Methicillin-resistenta Staphylococcus aureus (MRSA) har blivit en allt vanligare patogen inom sjukvården och i samhället. MRSA orsakar infektioner som inte kan behandlas med β-laktamantibiotika. För att förhindra spridning genomgår patienter och sjukvårdspersonal screening-tester. I dessa screening-tester ingår PCR-analys av mecA, nuc och/eller Sa442. MecA är lokaliserad på Staphylococcal Cromosomal Cassette mec (SCCmec) och används som en markör för MRSA medan nuc och Sa442 anger S. aureus. PCR-positiva isolat odlas ut på agarplattor efter anrikning i en selektiv buljong. Kolonier av S. aureus resistensbestäms mot cefoxitin som MRSA är resistent mot. Idag används dock PCR-analys av mecA som en referensmetod för diagnostisering av MRSA. Under det senaste årtiondet har även rapporterats fynd av stammar som enligt resistensmönster är MRSA men som har utfallit negativt för mecA i PCR. Under 2011 rapporterades en ny variant av SCCmec och en ny variant av mecA, mecALGA251. I denna studie har en realtids-PCR tagits fram för att identifiera den nya varianten, mecALGA251. Denna PCR användes för att undersöka 43 kliniska isolat, fyra cefoxitin-känsliga S. aureus från rutinen och tre referensstammar. De kliniska isolaten hade samlats in under perioden 2004 – 2011 och hade uppvisat cefoxitin-resistens men gett negativt resultat i mecA-PCR. Totalt 40 av de 43 cefoxitin-resistenta kliniska isolaten visades bära mecALGA251. Resistensbestämningar med diskdiffusion och E-test mot cefoxitin, oxacillin, cefuroxim och cefotaxim visade att denna typ av MRSA inte kan skiljas från klassisk MRSA. Resultaten visar att cefoxitin-resistenta S. aureus isolat som bär mecALGA251 finns bland skånska patienter. De visar också att det finns cefoxitin-resistent S. aureus som saknar både klassiska mecA och mecALGA251. Dessa stammar har inte studerats vidare i denna studie. / Methicillin-resistant Staphylococcus aure s (MRSA) has become a more frequent pathogen within health care facilities and the community. MRSA causes infections that can’t be treated with β-lactamantibiotics. To prevent the spread of MRSA, patients and medical personnel undergo screening-tests. In the screening-tests PCR-analysis of mecA, nuc and/or Sa442 is included. MecA is located at Staphylococcal Chromosomal Cassette mec (SCCmec) and is a marker for MRSA, whereas nuc and Sa442 state regular S. Aureus infections. PCR-positive isolates are grown on agar plates after enrichment in selective broth. Colonies of S. aureus are tested for cefoxitin susceptibility to which MRSA is resistant. PCR-analysis of mecA is the reference method that is being used today when MRSA is being diagnosed. During the last decade cefoxitin-resistant strains that lack mecA in the PCR has been reported. In 2011 a new variant of SCCmec and a new variant of mecA, mecALGA251 was reported. In this study a new real-time-PCR has been developed in order to identify mecALGA251. The new PCR protocol was being used to examine 43 clinical isolates, four cefoxitin-susceptible S. aureus from the routine and three reference strains were examined. The clinical isolates had been collected during the period 2004-2011 and were cefoxitin-resistant but lacked mecA. In total of 40 of the 43 cefoxitin-resistant was PCR positive for mecALGA251. Susceptibility testing with disk diffusion and E-test for cefoxitin, oxacillin, cefuroxime and cefotaxime showed that this type of MRSA can’t be distinguished from regular MRSA. The results showed that cefoxitin-resistantS. aureus isolates carrying mecALGA251 exist among patients in Skåne County. One cefoxitin-resistant S. aureus isolate lacked both classic mecA and mecALGA251, which indicates that other mechanisms may exist, however these results has not been further analysed in this study.

Methicillin Resistant Staphylococcus Aureus (MRSA) has been receiving significant attention, highlighting an increased risk of infectious transmission associated with athletic participation. As MRSA infections are becoming increasing virulent, athletic trainers are presented with immediate prevention challenges. While recommendations have been offered by the Centers for Disease Control and Prevention outlining basic prevention procedures, adherence to proposed guidelines and actual perception of the threat still pose the greatest hurdles to eradication of MRSA. Success in control and prevention of transmission of MRSA in athletic environments can be furthered by first investigating the perceptions of the problem in one of the first line of defense for athletes—their athletic trainers. Of particular importance are the perceptions of trainers' adherence to guidelines, perceptions of protocol standards, and relative threat of MRSA in the athletic environment. This study attempts to determine these perceptions and predict how athletic trainers will receive and adhere to standardized guidelines through written policy for MRSA prevention. Results reflect an increase in the awareness of MRSA as a threat to athletics since 2004. Overall positive perception of the development of guidelines and protocols specifically targeted to prevention of MRSA transmission in the athletic environment were also defined through this study. Athletic trainers surveyed expressed strong desire for additional training in procedures specific to reducing transmission of MRSA to prevent outbreaks. / Ph. D.

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious public health problem nationwide, threatening to develop into an epidemic. Many of these patients are presenting to their primary care clinics with skin and soft tissue infections (SSTIs). The CDC has reported that in 2005, MRSA was responsible for an estimated 94,000 life-threatening infections and 16,650 deaths. The purpose of this study is to estimate the incidence of CA-MRSA within a specific family practice in Florida and to identify epidemiologic factors, classify antibiotic susceptibility patterns, and evaluate patient education in regard to disease management and prevention. This study was a descriptive, epidemiologic, three-year retrospective medical record review of all wound cultured skin and soft tissue infections that presented to a family practice between January 2007 and December 2009. Sixty-two medical records met the inclusion and exclusion criteria for the study. Of these 62 SSTIs, 44 cultures grew one or more bacterial organisms. The incidence of CA-MRSA was 66% (n=29). The mean age of those with CA-MRSA was 40 years old, with a range from 7 to 90 years old. Sixty-two percent (n=18) were male and 38% (n=11) were female; additionally 69% (n=20) lived within a 10 mile radius from the family practice, while 31% (n=9) lived in a surrounding suburb. The most frequent race was Caucasian 83% (n=24), with African American at 10% (n=3) and Hispanics 7% (n=2). Risk factors associated with CA-MRSA was obesity 41% (n=10), diabetes mellitus 24% (n=7), and a previous history of MRSA infection 24% (n=7). Skin and soft tissue infections were diagnosed as either an abscess 62% (n=18), boil 24% (n=7), pustule 10% (n=3), or cellulitis 4% (n=1). CA-MRSA isolates were susceptible to trimethoprim-sulfamethoxazole 100% (n=29), doxycycline 93% (n=27), and rifampin 100% (n=14). Clindamycin susceptibility was 65% (n=15) with resistance at 30% (n=7) and 5% (n=1) intermediate. Both cephalexin and erythromycin were 100% resistant. Documentation in the medical record on wound care was found in 45% (n=13) of the records. The incidence of CA-MRSA SSTI was 66%, which identifies this suburban community at high risk for this bacterial infection. Risk factors associated with CA-MRSA included obesity (BMI > 30), history of previous MRSA infection, and diabetes mellitus. There were no clinical characteristics that helped distinguish MRSA infection from other bacterial SSTIs. Most SSTI were treated with incision and drainage and a susceptible antibiotic. Judicious use of antibiotics not only provides appropriate treatment, but is also critical in prevention of antibiotic resistance. Lastly, patient education in adequate hygiene is essential in preventing the spread of CA-MRSA

No description available.

Quaternary ammonium compounds (QACs) have long served as lead disinfectants in residential, industrial, and hospital settings. Their simple yet effective amphiphilic nature makes them an ideal class of compounds through which to explore antibacterial activity. We have developed novel multiQAC scaffolds through simple and cost-efficient syntheses, yielding hundreds of diverse compounds strategically designed to examine various aspects of antibacterial and anti-biofilm activity, as well as toxicity. Many of these bis-, tris-, and tetraQACs display antibacterial activity 10 to 100 times greater than conventional monoQACs, and are among the most potent biofilm eradicators to date. Through analyzing their activity against several strains, we have uncovered and provided further evidence for key tenets of amphiphilic QAC bioactivity: a balance of hydrophobic side chains with cationic head groups generates optimal antibacterial activity, though toxicity to eukaryotic cells needs to be mitigated. Given their ubiquitous nature and chemical robustness, the overuse of QACs has led to the development of QAC resistance genes that are spreading throughout the microbial world at an alarming rate. These resistant strains, when found in bacterial biofilms, are able to persist in the presence of lead commercial QAC disinfectants, warranting the development of next-generation biocides. Several of our scaffolds were designed with QAC resistance machinery in mind; thus, we utilized these compounds not only as antibacterial agents but also as chemical probes to better understand and characterize QAC-resistance in methicillin-resistant Staphylococcus aureus (MRSA). Our findings support previous postulations that triscationic QACs would retain potency against QAC-resistant strains. Furthermore, we have identified monocationic and aromatic moieties, as well as conformational rigidity, as being more prone to recognition by the resistance machinery. Using our chemical toolbox comprised of QACs of various charge state and scaffold, we explored both the mechanism and scope of QAC-resistance by examining their structure-resistance relationship. Our holistic findings have allowed us to better understand the dynamics of this system towards the design and development of next-generation QACs that will: (1) allow us to better probe the resistance machinery, and (2) remain efficacious against a variety of microbial pathogens. / Chemistry

Orientador: Carlos Eduardo Vergani / Banca: Ana Lucia Machado / Banca: Wander José da Silva / Resumo: Infecções causadas por bactérias resistentes a antibióticos são consideradas causa principal de mortalidade entre indivíduos imunocomprometidos e aproximadamente 50 % destas infecções tem sido relacionadas ao Staphylococcus aureus resistente à meticilina (MRSA). Estes microrganismos, quando presentes no biofilme das próteses dentarias, podem se dispersar nas secreções salivares e se disseminar pelo trato respiratório, causando pneumonia aspirativa. Assim, o presente estudo comparou a efetividade do hipoclorito de sódio a 1 %, digluconato de clorexidina a 2 % e irradiação por micro-ondas na desinfecção de próteses totais e corpos-de-prova circulares de resina acrílica para base de prótese (10 x 2 mm) contaminados com MRSA. Para isso, 36 próteses totais simuladas e 36 corpos-de-prova circulares foram confeccionados, esterilizados, inoculados com MRSA (107 ufc/mL) e incubados a 37 °C (por 24 e 48 h, respectivamente). Após incubação, próteses totais e corpos-de-prova foram distribuídos em 4 grupos de estudo (n=9): GC - não foi realizado nenhum método de desinfecção; GH - foi realizada a imersão em solução de hipoclorito de sódio a 1 % por 10 min; GCl -foi realizada a imersão em solução de digluconato de clorexidina a 2 % por 10 min; GM - foi realizada a desinfecção por irradiação em forno de micro-ondas a 650 W por 3 min. A efetividade dos procedimentos de desinfecção foi avaliada por meio de quantificação de colônias viáveis e da viabilidade celular. Para a quantificação de colônias viáveis, alíquotas de 25 μL da solução resultante das diluições seriadas (10-3 a 10-6 para GC e 100 a 10-3 para os grupos experimentais) foram semeadas em placas de Petri em duplicata e todas as placas foram incubadas por 48 h a 37 °C. As colônias foram quantificadas em ufc/mL. Para verificar a efetividade da desinfecção por micro-ondas em longo prazo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Infections caused by antibiotic-resistant bacteria have been recognized as a predominant risk factor for mortality in elderly patients and approximately 50 % of these infections have been related to methicillin-resistant Staphylococcus aureus (MRSA). This microorganism, when present in the denture biofilm, can be released into the oral fluids and aspirated into the lower respiratory tract, thus causing infections such as aspiration pneumonia. The present study compared the efficacy of 1 % sodium hypochlorite, 2 % chlorhexidine gluconate, and microwave irradiation in disinfecting simulated complete dentures and circular specimens of acrylic resin denture base material (10 x 2 mm) contaminated with MRSA. Thirty-six dentures and 36 specimens were made, sterilized, inoculated with MRSA (107 cfu/mL), and incubated at 37 °C (for 24 and 48 h, respectively). After incubation, dentures and specimens were divided into 4 groups of study (n=9): PC - positive control, consisting of dentures and specimens not disinfected; HY - soaking in 1 % sodium hypochlorite solution for 10 min; CHL - soaking in 2 % chlorhexidine gluconate solution for 10 min; and MW - irradiating by microwave for 3 min at 650W. The effectiveness of the disinfection procedures was assessed cell viability (quantification of viable cells and XTT reduction method). For quantification procedures, aliquots of suspensions were plated at dilutions (10-3 to 10-6 for PC and 100 to 10-3 for experimental groups) and incubated (37 °C/48 h). Colonies counts (cfu/mL) were quantified. Dentures disinfected were also incubated at 37 °C for 7 days to verify the long-term effectiveness of disinfection. The viability of cells in each group of specimens was evaluated by XTT reduction method. The results showed that all dentures and specimens from the PC groups showed substantial microbial growth. No evidence of microbial growth was observed... (Complete abstract click electronic access below) / Mestre

Seventy-four methicillin-resistant Staphylococcus aureus (MRSA) from two Malaysian hospitals were characterised by both phenotypic and genotypic techniques. These isolates were collected over an 18 year time period in the years, 1982, 1989, 1994 and 2000. All of the Malaysian MRSA isolates were found to be multiresistant and resistant to at least five different antimicrobial agents. Over 30% of them were non-typable by the International Basic Set of bacteriophages. The majority of the typable isolates were susceptible to the group III phages, especially phage 85. The majority of the isolates carried one to six plasmids. Only two isolates were plasmid free. The plasmid profiles of these isolates, other than the 1982 isolates, were very similar to each other. Contour-clamped homogeneous electric field (CHEF) gel electrophoresis was used to examine the genetic relatedness of the isolates. Twenty-six CHEF patterns were found among the isolates. These CHEF patterns were closely related to each other. The predominant CHEF pattern A was found in the 1982, 1989 and 1994 isolates. The CHEF patterns of the year 2000 isolates were different to CHEF pattern A, but still closely related. All of the isolates were found to carry the Allotype III SCCmec and have coagulase-gene type 24. Multilocus sequence typing was preformed on the isolates with CHEF pattern A collected in different years. These isolates were found to have either sequence type 239 (ST239), or its single locus variant. The predominant Malaysian clone belongs to the pandemic clone ST239-MRSA-III that is pandemic in Asian countries. (Enright, 2003, Ko et al., 2005). / A 1.5 kb cryptic plasmid found in Malaysian isolates was indistinguishable from a cryptic plasmid found in an Australian isolate. A 3.0 kb cryptic plasmid found in Malaysian isolates was undistinguishable from a 3.0 kb plasmid found in Singaporean isolates. Class II multiresistance plasmids of 28, 30.5 and 35 kb were commonly found together in many Malaysian MRSA isolates. Both the 28 and 30.5 kb plasmids encode resistance to the heavy-metals and nucleic acid-binding (NAB) compounds. The 35 kb plasmid carries heavy-metal and NAB resistance but also encodes β-lactamase. Structurally these three plasmids are almost identical and probably have the same origin. The differences observed between these plasmids is probably due to excision or partial deletion of the β-lactamase transposon of the original plasmid. The 28 kb plasmid is identical to the 28 kb plasmid of Singaporean and some Australian isolates. A 20 kb plasmid in Indonesian isolates was found to be closely related to these three plasmids. A conjugative plasmid, pWBG707, conferring trimethoprim-resistance was found in Malaysian isolates. It did not carry either of the two staphylococcal trimethoprim-resistance genes, dfrA and dfrD. (Lyon and Skurray, 1987, Dale et al., 1995b) It either encodes a novel resistance gene or the recently discovered dfrG gene. (Sekiguchi et al., 2005) pWBG707 was also found to mobilise a small 3.0 kb kanamycin-resistance plasmid during conjugation. / The mecR1 and mecI genes regulating the transcription of the methicillin-resistance gene, mecA, were also examined in the isolates. The Malaysian isolate, WBG7422, with the predominant CHEF pattern A has a nonsense mutation in its mecI gene that disables it. However, its mecR1 gene is intact. The eastern Australia MRSA (EA MRSA), WBG525, has a CHEF pattern that is closely related to the Malaysian predominant CHEF pattern A and its mecI gene has a mutation identical to the Malaysian isolate. Unlike the Malaysian isolate however, its mecR1 gene has a 166 bp deletion. Both WBG7422 and WBG525 express Class III heterogeneous methicillin resistance. However, WBG525 has more highly resistant cell in its population than WBG7422. The loss of aminoglycoside resistance, together with c. 114 kb of chromosomal DNA, was observed in some Malaysian isolates. The deleted segment was found to carry the aacA-aphD gene that encodes a bifunctional aminoglycoside-modifying enzyme conferring resistance to many of the aminoglycosides. The Malaysian isolates were compared with MRSA from different countries. These MRSA included 18 epidemic MRSA (EMRSA) from the United Kingdom, 15 Australian nosocomial MRSA, five classical MRSA, 22 community-acquired MRSA (CMRSA) from Australia and New Zealand and 46 nosocomial MRSAs from eight Asian-Pacific countries and South Africa. These Asian-Pacific countries were Australia, PR China, Hong Kong, Indonesia, Japan, Philippines, Singapore and Taiwan. / The CHEF patterns of most of the Asian-Pacific and South African isolates were closely related to the Malaysian isolates. Isolates from Singapore, Indonesia and Philippines were found to have an identical CHEF pattern to the Malaysian CHEF patterns A5. The Asian-Pacific and South African isolates, including the Malaysian isolates, were found to be closely related to EMRSA-1, -4 and -7. These EMRSA belong to the ST239-MRSA-III clone and are coagulase-gene type 24. The isolates from Japan were the only Asian-Pacific isolates not related to the other Asian-Pacific isolates and EMRSAs. EMRSA-1 and EA MRSA have the same 166 bp deletion in their mecR1 gene. Both of these strains have closely related CHEF patterns, the same sequence type, coagulase-gene type and SCCmec. These results indicate that these two strains belongs to the same clone and confirms the international spread of this clone in the early 1980s. However, the Malaysian isolates have CHEF patterns that are more closely related to EMRSA-4 than to EMRSA-1. Similar to the Malaysian isolates EMRSA-4 has an intact mecR1 gene. The CMRSA isolates were not related to any of the nosocomial MRSA. They also have very diverse genetic backgrounds but carry less diverse SCCmec allotypes. Most of the CMRSA carry either Allotype IV or V SCCmec These results show that the spread of Malaysian MRSA is due to a single clonal expansion. Infection control measures would have to have been more efficient if this clone was to have been contained. The Malaysian epidemic clone is the Asian pandemic clone, ST239-MRSA-III. The Malaysian isolates and EMRSA-4 probably share the same ancestor. / The presence of the same MRSA strain in Malaysian hospitals and in the hospitals of neighbouring countries indicates that the inter-hospital spread of an epidemic MRSA has occurred. This observation also suggests that the infection control measures in Malaysian hospitals have not been totally effective. The ineffectiveness of infection control has left Malaysian hospitals vulnerable to the future importation of new pandemic clones and/or highly virulent or resistant clones.

Source: Masters Abstracts International, Volume: 47-01, page: 0341. Adviser: Luis Ostrosky. Includes bibliographical references.

Dissertation (Ph.D.)--University of South Florida, 2006. / Title from PDF of title page. Document formatted into pages; contains 117 pages. Includes vita. Includes bibliographical references.

Made available in DSpace on 2014-06-11T19:28:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-03-28Bitstream added on 2014-06-13T20:18:32Z : No. of bitstreams: 1 altieri_kt_me_arafo.pdf: 598917 bytes, checksum: 3b01564a77c1be4471454056881b7028 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Infecções causadas por bactérias resistentes a antibióticos são consideradas causa principal de mortalidade entre indivíduos imunocomprometidos e aproximadamente 50 % destas infecções tem sido relacionadas ao Staphylococcus aureus resistente à meticilina (MRSA). Estes microrganismos, quando presentes no biofilme das próteses dentarias, podem se dispersar nas secreções salivares e se disseminar pelo trato respiratório, causando pneumonia aspirativa. Assim, o presente estudo comparou a efetividade do hipoclorito de sódio a 1 %, digluconato de clorexidina a 2 % e irradiação por micro-ondas na desinfecção de próteses totais e corpos-de-prova circulares de resina acrílica para base de prótese (10 x 2 mm) contaminados com MRSA. Para isso, 36 próteses totais simuladas e 36 corpos-de-prova circulares foram confeccionados, esterilizados, inoculados com MRSA (107 ufc/mL) e incubados a 37 °C (por 24 e 48 h, respectivamente). Após incubação, próteses totais e corpos-de-prova foram distribuídos em 4 grupos de estudo (n=9): GC - não foi realizado nenhum método de desinfecção; GH - foi realizada a imersão em solução de hipoclorito de sódio a 1 % por 10 min; GCl -foi realizada a imersão em solução de digluconato de clorexidina a 2 % por 10 min; GM - foi realizada a desinfecção por irradiação em forno de micro-ondas a 650 W por 3 min. A efetividade dos procedimentos de desinfecção foi avaliada por meio de quantificação de colônias viáveis e da viabilidade celular. Para a quantificação de colônias viáveis, alíquotas de 25 μL da solução resultante das diluições seriadas (10-3 a 10-6 para GC e 100 a 10-3 para os grupos experimentais) foram semeadas em placas de Petri em duplicata e todas as placas foram incubadas por 48 h a 37 °C. As colônias foram quantificadas em ufc/mL. Para verificar a efetividade da desinfecção por micro-ondas em longo prazo... / Infections caused by antibiotic-resistant bacteria have been recognized as a predominant risk factor for mortality in elderly patients and approximately 50 % of these infections have been related to methicillin-resistant Staphylococcus aureus (MRSA). This microorganism, when present in the denture biofilm, can be released into the oral fluids and aspirated into the lower respiratory tract, thus causing infections such as aspiration pneumonia. The present study compared the efficacy of 1 % sodium hypochlorite, 2 % chlorhexidine gluconate, and microwave irradiation in disinfecting simulated complete dentures and circular specimens of acrylic resin denture base material (10 x 2 mm) contaminated with MRSA. Thirty-six dentures and 36 specimens were made, sterilized, inoculated with MRSA (107 cfu/mL), and incubated at 37 °C (for 24 and 48 h, respectively). After incubation, dentures and specimens were divided into 4 groups of study (n=9): PC - positive control, consisting of dentures and specimens not disinfected; HY - soaking in 1 % sodium hypochlorite solution for 10 min; CHL - soaking in 2 % chlorhexidine gluconate solution for 10 min; and MW - irradiating by microwave for 3 min at 650W. The effectiveness of the disinfection procedures was assessed cell viability (quantification of viable cells and XTT reduction method). For quantification procedures, aliquots of suspensions were plated at dilutions (10-3 to 10-6 for PC and 100 to 10-3 for experimental groups) and incubated (37 °C/48 h). Colonies counts (cfu/mL) were quantified. Dentures disinfected were also incubated at 37 °C for 7 days to verify the long-term effectiveness of disinfection. The viability of cells in each group of specimens was evaluated by XTT reduction method. The results showed that all dentures and specimens from the PC groups showed substantial microbial growth. No evidence of microbial growth was observed... (Complete abstract click electronic access below)