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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
161

Lipid Modified Polymers for Transfection of Human CRL Fibroblasts, and for siRNA Mediated MDR Reversal in Melanoma Cancer Therapy

Abbasi Dezfouli, Meysam Unknown Date
No description available.
162

Analysis of a multidrug resistant acinetobacter SPP. outbreak in the intensive care unit of King Edward VIII Hospital.

Deedat, Fathima. January 2000 (has links)
The study arose out of a need to investigate and control a nosocomial outbreak caused by multidrug resistant Acinetobacter spp in the fifteen-bed intensive care unit of King Edward VIII Hospital. Following the discovery of the index case, four other patients were found to have a similar strain of Acinetobacter spp. All fifteen patients in the ward were subsequently screened for the organism. Forty-seven isolates were obtained from 12 patients. Eight of the patients were infected with the organism and six of these eight patients subsequently died. Swabs from the ward environment were also screened for the organism, which was found in patients' baths, suction water and urine collection jars. The outbreak was aborted by the use of strict infection control techniques. Minimum inhibitory concentrations (MICs) of 20 of the 47 isolates were determined for the following antimicrobials: imipenem, ciprofloxacin, gentamicin, amikacin, netilmycin,cefotaxime, ceftazidime and tetracycline. The same 20 isolates were further typed using ribotyping. Seven different antibiogram patterns were obtained using the MIC data. The majority of isolates (11) fit into a Single type, and showed resistance to all drugs tested, except for susceptibility to tetracycline and netilmycin only. Ribotyping revealed 5 different types. There were 9 isolates of ribotype a, 2 of ribotype b, 3 of ribotype c, 5 of ribotype d and 1 of ribotype e. In conclusion, this study describes a nosocomial outbreak with a multidrug resistant Acinetobacter spp. in an intensive care unit. The results showed that there was no correlation between the two typing methods used, ribotyping was more discriminatory than antibiogram types, with the majority of strains belonging to two different ribotypes. / Thesis (M.Med.)-University of Natal, Durban, 2000.
163

Développement de biomarqueur Sentinelle en réponse à la pollution aquatique à partir de l'expression de protéines de phénotype "Multidrug Resistance" dans les érythrocytes de la truite Salmo trutta fario

Valton, Emeline 19 October 2012 (has links) (PDF)
La pollution croissante des milieux aquatiques nécessite la mise au point de nouvelles technologies permettant d'optimiser la surveillance de la qualité de l'eau. Dans ce contexte, nous avons développé un biomarqueur de susceptibilité du degré de la pollution globale des milieux aquatiques intitulé " Sentinelle ". Le principe du biomarqueur Sentinelle est basé sur le niveau de coexpression de deux protéines " Multidrug Resistance " (MDR), la protéine ABCG2-like et la P-gp, dans les érythrocytes de la truite Salmo trutta fario. Le biomarqueur sentinelle a été validé en conditions in vitro grâce au développement des cultures primaires d'érythrocytes de truite. Après l'exposition des globules rouges de truites à des concentrations croissantes d'un polluant modèle, le Benzo-a-pyrène, l'expression de la protéine ABCG2-like et de la P-gp augmente d'une manière dose dépendante. Le biomarqueur Sentinelle a ensuite été validé en milieu naturel sur des truites fario en provenance de différents cours d'eau d'Auvergne. En milieu naturel, les deux protéines MDR sont exprimées différemment dans les érythrocytes de truites fario selon le degré de contamination du cours d'eau. En effet, dans une rivière où la pollution est faible voire nulle, seule la protéine ABCG2-like est exprimée, alors que dans une rivière présentant une contamination plus importante, la P-gp et l'ABCG2-like sont toutes les deux coexprimées par une réponse de type relais. Les expériences menées en conditions in vitro et en milieu naturel, laissent supposer que la protéine ABCG2-like assure une fonction de garde alors que la P-gp assurerait une fonction de protection défensive. En conséquence, selon le niveau d'expression de la protéine de garde et de la protéine de défense, le degré de contamination de la rivière pourrait être évalué. L'intérêt de l'utilisation du biomarqueur Sentinelle a aussi été validé sur des Salmonidés en provenance de pisciculture. Ce nouvel outil biologique apporte des informations plus intégratives et plus précoces sur la qualité des milieux aquatiques, informations essentielles pour une meilleure gestion des ressources en eau.
164

Lipid Modified Polymers for Transfection of Human CRL Fibroblasts, and for siRNA Mediated MDR Reversal in Melanoma Cancer Therapy

Abbasi Dezfouli, Meysam 11 1900 (has links)
Gene delivery for therapeutic purposes is quickly emerging as the best potential treatment option for inherited genetic diseases and cancer. Viral gene carriers have been the choice for this purpose due to their high efficiency, but harmful immunogenic and oncogenic host reactions have limited their in vivo use. Cationic polymers provide a safe alternative to viral carriers as they can be engineered to reduce immunogenic and toxic responses and serve therapeutic purposes in the body. Due to their strong positive charge, they are able to compact the negatively charged nucleotides to small nano-sized particles appropriate for cellular uptake. Additionally, they efficiently encapsulate the highly sensitive nucleotides, and protect them against degradation by the nucleases present at the physiological milieu. In this thesis work, I have used a novel approach for gene delivery by combining the critical properties of a cationic polymer (i.e., nucleotide condensing ability) with that of a fatty acid (i.e., lipid membrane compatibility). The resulting lipid modified polymer increased delivery of our gene of interest into target cells and resulted in increased siRNA delivery for cancer gene therapy. / Biomedical Sciences
165

Characterization of ABC transporters in both mammalian cells (ABCG2, ABCC2) and Plasmodium falciparum (Pgh1)

Leimanis, Mara L. January 1900 (has links)
Thesis (Ph.D.). / Written for the Institute of Parasitology. Title from title page of PDF (viewed 2008/02/12). Includes bibliographical references.
166

Studium inhibice ABC lékových efluxních transportérů vybranými inhibitory tyrozinkináz pomocí akumulačních metod s cytostatickými substráty / Study of ABC drug efflux transporter inhibition by selected tyrosine kinase inhibitors using accumulation methods with cytostatic substrates

Suchá, Simona January 2018 (has links)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Simona Suchá Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Study of ABC drug efflux transporter inhibition by selected tyrosine kinase inhibitors using accumulation methods with cytostatic substrates ATP-binding cassette (ABC) drug efflux transporters are transmembrane proteins that utilize the energy from ATP hydrolysis to drive transport of endogenous and exogenous compounds out of the cell. The overexpression of ABC transporters plays a crucial role in the development of multidrug resistance (MDR), a phenomenon responsible for the failure of chemotherapy. Tyrosine kinase inhibitors (TKI) represent novel beneficial therapeutic approach in cancer treatment. TKI block tyrosine kinases which regulate important cellular processes. Deregulation of these enzymes can lead to various types of cancers. In the present work, we investigated interaction potential of selected TKI (alectinib, brivanib, osimertinib, selumetinib) in MDCKII parent cell line and those transduced with human efflux transporters ABCB1, ABCC1 and ABCG2. Using the accumulation studies, we determined the amount of accumulated model substrates (daunorubicin, mitoxantrone) and evaluated the inhibitory effect of...
167

Associação entre polimorfismos no gene da glicoproteína-P (PgP) e resistência múltipla a anti-helmínticos em Haemonchus contortus e identificação de fatores de risco relacionados

Mello, Suelen Scarpa de 28 February 2014 (has links)
Made available in DSpace on 2016-06-02T20:21:35Z (GMT). No. of bitstreams: 1 6279.pdf: 1318446 bytes, checksum: 3ea93c315d61578a7b50605db41ae915 (MD5) Previous issue date: 2014-02-28 / Financiadora de Estudos e Projetos / Among the sheep parasites, Haemonchus contortus is the most prevalent and pathogenic nematode in tropical areas, causing huge economic losses. Alterations in the gene encoding the membrane P-glycoprotein (PgP) have been associated with multidrug resistance. The goal of this study was to identify single nucleotide polymorphisms (SNPs) on the PgP gene in H. contortus by comparing two isolates with different status of anthelmintic resistance. For this purpose, two ewes were experimentally infected, one with a susceptible H. contortus isolate (McMaster, Australia) and the other with a multidrug-resistant isolate (Embrapa2010, Brazil). Feces from infected animals were submitted to coproculture in order to obtain larvae and, after the slaughter of the ewes, adults H. contortus were collected from abomasum and individually submitted to DNA extraction. For the evaluation of potential molecular markers of geographic isolation, larvae originating from coprocultures of two other Brazilian isolates (Bahia and Pernambuco), with no determined resistance status, were also submitted to DNA extraction. After PCR amplification of DNA extracted from adults H. contortus, a fragment of the PgP gene was sequenced and six SNPs (position described in relation to the sequence of contig 004690 from the Wellcome Trust Sanger Institute) were identified : 508 (A> G in exon), 580 (G> A), 601 (G> C), 800 (G> A), 845 (G> T), and 878 (G> T), the last five in introns. The Fisher's exact test, applying the Bonferroni correction, revealed a significant association (P<0.01) between 508, 580, 601, 845 and 878 SNPs and the state of resistance to anthelmintics. Among these SNPs, 601 and 878 are in linkage disequilibrium and form the CC haplotype associated (P < 0.05) to resistance and the GG haplotype associated to susceptibility. Considering that there is considerable genetic differentiation between different continental areas in H. contortus, the frequencies of the SNPs were compared between the Brazilian isolates and the susceptible Australian one. It was found that the SNPs 580 and 845 may be associated with geographic isolation. We conclude that the 508, 601 and 878 SNPs in the P-glycoprotein gene can be molecular markers for multiple resistance to anthelmintics, and that the 580 and 845 SNPs can be candidate markers of geographical isolation in H. contortus. We developed software for Risk Analysis Development of Parasitic Resistance to Anthelmintics in Sheep (SARA) which will provide information that may guide the rational use of anthelmintics. / Among the sheep parasites, Haemonchus contortus is the most prevalent and pathogenic nematode in tropical areas, causing huge economic losses. Alterations in the gene encoding the membrane P-glycoprotein (PgP) have been associated with multidrug resistance. The goal of this study was to identify single nucleotide polymorphisms (SNPs) on the PgP gene in H. contortus by comparing two isolates with different status of anthelmintic resistance. For this purpose, two ewes were experimentally infected, one with a susceptible H. contortus isolate (McMaster, Australia) and the other with a multidrug-resistant isolate (Embrapa2010, Brazil). Feces from infected animals were submitted to coproculture in order to obtain larvae and, after the slaughter of the ewes, adults H. contortus were collected from abomasum and individually submitted to DNA extraction. For the evaluation of potential molecular markers of geographic isolation, larvae originating from coprocultures of two other Brazilian isolates (Bahia and Pernambuco), with no determined resistance status, were also submitted to DNA extraction. After PCR amplification of DNA extracted from adults H. contortus, a fragment of the PgP gene was sequenced and six SNPs (position described in relation to the sequence of contig 004690 from the Wellcome Trust Sanger Institute) were identified : 508 (A> G in exon), 580 (G> A), 601 (G> C), 800 (G> A), 845 (G> T), and 878 (G> T), the last five in introns. The Fisher's exact test, applying the Bonferroni correction, revealed a significant association (P<0.01) between 508, 580, 601, 845 and 878 SNPs and the state of resistance to anthelmintics. Among these SNPs, 601 and 878 are in linkage disequilibrium and form the CC haplotype associated (P < 0.05) to resistance and the GG haplotype associated to susceptibility. Considering that there is considerable genetic differentiation between different continental areas in H. contortus, the frequencies of the SNPs were compared between the Brazilian isolates and the susceptible Australian one. It was found that the SNPs 580 and 845 may be associated with geographic isolation. We conclude that the 508, 601 and 878 SNPs in the P-glycoprotein gene can be molecular markers for multiple resistance to anthelmintics, and that the 580 and 845 SNPs can be candidate markers of geographical isolation in H. contortus. We developed software for Risk Analysis Development of Parasitic Resistance to Anthelmintics in Sheep (SARA) which will provide information that may guide the rational use of anthelmintics. / Entre os parasitas de ovinos, Haemonchus contortus é o nematoide mais prevalente e patogênico em áreas tropicais, causando grandes perdas econômicas. As alterações no gene que codifica a glicoproteína-P de membrana (PgP) foram associadas com a resistência a múltiplas drogas. Assim, o objetivo deste estudo foi identificar polimorfismos de nucleotídeo único (SNPs) no gene PgP em H. contortus por comparação de dois isolados com diferentes status de resistência a anti-helmínticos. Para esse fim, uma ovelha foi infectada experimentalmente com o isolado suscetível (McMaster, Austrália) e outra ovelha com o isolado multirresistente (Embrapa2010, Brasil) de H. contortus. Fezes dos animais infectados foram submetidas à coprocultura para a obtenção de larvas e, após o abate das ovelhas, H. contortus adultos foram colhidos do abomaso e individualmente submetidos à extração de DNA. Para a avaliação de possíveis marcadores moleculares de isolamento geográfico, larvas oriundas de coprocultura de dois outros isolados brasileiros (Bahia e Pernambuco), com status de resistência não determinado, também foram submetidas à extração de DNA. Após amplificação por PCR do DNA extraído de H. contortus adultos, um fragmento do gene PgP foi sequenciado e foram identificados seis SNPs (posição descrita em relação à sequência do contig 004690 do Wellcome Trust Sanger Institute): 508 (A>G, em éxon), 580 (G>A), 601 (G>C), 800 (G>A), 845 (G>T) e 878 (G>T), os cinco últimos em íntrons. O teste exato de Fisher, por meio da correção de Bonferroni, revelou associação significativa (P<0,01) entre os SNPs 508, 580, 601, 845 e 878 e o estado de resistência a anti-helmínticos. Desses SNPs, o 601 e o 878 estão em desequilíbrio de ligação e formam os haplótipos CC, que está associado (P<0,05) à resistência, e GG, à suscetibilidade. Levando em consideração que em H. contortus há considerável diferenciação genética entre áreas continentais distintas, as frequências dos SNPs foram comparadas entre os isolados brasileiros e o isolado suscetível australiano e foi verificado que os SNPs 580 e 845 podem estar associados ao isolamento geográfico. Conclui-se que os SNPs 508, 601 e 878 no gene de glicoproteína-P podem ser marcadores moleculares para a resistência múltipla a anti-helmínticos, e os SNPs 580 e 845 candidatos a marcadores de isolamento geográfico em H. contortus. Nós desenvolvemos um software para Análise de Risco de Desenvolvimento de Resistência Parasitária a Anti- Helmínticos em Ovinos (SARA) que fornecerá informações que poderão orientar a utilização racional de anti-helmínticos. / Entre os parasitas de ovinos, Haemonchus contortus é o nematoide mais prevalente e patogênico em áreas tropicais, causando grandes perdas econômicas. As alterações no gene que codifica a glicoproteína-P de membrana (PgP) foram associadas com a resistência a múltiplas drogas. Assim, o objetivo deste estudo foi identificar polimorfismos de nucleotídeo único (SNPs) no gene PgP em H. contortus por comparação de dois isolados com diferentes status de resistência a anti-helmínticos. Para esse fim, uma ovelha foi infectada experimentalmente com o isolado suscetível (McMaster, Austrália) e outra ovelha com o isolado multirresistente (Embrapa2010, Brasil) de H. contortus. Fezes dos animais infectados foram submetidas à coprocultura para a obtenção de larvas e, após o abate das ovelhas, H. contortus adultos foram colhidos do abomaso e individualmente submetidos à extração de DNA. Para a avaliação de possíveis marcadores moleculares de isolamento geográfico, larvas oriundas de coprocultura de dois outros isolados brasileiros (Bahia e Pernambuco), com status de resistência não determinado, também foram submetidas à extração de DNA. Após amplificação por PCR do DNA extraído de H. contortus adultos, um fragmento do gene PgP foi sequenciado e foram identificados seis SNPs (posição descrita em relação à sequência do contig 004690 do Wellcome Trust Sanger Institute): 508 (A>G, em éxon), 580 (G>A), 601 (G>C), 800 (G>A), 845 (G>T) e 878 (G>T), os cinco últimos em íntrons. O teste exato de Fisher, por meio da correção de Bonferroni, revelou associação significativa (P<0,01) entre os SNPs 508, 580, 601, 845 e 878 e o estado de resistência a anti-helmínticos. Desses SNPs, o 601 e o 878 estão em desequilíbrio de ligação e formam os haplótipos CC, que está associado (P<0,05) à resistência, e GG, à suscetibilidade. Levando em consideração que em H. contortus há considerável diferenciação genética entre áreas continentais distintas, as frequências dos SNPs foram comparadas entre os isolados brasileiros e o isolado suscetível australiano e foi verificado que os SNPs 580 e 845 podem estar associados ao isolamento geográfico. Conclui-se que os SNPs 508, 601 e 878 no gene de glicoproteína-P podem ser marcadores moleculares para a resistência múltipla a anti-helmínticos, e os SNPs 580 e 845 candidatos a marcadores de isolamento geográfico em H. contortus. Nós desenvolvemos um software para Análise de Risco de Desenvolvimento de Resistência Parasitária a Anti- Helmínticos em Ovinos (SARA) que fornecerá informações que poderão orientar a utilização racional de anti-helmínticos.
168

Ocorrência de cepas de Escherichia coli que apresentam o gene de Shiga toxina em queijo mussarela produzido artesanalmente

Cardoso, Patrícia Alves [UNESP] 05 June 2009 (has links) (PDF)
Made available in DSpace on 2014-06-11T19:27:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-06-05Bitstream added on 2014-06-13T20:35:49Z : No. of bitstreams: 1 cardoso_pa_me_jabo.pdf: 283666 bytes, checksum: 9d4635249701354d18b4a667c65f5861 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo deste estudo foi investigar a ocorrência de cepas de Escherichia coli produtoras de Shiga toxina (STEC) em queijos mussarela produzidos artesanalmente. Foram analisadas 59 amostras de queijo, produzidas no Vale do Jequitinhonha (Nordeste de Minas Gerais, Brasil). Isolando-se 147 cepas de E. coli e através da técnica de PCR, foram investigadas a presença dos genes da Shiga toxina (stx 1 e stx 2) e da intimina (eae). Dezesseis cepas bacterianas (10,8%) apresentaram o gene stx ( todas portavam o gene stx 1) e 13 delas também se mostraram eae positivas. Os isolados de E. coli foram também examinados para a detecção dos genes codificadores de adesinas (pap, sfa e afa). Não foram identificados nenhum desses genes.Todas as cepas STEC isoladas foram pesquisadas para resistência a 12 agentes antimicrobianos. As resistências predominantes detectadas foram de 37,5% para estreptomicina, 37,5% para a tetraciclina, 31,2% para a ampicilina e 31,2% para a amicacina. A resistência a múltiplas drogas foi encontrada em 5 cepas (31,2%). A presença dos genes codificadores dos fatores de virulência indica que o queijo mussarela produzido artesanalmente pode representar um risco à saúde dos consumidores. / The aim of the present study was to investigate the occurrence of Escherichia coli strains presenting the Shiga toxin gene (probably STEC strains) in mussarela cheese produced by artesanal method in the Jequitinhonha Valey (Northeast of Minas Gerais State, Brazil). Fifty-nine cheese samples were analyzed and a hundred forty seven strains of E. coli were isolated. Using the PCR method the strains were screened for the Shiga toxin (stx 1 and stx 2) and the intimin (eae) genes. Sixteen isolates (10,8%) carried the stx gene (all of them showed the stx 1 gene ) and thirteen also presented the eae gene. Using the same method the strains were screened for the presence of pap, afa, and sfa genes, adhesin genes caracteristics of the E. coli extraintestinal pathogenic strains (ExPEC). None of them showed these adhesin genes and could not be classified as an ExPEC strains. The susceptibility of the probably STEC strains to twelve antimicrobial drugs were evaluated. The most important resistance was detected to the streptomycin (37,5%), tetracycline (37,5%), ampicillin (31,2%) and amikacin (31,2%). The multidrug resistance was detected in 5 isolates (31,2%). The presence of coding genes for virulence factors in the E. coli isolates recovered from mussarela cheese produced by artesanal method could represent a risk for the human health.
169

Caracterização fenotípica e similiaridade genética de Pseudomonas aeruginosa provenientes de efluentes hospitalares e água superficial do igarapé do Mindu/Manaus - AM

Magalhães, Mary Joyce Targino Lopes 26 July 2013 (has links)
Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-07-31T15:27:26Z No. of bitstreams: 1 Dissertação- Mary Joyce Targino Lopes Magalhães.pdf: 1466870 bytes, checksum: 3202ba13f65ece33fb3c071651d2a444 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-08-04T15:21:41Z (GMT) No. of bitstreams: 1 Dissertação- Mary Joyce Targino Lopes Magalhães.pdf: 1466870 bytes, checksum: 3202ba13f65ece33fb3c071651d2a444 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-08-04T15:28:13Z (GMT) No. of bitstreams: 1 Dissertação- Mary Joyce Targino Lopes Magalhães.pdf: 1466870 bytes, checksum: 3202ba13f65ece33fb3c071651d2a444 (MD5) / Made available in DSpace on 2015-08-04T15:28:41Z (GMT). No. of bitstreams: 1 Dissertação- Mary Joyce Targino Lopes Magalhães.pdf: 1466870 bytes, checksum: 3202ba13f65ece33fb3c071651d2a444 (MD5) Previous issue date: 2013-07-26 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / Pseudomonas aeruginosa is a leading bacterial cause of nosocomial infections. Possessing aquatic habitats, it presents a good indicator of water contamination. To verify that this bacterial species represents a potential source of contamination to the Mindu stream, we must carry out and analyze the following objectives: first, identify isolates of P. aeruginosa in samples of hospital effluent surface water obtained from the Mindu stream and second, to check whether the strains posses virulence factors as related mobility scourge, "twitching motility", biofilm formation and antimicrobial resistance, and finnaly, to assess the genetic similarity between isolates. Method: To identify the microbiological and biochemical composition, the 16S rRNA gene sequencing was used. For phenotype characterization we performed the following tests: first, we tested for mobility, using the scourge test "twithing motility", second we tested the biofilm formation and profile of antimicrobial resistance using the disk diffusion technique, the genetic similarity among isolates found was determined by PFGE. Results: We identified 17 isolates of P. aeruginosa in effluent water from the hospital. 8 isolates of the same species were in the surface water of the Mindu stream. The strains tested with mobility scourge; 100 % of the strains were found in water Mindu and 88 % of the strains were found in the hospital´s effluent samples were positive for " twitching motility ". All of the 25 isolates studied showed biofilm formation and more than 70 % of the strains found in hospital´s effluent water (raw and treated) had the phenotype of multidrug resistance.100 % of P. aeruginosa isolates found showed resistance to Ampicillin and 50 % of the strains were intermediately resistant to ceftriaxone. Among all the samples, there was genetic similarity; in the hospital´s effluent water and the hospital´s treated sewage, samples found in different seasonal periods, and among isolates found in hospital effluent and surface water of Mindu. Conclusion: The presence of P. aeruginosa containing virulence factors in surface water samples is indicative of the spread of nosocomial origin of microorganisms in the aquatic environment studied. There is strong evidence that the system of sewage treatment in the study is not efficient. Contaminants from P. aeruginosa containing multidrug resistance were in the samples of treated effluent water, and showed high genetic similarity among isolates of P. aeruginosa derived from raw wastewater. Therefore, it is necessary for action and more oversight on the part of health surveillance agencies with health services so that they meet the requirements of the laws in force in order to preserve the environment and people's health. / Pseudomonas aeruginosa é uma das principais bactérias causadora de infecções hospitalares, e por possuir habitat comumente aquático apresenta-se como boa indicadora de contaminação de águas, para verificar se essa espécie bacteriana representa uma fonte de contaminação em potencial para o igarapé do Mindu, o estudo se propôs a analisar os seguintes objetivos: identificar isolados de P. aeruginosa em amostras de efluentes hospitalares e água superficial do igarapé do Mindu; verificar se as cepas encontradas apresentavam fatores de virulência como, mobilidade ligada ao flagelo, “twitching motility”, formação de biofilme e resistência aos antimicrobianos, e avaliar a similaridade genética entre os isolados. Metodologia: Para a identificação microbiológica foram realizados testes bioquímicos e o sequenciamento do gene 16S rRNA dos isolados encontrados; Para a caracterização fenotípica foram realizados os seguintes testes: teste de mobilidade ligada ao flagelo, teste “twithing motility”, teste de formação de biofilme e perfil de resistência aos antimicrobianos pela técnica de disco-difusão; A similaridade genética entre os isolados de encontrados no estudo foi determinada por PFGE. Resultados: Foram identificados 17 isolados de P. aeruginosa em efluentes hospitalares e 8 isolados da mesma espécie na água superficial do igarapé do Mindu; Todas as cepas estudadas apresentaram mobilidade ligada ao flagelo; 100% das cepas encontradas na água do Mindu e 88% das cepas encontradas em amostras de efluentes hospitalares apresentaram “twitching motility” positivo; todos os 25 isolados estudados apresentaram formação de biofilme; mais de 70% das cepas encontradas nos efluentes hospitalares (brutos e tratados) apresentaram o fenótipo da multirresistência e 100% dos isolados de P aeruginosa encontrados na água do Mindu apresentaram resistência à Ampicilina e 50% dessas cepas apresentaram resistência intermediária a Ceftriaxona; houve similaridade genética entre isolados encontrados em efluente hospitalar bruto e efluente hospitalar tratado, entre isolados encontrados em diferentes períodos sazonais e entre isolados encontrados em efluentes hospitalares e água superficial do Mindu. Conclusão: A presença de P. aeruginosa contendo fatores de virulência, em amostras de água superficial, é indicativo de disseminação de microrganismos de origem nosocomial no ambiente aquático estudado. Há fortes indícios de que o sistema de tratamento de efluentes do estudo não está sendo eficiente, já que foram encontradas cepas de P. aeruginosa contendo fatores de virulência, inclusive a multirresistência em amostras do efluente tratado; e foi observada alta similaridade genética entre isolados de P. aeruginosa oriundos de efluente bruto e efluente tratado. Por isso, se faz necessário maior fiscalização por parte dos órgãos de vigilância sanitária junto aos serviços de saúde para que sejam cumpridas as exigências das legislações vigentes a fim de preservar o meio ambiente e a saúde da população.
170

Isolamento e caracterização de Campylobacter spp. em carcaças de frango e ovos comerciais / Isolation and characterization of Campylobacter spp. in chicken and commercial eggs

Oliveira, Julierme José de 26 February 2014 (has links)
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No. of bitstreams: 2 Dissertação - Julierme José de Oliveira - 2014.pdf: 1186561 bytes, checksum: ee682becf7a3d3796f34ba3239320262 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-02-26 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Thermophilic species of Campylobacter spp. are often cited as the most common etiologic agent in cases of foodborne illnesses in several countries. When ingested even in small quantities (400 to 500 cells) this organism, through the mechanisms of pathogenicity, the host develops infectious gastroenteritis known as campylobacteriosis. This disease is closely related to the intake and improper handling of contaminated food, especially poultry, and birds the main reservoir of the microorganism. Poor knowledge climbs the role of eggs in the transmission of campylobacteriosis, based on the annual consumption of the food matrix, research clarifying all variables involving this food are essential. This study aimed to verify the occurrence of Campylobacter spp., identify thermophilic species involved and assess the level of antimicrobial susceptibility of isolates from chicken carcasses and eggs obtained from commercial establishments in the city of Goiânia. A total of 100 chicken carcasses and 100 dozens of eggs were evaluated using enzyme- linked immunosorbent assay for screening miniVIDAS ® with subsequent bacterial isolation and identification of samples by biotyping . By means of the methods employed observed that Campylobacter spp. was identified in 7% (7 /100) of chicken carcasses in 1% (1/100 ) of samples of eggs, 100 % of these were confirmed as belonging to the bacterial genus . The species with the highest number of isolates were Campylobacter jejuni (50 %), followed by Campylobacter coli ( 25 % ) and Campylobacter lari ( 25 % ). For antimicrobial resistance results were that 100% of the isolates were resistant to erythromycin, as well as ciprofloxacin and tetracyclines. For ceftriaxone and nalidixic acid resistance values were 91.7 % resistant. Chloramphenicol was 50% of resistant isolates, showed 33.3% sensitivity and 16.7% intermediate resistance. It was also observed that 100% of the isolates showed multidrug profile. Through the results obtained it is concluded that thermophilic Campylobacter are present in chicken carcasses and marketed in Goiânia eggs, whose distribution becomes relevant from the point of view of exposure to consumers. Campylobacter jejuni was the most frequent species. The six districts surveyed had bacteria of the genus in at least one category. Isolates when screened for response to antimicrobial proved multiresistant, denoting the importance of research as a way to support the election of bases and principles for the antibiotic treatment prescribed treatments. Poultry foods surveyed in this study and considered positive for the pathogen constitute risk to the consumer. / As espécies termotolerantes de Campylobacter spp. são frequentemente apontadas como o agente etiológico mais comum nos casos de doenças veiculadas por alimentos em diversos países. Ao ser ingerido mesmo em pequenas quantidades (de 400 a 500 células), esse microrganismo, por meio dos mecanismos de patogenicidade, desenvolve gastroenterite infecciosa no hospedeiro, conhecida como campilobacteriose. Esta doença está intimamente relacionada à ingestão e manipulação incorreta de alimentos contaminados, especialmente carne de frango, sendo as aves o principal reservatório. Pouco se sabe sobre o papel desempenhado pelos ovos na transmissão de campilobacteriose, tendo como base o consumo anual dessa matriz alimentar, pesquisas que esclareçam todas as variáveis envolvendo esse alimento são de fundamental importância. Pelo exposto, este estudo teve como objetivo verificar a ocorrência de Campylobacter spp., identificar as espécies termotolerantes circulantes na cidade de Goiânia, vinculadas aos estabelecimentos comercializadores de alimentos, relacionados à alta densidade populacional, localizados em distritos comerciais e avaliar o nível de suscetibilidade a antimicrobianos dos isolados de carcaças de frango e ovos provenientes destes estabelecimentos. Um total de 100 carcaças de frango e 100 dúzias de ovos foram avaliados utilizando como triagem o ensaio imunoenzimatico miniVIDAS® com posterior isolamento bacteriano das amostras e identificação por meio da biotipificação. Por meio dos métodos empregados observou-se que Campylobacter termotolerantes foi identificada em 7% (7/100) das carcaças de frango e em 1% (1/100) das amostras de ovos. A espécie com maior número de isolados foi Campylobacter jejuni (50%), seguidos por Campylobacter coli (25%) e Campylobacter lari (25%). Para a resistência à antimicrobianos os resultados reportaram que 100% dos isolados são resistentes à eritromicina, assim como à ciprofloxacina e tetraciclina. Para ácido nalidixico e ceftriaxona, os valores de resistência foram da ordem de 91,7%. Cloranfenicol foi resistente em 50% dos isolados, apresentou sensibilidade a 33,3% e resistência intermediária a 16,7%. Observou-se ainda que 100% dos isolados apresentaram perfil de multirresistência. Pelos resultados obtidos conclui-se que Campylobacter termotolerantes estão presentes em carcaças de frango e ovos comercializados na cidade de Goiânia, cuja distribuição torna-se relevante do ponto de vista da exposição aos consumidores. Campylobacter jejuni foi a espécie mais frequente. Os seis distritos pesquisados apresentaram bactérias do gênero pelo menos em um estabelecimento. Os isolados quando pesquisados quanto a resposta a antimicrobianos mostraram-se multirresistentes, denotando a importância desta investigação como forma de respaldar a eleição de bases e princípios para a antibioticoterapia para tratamentos prescritos. Os alimentos avícolas pesquisados neste estudo e considerados positivos para o patógeno constituem risco ao consumidor.

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