infectious diseases in general and rising antibiotic resistant microorganisms in particular. Alternative strategies are desirable, and antimicrobial peptides (AMP) represent attractive control agents. Mexican avocado (Persea americana var. drymifolia) is used in traditional medicine; however, the AMP production has not been reported in this plant. We obtained a cDNA library from avocado fruit and clone PaDef was identified, which has a cDNA (249 bp) encoding a protein (78 aa) homologous with plant defensins (>80%). We expressed the defensin PaDef cDNA (pBME3) in the bovine endothelial cell line BVE-E6E7. Polyclonal and clonal populations were obtained and their activity was evaluated against Escherichia coli, Staphylococcus aureus, and Candida albicans. E. coli viability was inhibited with 100 μg/mL of total protein from clones (>55%). Also, S. aureus viability was inhibited from 50 μg/mL total protein (27-38%) but was more evident at 100 μg/mL (52-65%). This inhibition was higher than the effect showed by polyclonal population (~23%). Finally, we did not detect activity against C. albicans. These results are the first report that shows antimicrobial activity of a defensin produced by avocado and suggest that this AMP could be used in the control of pathogens.

  5. Phenol soluble modulin (PSM) variants of community-associated methicillin-resistant Staphylococcus aureus (MRSA) captured using mass spectrometry-based molecular networking.

    Science.gov (United States)

    Gonzalez, David J; Vuong, Lisa; Gonzalez, Isaiah S; Keller, Nadia; McGrosso, Dominic; Hwang, John H; Hung, Jun; Zinkernagel, Annelies; Dixon, Jack E; Dorrestein, Pieter C; Nizet, Victor

    2014-05-01

    Molecular genetic analysis indicates that the problematic human bacterial pathogen methicillin-resistant Staphylococcus aureus possesses more than 2000 open reading frames in its genome. This number of potential gene products, coupled with intrinsic mechanisms of posttranslational modification, endows methicillin-resistant Staphylococcus aureus with a highly complex biochemical repertoire. Recent proteomic and metabolomic advances have provided methodologies to better understand and characterize the biosynthetic factors released by microbial organisms. Here, the emerging tool of mass spectrometry-based molecular networking was used to visualize and map the repertoire of biosynthetic factors produced by a community-associated methicillin-resistant Staphylococcus aureus strain representative of the epidemic USA300 clone. In particular, the study focused on elucidating the complexity of the recently discovered phenol soluble modulin family of peptides when placed under various antibiotic treatment stresses. Novel PSM truncated variant peptides were captured, and the type of variants that were clustered by the molecular networks platform changed in response to the different antibiotic treatment conditions. After discovery, a group of the peptides were selected for functional analysis in vitro. The peptides displayed bioactive properties including the ability to induce proinflammatory responses in human THP-1 monocytes. Additionally, the tested peptides did not display antimicrobial activity as previously reported for other phenol soluble modulin truncated variants. Our findings reveal that the PSM family of peptides are quite structurally diverse, and suggest a single phenol soluble modulin parent peptide can functionally spawn differential bioactivities in response to various external stimuli.

  6. Molecular typing of toxic shock syndrome toxin-1- and Enterotoxin A-producing methicillin-sensitive Staphylococcus aureus isolates from an outbreak in a neonatal intensive care unit.

    Science.gov (United States)

    Layer, Franziska; Sanchini, Andrea; Strommenger, Birgit; Cuny, Christiane; Breier, Ann-Christin; Proquitté, Hans; Bührer, Christoph; Schenkel, Karl; Bätzing-Feigenbaum, Jörg; Greutelaers, Benedikt; Nübel, Ulrich; Gastmeier, Petra; Eckmanns, Tim; Werner, Guido

    2015-10-01

    Outbreaks of Staphylococcus aureus are common in neonatal intensive care units (NICUs). Usually they are documented for methicillin-resistant strains, while reports involving methicillin-susceptible S. aureus (MSSA) strains are rare. In this study we report the epidemiological and molecular investigation of an MSSA outbreak in a NICU among preterm neonates. Infection control measures and interventions were commissioned by the Local Public Health Authority and supported by the Robert Koch Institute. To support epidemiological investigations molecular typing was done by spa-typing and Multilocus sequence typing; the relatedness of collected isolates was further elucidated by DNA SmaI-macrorestriction, microarray analysis and bacterial whole genome sequencing. A total of 213 neonates, 123 healthcare workers and 205 neonate parents were analyzed in the period November 2011 to November 2012. The outbreak strain was characterized as a MSSA spa-type t021, able to produce toxic shock syndrome toxin-1 and Enterotoxin A. We identified seventeen neonates (of which two died from toxic shock syndrome), four healthcare workers and three parents putatively involved in the outbreak. Whole-genome sequencing permitted to exclude unrelated cases from the outbreak and to discuss the role of healthcare workers as a reservoir of S. aureus on the NICU. Genome comparisons also indicated the presence of the respective clone on the ward months before the first colonized/infected neonates were detected.

  7. Typing of Panton-Valentine Leukocidin-encoding Phages and lukSF-PV Gene Sequence Variation in Staphylococcus aureus from China

    Directory of Open Access Journals (Sweden)

    Huanqiang Zhao

    2016-08-01

    Full Text Available Panton-Valentine leucocidin (PVL, encoded by lukSF-PV genes, a bi-component and pore-forming toxin, is carried by different staphylococcal bacteriophages. The prevalence of PVL in Staphylococcus aureus (S. aureus have been reported around the globe. However, the data on PVL-encoding phage types, lukSF-PV gene variation and chromosomal phage insertion sites for PVL-positive S. aureus are limited, especially in China. In order to obtain a more complete understanding of the molecular epidemiology of PVL-positive S. aureus, an integrated and modified PCR-based scheme was applied to detect the PVL-encoding phage types. Phage insertion locus and the lukSF-PV variant were determined by PCR and sequencing. Meanwhile, the genetic background was characterized by staphylococcal cassette chromosome mec (SCCmec typing, staphylococcal protein A (spa gene polymorphisms typing, pulsed-field gel electrophoresis (PFGE typing, accessory gene regulator (agr locus typing and multilocus sequence typing (MLST. Seventy eight (78/1175, 6.6% isolates possessed the lukSF-PV genes and 59.0% (46/78 of PVL-positive strains belonged to CC59 lineage. Eight known different PVL-encoding phage types were detected, and Φ7247PVL/ΦST5967PVL (n=13 and ΦPVL (n=12 were the most prevalent among them. While 25 (25/78, 32.1% isolates, belonging to ST30 and ST59 clones, were unable to be typed by the modified PCR-based scheme. Single nucleotide polymorphisms (SNPs were identified at five locations in the lukSF-PV genes, two of which were non-synonymous. Maximum-likelihood tree analysis of attachment sites sequences detected six SNP profiles for attR and eight for attL, respectively. In conclusion, the PVL-positive S. aureus mainly harbored Φ7247PVL/ΦST5967PVL and ΦPVL in the regions studied. lukSF-PV gene sequences, PVL-encoding phages and phage insertion locus generally varied with lineages. Moreover, PVL-positive clones that have emerged worldwide likely carry distinct phages.

  8. AN APPROACH FOR CLONE DETECTION IN DOCUMENTATION REUSE

    Directory of Open Access Journals (Sweden)

    D. V. Lutsiv

    2014-07-01

    Full Text Available The paper focuses on the searching method for repetitions in DocBook/DRL or plain text documents. An algorithm has been designed based on software clone detection. The algorithm supports filtering results: clones are rejected if clone length in the group is less than 5 symbols, intersection of clone groups is eliminated, meaningfulness clones are removed, the groups containing clones consisting only of XML are eliminated. Remaining search is supported: found clones are extracted from the documentation, and clone search is repeated. One step is proved to be enough. Adaptive reuse technique of Paul Bassett – Stan Jarzabek has been implemented. A software tool has been developed on the basis of the algorithm. The tool supports setting parameters for repetitions detection and visualization of the obtained results. The tool is integrated into DocLine document development environment, and provides refactoring of documents using found clones. The Clone Miner clone detection utility is used for clones search. The method has been evaluated for Linux Kernel Documentation (29 documents, 25000 lines. Five semantic kinds of clones have been selected: terms (abbreviations, one word and two word terms, hyperlinks, license agreements, functionality description, and code examples. 451 meaningful clone groups have been found, average clone length is 4.43 tokens, and average number of clones in a group is 3.56.

  9. Universal Quantum Cloning Machines for Two Identical Mixed Qubits

    Institute of Scientific and Technical Information of China (English)

    YANG Shuai; ZHAO Mei-Sheng; LIU Nai-Le; CHEN Zeng-Bing

    2007-01-01

    We present a series of universal quantum cloning machines for two identical mixed qubits. Every machine is optimal in the sense that it achieves the optimal bound of the single copy shrinking factor. Unlike in the case of pure state cloning, the single copy shrinking factor does not uniquely determine the cloning map in the case of mixed state cloning.

  10. Public perceptions of farm animal cloning in Europe

    DEFF Research Database (Denmark)

    Lassen, Jesper

    This report presents a picture of European opinion on farm animal cloning. In the report, both agricultural and biomedical applications of farm animal cloning are considered. With the arrival of Dolly, animal cloning became an integral part of the biotech debate, but this debate did not isolate...... animal cloning as a single issue....

  11. Cloning: Past, Present, and the Exciting Future. Breakthroughs in Bioscience.

    Science.gov (United States)

    Di Berardino, Marie A.

    This document explores the history of cloning by focusing on Dolly the Sheep, one of the first large animal clonings. The disadvantages and advantages of transgenic clones are discussed as well as the future implications of cloning from the perspective of human health. (Contains 10 resources.) (YDS)

  12. [Ethical considerations on human cloning. A psychoanalytic perspective].

    Science.gov (United States)

    Alvarez, A

    2000-01-01

    A brief review of ethical issues related to two types of human cloning is presented: cloning embryonic cells not intended to culminate in the birth of a new individual and cloning human beings. Advantages and objections related to both types of human cloning are analyzed from an ethical point of view. Repercussions on individuals born by the technique of cloning are discussed from a psychoanalytical perspective. It can be concluded that cloning embryonic cells could be admissible, while not cloning considered as a reproductive option.

  13. Contribution of coagulases towards Staphylococcus aureus disease and protective immunity.

    Directory of Open Access Journals (Sweden)

    Alice G Cheng

    Full Text Available The bacterial pathogen Staphylococcus aureus seeds abscesses in host tissues to replicate at the center of these lesions, protected from host immune cells via a pseudocapsule. Using histochemical staining, we identified prothrombin and fibrin within abscesses and pseudocapsules. S. aureus secretes two clotting factors, coagulase (Coa and von Willebrand factor binding protein (vWbp. We report here that Coa and vWbp together are required for the formation of abscesses. Coa and vWbp promote the non-proteolytic activation of prothrombin and cleavage of fibrinogen, reactions that are inhibited with specific antibody against each of these molecules. Coa and vWbp specific antibodies confer protection against abscess formation and S. aureus lethal bacteremia, suggesting that coagulases function as protective antigens for a staphylococcal vaccine.

  14. Staphylococcus aureus biofilms: recent developments in biofilm dispersal.

    Science.gov (United States)

    Lister, Jessica L; Horswill, Alexander R

    2014-01-01

    Staphylococcus aureus is a major cause of nosocomial and community-acquired infections and represents a significant burden on the healthcare system. S. aureus attachment to medical implants and host tissue, and the establishment of a mature biofilm, play an important role in the persistence of chronic infections. The formation of a biofilm, and encasement of cells in a polymer-based matrix, decreases the susceptibility to antimicrobials and immune defenses, making these infections difficult to eradicate. During infection, dispersal of cells from the biofilm can result in spread to secondary sites and worsening of the infection. In this review, we discuss the current understanding of the pathways behind biofilm dispersal in S. aureus, with a focus on enzymatic and newly described broad-spectrum dispersal mechanisms. Additionally, we explore potential applications of dispersal in the treatment of biofilm-mediated infections.

  15. Genetically enhanced cows resist intramammary Staphylococcus aureus infection.

    Science.gov (United States)

    Wall, Robert J; Powell, Anne M; Paape, Max J; Kerr, David E; Bannerman, Douglas D; Pursel, Vernon G; Wells, Kevin D; Talbot, Neil; Hawk, Harold W

    2005-04-01

    Mastitis, the most consequential disease in dairy cattle, costs the US dairy industry billions of dollars annually. To test the feasibility of protecting animals through genetic engineering, transgenic cows secreting lysostaphin at concentrations ranging from 0.9 to 14 micrograms/ml [corrected] in their milk were produced. In vitro assays demonstrated the milk's ability to kill Staphylococcus aureus. Intramammary infusions of S. aureus were administered to three transgenic and ten nontransgenic cows. Increases in milk somatic cells, elevated body temperatures and induced acute phase proteins, each indicative of infection, were observed in all of the nontransgenic cows but in none of the transgenic animals. Protection against S. aureus mastitis appears to be achievable with as little as 3 micrograms/ml [corrected] of lysostaphin in milk. Our results indicate that genetic engineering can provide a viable tool for enhancing resistance to disease and improve the well-being of livestock.

  16. Repurposing the antihistamine terfenadine for antimicrobial activity against Staphylococcus aureus.

    Science.gov (United States)

    Perlmutter, Jessamyn I; Forbes, Lauren T; Krysan, Damian J; Ebsworth-Mojica, Katherine; Colquhoun, Jennifer M; Wang, Jenna L; Dunman, Paul M; Flaherty, Daniel P

    2014-10-23

    Staphylococcus aureus is a rapidly growing health threat in the U.S., with resistance to several commonly prescribed treatments. A high-throughput screen identified the antihistamine terfenadine to possess, previously unreported, antimicrobial activity against S. aureus and other Gram-positive bacteria. In an effort to repurpose this drug, structure-activity relationship studies yielded 84 terfenadine-based analogues with several modifications providing increased activity versus S. aureus and other bacterial pathogens, including Mycobacterium tuberculosis. Mechanism of action studies revealed these compounds to exert their antibacterial effects, at least in part, through inhibition of the bacterial type II topoisomerases. This scaffold suffers from hERG liabilities which were not remedied through this round of optimization; however, given the overall improvement in activity of the set, terfenadine-based analogues provide a novel structural class of antimicrobial compounds with potential for further characterization as part of the continuing process to meet the current need for new antibiotics.

  17. Local inflammation exacerbates the severity of Staphylococcus aureus skin infection.

    Directory of Open Access Journals (Sweden)

    Christopher P Montgomery

    Full Text Available Staphylococcus aureus is the leading cause of skin infections. In a mouse model of S. aureus skin infection, we found that lesion size did not correlate with bacterial burden. Athymic nude mice had smaller skin lesions that contained lower levels of myeloperoxidase, IL-17A, and CXCL1, compared with wild type mice, although there was no difference in bacterial burden. T cell deficiency did not explain the difference in lesion size, because TCR βδ (-/- mice did not have smaller lesions, and adoptive transfer of congenic T cells into athymic nude mice prior to infection did not alter lesion size. The differences observed were specific to the skin, because mortality in a pneumonia model was not different between wild type and athymic nude mice. Thus, the clinical severity of S. aureus skin infection is driven by the inflammatory response to the bacteria, rather than bacterial burden, in a T cell independent manner.

  18. Botryomycosis Due to Staphylococcus Aureus-A Case Report

    Directory of Open Access Journals (Sweden)

    Manjula A.Vagarali

    2012-04-01

    Full Text Available Objectives: To study Staphylococcus aureus as the causative organism of botryomycosis. Background: the botryomycosis is a chronic purulent granulomatous lesion of the skin, subcutaneous tissue and visceral organs caused by several bacterial species. This condition clinically and histopathologically resembles with that of mycetoma and Actinomycosis. Method: A 51 year old male presented to us with swelling over medial aspect of the right foot with multiple sinuses. He gave a history of trauma 3 years back at the same site. The sample was examined directly by KOH preparation and grams stain. The culture was put up on blood, chocolate, lowenstein Jensen (LJ and sabouraud dextrose agar (SDA media. Fungal culture was negative. Result: Staphylococcus aureus was isolated in aerobic culture. Conclusion: the patient with botryomycosis caused by Staphylococcus aureus was subsequently treated with antibiotics and he recovered completely.

  19. Synergistic antibacterial activity of Curcumin with antibiotics against Staphylococcus aureus.

    Science.gov (United States)

    Teow, Sin-Yeang; Ali, Syed Atif

    2015-11-01

    This study evaluated the synergistic antibacterial activity of Curcumin with 8 different antibiotic groups. Two reference, one clinical and ten environmental strains of Staphylococcus aureus (S. aureus) were tested. Disc diffusion assay with 25 μg/mL Curcumin demonstrated synergism in combination with a majority of tested antibiotics against S. aureus. However, checkerboard micro dilution assay only showed synergism, fractional inhibitory concentration index (FICI) indifferent interactions but no antagonism was observed. In time-kill curve, appreciable reduction of bacterial cells was also observed in combination therapy (Curcumin + antibiotics) compared to monotherapy (Curcumin or antibiotic(s) alone). The antibiotics with higher synergistic interaction with Curcumin are arranged in a decreasing order: Amikacin > Gentamicin > Ciprofloxacin.

  20. Prevalence and Trends of Staphylococcus aureus Bacteraemia in Hospitalized Patients in South Africa, 2010 to 2012: Laboratory-Based Surveillance Mapping of Antimicrobial Resistance and Molecular Epidemiology.

    Directory of Open Access Journals (Sweden)

    Olga Perovic

    Full Text Available We aimed to obtain an in-depth understanding on recent antimicrobial resistance trends and molecular epidemiology trends of S. aureus bacteraemia (SAB.Thirteen academic centres in South Africa were included from June 2010 until July 2012. S. aureus susceptibility testing was performed on the MicroScan Walkaway. Real-time PCR using the LightCycler 480 II was done for mecA and nuc. SCCmec and spa-typing were finalized with conventional PCR. We selected one isolate per common spa type per province for multilocus sequence typing (MLST.S. aureus from 2709 patients were included, and 1231 (46% were resistant to methicillin, with a significant decline over the three-year period (p-value = 0.003. Geographical distribution of MRSA was significantly higher in Gauteng compared to the other provinces (P<0.001. Children <5 years were significantly associated with MRSA with higher rates compared to all other age groups (P = 0.01. The most prevalent SCCmec type was SCCmec type III (531 [41%] followed by type IV (402 [31%]. Spa-typing discovered 47 different spa-types. The five (87% most common spa-types were t037, t1257, t045, t064 and t012. Based on MLST, the commonest was ST612 clonal complex (CC8 (n = 7 followed by ST5 (CC5 (n = 4, ST36 (CC30 (n = 4 and ST239 (CC8 (n = 3.MRSA rate is high in South Africa. Majority of the isolates were classified as SCCmec type III (41% and type IV (31%, which are typically associated with hospital and community- acquired infections, respectively. Overall, this study reveals the presence of a variety of hospital-acquired MRSA clones in South Africa dominance of few clones, spa 037 and 1257. Monitoring trends in resistance and molecular typing is recommended to detect changing epidemiological trends in AMR patterns of SAB.

  1. Prevalence and Trends of Staphylococcus aureus Bacteraemia in Hospitalized Patients in South Africa, 2010 to 2012: Laboratory-Based Surveillance Mapping of Antimicrobial Resistance and Molecular Epidemiology

    Science.gov (United States)

    Perovic, Olga; Iyaloo, Samantha; Kularatne, Ranmini; Lowman, Warren; Bosman, Noma; Wadula, Jeannette; Seetharam, Sharona; Duse, Adriano; Mbelle, Nontombi; Bamford, Colleen; Dawood, Halima; Mahabeer, Yesholata; Bhola, Prathna; Abrahams, Shareef; Singh-Moodley, Ashika

    2015-01-01

    Introduction We aimed to obtain an in-depth understanding on recent antimicrobial resistance trends and molecular epidemiology trends of S. aureus bacteraemia (SAB). Methods Thirteen academic centres in South Africa were included from June 2010 until July 2012. S. aureus susceptibility testing was performed on the MicroScan Walkaway. Real-time PCR using the LightCycler 480 II was done for mecA and nuc. SCCmec and spa-typing were finalized with conventional PCR. We selected one isolate per common spa type per province for multilocus sequence typing (MLST). Results S. aureus from 2709 patients were included, and 1231 (46%) were resistant to methicillin, with a significant decline over the three-year period (p-value = 0.003). Geographical distribution of MRSA was significantly higher in Gauteng compared to the other provinces (P<0.001). Children <5 years were significantly associated with MRSA with higher rates compared to all other age groups (P = 0.01). The most prevalent SCCmec type was SCCmec type III (531 [41%]) followed by type IV (402 [31%]). Spa-typing discovered 47 different spa-types. The five (87%) most common spa-types were t037, t1257, t045, t064 and t012. Based on MLST, the commonest was ST612 clonal complex (CC8) (n = 7) followed by ST5 (CC5) (n = 4), ST36 (CC30) (n = 4) and ST239 (CC8) (n = 3). Conclusions MRSA rate is high in South Africa. Majority of the isolates were classified as SCCmec type III (41%) and type IV (31%), which are typically associated with hospital and community- acquired infections, respectively. Overall, this study reveals the presence of a variety of hospital-acquired MRSA clones in South Africa dominance of few clones, spa 037 and 1257. Monitoring trends in resistance and molecular typing is recommended to detect changing epidemiological trends in AMR patterns of SAB. PMID:26719975

  2. Human scFvs That Counteract Bioactivities of Staphylococcus aureus TSST-1

    Science.gov (United States)

    Rukkawattanakul, Thunchanok; Sookrung, Nitat; Seesuay, Watee; Onlamoon, Nattawat; Diraphat, Pornphan; Chaicumpa, Wanpen; Indrawattana, Nitaya

    2017-01-01

    Some Staphylococcus aureus isolates produced toxic shock syndrome toxin-1 (TSST-1) which is a pyrogenic toxin superantigen (PTSAg). The toxin activates a large fraction of peripheral blood T lymphocytes causing the cells to proliferate and release massive amounts of pro-inflammatory cytokines leading to a life-threatening multisystem disorder: toxic shock syndrome (TSS). PTSAg-mediated-T cell stimulation circumvents the conventional antigenic peptide presentation to T cell receptor (TCR) by the antigen-presenting cell (APC). Instead, intact PTSAg binds directly to MHC-II molecule outside peptide binding cleft and simultaneously cross-links TCR-Vβ region. Currently, there is neither specific TSS treatment nor drug that directly inactivates TSST-1. In this study, human single chain antibodies (HuscFvs) that bound to and neutralized bioactivities of the TSST-1 were generated using phage display technology. Three E. coli clones transfected with TSST-1-bound phages fished-out from the human scFv library using recombinant TSST-1 as bait expressed TSST-1-bound-HuscFvs that inhibited the TSST-1-mediated T cell activation and pro-inflammatory cytokine gene expressions and productions.Computerized simulation, verified by mutations of the residues of HuscFv complementarity determining regions (CDRs),predicted to involve in target binding indicated that the HuscFvs formed interface contact with the toxin residues important for immunopathogenesis. The HuscFvs have high potential for future therapeutic application. PMID:28218671

  3. Clonal complex 398 methicillin susceptible Staphylococcus aureus: a frequent unspecialized human pathogen with specific phenotypic and genotypic characteristics.

    Directory of Open Access Journals (Sweden)

    Tomasz Chroboczek

    Full Text Available Clonal complex 398 livestok-associated-MRSA (CC398 LA-MRSA clone is described as a major animal pathogen that can also colonize and infect humans. CC398 methicillin susceptible Staphylococcus aureus (CC398 MSSA is less described. We identified 126 CC398 MSSA strains of human origin within 6380 S. aureus isolates gathered between 2009 and 2011, from the French National Reference Centre for Staphylococci. They were characterized using antimicrobial susceptibility testing, spa typing, DNA microarrays (Identibac S. aureus Genotyping ®, Alere, CC398-specific sequence PCR, ermT (encoding macrolides résistance PCR. Fifty-three CC398 LA-MRSA collected from French pigs and veal were used as comparators, and phylogenetic relations between human CC398 MSSA and animal CC398 MRSA populations were explored on the basis of spa-typing and DNA microarrays. CC398 MSSA were able to induce a large spectrum of infections (especially skin, bloodstream, and pneumonias. The prevalence rate of this clone was high in MSSA population, i.e., 24.7% in a local prospective study on nasal colonization, and 7.5% in a national prospective study on infective endocarditis. CC398 MSSA isolates were frequently (89% erythromycin resistant, due to the presence of the ermT gene, a gene not detected in erythromycin resistant CC398 LA-MRSA strains. Expression of staphylococcal complement inhibitor (scn and the chemotaxis inhibitory protein (chp, was also specific to this population. The CC398 MRSA signature included also a panel of antibiotic resistance genes, especially a type IV or V cassette mec and tetM. CC398 MSSA and CC398 LA-MRSA populations were closely related based on spa-typing and DNA microarrays, with the MRSA strains forming the most derived lineage in phylogenic trees. Both MSSA and MRSA populations may come from common ancestors, which would have evolved in the settings of different selective pressures, explaining the acquisition of ermT, chp and scn for MSSA, and

  4. Quality control of direct molecular diagnostics for methicillin-resistant Staphylococcus aureus.

    NARCIS (Netherlands)

    A.F. van Belkum (Alex); H.G.M. Niesters (Bert); W.G. MacKay (William); W.B. van Leeuwen (Willem)

    2007-01-01

    textabstractTen samples containing various amounts of methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible S. aureus, methicillin-resistant Staphylococcus epidermidis (MRSE), and combinations thereof were distributed to 51 laboratories for molecular diagnostics testing. Sample

  5. Quality control of direct molecular diagnostics for methicillin-resistant Staphylococcus aureus

    NARCIS (Netherlands)

    van Belkum, Alex; Niesters, Hubert G M; MacKay, William G; van Leeuwen, Willem B

    2007-01-01

    Ten samples containing various amounts of methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible S. aureus, methicillin-resistant Staphylococcus epidermidis (MRSE), and combinations thereof were distributed to 51 laboratories for molecular diagnostics testing. Samples containing

  6. Staphylococcus aureus adherence to Candida albicans hyphae is mediated by the hyphal adhesin Als3p

    NARCIS (Netherlands)

    Peters, Brian M.; Ovchinnikova, Ekaterina S.; Krom, Bastiaan P.; Schlecht, Lisa Marie; Zhou, Han; Hoyer, Lois L.; Busscher, Henk J.; van der Mei, Henny C.; Jabra-Rizk, Mary Ann; Shirtliff, Mark E.

    2012-01-01

    The bacterium Staphylococcus (St.) aureus and the opportunistic fungus Candida albicans are currently among the leading nosocomial pathogens, often co-infecting critically ill patients, with high morbidity and mortality. Previous investigations have demonstrated preferential adherence of St. aureus

  7. Prevalence of infective endocarditis in patients with Staphylococcus aureus bacteraemia: the value of screening with echocardiography

    DEFF Research Database (Denmark)

    Rasmussen, Rasmus V; Høst, Ulla; Arpi, Magnus;

    2011-01-01

    Staphylococcus aureus infective endocarditis (IE) is a critical medical condition associated with a high morbidity and mortality. In the present study, we prospectively evaluated the importance of screening with echocardiography in an unselected S. aureus bacteraemia (SAB) population....

  8. Mupirocin prophylaxis against nosocomial Staphylococcus aureus infections in nonsurgical patients: a randomized study

    NARCIS (Netherlands)

    M.C. Vos (Margreet); A. Ott (Alewijn); A. Voss (Andreas); J.A.J.W. Kluytmans (Jan); C.M.J.E. Vandenbroucke-Grauls (Christina); M.H.M. Meester (Marlene); P.H.J. van Keulen (Peter); H.A. Verbrugh (Henri); H.F.L. Wertheim (Heiman)

    2004-01-01

    textabstractBACKGROUND: Staphylococcus aureus nasal carriage is a major risk factor for nosocomial S. aureus infection. Studies show that intranasal mupirocin can prevent nosocomial surgical site infections. No data are available on the efficacy of mupirocin in nonsurgical patients

  9. The Significance of Nasal Carriage of Staphylococcus Aureus and the Incidence of Postoperative Wound Infection

    NARCIS (Netherlands)

    R.P. Wenzel (Richard); T. M. Perl

    1995-01-01

    textabstractStaphylococcus aureus infections are associated with considerable morbidity and, in certain situations, mortality. The association between the nasal carriage of S. aureus and subsequent infection has been comprehensively established in a variety of clinical settings, in particular, patie

  10. Cross-Talk between Staphylococcus aureus and Other Staphylococcal Species via the agr Quorum Sensing System

    DEFF Research Database (Denmark)

    Canovas de la Nuez, Jaime; Baldry, Mara; Bojer, Martin S;

    2016-01-01

    between staphylococci and S. aureus, and show that this interaction may eventually lead to the identification of new anti-virulence candidates to target S. aureus infections. Here we show that culture supernatants of 37 out of 52 staphylococcal isolates representing 17 different species inhibit S. aureus......-inducing peptides (AIPs) sensed by AgrC, a two component histidine kinase. agr loci are found also in other staphylococcal species and for Staphylococcus epidermidis, the encoded AIP represses expression of agr regulated virulence genes in S. aureus. In this study we aimed to better understand the interaction....... To assess impact on S. aureus virulence, we co-inoculated S. aureus and S. schleiferi in vivo in the Galleria mellonella wax moth larva, and found that expression of key S. aureus virulence factors was abrogated. Our data show that the S. aureus agr locus is highly responsive to other staphylococcal species...

  11. Highly sensitive detection of Staphylococcus aureus directly from patient blood.

    Directory of Open Access Journals (Sweden)

    Padmapriya P Banada

    Full Text Available BACKGROUND: Rapid detection of bloodstream infections (BSIs can be lifesaving. We investigated the sample processing and assay parameters necessary for highly-sensitive detection of bloodstream bacteria, using Staphylococcus aureus as a model pathogen and an automated fluidic sample processing-polymerase chain reaction (PCR platform as a model diagnostic system. METHODOLOGY/PRINCIPAL FINDINGS: We compared a short 128 bp amplicon hemi-nested PCR and a relatively shorter 79 bp amplicon nested PCR targeting the S. aureus nuc and sodA genes, respectively. The sodA nested assay showed an enhanced limit of detection (LOD of 5 genomic copies per reaction or 10 colony forming units (CFU per ml blood over 50 copies per reaction or 50 CFU/ml for the nuc assay. To establish optimal extraction protocols, we investigated the relative abundance of the bacteria in different components of the blood (white blood cells (WBCs, plasma or whole blood, using the above assays. The blood samples were obtained from the patients who were culture positive for S. aureus. Whole blood resulted in maximum PCR positives with sodA assay (90% positive as opposed to cell-associated bacteria (in WBCs (71% samples positive or free bacterial DNA in plasma (62.5% samples positive. Both the assays were further tested for direct detection of S. aureus in patient whole blood samples that were contemporaneous culture positive. S. aureus was detected in 40/45 of culture-positive patients (sensitivity 89%, 95% CI 0.75-0.96 and 0/59 negative controls with the sodA assay (specificity 100%, 95% CI 0.92-1. CONCLUSIONS: We have demonstrated a highly sensitive two-hour assay for detection of sepsis causing bacteria like S. aureus directly in 1 ml of whole blood, without the need for blood culture.

  12. Staphylococcus aureus resistance to topical antimicrobials in atopic dermatitis*

    Science.gov (United States)

    Bessa, Giancarlo Rezende; Quinto, Vanessa Petry; Machado, Daiane Corrêa; Lipnharski, Caroline; Weber, Magda Blessmann; Bonamigo, Renan Rangel; D'Azevedo, Pedro Alves

    2016-01-01

    Background Topical antimicrobial drugs are indicated for limited superficial pyodermitis treatment, although they are largely used as self-prescribed medication for a variety of inflammatory dermatoses, including atopic dermatitis. Monitoring bacterial susceptibility to these drugs is difficult, given the paucity of laboratory standardization. Objective To evaluate the prevalence of Staphylococcus aureus topical antimicrobial drug resistance in atopic dermatitis patients. Methods We conducted a cross-sectional study of children and adults diagnosed with atopic dermatitis and S. aureus colonization. We used miscellaneous literature reported breakpoints to define S. aureus resistance to mupirocin, fusidic acid, gentamicin, neomycin and bacitracin. Results A total of 91 patients were included and 100 S. aureus isolates were analyzed. All strains were methicillin-susceptible S. aureus. We found a low prevalence of mupirocin and fusidic acid resistance (1.1% and 5.9%, respectively), but high levels of neomycin and bacitracin resistance (42.6% and 100%, respectively). Fusidic acid resistance was associated with more severe atopic dermatitis, demonstrated by higher EASI scores (median 17.8 vs 5.7, p=.009). Our results also corroborate the literature on the absence of cross-resistance between the aminoglycosides neomycin and gentamicin. Conclusions Our data, in a southern Brazilian sample of AD patients, revealed a low prevalence of mupirocin and fusidic acid resistance of S. aureus atopic eczema colonizer strains. However, for neomycin and bacitracin, which are commonly used topical antimicrobial drugs in Brazil, high levels of resistance were identified. Further restrictions on the use of these antimicrobials seem necessary to keep resistance as low as possible. PMID:27828633

  13. Staphylococcus aureus in locally produced white cheese in Tirana market

    Directory of Open Access Journals (Sweden)

    ELVIRA BELI

    2014-06-01

    Full Text Available Cheese has nutritional value, its consumption is very common in Albania, but is also excellent medium for bacterial growth, source of bacterial infection, particularly when it is produced from raw poor quality or unpasteurized milk. Microbial safety of cheeses may be enhanced by usage good quality raw milk, pasteurized milk, following GMP in aim to prevent cross-contamination. The aim of this study was to evaluate the presence and amount of Staphylococcus aureus in white cheeses, as an Albanian traditional product. Totally 120 samples of white cheese, produced in small big plant at different Albanian district, by raw milk or pasteurized milk, were collected from Tirana market. All samples were tested by phosphatase test to determine whether raw milk or pasteurized milk it was used for cheese production. 53/120 samples (44% resulted produced by pasteurized milk, 67/120 samples (56 % resulted produced by raw milk. The S. aureus was isolated in Baird Parker agar, and submitted to coagulase and API-staph test. Out of 120 cheese samples, 47 showed contamination by S. aureus coagulase-positive corresponding to 39.16%, otherwise 58 out of 120, 48.33 % of cheeses samples being contaminated with coagulase-negative strain of S. aureus. The occurrence S. aureus coagulase-positive in cheeses produced by pasteurized milk and raw milk it was respectively 7/53 (13.2 % and 40/67 (59.7%. 10% of the samples had high levels 105- 106cfu/g of S. aureus coagulase-positive, suggested that white cheese, may represent a health risk for the consumers

  14. Nonlocal quantum cloning via quantum dots trapped in distant cavities

    Institute of Scientific and Technical Information of China (English)

    Yu Tao; Zhu Ai-Dong; Zhang Shou

    2012-01-01

    A scheme for implementing nonlocal quantum cloning via quantum dots trapped in cavities is proposed.By modulating the parameters of the system,the optimal 1 → 2 universal quantum cloning machine,1 → 2 phase-covariant cloning machine,and 1 → 3 economical phase-covariant cloning machine are constructed.The present scheme,which is attainable with current technology,saves two qubits compared with previous cloning machines.

  15. Whole genome comparison of donor and cloned dogs

    OpenAIRE

    Kim, Hak-Min; Cho, Yun Sung; Kim, Hyunmin; Jho, Sungwoong; Son, Bongjun; Choi, Joung Yoon; Kim, Sangsoo; Lee, Byeong Chun; Bhak, Jong; Jang, Goo

    2013-01-01

    Cloning is a process that produces genetically identical organisms. However, the genomic degree of genetic resemblance in clones needs to be determined. In this report, the genomes of a cloned dog and its donor were compared. Compared with a human monozygotic twin, the genome of the cloned dog showed little difference from the genome of the nuclear donor dog in terms of single nucleotide variations, chromosomal instability, and telomere lengths. These findings suggest that cloning by somatic ...

  16. Cloning and sequencing genes related to preeclampsia

    Institute of Scientific and Technical Information of China (English)

    SHI Juan-zi; LIU Yan-fang; YAO Yuan-qing; YAN Wei; ZHU Feng; ZHAO Zhong-liang

    2001-01-01

    To clone genes specifically expressed in the placenta of patients with preeclampsia, and to explain the mechanism in the etiopathology ofpreeclampsia. Methods: The placentae ofpreeclamptic and normotensive subjects with pregnancy were used as models, and the cDNA Library was constructed and 20 differentially expressed fragments were cloned after a new version of PCR-based subtractive hybridization. The false positive clones were identified by reverse dot blot analysis. With one of the obtained gene taken as the probe, the placentas of 10 normal pregnant women and 10 preeclamptic patients were studied by using dot hybridization methods. Results: Six false positive clones were identified by reverse dot blot, and the rest 14 clones were identified as preeclampsia-related genes. These clones were sequenced, and analyzed with BLAST analysis system. Eleven of 14 clones were genes already known, among which one belongs to necdin family; the rest 3 were identified as novel genes. These 3 genes were acknowledged by GenBank, with the accession numbers AF232216, AF232217, AF233648. The results of dot hybridization using necdin gene as probe were as follows: (1) There was this mRNA in the placental tissues of normal pregnancy as well as in that ofpreeclampsia.(2) The intensity of transcription of this mRNA in the placental tissues of preeclampsia increased significantly compared with that of the normal pregnancy (P<0.05). Conclusions: This study for the first time reported this group of genes, especially necdin-expressing gene, which are related to the etiopathology of preeclampsia. In addition, the overtranscription ofnecdin gene has been found in preeclampsia. It is helpful in further studies of the etiology ofpreeclampsia.

  17. [recovery Of Staphylococcus Aureus After Acid Injury In Milk Products].

    OpenAIRE

    Assis, E M; CARVALHO, E.P. de; E.R. Asquieri; Robbs, P G

    2015-01-01

    The growth behavior of Staphylococcus aureus in fresh Cheese (Minas and Muzzarella) during their shelf-life was studied. The possible injury of this microorganism caused by the increasing acidity was also investigated. Raw milk was inoculated with 10(6) cells/ml (S. aureus FRIA-100) and the cheese production was performed according to normal procedures. Minas and muzzarella cheese were stored at 7 degrees C for 40 and 60 days, respectively. At 2-3 days intervals, the following analysis were p...

  18. CHARACTERISATION OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS ISOLATES FROM SHINGLES PATIENTS

    Directory of Open Access Journals (Sweden)

    Jasmine R. et al.

    2011-11-01

    Full Text Available Even after treating Shingles patients with antiviral drugs, they are found to suffer from secondary bacterial infections. With this background as a guide, we undertook an investigation to isolate the bacterial pathogens from the pus of Shingles patients. Among the isolates obtained during the one year study period, Staphylococcus aureus sp. was found to be multi drug resistant and hence it was chosen for the study. The antibiogram pattern of the methicillin resistant S. aureus was obtained, since this could serve as a tool for suggesting useful drugs.

  19. Staphylococcus aureus sternal osteomyelitis: a rare cause of chest pain

    Directory of Open Access Journals (Sweden)

    Kaur M

    2015-10-01

    Full Text Available Chest pain is a common presenting symptom with a broad differential. Life-threatening cardiac and pulmonary etiologies of chest pain should be evaluated first. However, it is critical to perform a thorough assessment for other sources of chest pain in order to limit morbidity and mortality from less common causes. We present a rare case of a previously healthy 45 year old man who presented with focal, substernal, reproducible chest pain and Staphylococcus aureus bacteremia who was later found to have primary Staphylococcus aureus sternal osteomyelitis.

  20. Staphylococcus aureus Transcriptome Architecture

    DEFF Research Database (Denmark)

    Mäder, Ulrike; Nicolas, Pierre; Depke, Maren;

    2016-01-01

    to their dependence on the RNA polymerase sigma factors SigA or SigB, and allow identification of new potential targets for several known transcription factors. In particular, this study revealed a relatively low abundance of antisense RNAs in S. aureus, where they overlap only 6% of the coding genes, and only 19...... antisense RNAs not co-transcribed with other genes were found. Promoter analysis and comparison with Bacillus subtilis links the small number of antisense RNAs to a less profound impact of alternative sigma factors in S. aureus. Furthermore, we revealed that Rho-dependent transcription termination...

  1. Synthetic peptide inhibitors of DNA replication in Staphylococcus aureus

    DEFF Research Database (Denmark)

    Løbner-Olesen, Anders; Kjelstrup, Susanne

    of clinically important pathogens and is essential for bacterial proliferation. The bacterial replication apparatus fulfill the requirements for a good drug target. The replisome of S. aureus consists of 5 different subunits (2, PolC2, 4, δ and δ`) who’s organization depends on multiple protein-protein...... interactions. Centrally in the replisome is the -clamp where to multiple proteins binds through a conserved motif. We have identified the protein-protein interactions in the replisome of S. aureus by use of a bacterial two-hybrid system. A reverse bacterial two-hybrid system (R-BTH) based on Pyr...

  2. Response of Staphylococcus Aureus to a Spaceflight Analogue

    Science.gov (United States)

    Castro, S. L.; Ott, C. M.

    2010-01-01

    The decreased gravity of the spaceflight environment creates quiescent, low fluid shear conditions. This environment can impart considerable effects on the physiology of microorganisms as well as their interactions with potential hosts. Using the rotating wall vessel (RWV), as a spaceflight analogue, the consequence of low fluid shear culture on microbial pathogenesis has provided a better understanding of the risks to the astronaut crew from infectious microorganisms. While the outcome of low fluid shear culture has been investigated for several bacterial pathogens, little has been done to understand how this environmental factor affects Staphylococcus aureus. S. aureus is an opportunistic human pathogen which presents a high level of infection risk to the crew, as it has been isolated from both the space shuttle and International Space Station. Given that approximately forty percent of the population are carriers of the bacteria, eradication of this organism from in flight environments is impractical. These reasons have lead to us to assess the response of S. aureus to a reduced fluid shear environment. Culture in the RWV demonstrated that S. aureus grown under the low-shear condition had lower cell concentrations after 10 hours when compared to the control culture. Furthermore, the low-shear cultured bacteria displayed a reduction in carotenoid production, pigments responsible for their yellow/gold coloration. When exposed to various environmental stressors, post low-shear culture, a decrease in the ability to survive oxidative assault was observed compared to control cultures. The low fluid shear environment also resulted in a decrease in hemolysin secretion, a staphylococcal toxin responsible for red blood cell lysis. When challenged by the immune components present in human whole blood, low-shear cultured S. aureus demonstrated significantly reduced survival rates as compared to the control culture. Assays to determine the duration of these alterations

  3. Identification of the ClpX Regulon in Staphylococcus aureus

    DEFF Research Database (Denmark)

    Jelsbak, Lotte; Thomsen, Line Elnif; Ingmer, Hanne;

    Staphyloccous aureus is a major human pathogen capable of causing a wide spectrum of infections ranging from superficial wound infections to life-threatening endocarditis and toxic shock syndrome. Essential for S. aureus virulence is a large number of cell-surface-associated proteins and secreted...... proteins. Results from our group have shown that the ClpXP proteolytic complex and the ClpX chaperone play central roles in regulating expression of many of these factors (2;3). By using DNA microarrays to compare transcription of strain 8325-4 (wt) and the isogenic ¿clpX strain during the transition phase...

  4. Disruption of Methicillin-resistant Staphylococcus aureus Biofilms with Enzymatic Therapeutics

    Science.gov (United States)

    2015-04-29

    NAVAL MEDICAL RESEARCH UNIT SAN ANTONIO Disruption of Methicillin-resistant Staphylococcus aureus Biofilms with Enzymatic...fold lower than that needed to thoroughly disrupt biofilms in the current investigation. A previous study of α-amylase applied to S. aureus biofilms...Staphylococcus aureus biofilms. Open Microbiology Journal, 2011. 5: p. 21-31. 36. Wu, J.A., et al., Lysostaphin disrupts Staphylococcus aureus and

  5. Minimum inhibitory concentration of ciprofloxacin in combination with hexahydroquinoline derivatives against Staphylococcus aureus

    OpenAIRE

    F Amin Harati; M Amini; Shahverdi AR; Pourmand, MR; Yousefi, M.

    2012-01-01

    Background: Staphylococcus aureus is the most common pathogen responsible for skin and soft tissue infections worldwide. Methicillin-resistant S. aureus is a major cause of both nosocomial and community acquired infections. The emergence of antimicrobial-resistant S. aureus is of global concern. Fluoroquinolone antimicrobials including ciprofloxacin, levofloxacin, and moxifloxacin are used to treat skin and soft tissue infections due to S. aureus. Emergence of ciprofloxacin resistance has inc...

  6. Changes in the Staphylococcus aureus Transcriptome during Early Adaptation to the Lung

    OpenAIRE

    Chaffin, Donald O.; Destry Taylor; Skerrett, Shawn J.; Craig E Rubens

    2012-01-01

    Staphylococcus aureus is a common inhabitant of the human nasopharynx. It is also a cause of life-threatening illness, producing a potent array of virulence factors that enable survival in normally sterile sites. The transformation of S. aureus from commensal to pathogen is poorly understood. We analyzed S. aureus gene expression during adaptation to the lung using a mouse model of S. aureus pneumonia. Bacteria were isolated by bronchoalveolar lavage after residence in vivo for up to 6 hours....

  7. Piglets born from handmade cloning, an innovative cloning method without micromanipulation

    DEFF Research Database (Denmark)

    Du, Y.; Kragh, P.M.; Zhang, Y.

    2007-01-01

    Porcine handmade cloning (HMC), a simplified alternative of micromanipulation based traditional cloning (TC) has been developed in multiple phases during the past years, but the final evidence of its biological value, births of piglets was missing. Here we report the first births of healthy piglets...

  8. Update on the First Cloned Dog and Outlook for Canine Cloning.

    Science.gov (United States)

    Jang, Goo; Lee, ByeongChun

    2015-10-01

    As man's best friend, dogs have an important position in human society. Ten years ago, we reported the first cloned dog, and his birth has raised various scientific issues, such as those related to health, reproduction, and life span. He has developed without any unique health issues. In this article, we summarize and present perspectives on canine cloning.

  9. Cloning mice and men: prohibiting the use of iPS cells for human reproductive cloning.

    Science.gov (United States)

    Lo, Bernard; Parham, Lindsay; Alvarez-Buylla, Arturo; Cedars, Marcelle; Conklin, Bruce; Fisher, Susan; Gates, Elena; Giudice, Linda; Halme, Dina Gould; Hershon, William; Kriegstein, Arnold; Kwok, Pui-Yan; Wagner, Richard

    2010-01-08

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation.

  10. Cloning Mice and Men: Prohibiting the Use of iPS Cells for Human Reproductive Cloning

    OpenAIRE

    Lo, Bernard; Parham, Lindsay; Alvarez-Buylla, Arturo; Cedars, Marcelle; Conklin, Bruce; Fisher, Susan; Gates, Elena; Giudice, Linda; Halme, Dina Gould; Hershon, William; Kriegstein, Arnold; Kwok, Pui-Yan; Wagner, Richard

    2010-01-01

    The use of iPSCs and tetraploid complementation for human reproductive cloning would raise profound ethical objections. Professional standards and laws that ban human reproductive cloning by somatic cell nuclear transfer should be revised to also forbid it by other methods, such as iPSCs via tetraploid complementation.

  11. Familial Clustering of Staphylococcus aureus Bacteremia in First-Degree Relatives

    DEFF Research Database (Denmark)

    Oestergaard, Louise B.; Christiansen, Mia N.; Schmiegelow, Michelle D.

    2016-01-01

    BACKGROUND: A genetic predisposition to Staphylococcus aureus bacteremia has been demonstrated in animals, suggesting that genetic differences might influence susceptibility to S aureus in humans. OBJECTIVE: To determine whether a history of S aureus bacteremia in first-degree relatives increases...

  12. Long-term cortisol levels are not associated with nasal carriage of Staphylococcus aureus

    NARCIS (Netherlands)

    L. Manenschijn (Laura); A.M. Jetten; W.J.B. van Wamel (Willem); M. Tavakol (Mehri); E.L.T. van den Akker (Erica); J.W. Koper (Jan); A.F. van Belkum (Alex); E.F.C. van Rossum (Liesbeth)

    2012-01-01

    textabstractStaphylococcus aureus (S. aureus) colonizes the anterior nares in part of the population and the persistent carrier state is associated with increased infection risk. Knowledge concerning the determinants of S. aureus nasal carriage is limited. Previously, we found that glucocorticoid re

  13. Genotypic characterisation of Staphylococcus aureus isolates causing bacteraemia at Tygerberg hospital, western cape province, South Africa

    NARCIS (Netherlands)

    Orth, H.; Salaam-Dreyer, Z.; Makgotlho, E.; Sinha, B.; Wasserman, E.

    2011-01-01

    Objectives: There is a paucity of studies on the genotypic characterisation of invasive S. aureus strains and the incidence of communityacquired methicillin resistant S. aureus (CA-MRSA) infections in South Africa. In this study we characterized S. aureus isolates from bacteraemia episodes using mol

  14. Phenotypic and genotypic antimicrobial resistance traits of foodborne Staphylococcus aureus isolates from Shanghai

    Science.gov (United States)

    Staphylococcus aureus is a recognized pathogen in humans, which causes nosocomial infections and food poisoning. The transmission of antibiotic resistant S. aureus (ARSA), especially methicillin-resistant S. aureus (MRSA), between food products and humans has become a serious problem. Hence, it is n...

  15. Antibiotic-mediated selection of quorum-sensing-negative Staphylococcus aureus

    DEFF Research Database (Denmark)

    Paulander, Wilhelm Erik Axel; Varming, Anders Nissen; Bæk, Kristoffer Torbjørn;

    2012-01-01

    -acquired S. aureus infections and suggest that the adaptability of S. aureus to antibiotics involves the agr locus. IMPORTANCE: Staphylococcus aureus is the most frequently isolated pathogen in intensive care units and a common cause of nosocomial infections, resulting in a high degree of morbidity...

  16. Elephant grass clones for silage production

    Directory of Open Access Journals (Sweden)

    Rerisson José Cipriano dos Santos

    2013-02-01

    Full Text Available Ensiling warm-season grasses often requires wilting due to their high moisture content, and the presence of low-soluble sugars in these grasses usually demands the use of additives during the ensiling process. This study evaluated the bromatological composition of the fodder and silage from five Pennisetum sp. clones (IPA HV 241, IPA/UFRPE Taiwan A-146 2.114, IPA/UFRPE Taiwan A-146 2.37, Elephant B, and Mott. The contents of 20 Polyvinyl chloride (PVC silos, which were opened after 90 days of storage, were used for the bromatological analysis and the evaluation of the pH, nitrogen, ammonia, buffer capacity, soluble carbohydrates, and fermentation coefficients. The effluent losses, gases and dry matter recovery were also calculated. Although differences were observed among the clones (p < 0.05 for the concentrations of dry matter, insoluble nitrogen in acid detergents, insoluble nitrogen in neutral detergents, soluble carbohydrates, fermentation coefficients, and in vitro digestibility in the forage before ensiling, no differences were observed for most of these variables after ensiling. All of the clones were efficient in the fermentation process. The IPA/UFRPE TAIWAN A-146 2.37 clone, however, presented a higher dry matter concentration and the best fermentation coefficient, resulting in a better silage quality, compared to the other clones.

  17. Consensus maps of cloned plant cuticle genes

    Institute of Scientific and Technical Information of China (English)

    Eviatar; Nevo

    2010-01-01

    Plant cuticle,which covers the plant surface,consists of waxes and cutins,and is associated with plant drought,cold,and salt resistance.Hitherto,at least 47 genes participating in the formation of plant cuticle have been cloned from Arabidopsis thaliana,Oryza sativa,Zea mays,Ricinus communis,Brassica napus,and Medicago truncatula;and about 85% of them encode proteins sharing above 50% identities with their rice homologous sequences.These cloned cuticle genes were mapped in silico on different chromosomes of rice and Arabidopsis,respectively.The mapping results revealed that plant cuticle genes were not evenly distributed in both genomes.About 40% of the mapped cuticle genes were located on chromosome 1 in Arabidopsis,while 20% of the mapped cuticle genes were located on chromosome 2 but none on chromosome 12 in rice.Some cloned plant cuticle genes have several rice homologous sequences,which might be produced by chromosomal segment duplication.The consensus map of cloned plant cuticle genes will provide important clues for the selection of candidate genes in a positional cloning of an unknown cuticle gene in plants.

  18. Pre-weaning performance and health of pigs born to cloned (fetal cell derived) swine versus non-cloned swine.

    Science.gov (United States)

    Martin, M; Adams, C; Wiseman, B

    2004-07-01

    The objective of this study was to compare the pre-weaning performance of pigs derived from cloned versus non-cloned parents. Five cloned gilts and one cloned boar were used to produce five litters of pigs. One of five cloned females and the cloned boar were derived from two genetically unmanipulated fetal fibroblast cell lines. The remaining female clones were derived from a fetal fibroblast cell line in which random insertion of a alpha-1,3-galactosyltransferase gene targeting construct had occurred. Fetal cell lines had similar genetic backgrounds and were derived from three different fetuses in three different litters. Five litters of pigs were also generated from matings between two non-cloned boars and five non-cloned gilts. The mean gestation length, mean litter size, mean birth and weaning weights for male and female pigs were similar for litters derived from cloned parents versus non-cloned parents. The proportions of pigs born live and pigs that survived to weaning were also similar for pigs born to cloned as compared to non-cloned parents. In summary, matings between cloned swine derived from fetal fibroblast cell lines yielded litters of pigs that were similar in the number born, piglet birth weight and perinatal and pre-weaning mortality to litters produced by non-cloned swine.

  19. Prevalence and diversity of enterotoxin genes with genetic background of Staphylococcus aureus isolates from different origins in China.

    Science.gov (United States)

    Chao, Guoxiang; Bao, Guangyu; Cao, Yongzhong; Yan, Wenguang; Wang, Yan; Zhang, Xiaorong; Zhou, Liping; Wu, Yantao

    2015-10-15

    Staphylococcal enterotoxins (SE) induce toxin-mediated diseases, such as food poisoning. In the present study, 568 isolates from different sources were tested for the prevalence of 18 SE genes and performed spa typing. In addition, we characterized the relationships between the distribution of SE genes and molecular clones based on multilocus sequence typing (MLST), spa and staphylococcal cassette chromosome mec (SCCmec) typing in selected 250 isolates. Approximately 54.40% of the isolates from different sources harbored one or more SE genes forming 120 distinct gene profiles. Seven genes, sea, seb, seg, seo, sem, seq, and sel were more frequently detected. The distributions of the SE genes among the isolates from human, animals, and foodborne origins were highly different with isolates from environments (P0.05). We identified two important gene clusters, sea-sek-seq, which is closely related to hospital-acquired (HA) methicillin-resistant Staphylococcus aureus (MRSA)-III, and the egc cluster, which accounts for nearly half of all genes. Approximately 71% isolates could be typed by spa, yielding 103 spa types, of which 18 spa types were primary types. In clonal complex (CC) 239, an important Asian HA-MRSA-III clone from humans, nearly all isolates harbored complete or partial sea-sek-seq cluster; the main spa types were t030 and t037. In CC630, an important new community-associated (CA) MRSA-V CC in China, only sporadic SE genes, three main spa types, t4549, t2196, and t377 were observed. The egc cluster coexisting with other genes was present in isolates of CC5, CC9, CC1281, CC1301, CC30 and sequence type (ST) 25, but completely absent in isolates of CC239, CC59, CC7, and CC88. The results illustrate the genetic clonal diversity and the identity of S. aureus isolates from different sources with respect to SE genes and highlight a correlation between SE genes or gene clusters and CCs, spa, and MRSA clones. The foodborne and human origin isolates were the main

  20. MOLECULAR CLONING OF HUMAN NEUROTROPHIN-4 GENE

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective Cloning and sequencing of the human neurotrophin-4(hNT-4) gene.Methods With the chromosomal DNA of human blood lymphocytes as template,hNT-4 coding genes were amplified by polymerase chain reaction(PCR) and recombinated into phage vector pGEM-T Easy,which were sequenced by using Sanger's single stranded DNA terminal termination method.Results The sequence of the cloned gene is completely the same as that reported in the literature(GenBank data base,M86528).Conclusion This study successfully cloning and sequenced the gene of mhNT-4,and it would be convenient for us to study the expression of mhNT-4 in eukaryote,and to continue the research on the gene therapy of Alzheimer's disease intensively.This study indicate that the hNT-4 is conservative in different races and individuals.

  1. Simplified cryopreservation of porcine cloned blastocysts

    DEFF Research Database (Denmark)

    Du, Yutao; Zhang, Yunhai; Li, Juan

    2007-01-01

    )â€"handmade cloning (HMC)â€"to establish a simplified and efficient cryopreservation system for porcine cloned embryos. In Experiment 1, zonae pellucidae of oocytes were partially digested with pronase, followed by centrifugation to polarize lipid particles. Ninety percent (173/192) oocytes were successfully...... delipated in this way. Parthenogenetic activation (PA) after complete removal of zona resulted in similar blastocyst rates in delipated vs. control oocytes (28 ± 7% vs. 28 ± 5%, respectively). Subsequent vitrification of produced blastocysts with the Cryotop technique resulted in higher survival rates......). Our results prove that porcine embryos produced from delipated oocytes by PA or HMC can be cryopreserved effectively by ultrarapid vitrification. Further experiments are required to assess the in vivo developmental competence of the cloned-vitrified embryos  ...

  2. Bac clones generated from sheared dna

    Energy Technology Data Exchange (ETDEWEB)

    Osoegawa, Kazutoyo; Vessere, Gery M.; Shu, Chung Li; Hoskins,Roger A.; Abad, Jose P.; de Pablos, Beatriz; Villasante, Alfredo; deJong, Pieter J.

    2006-08-09

    BAC libraries generated from restriction-digested genomic DNA display representational bias and lack some sequences. To facilitate completion of genome projects, procedures have been developed to create BACs from DNA physically sheared to create fragments extending up to 200kb. The DNA fragments were repaired to create blunt ends and ligated to a new BAC vector. This approach has been tested by generating BAC libraries from Drosophila DNA, with insert lengths of 50 kb to 150 kb. The libraries lack chimeric clone problems as determined by mapping paired BAC-end sequences of one library to the D. melanogaster genome sequence. The utility of ''sheared'' libraries was demonstrated by closure of a previous clone gap and by isolation of clones from telomeric regions, which were notably absent from previous Drosophila BAC libraries.

  3. THE CLONING OF HUMAN NEUROTROPHIN-3 GENE

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    In the present study, we have cloned the gene of human neurotrophin-3 (hNT-3) from the genomic DNA of white blood cells (WBC) by polymerase chain reaction (PCR). The amplification products were cloned into pUC19 and sequenced. Genomic sequence comparison of the cloned fragment and the reported hNT-3 (GenBank M61180) reveals 7 base differences: 1 in the signal peptide, 3 in the prepro peptide, and 3 in the mature hNT-3. Except the 2 varied bases (16th, T to G; 285th, A to C) in the signal peptide and pro-sequence resulted in the change of their encoded amino-acids (Tyr→Asp; Gln→His), the other varied bases have no influence on their respective encoded amino-acids, and all the changes have no influence on the open reading frame (ORF) of the hNT-3.

  4. Dogs cloned from adult somatic cells.

    Science.gov (United States)

    Lee, Byeong Chun; Kim, Min Kyu; Jang, Goo; Oh, Hyun Ju; Yuda, Fibrianto; Kim, Hye Jin; Hossein, M Shamim; Shamim, M Hossein; Kim, Jung Ju; Kang, Sung Keun; Schatten, Gerald; Hwang, Woo Suk

    2005-08-04

    Several mammals--including sheep, mice, cows, goats, pigs, rabbits, cats, a mule, a horse and a litter of three rats--have been cloned by transfer of a nucleus from a somatic cell into an egg cell (oocyte) that has had its nucleus removed. This technology has not so far been successful in dogs because of the difficulty of maturing canine oocytes in vitro. Here we describe the cloning of two Afghan hounds by nuclear transfer from adult skin cells into oocytes that had matured in vivo. Together with detailed sequence information generated by the canine-genome project, the ability to clone dogs by somatic-cell nuclear transfer should help to determine genetic and environmental contributions to the diverse biological and behavioural traits associated with the many different canine breeds.

  5. Human reproductive cloning and reasons for deprivation.

    Science.gov (United States)

    Jensen, D A

    2008-08-01

    Human reproductive cloning provides the possibility of genetically related children for persons for whom present technologies are ineffective. I argue that the desire for genetically related children is not, by itself, a sufficient reason to engage in human reproductive cloning. I show this by arguing that the value underlying the desire for genetically related children implies a tension between the parent and the future child. This tension stems from an instance of a deprivation and violates a general principle of reasons for deprivation. Alternative considerations, such as a right to procreative autonomy, do not appear helpful in making the case for human reproductive cloning merely on the basis of the desire for genetically related children.

  6. Cloning for human reproduction: one American perspective.

    Science.gov (United States)

    Chester, R

    2001-09-01

    The author, an American law professor, believes that whole-body cloning of adult humans will be possible in the near future. He does not believe the procedure should be banned when used as a form of assisted reproduction, but that it should be regulated by the government to ensure proper testing and application. After raising a number of scientific, ethical, religious and legal issues, Professor Chester addresses parentage in light of both old and new concepts of the 'family.' Finally, he focuses on the problem of women as surrogate mothers of clones, arguing in the process that the surrogate, having no real genetic tie to the clone, would have less of a claim to parentage than at least some of the surrogates currently gestating foetuses.

  7. Human cloning: three mistakes and an alternative.

    Science.gov (United States)

    Baylis, Françoise

    2002-06-01

    The current debate on the ethics of cloning humans is both uninspired and uninspiring. In large measure this is because of mistakes that permeate the discourse, including the mistake of thinking that cloning technology is strictly a reproductive technology when it is used to create whole beings. As a result, the challenge this technology represents regarding our understanding of ourselves and the species to which we belong typically is inappropriately downplayed or exaggerated. This has meant that important (albeit disquieting) societal issues and species-type concerns have not been fully explored. This paper, intended as a corrective, suggests that we take an alternate view of human cloning as both an enhancement and a reproductive technology. This proposed shift in the framework for analysis counters the current narrow framing of the issues and introduces new questions about the prospect of modifying the species.

  8. Threat of multidrug resistant Staphylococcus aureus in Western Nepal

    DEFF Research Database (Denmark)

    Bhatta, Dharm R.; Cavaco, Lina; Nath, Gopal;

    2015-01-01

    ObjectiveTo determine the prevalence of methicillin resistant Staphylococcus aureus (MRSA) and antimicrobial susceptibility patterns of the isolates from Manipal Teaching Hospital, Pokhara, Nepal. MethodsThis study was conducted over a period of 11 months (September 2012–August 2013) at the Manip...

  9. Methicillin-Resistant Staphylococcus aureus Colonization among Medical Residents

    Directory of Open Access Journals (Sweden)

    Pascale Trépanier

    2013-01-01

    Full Text Available BACKGROUND: Medical residents may be at risk of becoming colonized by methicillin-resistant Staphylococcus aureus (MRSA during their training. The occupational risk of this specific population is unknown. Furthermore, there are no data regarding MRSA colonization among health care professionals in Quebec.

  10. Multilocus sequence typing of Staphylococcus aureus with DNA array technology

    NARCIS (Netherlands)

    W.B. van Leeuwen (Willem); C. Jay (Corinne); S.V. Snijders (Susan); N. Durin (Nathalia); B. Lacroix (Bruno); H.A. Verbrugh (Henri); M.C. Enright (Mark); A. Troesch (Alain); A.F. van Belkum (Alex)

    2003-01-01

    textabstractA newly developed oligonucleotide array suited for multilocus sequence typing (MLST) of Staphylococcus aureus strains was analyzed with two strain collections in a two-center study. MLST allele identification for the first strain collection fully agreed with conventiona

  11. Staphylococcus aureus colonization related to severity of hand eczema.

    Science.gov (United States)

    Mernelius, S; Carlsson, E; Henricson, J; Löfgren, S; Lindgren, P-E; Ehricht, R; Monecke, S; Matussek, A; Anderson, C D

    2016-08-01

    Knowledge on Staphylococcus aureus colonization rates and epidemiology in hand eczema is limited. The aim of this study was to clarify some of these issues. Samples were collected by the "glove juice" method from the hands of 59 patients with chronic hand eczema and 24 healthy individuals. Swab samples were taken from anterior nares and throat from 43 of the 59 patients and all healthy individuals. S. aureus were spa typed and analysed by DNA-microarray-based genotyping. The extent of the eczema was evaluated by the hand eczema extent score (HEES). The colonization rate was higher on the hands of hand eczema patients (69 %) compared to healthy individuals (21 %, p eczema (HEES ≥ 13) had a significantly higher S. aureus density on their hands compared to those with milder eczema (HEES = 1 to 12, p = 0.004). There was no difference between patients and healthy individuals regarding colonization rates in anterior nares or throat. spa typing and DNA-microarray-based genotyping indicated certain types more prone to colonize eczematous skin. Simultaneous colonization, in one individual, with S. aureus of different types, was identified in 60-85 % of the study subjects. The colonization rate and density indicate a need for effective treatment of eczema and may have an impact on infection control in healthcare.

  12. Menaquinone biosynthesis potentiates haem toxicity in Staphylococcus aureus.

    Science.gov (United States)

    Wakeman, Catherine A; Hammer, Neal D; Stauff, Devin L; Attia, Ahmed S; Anzaldi, Laura L; Dikalov, Sergey I; Calcutt, M Wade; Skaar, Eric P

    2012-12-01

    Staphylococcus aureus is a pathogen that infects multiple anatomical sites leading to a diverse array of diseases. Although vertebrates can restrict the growth of invading pathogens by sequestering iron within haem, S. aureus surmounts this challenge by employing high-affinity haem uptake systems. However, the presence of excess haem is highly toxic, necessitating tight regulation of haem levels. To overcome haem stress, S. aureus expresses the detoxification system HrtAB. In this work, a transposon screen was performed in the background of a haem-susceptible, HrtAB-deficient S. aureus strain to identify the substrate transported by this putative pump and the source of haem toxicity. While a recent report indicates that HrtAB exports haem itself, the haem-resistant mutants uncovered by the transposon selection enabled us to elucidate the cellular factors contributing to haem toxicity. All mutants identified in this screen inactivated the menaquinone (MK) biosynthesis pathway. Deletion of the final steps of this pathway revealed that quinone molecules localizing to the cell membrane potentiate haem-associated superoxide production and subsequent oxidative damage. These data suggest a model in which membrane-associated haem and quinone molecules form a redox cycle that continuously generates semiquinones and reduced haem, both of which react with atmospheric oxygen to produce superoxide.

  13. Putative link between Staphylococcus aureus bacteriophage serotype and community association.

    Science.gov (United States)

    Mohamed, D H; Saberesheikh, S; Kearns, A M; Saunders, N A

    2012-07-01

    Methicillin-resistant Staphylococcus aureus (MRSA) from humans can be broadly separated into 3 groups: healthcare-associated (HA), community-associated (CA), and livestock-associated (LA) MRSA. Initially based on epidemiological features, division into these classes is becoming increasingly problematic. The sequencing of S. aureus genomes has highlighted variations in their accessory components, which likely account for differences in pathogenicity and epidemicity. In particular, temperate bacteriophages have been regarded as key players in bacterial pathogenesis. Bacteriophage-associated Panton-Valentine leukocidin genes (luk-PV) are regarded as epidemiological markers of the CA-MRSA due to their high prevalence in CA strains. This paper describes the development and application of a partial composite S. aureus virulence-associated gene microarray. Epidemic, pandemic, and sporadic lineages of UK HA and CA S. aureus were compared. Phage structural genes linked with CA isolates were identified and in silico analysis revealed these to be correlated with phage serogroup. CA strains predominantly carried a PVL-associated phage either of the A or Fb serogroup, whilst HA strains predominantly carried serogroup Fa or B phages. We speculate that carriage of a serogroup A/Fb PVL-associated phage rather than the luk-PV genes specifically is correlated with CA status.

  14. Phenotype switching is a natural consequence of Staphylococcus aureus replication.

    Science.gov (United States)

    Edwards, Andrew M

    2012-10-01

    The pathogen Staphylococcus aureus undergoes phenotype switching in vivo from its normal colony phenotype (NCP) to a slow-growing, antibiotic-resistant small-colony-variant (SCV) phenotype that is associated with persistence in host cells and tissues. However, it is not clear whether phenotype switching is the result of a constitutive process that is selected for under certain conditions or is triggered by particular environmental stimuli. Examination of cultures of diverse S. aureus strains in the absence of selective pressure consistently revealed a small gentamicin-resistant SCV subpopulation that emerged during exponential-phase NCP growth and increased in number until NCP stationary phase. Treatment of replicating bacteria with the antibiotic gentamicin, which inhibited NCP but not SCV replication, resulted in an initial decrease in SCV numbers, demonstrating that SCVs arise as a consequence of NCP replication. However, SCV population expansion in the presence of gentamicin was reestablished by selection of phenotype-stable SCVs and subsequent SCV replication. In the absence of selective pressure, however, phenotype switching was bidirectional and occurred at a high frequency during NCP replication, resulting in SCV turnover. In summary, these data demonstrate that S. aureus phenotype switching occurs via a constitutive mechanism that generates a dynamic, antibiotic-resistant subpopulation of bacteria that can revert to the parental phenotype. The emergence of SCVs can therefore be considered a normal part of the S. aureus life cycle and provides an insurance policy against exposure to antibiotics that would otherwise eliminate the entire population.

  15. Surveillance van meticilline resistente Staphylococcus aureus in Nederland in 1990

    NARCIS (Netherlands)

    Frenay HME; van Leeuwen WJ; van Klingeren B; Rost JA; Schot CS

    1991-01-01

    Follow-up studies on the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Dutch hospitals were continued in 1990. The number of MRSA-isolates in 1990 compared to 1989 is approximately the same. Phage-type pattern and antibiogram were determined for 168 MRSA-isolates from 42 hosp

  16. Staphylococcus aureus with reduced susceptibility to vancomycin in healthcare settings.

    Science.gov (United States)

    Spagnolo, A M; Orlando, P; Panatto, D; Amicizia, D; Perdelli, F; Cristina, M L

    2014-12-01

    Glycopeptide resistance in Staphylococcus aureus is a source of great concern because, especially in hospitals, this class of antibiotics, particularly vancomycin, is one of the main resources for combating infections caused by methicillin-resistant Staphylococcus aureus strains (MRSA). Reduced susceptibility to vancomycin (VISA) was first described in 1996 in Japan; since then, a phenotype with heterogeneous resistance to vancomycin (h-VISA) has emerged. H-VISA isolates are characterised by the presence of a resistant subpopulation, typically at a rate of 1 in 10(5) organisms, which constitutes the intermediate stage betweenfully vancomycin-susceptible S. aureus (VSSA) and VISA isolates. As VISA phenotypes are almost uniformly cross-resistant to teicoplanin, they are also called Glycopeptides-intermediate Staphylococcus aureus strains (GISA) and, in the case of heterogeneous resistance to glycopeptides, h-GISA. The overall prevalence of h-VISA is low, accounting for approximately 1.3% of all MRSA isolates tested. Mortality due to h-GISA infections is very high (about 70%), especially among patients hospitalised in high-risk departments, such as intensive care units (ICU). Given the great clinical relevance of strains that are heteroresistant to glycopeptides and the possible negative impact on treatment choices, it is important to draw up and implement infection control practices, including surveillance, the appropriate use of isolation precautions, staff training, hand hygiene, environmental cleansing and good antibiotic stewardship.

  17. Superantigens in Staphylococcus aureus isolated from prosthetic joint infection

    Science.gov (United States)

    Kim, Choon K.; Karau, Melissa J.; Greenwood-Quaintance, Kerryl E.; Tilahun, Ashenafi Y.; David, Chella S.; Mandrekar, Jayawant N.; Patel, Robin; Rajagopalan, Govindarajan

    2014-01-01

    Staphylococcus aureus is a common cause of prosthetic joint infection (PJI). The prevalence of superantigens (SAgs) among PJI-associated S. aureus is unknown. Eighty-four S. aureus isolates associated with PJI isolated between 1999 and 2006 were studied. SAg genes, sea, seb, sec, sed, see, seg, seh, sei and tst, were assayed by PCR. Seventy-eight (92.9%) isolates carried at least one SAg gene studied, with 61 (72.6%) harboring more than one. seg was most commonly (70.2%) and seh was least frequently (4.8%) detected. tst-positive isolates were associated with early infection and increased ESR at diagnosis (P = 0.006 and P = 0.021, respectively). seg and sei were associated with methicillin resistance (P = 0.008 and 0.002, respectively). SAg genes are prevalent in S. aureus causing PJI; a majority of PJI-associated isolates produce biologically active SAgs in both planktonic and biofilm growth modes. PMID:25619753

  18. Pulsed-field gel electrophoresis typing of Staphylococcus aureus isolates

    Science.gov (United States)

    Pulsed-field gel electrophoresis (PFGE) is the most applied and effective genetic typing method for epidemiological studies and investigation of foodborne outbreaks caused by different pathogens, including Staphylococcus aureus. The technique relies on analysis of large DNA fragments generated by th...

  19. An Interdisciplinary Experiment: Azo-Dye Metabolism by "Staphylococcus Aureus"

    Science.gov (United States)

    Brocklesby, Kayleigh; Smith, Robert; Sharp, Duncan

    2012-01-01

    An interdisciplinary and engaging practical is detailed which offers great versatility in the study of a qualitative and quantitative metabolism of azo-dyes by "Staphylococcus aureus". This practical has broad scope for adaptation in the number and depth of variables to allow a focused practical experiment or small research project. Azo-dyes are…

  20. Increased risk of arterial thromboembolic events after Staphylococcus aureus bacteremia

    DEFF Research Database (Denmark)

    Mejer, N; Gotland, N; Uhre, M L;

    2015-01-01

    OBJECTIVES: An association between infection and arterial thromboembolic events (ATE) has been suggested. Here we examined the risk of myocardial infarction (MI), stroke and other ATE after Staphylococcus aureus bacteremia (SAB). METHODS: Danish register-based nation-wide observational cohort study...

  1. Staphylococcus aureus causing tropical pyomyositis, Amazon Basin, Peru.

    NARCIS (Netherlands)

    Garcia, C.; Hallin, M.; Deplano, A.; Denis, O.; Sihuincha, M.; Groot, R. de; Gotuzzo, E.; Jacobs, J.

    2013-01-01

    We studied 12 Staphylococcus aureus isolates causing tropical pyomyositis in the Amazon Basin of Peru. All isolates were methicillin-susceptible; 11 carried Panton-Valentine leukocidin-encoding genes, and 5 belonged to multilocus sequence type 25 and possessed an extensive set of enterotoxins. Our f

  2. Diabetes and risk of community-acquired Staphylococcus aureus bacteremia

    DEFF Research Database (Denmark)

    Smit, Jesper; Søgaard, Mette; Schønheyder, Henrik Carl;

    2016-01-01

    OBJECTIVE: Patients with diabetes may experience higher risk of Staphylococcus aureus bacteremia (SAB) than patients without diabetes due to decreased immunity or coexisting morbidities. We investigated the risk of community-acquired (CA) SAB in persons with and without diabetes. DESIGN: Using...

  3. Transmissibility of Livestock-associated Methicillin-Resistant Staphylococcus aureus

    NARCIS (Netherlands)

    Hetem, D.J.; Bootsma, M.C.J.; Troelstra, A.; Bonten, M.J.M.

    2013-01-01

    Previous findings have suggested that the nosocomial transmission capacity of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is lower than that of other MRSA genotypes. We therefore performed a 6-month (June 1–November 30, 2011) nationwide study to quantify the single-adm

  4. Staphylococcus aureus α toxin potentiates opportunistic bacterial lung infections.

    Science.gov (United States)

    Cohen, Taylor S; Hilliard, Jamese J; Jones-Nelson, Omari; Keller, Ashley E; O'Day, Terrence; Tkaczyk, Christine; DiGiandomenico, Antonio; Hamilton, Melissa; Pelletier, Mark; Wang, Qun; Diep, Binh An; Le, Vien T M; Cheng, Lily; Suzich, JoAnn; Stover, C Kendall; Sellman, Bret R

    2016-03-01

    Broad-spectrum antibiotic use may adversely affect a patient's beneficial microbiome and fuel cross-species spread of drug resistance. Although alternative pathogen-specific approaches are rationally justified, a major concern for this precision medicine strategy is that co-colonizing or co-infecting opportunistic bacteria may still cause serious disease. In a mixed-pathogen lung infection model, we find that the Staphylococcus aureus virulence factor α toxin potentiates Gram-negative bacterial proliferation, systemic spread, and lethality by preventing acidification of bacteria-containing macrophage phagosomes, thereby reducing effective killing of both S. aureus and Gram-negative bacteria. Prophylaxis or early treatment with a single α toxin neutralizing monoclonal antibody prevented proliferation of co-infecting Gram-negative pathogens and lethality while also promoting S. aureus clearance. These studies suggest that some pathogen-specific, antibody-based approaches may also work to reduce infection risk in patients colonized or co-infected with S. aureus and disparate drug-resistant Gram-negative bacterial opportunists.

  5. Genetic Diversity of Staphylococcus aureus in Buruli Ulcer

    NARCIS (Netherlands)

    Amissah, Nana Ama; Glasner, Corinna; Ablordey, Anthony; Tetteh, Caitlin S.; Kotey, Nana Konama; Prah, Isaac; van der Werf, Tjip; Rossen, John W.; van Dijl, Jan Maarten; Stienstra, Ymkje

    2015-01-01

    Background Buruli ulcer (BU) is a necrotizing skin disease caused by Mycobacterium ulcerans. Previous studies have shown that wounds of BU patients are colonized with M. ulcerans and several other microorganisms, including Staphylococcus aureus, which may interfere with wound healing. The present st

  6. Strain Discrimination of Staphylococcus aureus Using Superantigen Profiles.

    Science.gov (United States)

    Tsen, Hau-Yang; Li, Sheng-Chih; Chiang, Yu-Cheng; Tsai, Shuo-Wen

    2016-01-01

    Staphylococcus aureus is one of the major bacterial species that may cause clinical infection and food-poisoning cases. Strains of this species may produce a series of superantigens (SAgs). Due to the importance of staphylococcal infections, reliable methods for the discrimination of strains of this species are important. Such data may allow us to trace the infection origins and be used for epidemiological study. For strain discrimination, genotyping methods, such as pulsed-field gel electrophoresis (PFGE), random amplified polymorphic DNA (RAPD), and multi-locus sequence typing (MLST), etc., could be used. Recently, toxin gene profiles, which can be used for the elucidation of the genetic and pathogenic relatedness between strains, also have been used to improve the strain discrimination. For S. aureus, as more SAg genes were discovered, the SAg profiles become more useful for the strain discrimination of S. aureus. In this chapter, a method for the discrimination of S. aureus strains using superantigen profiles will be described in detail.

  7. Efficacy of extended cefquinome treatment of clinical Staphylococcus aureus mastitis

    NARCIS (Netherlands)

    Swinkels, J. M.; Cox, P.; Schukken, Y. H.; Lam, T. J G M

    2013-01-01

    Clinical Staphylococcus aureus mastitis is difficult to cure. Extended antimicrobial treatment is often advocated as a practical approach to improve cure rates; however, scientific evidence of this hypothesis is lacking. A multi-centered, nonblinded, randomized, positive-controlled clinical trial wa

  8. spa typing for epidemiological surveillance of Staphylococcus aureus

    NARCIS (Netherlands)

    Hallin, Marie; Friedrich, Alexander W; Struelens, Marc J; Caugant, Dominique A.

    2009-01-01

    The spa typing method is based on sequencing of the polymorphic X region of the protein A gene (spa), present in all strains of Staphylococcus aureus. The X region is constituted of a variable number of 24-bp repeats flanked by well-conserved regions. This single-locus sequence-based typing method c

  9. Low efficacy of tobramycin in experimental Staphylococcus aureus endocarditis

    DEFF Research Database (Denmark)

    Lerche, C. J.; Christophersen, L. J.; Trøstrup, H.;

    2015-01-01

    The empiric treatment of infective endocarditis (IE) varies widely and, in some places, a regimen of penicillin in combination with an aminoglycoside is administered. The increasing incidence of Staphylococcus aureus IE, poor tissue penetration by aminoglycosides and low frequency of penicillin...

  10. Staphylococcus aureus redirects central metabolism to increase iron availability.

    Directory of Open Access Journals (Sweden)

    David B Friedman

    2006-08-01

    Full Text Available Staphylococcus aureus pathogenesis is significantly influenced by the iron status of the host. However, the regulatory impact of host iron sources on S. aureus gene expression remains unknown. In this study, we combine multivariable difference gel electrophoresis and mass spectrometry with multivariate statistical analyses to systematically cluster cellular protein response across distinct iron-exposure conditions. Quadruplicate samples were simultaneously analyzed for alterations in protein abundance and/or post-translational modification state in response to environmental (iron chelation, hemin treatment or genetic (Deltafur alterations in bacterial iron exposure. We identified 120 proteins representing several coordinated biochemical pathways that are affected by changes in iron-exposure status. Highlighted in these experiments is the identification of the heme-regulated transport system (HrtAB, a novel transport system which plays a critical role in staphylococcal heme metabolism. Further, we show that regulated overproduction of acidic end-products brought on by iron starvation decreases local pH resulting in the release of iron from the host iron-sequestering protein transferrin. These findings reveal novel strategies used by S. aureus to acquire scarce nutrients in the hostile host environment and begin to define the iron and heme-dependent regulons of S. aureus.

  11. Natural Population Dynamics and Carriage of Staphylococcus aureus

    NARCIS (Netherlands)

    D.C. Melles (Damian)

    2008-01-01

    textabstractStaphylococcus aureus is a major human pathogen capable of causing a wide range of infections, from relatively mild skin infections such as folliculitis and furunculosis to life-threatening conditions, including sepsis, deep abscesses, pneumonia, osteomyelitis, and infective endocarditis

  12. Methicillin‐resistant Staphylococcus aureus and the media.

    Science.gov (United States)

    Perencevich, Eli N; Treise, Debbie M

    2010-11-01

    How the media communicate and how the scientific community influences the media are important factors to consider in the public health response to emerging pathogens, including methicillin-resistant Staphylococcus aureus. Social representation theory suggests that the media link "the threatening" to commonplace "anchor representations" which can serve to educate or to create fear.

  13. No decrease in susceptibility to NVC-422 in multiple-passage studies with methicillin-resistant Staphylococcus aureus, S. aureus, Pseudomonas aeruginosa, and Escherichia coli.

    Science.gov (United States)

    D'Lima, Louisa; Friedman, Lisa; Wang, Lu; Xu, Ping; Anderson, Mark; Debabov, Dmitri

    2012-05-01

    Twenty-five serial passages of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus and 50 passages of methicillin-resistant Staphylococcus aureus resulted in no significant increase in NVC-422 MICs, while ciprofloxacin MICs increased 256-fold for E. coli and 32-fold for P. aeruginosa and S. aureus. Mupirocin, fusidic acid, and retapamulin MICs for MRSA increased 64-, 256-, and 16-fold, respectively. No cross-resistance to NVC-422 was observed with mupirocin-, fusidic acid-, and retapamulin-resistant strains.

  14. Information cloning of harmonic oscillator coherent states

    Indian Academy of Sciences (India)

    N D Hari Dass; Pradeep Ganesh

    2002-08-01

    We show that in the case of unknown harmonic oscillator coherent statesit is possible to achieve what we call perfect information cloning. By this we mean that it is still possible to make arbitrary number of copies of a state which has exactly the same information content as the original unknown coherent state. By making use of this perfect information cloning it would be possible to estimate the original state through measurements and make arbitrary number of copies of the estimator. We define the notion of a measurement fidelity and calculate it for our case as well as for the Gaussian cloners.

  15. Photonic Programmable Tele-Cloning Network

    Science.gov (United States)

    Li, Wei; Chen, Ming-Cheng

    2016-06-01

    The concept of quantum teleportation allows an unknown quantum states to be broadcasted and processed in a distributed quantum network. The quantum information injected into the network can be diluted to distant multi-copies by quantum cloning and processed by arbitrary quantum logic gates which were programed in advance in the network quantum state. A quantum network combines simultaneously these fundamental quantum functions could lead to new intriguing applications. Here we propose a photonic programmable telecloning network based on a four-photon interferometer. The photonic network serves as quantum gate, quantum cloning and quantum teleportation and features experimental advantage of high brightness by photon recycling.

  16. Cloning arbuscule-related genes from mycorrhizas

    DEFF Research Database (Denmark)

    Burleigh, Stephen

    2000-01-01

    Until recently little was known about the identity of the genes expressed in the arbuscules of mycorrhizas, due in part to problems associated with cloning genes from the tissues of an obligate symbiont. However, the combination of advanced molecular techniques, innovative use of the materials...... available and fortuitous cloning has resulted in the recent identification of a number of arbuscule-related genes. This article provides a brief summary of the genes involved in arbuscule development, function and regulation, and the techniques used to study them. Molecular techniques include differential...

  17. Complex network perspective on structure and function of Staphylococcus aureus metabolic network

    Indian Academy of Sciences (India)

    L Ying; D W Ding

    2013-02-01

    With remarkable advances in reconstruction of genome-scale metabolic networks, uncovering complex network structure and function from these networks is becoming one of the most important topics in system biology. This work aims at studying the structure and function of Staphylococcus aureus (S. aureus) metabolic network by complex network methods. We first generated a metabolite graph from the recently reconstructed high-quality S. aureus metabolic network model. Then, based on `bow tie' structure character, we explain and discuss the global structure of S. aureus metabolic network. The functional significance, global structural properties, modularity and centrality analysis of giant strong component in S. aureus metabolic networks are studied.

  18. Whole genome comparison of donor and cloned dogs.

    Science.gov (United States)

    Kim, Hak-Min; Cho, Yun Sung; Kim, Hyunmin; Jho, Sungwoong; Son, Bongjun; Choi, Joung Yoon; Kim, Sangsoo; Lee, Byeong Chun; Bhak, Jong; Jang, Goo

    2013-10-21

    Cloning is a process that produces genetically identical organisms. However, the genomic degree of genetic resemblance in clones needs to be determined. In this report, the genomes of a cloned dog and its donor were compared. Compared with a human monozygotic twin, the genome of the cloned dog showed little difference from the genome of the nuclear donor dog in terms of single nucleotide variations, chromosomal instability, and telomere lengths. These findings suggest that cloning by somatic cell nuclear transfer produced an almost identical genome. The whole genome sequence data of donor and cloned dogs can provide a resource for further investigations on epigenetic contributions in phenotypic differences.

  19. Methicillin resistant S. aureus in human and bovine mastitis.

    Science.gov (United States)

    Holmes, Mark A; Zadoks, Ruth N

    2011-12-01

    Staphylococcus aureus is a ubiquitous organism that causes a variety of diseases including mastitis in cattle and humans. High-level resistance of S. aureus to β-lactams conferred by a mecA gene encoding a modified penicillin binding protein (PBP2a) was first observed in the early 1960's. These methicillin resistant S. aureus (MRSA) have been responsible for both hospital acquired infections (HA-MRSA) and, more recently, community acquired MRSA (CA-MRSA). A small number of human MRSA mastitis cases and outbreaks in maternity or neonatal units have been reported which are generally the result of CA-MRSA. The establishment of the sequence type 398 (ST398) in farm animals, primarily pigs, in the early 2000's has provided a reservoir of infection for humans and dairy cattle, particularly in continental Europe, described as livestock-associated MRSA (LA-MRSA). Prior to the emergence of ST398 there were sporadic reports of MRSA in bovine milk and cases of mastitis, often caused by strains from human associated lineages. Subsequently, there have been several reports describing bovine udder infections caused by ST-398 MRSA. Recently, another group of LA-MRSA strains was discovered in humans and dairy cattle in Europe. This group carries a divergent mecA gene and includes a number of S. aureus lineages (CC130, ST425, and CC1943) that were hitherto thought to be bovine-specific but are now also found as carriage or clinical isolates in humans. The emergence of MRSA in dairy cattle may be associated with contact with other host species, as in the case of ST398, or with the exchange of genetic material between S. aureus and coagulase negative Staphylococcus species, which are the most common species associated with bovine intramammary infections and commonly carry antimicrobial resistance determinants.

  20. Heme Recognition By a Staphylococcus Aureus IsdE

    Energy Technology Data Exchange (ETDEWEB)

    Grigg, J.C.; Vermeiren, C.L.; Heinrichs, D.E.; Murphy, M.E.P.

    2009-06-03

    Staphylococcus aureus is a Gram-positive bacterial pathogen and a leading cause of hospital acquired infections. Because the free iron concentration in the human body is too low to support growth, S. aureus must acquire iron from host sources. Heme iron is the most prevalent iron reservoir in the human body and a predominant source of iron for S. aureus. The iron-regulated surface determinant (Isd) system removes heme from host heme proteins and transfers it to IsdE, the cognate substrate-binding lipoprotein of an ATP-binding cassette transporter, for import and subsequent degradation. Herein, we report the crystal structure of the soluble portion of the IsdE lipoprotein in complex with heme. The structure reveals a bi-lobed topology formed by an N- and C-terminal domain bridged by a single {alpha}-helix. The structure places IsdE as a member of the helical backbone metal receptor superfamily. A six-coordinate heme molecule is bound in the groove established at the domain interface, and the heme iron is coordinated in a novel fashion for heme transporters by Met{sup 78} and His{sup 229}. Both heme propionate groups are secured by H-bonds to IsdE main chain and side chain groups. Of these residues, His{sup 299} is essential for IsdE-mediated heme uptake by S. aureus when growth on heme as a sole iron source is measured. Multiple sequence alignments of homologues from several other Gram-positive bacteria, including the human pathogens pyogenes, Bacillus anthracis, and Listeria monocytogenes, suggest that these other systems function equivalently to S. aureus IsdE with respect to heme binding and transport.