Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-07-28T13:08:05Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese_EDUARDA_versãoFinalDigital.pdf: 2657877 bytes, checksum: 2ce4a0aa738737b2947e45ed5fb9e912 (MD5) / Made available in DSpace on 2016-07-28T13:08:05Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Tese_EDUARDA_versãoFinalDigital.pdf: 2657877 bytes, checksum: 2ce4a0aa738737b2947e45ed5fb9e912 (MD5) Previous issue date: 2016-02-29 / FACEPE / Devido à crescente resistência de isolados clínicos de Staphylococcus spp. a antimicrobianos e a consequente diminuição de opções terapêuticas eficazes, este estudo investigou a influência in vitro de sub-Concentrações Inibitórias Mínimas (sub-CIM) de oxacilina e tigeciclina, isoladamente e em combinação, no crescimento bacteriano e na expressão dos genes mecA, ica e tst em isolados clínicos de Staphylococcus spp.Para realização da primeira etapa deste estudo 45 isolados,identificados quanto a espécie de Staphylococcus spp. e susceptibilidade pelo Vitek2, foram obtidos de dois hospitais públicos de Recife-PE e avaliados pelas técnicas de MALDI-TOF MS e sequenciamento do rDNA 16S para confirmação das espécies bacterianas. A relação clonal dos isolados de Staphylococcus spp. foi determinada pela análise da Região Intergênica 16S-23S para posterior determinação do tipo de SCCmec e do perfil de virulência. O MALDI-TOF MS e sequenciamento do 16S se mostraram mais concordantes na identificação das espécies bacterianas quando comparadas ao Vitek2, sendo identificados isolados de Staphylococcus aureus, S. epidermidis, S.hominis, S. haemolyticus e S. saprophyticus. Trinta e um ribotipos foram determinados, demonstrando grande variabilidade genética. Os ribotipos foram encontrados dispersos nos diferentes setores dos dois hospitais investigados. A maioria dos isolados apresentaram SCCmec tipo IV (51%), seguido pelo tipo III (26%), tipo II (16%) e tipo V (7%). Foi possível encontrar nove perfis de virulência, onde o gene luk foi observado em 80% dos isolados, seguido de 51% para o gene ica, 35,5% para o hlg e 33,3% para o tstratificando a crescente evidência da presença de isolados tradicionalmente comunitários no ambiente hospitalar. Na segunda fase do estudo quatro isolados clínicos com diferentes tipos de SCCmece baixa relação clonal foram analisados in vitro para a determinação da CIM frente a oxacilina e tigeciclina. A influência in vitro destes antimicrobianos no crescimento bacteriano foi avaliada isoladamente e em associação, assim como a expressão dos genes mecA, ica e tst por RT-qPCR. O ensaioin vitro dos diferentes isolados de Staphylococcus spp. frente a tigeciclina associada à oxacilina resultou em inibição de crescimento bacteriano maior em relação a utilização isolada dos antimicrobianos, além de não influenciar ou diminuir a expressão de todos os genes analisados, dando suporte à ideia de que o uso associado da tigeciclina e oxacilina no tratamento de infecções por isolados MRS portadores dos genes tst e ica possivelmente poderiam promover benefícios na modulação destes genes. / Due to the increasing resistance of clinical isolates of Staphylococcus spp. to antimicrobial and the consequent decrease of effective therapeutic options, this study determined the in vitro influence of sub-minimum inhibitory concentrations (sub-MIC) of oxacillin and tigecycline, alone and in combination, on bacterial growth and expression ofvirulence genes in clinical isolates of Staphylococcus spp. To perform the first stage of this study 45 isolates of Staphylococcus sppwere obtained from two public hospitals of Recife-PE and evaluated by MALDI-TOF MS techniques and 16S sequencing for confirmation of bacterial species. The clonal profile Staphylococcus spp. isolates was determined by PCR-Ribotyping for subsequent determination of the type of SCCmec and virulence profile. The MALDI-TOF MS and sequencing of 16S were more effective in the identification of bacterial species when compared to Vitek2, being identified Staphylococcus aureus, S. epidermidis, S.hominis, S. haemolyticus e S. saprophyticus.Thirty-one ribotypes could be determined among the strains tested, with most harboring the SCCmec type IV (51%), followed by type III (26%), type II (16%) and type V (7%).%). 80% of the isolates presented the luk gene, 51% icaAD, 35.5% hlg and 33.3% tst. These results demonstrate a wide variability and dispersion of the isolates analysed, in different sectors of the hospitals confirming the growing evidence of the presence of traditionally community isolates in the hospital environment. In the second phase of the study, four clinical isolates were selected according to different types of SCCmec were analyzed in vitro to determine the MIC front oxacillin and tigecycline. The antimicrobial effect in vitro tests on bacterial growth was evaluated separately and in combination, as well as the expression of the mecA, ica and tst genes by RT-qPCR. Subjecting the different Staphylococcus spp. isolates in vitro to tigecycline in association with oxacillin resulted in inhibition of bacterial growth that was more potent than subjecting them to the antimicrobials separately, and this also did not influence or decrease the expression of any of the genes analyzed. The present study supports the idea that the associated use of tigecycline and oxacillin for treating infections caused by methicillin-resistant Staphylococcus that is positive for tst and ica could promote possible benefits in modulating these genes.

Evidence exists that vitamin D has a potential antimicrobial activity and its deficiency has deleterious effects on general well-being and longevity. Vitamin D may reduce the risk of infection through multiple mechanisms. Vitamin D boosts innate immunity by modulating production of anti-microbial peptides (AMPs) and cytokine response. Vitamin D and its analogues via these mechanisms are playing an increasing role in the management of atopic dermatitis, psoriasis, vitiligo, acne and rosacea. Vitamin D may reduce susceptibility to infection in patients with atopic dermatitis and the ability to regulate local immune and inflammatory responses offers exciting potential for understanding and treating chronic inflammatory dermatitides. Moreover, B and T cell activation as well as boosting the activity of monocytes and macrophages also contribute to a potent systemic anti-microbial effect. The direct invasion by pathogenic organisms may be minimized at sites such as the respiratory tract by enhancing clearance of invading organisms. A vitamin D replete state appears to benefit most infections, with the possible noteworthy exception of Leishmaniasis. Antibiotics remain an expensive option and misuse of these agents results in significant antibiotic resistance and contributes to escalating health care costs. Vitamin D constitutes an inexpensive prophylactic option and possibly therapeutic product either by itself or as a synergistic agent to traditional antimicrobial agents. This review outlines the specific antimicrobial properties of vitamin D in combating a wide range of organisms. We discuss the possible mechanisms by which vitamin D may have a therapeutic role in managing a variety of infections.

Background: Antibiotic stewardship (ABS) is a set of strategies to optimize the use of antibiotics with the goal of reducing antibiotic resistance, improving patient outcomes and decreasing unnecessary costs. ABS affects all venues of patient care, including outpatient, inpatient, and long-term care. While many strategies for ABS exist and best practice continues to evolve, successful ABS programs utilize a multidisciplinary approach. Nurse practitioners (NPs) play an essential role in health care education and represent a valuable potential resource for ABS efforts. The purpose of this study is to describe the knowledge, attitudes, and perceptions of NPs towards ABS.Methods: A convenience sample of NPs attending the American Association of Nurse Practitioners annual conference was given a modified descriptive survey developed for use in a previous study conducted at a university-affiliated hospital in Florida. Descriptive statistics were used to assess normality. Chi-Square test of independence was used to test differences categorical scores by NP setting, gender, and level of education. Pearson r correlation was completed to measure the relationship between age and years in practice.Results: Two hundred NPs completed the questionnaire (88% female; 70% Master’s degree). The range of experience was 0-45 years (mean 11 years). Most NPs worked in a private office (23%) or community setting (29%). Factors affecting the decisions of antibiotic prescriptions included patient condition (79%) and patient cost (58%). NPs in this study also based their antibiotic decisions on the antibiogram (63%) in their setting, while 56% indicated they start with broad spectrum and tailor antibiotic choices when culture results are received. NPs reported understanding that inappropriate use of antibiotics causes resistance (97%), harms the patient (97%), and optimum antibiotic use will reduce resistance (94%). Participants also recognized that strong knowledge of antibiotics was important for their job (94%) and felt confident in their use of antibiotics (86%). However, while 94% of respondents somewhat or strongly agreed that antibiotics are overused nationally, only 62% thought antibiotics were overused in their health care setting. Conclusion: In this study, most NPs reported that antibiotic resistance is a problem and antibiotics are overused nationally. Fewer believe that antibiotic resistance is a problem locally and fewer still that they, personally, contribute to the problem. NPs recognize that knowledge about antibiotics is important to their career and would like more education about antibiotics and feedback about their antibiotic choices. Finding effective ways to provide this education could change practice and improve antibiotic use.

Post process application of antimicrobials is one method to control recontamination of Listeria monocytogenes on cooked RTE meat products. This study evaluated the antilisterial effect of externally applied solutions of nisin, buffered solution of sodium citrate supplemented with sodium diacetate (SCSD) and a combined solution of the two antimicrobials on commercial beef frankfurters formulated without antimicrobials. Autoclaved frankfurters were inoculated (104 -105 CFU/g), dipped (5 min, 25 ± 2°C) in treatments consisting of 2000; 4000;or 6000 IU/ml nisin, and buffered solutions of 2.5; 3.0; or 3.5% SCSD (Study I), and 6000 IU/ml nisin followed by 3.5% SCSD, or 3.5% SCSD followed by 6000 IU/ml nisin, or a combined solution containing both 6000 IU/ml nisin and 3.5% SCSD(Study II). Treated hot dogs were vacuum packaged and stored at 4 and 10°C. L. monocytogenes counts were determined on Modified Oxford Agar on 0, 14, 28, and 42 days at 4°C and 0, 4, 8, 12, 16, and 20 days at 10°C. Nisin (6000 IU/ml) initially reduced L. monocytogenes population by 2.1 and 2.5 logs (at 4 and 10°C, respectively on day 0), buffered SCSD (3.5%) by 1.1 and 0.2 logs (at 4 and 10°C, respectively on day 0) and the combined solution by 1.7 and 2.0 logs (at 4 and 10°C, respectively on day 0). The combined solution was the most effective treatment compared to nisin and buffered SCSD, used in sequence or alone, since it inhibited the growth of L. monocytogenes during 28 days at 4°C. The buffered SCSD was effective against L. monocytogenes at 4°C but not at 10°C. Further research is needed to investigate the effectiveness of buffered SCSD with other antimicrobials. The results of this study may be useful for further research on combinations of antimicrobials.

No description available.

The effect of pressure on the log reduction of six strains of E. coli O157:H7 and five serovars of Salmonella enterica were investigated in tryptic soy broth, sterile distilled water and commercially sterile orange and apple juice. Samples were subjected to high hydrostatic pressure (HHP) at 300 and 550 MPa for 2 minutes at 6°C, and then held for 24 hours at 4°C following treatment. E. coli O157:H7 strain E009 was the most pressure resistant, having a decrease of only 0.77 log10 CFU/ml directly after pressurization in TSB. S. Agona was the most pressure resistant Salmonella serovar tested with a decrease of 3.79 log¬10 CFU/ml in TSB at 550 MPa. The two most pressure resistant cultures were then used in a subsequent study using HHP in conjunction with antimicrobials (dimethyl dicarbonate [DMDC] at 62.5 and 125 ppm, hydrogen peroxide at 150 and 300 ppm, cinnamic acid, potassium salt at 125 and 250 ppm, potassium sorbate [KS] at 500 and 1000 ppm and sodium benzoate [NaB] at 500 and 1000). For both E. coli O157:H7 and Salmonella, the most effective antimicrobial was DMDC having a 5.79 and 5.96 log10 CFU/ml decrease directly following pressurization, respectively. Other treatments that were significantly different from the samples treated with no antimicrobial were hydrogen peroxide, and NaB at 500 ppm for E. coli O157:H7 and a treatment of NaB at 1000 ppm for S. Agona. After 24 hours at 4°C, S. Agona samples with added antimicrobials had a close to or above 5-log10 CFU/ml reduction. DMDC should be further investigated as an antimicrobial agent that can work in conjunction with HHP. / Master of Science

Beta lactam resistance producing isolates of Enterobacteriacea and non-Enterobacteriacea have emerged since more than seventy years ago (Abraham and Chain, 1940). They are known to cause both community and hospital-acquired infections. Resistance against carbapenem is primarily mediated by the production of enzymes that destroy the beta lactam antimicrobials, which are produced by these isolates involving the expression of serine and metalobetalactamase genes KPC, IMP, VIM, NDM-1 and OXA-48. Quinolone resistance is predominanty mediated by mutations in the qnrA, qnrB, qnrS, and aac-6-Ib genes. Carbapenemase-producing organisms especially Klebsiella pneumoniae carbapenemases (KPCs) emerged as important pathogens especially among critically ill patients causing significant morbidity and mortality. This study aims to determine the prevalence and types of 15 quinolone resistance and carbapenemases genes among different isolates from patients admitted to the University Hospital of Leipzig over a period of ten months. During the period from January 2011 through October 2011, a total of 50 carbapenemases isolates were recovered from patients of the University Hospital of Leipzig/ Germany. The isolates were identified by biochemical tests and their susceptibility to antimicrobials was determined by the microbroth dilution method according to ISO standard. The KPC, IMP, VIM, OXA-48, NDM-1, and aac-6-Ib genes as well as qnrA, qnrB, and qnrS genes were detected by multiplex PCR, respectively. Results showed that KPC gene was detected in 82% of the isolates while 8% were KPC negative. The qnrA, qnrS, IMP, NDM-1, and OXA-48 genes were not detected in any of the isolates while qnrB and VIM genes were found in 2%. On the other hand, aac-6-Ib gene was the most prevalent gene among the study isolates and composed a percentage of 96%. Results also showed that KPC, and aac-6-Ib genes were detected in isolates collected from urine, blood, wounds, swabs, sputum, tracheal secretions, biopsies, and anal smears, while VIM gene was detected in one isolate collected from blood. The qnrB gene was found in one isolate collected from urine specimen. The wide spread of carbapenem and quinolone resistance-producing organisms is a critical problem that complicates the treatment of infections resulting from these organisms. Necessary measures must, therefore, be taken to limit their spread, which include appropriate antibiotic treatment, control of hospital infections, observe of personal hygiene, and the use of appropriate methods of sterilization and disinfection to prevent the dissemination of these organisms. Keywords: Resistance, carbapenemases, QRDR, multiplex PCR, antimicrobials

No description available.

Made available in DSpace on 2015-09-25T12:23:10Z (GMT). No. of bitstreams: 1 TaniaDeSouzaFarias.pdf: 409062 bytes, checksum: 154f204aa9cc212ec7064f63209a81ee (MD5) Previous issue date: 2007-02-28 / The discovery of antimicrobials provided a significant reduction in infections and promoted a significant improvement in the population s quality of life. However, this reality provoked an inadequate use of these medicines, and consequently, the appearance of lineages of bacteria resistant to the action of antibiotics. Clinical usage of antimicrobials in hospitals brings individual and collective consequences because it affects the individual who uses this pharmaceutical and the microbiology of the hospital atmosphere. Thus, the main objective was to get to know the patterns of antimicrobial usage in the public hospital. A transversal study was performed, in which all prescriptions of the patients in the different clinics of the hospital who used an antimicrobial agent during internment were analyzed. Therapeutic use of antimicrobials represented 89% of the cases studied in pediatrics. Prophylactic usage occurred in all of the maternity and surgery cases, and in 81% of the clinical oncology cases. Antimicrobial consumption was primarily prophylactic, being first generation cephalosporin the most prescribed pharmaceutical. Therapeutic usage occurred principally in pediatrics and the most prescribed medications were penicillin, gentamicine and chloramphenicol. Empiric therapeutics was predominant. Inadequate use of chloramphenicol and the need for controlling measures in the usage of antimicrobials were observed in the different clinics. / A descoberta dos antimicrobianos proporcionou a redução significativa das infecções e promoveu uma melhora significativa na qualidade de vida da população. Entretanto, essa realidade ocasionou o uso inadequado desses medicamentos e conseqüentemente o surgimento de cepas de bactérias resistentes à ação dos antibióticos. O uso clínico de antimicrobianos em hospitais tem conseqüências individuais e coletivas, pois afeta o indivíduo que faz uso desse fármaco, e a microbiota do ambiente hospitalar. Desse modo, o objetivo desse trabalho foi conhecer os padrões de uso de antimicrobianos em um hospital público. Realizou-se um estudo transversal, no qual foram analisadas todas as prescrições dos pacientes das diferentes clínicas do hospital que fizeram uso algum agente antimicrobiano durante a internação. O uso terapêutico de antimicrobianos representou 89% dos casos estudados na pediatria. O uso profilático ocorreu em todos os casos da maternidade e da Ala cirúrgica, e em 81% da clínica oncológica. O consumo de antimicrobianos foi principalmente profilático sendo as cefalosporinas de primeira geração os fármacos mais prescritos. O uso terapêutico ocorreu principalmente na pediatria e os fármacos mais prescritos foram penicilinas, gentamicina e cloranfenicol. A terapêutica empírica foi predominante. Observou-se o uso inadequado de cloranfenicol nas diferentes clínicas e a necessidade de medidas de controle do uso de antimicrobianos.

This aim of the research was to explore pharmacist prescribing (PP) with a focus on antimicrobials, in hospitals in Scotland. A mixed-methods approach was used to collect, generate and synthesise data. A systematic review of peer-reviewed published literature on evidence-based roles for the pharmacist as part of an antimicrobial multidisciplinary team, identified roles for pharmacists within the teams but limited evidence relating to outcomes associated with these roles. Six qualitative focus groups, with 37 hospital pharmacists in 5 Scottish Health Boards, contextualised perceptions of barriers to, and facilitators of, implementation of PP in hospitals. Key themes were: perceived lack of pharmacy management support to take on a prescribing role and little strategic attention paid to PP implementation and sustainability. These issues were discussed in relation to PP in general and not only for antimicrobials. Participants perceived successful implementation of PP to be associated with factors including ward type and patient’s clinical condition. None of the pharmacists were prescribing antimicrobials and consequently further studies focused on PP in general. A scoping exercise, utilising various sources of information, reinforced findings from Phase 1; it highlighted the absence of any national or Health Board frameworks to support implementation of PP in secondary care in Scotland. Consensus-based research was undertaken, therefore, to provide guidance to facilitate service redesign involving PP in secondary care in Scotland. A Delphi approach undertaken with 40 experts, mainly in strategic posts, resulted in a high level of agreement in areas relating to succession planning, rather than role development; more variability was obtained in areas relating to future orientation of service, competencies required by prescribers and potential development of non-medical prescribing teams. The guidance was developed into a self-assessment toolkit providing an analytical strategy for implementation and role development of PP in secondary care. While the results and conclusions generated through this research need to be interpreted with caution, the data generated is an original contribution to the evidence base relating to PP.