Clinical S. aureus strains were gathered from four hospitals, two in Turkey (Hacettepe hospital 200 strains and Ankara Hospital 106 strains) and the other two from Libya (Aljalla Hospital 88 strains and Jamahyria Hospital 62 strains). The clinical specimens were collected form different sources including blood, urine, wound, pus, burn, sputum, semen, catheter and aspiration. Patients were aged between 0 to 84 years and from both sexes. Resistance to Methicillin was determined by measuring the Oxacillin MIC / this was done by using the oxacillin E-test, with resistance defined as an MIC of &gt / 2 &micro / g ml. In this study all isolates displayed an Oxacillin MIC of &amp / #8805 / 256&micro / g/ml. The MRSA strains were (56%) in Turkish hospitals, and (59%) in Libyan hospitals. The percentage of the VRSA and VISA in Libyan hospitals was (7%) and (26%) respectively, although the percentage of VRSA in Turkish hospitals was only 2% and there were no intermediately susceptible Staphylococcus aureus (VISA). Besides the MRSA isolates, Coagulase Negative Staphylococcus showing Methicillin resistance was collected from clinical isolates in thirteen patients in Turkish hospitals. In both countries, the majority MRSA isolates were multiresistant to more than five classes of antibiotics including / Ampicillin, Amoxicillin, Tetracycline, Erythromycin and Ciprofloxacin. Most of the MRSA isolates were from blood (68%), wounds (57%) and pus (50%).The results of genetic investigations indicated that the mecA gene was present in the majority of isolates in both countries / the community acquired MRSA type (ccr-BIV) was present in three samples out of thirty in Turkish hospitals and in one case out of twenty in Libyan hospitals / There was no case out of fifty specimens that carry the hospitals acquired MRSA type (ccr-BI, II, III) in both countries. Besides the Methicillin resistance gene, the incidence of Tetracycline resistance gene was quite high (tetM and tetK 50%) in Turkish hospitals isolates, and the prevalence of Panton-Valentine Leukocidin gene was high (PVL 70%) in Libyan hospitals specimens.

Submitted by Etelvina Domingos (etelvina.domingos@ufpe.br) on 2015-04-09T19:51:49Z No. of bitstreams: 2 DISSERTAÇÃO Fábio José dos Santos Aguiar.pdf: 859548 bytes, checksum: 26707ed652529cbd3f3d6a6142b5b0dd (MD5) license_rdf: 9 bytes, checksum: 42dd12a06de379d3ffa39b67dc9c7aff (MD5) / Made available in DSpace on 2015-04-09T19:51:49Z (GMT). No. of bitstreams: 2 DISSERTAÇÃO Fábio José dos Santos Aguiar.pdf: 859548 bytes, checksum: 26707ed652529cbd3f3d6a6142b5b0dd (MD5) license_rdf: 9 bytes, checksum: 42dd12a06de379d3ffa39b67dc9c7aff (MD5) Previous issue date: 2014 / O objetivo deste estudo foi avaliar a ação sinérgica entre o ácido úsnico extraído de Cladonia substellata Vainio e cinco agentes antimicrobianos (ciprofloxacino, gentamicina, oxacilina e penicilina) sobre dez cepas de Staphylococcus aureus com fenótipo de resistência previamente definido. Cinco destas cepas de S. aureus (ATCC 33591, AM 13, AM 18, AM 20, AM 21) apresentaram resistência a todos os agentes antimicrobianos avaliados e desta forma foram selecionadas para o estudo do sinergismo entre o ácido úsnico e os agentes antimicrobianos através do método do tabuleiro de xadrez (checkerboard method). Os critérios utilizados para avaliar a atividade sinérgica foram definidos pelo Índice da Concentração Inibitória Fracionada (FICI). Todas as cepas de S. aureus foram suscetíveis ao ácido úsnico, determinado pelo método de microdiluição. O FICI variou de 0,25 – 1,0, sugerindo uma interação sinérgica frente as cepas de S. aureus MRSA. A associação do ácido úsnico com o ciprofloxacino apresentou efeito sinérgico sobre todas as cepas S. aureus MRSA. A oxacilina apresentou sinergismo em associação com ácido úsnico sobre as cepas de S. aureus ATCC 33591, AM 13, AM18 e AM24 e exibiu os menores valores FICI. A associação do ácido úsnico com a gentamicina foi sinérgica sobre as cepas AM18, AM21 e AM24. A associação do ácido úsnico com a penicilina apresentou-se indiferente para todas as cepas exceto para S. aureus AM13. Este estudo demonstrou que o ácido úsnico, quando associado à antimicrobianos fluoroquinolônicos, betalactâmicos e aminoglicosídeos pode agir sinergicamente, inibindo cepas de S. aureus MRSA.

Optical sensor systems that integrate Long-Period-Gratings (LPG) as the detection arm have been proven to be highly sensitive and reliable in many applications. With increasing public recognition of threats from bacteria-induced diseases and their potential outbreak among densely populated communities, an intrinsic, low-cost biosensor device that can perform quick and precise identification of the infection type is in high demand to respond to such challenging situations and control the damage those diseases could possibly cause. This dissertation describes the development of a biosensor platform that utilizes polymer thin films, known as ionic self-assembled multilayer (ISAM) films, to be the sensitivity- enhancing medium between an LPG fiber and specific, recognition layer. With the aid of cross- linking reactions, monoclonal antibodies (IgG) or DNA probes are immobilized onto the surface of the ISAM-coated fiber, which form the core component of the biosensor. By immersing such biosensor fiber into a sample suspension, the immobilized antibody molecules will bind the specific antigen and capture the target cells or cell fragments onto the surface of the fiber sensor, resulting in increasing the average thickness of the fiber cladding and changing the refractive index of the cladding. This change occurring at the surface of the fiber results in a decrease of optical power emerging from the LPG section of the fiber. By comparing the transmitted optical power before and after applying the sample suspension, we are able to determine whether or not certain bacterial species have attached to the surface of the fiber, and as a consequence, we are able to determine whether or not the solution contains the targeted bacteria. This platform has the potential for detection of a wide range of bacteria types. In our study, we have primarily investigated the sensitivity and specificity of the biosensor to methicillin- resistant Staphlococcus aureus (MRSA). The data we obtained have shown a sensitive threshold at as low as 102 cfu/ml with pure culture samples. A typical MRSA antibody-based biosensor assay with MRSA sample at this concentration has shown optical power reduction of 21.78%. In a detailed study involving twenty-six bacterial strains possessing the PBP2a protein that enables antibiotic resistance and sixteen strains that do not, the biosensor system was able to correctly identify every sample in pure culture samples at concentration of 104 cfu/ml. Further studies have also been conducted on infected mouse tissues and clinical swab samples from human ears, noses, and skin, and in each case, the system was in full agreement with the results of standard culture tests. However, the system is not yet able to correctly distinguish MRSA and non-MRSA infections in clinical swab samples taken from infected patient wounds. It is proposed that nonspecific binding due to insufficient blocking methods is the key issue. Other bacterial strains, such as Brucella and Francisella tularensis have also been studied using a similar biosensor platform with DNA probes and antibodies, respectively, and the outcomes are also promising. The Brucella DNA biosensor is able to reflect the existence of 3 Brucella strains at 100 cfu/ml with an average of 12.2% signal reduction, while negative control samples at 106cfu/ml generate an average signal reduction of -2.1%. Similarly, the F. tularensis antibodies biosensor has shown a 25.6% signal reduction to LVS strain samples at 100 cfu/ml, while for negative control samples at the same concentration, it only produces a signal reduction of 0.05%. In general, this biosensor platform has demonstrated the potential of detecting a wide range of bacteria in a rapid and relatively inexpensive manner. / Ph. D.