Thesis (Ph.D.)--University of North Carolina at Charlotte, 2005. / Includes bibliographical references (leaves 105-118).

München, Techn. Univ., Diss., 2002.

Introduction: Each year in the United States, Salmonella enterica infections cause an estimated 1.2 million illnesses that result in 19,000 hospitalizations and 390 deaths. Illnesses occur sporadically throughout the year, but might also occur as part of an outbreak. Outbreaks are most commonly linked to a food source, but contact with live animals can also result in human outbreaks of illness. Methods: Outbreaks of Salmonella reported to the Centers for Disease Control and Prevention through National Outbreak Reporting System (NORS) from 2009–2014 with a primary mode of transmission listed as animal contact or food were analyzed to characterize the demographics of zoonotic outbreaks and examine how they differ from foodborne outbreaks. Missing data for age or sex categories were recoded as age or sex unknown. Chi-square tests were conducted to compare proportions of categorical variables. Logistic regression was used to calculate odds ratios for age, sex, health outcomes and multistate exposure. Wilcoxon rank-sum tests were used to compare medians for outbreak size and duration. Analyses were conducted using SAS 9.3. Results: During 2009–2014, a total of 484 outbreaks were reported through NORS; of these, 99 (20.5%) resulted from Salmonella transmission through animal contact and 385 (79.5%) resulted from foodborne transmission. These outbreaks resulted in 3,604 (19.8%) and 13,568 (80.2%) illnesses, respectively. A higher proportion of outbreak-associated illnesses among children aged(15.2% vs. 1.4%, p Conclusions: Outbreaks of Salmonella resulting from animal contact frequently have characteristics that are distinct from food outbreaks. Findings are consistent with reports in the literature where young children are disproportionately affected by animal contact outbreaks. Animal contact outbreaks might have a higher proportion of hospitalizations, emergency room visits, and healthcare provider visits. Animal contact outbreaks might also be longer in duration and are more likely to be multistate. Future investigations of multistate Salmonella outbreaks that are consistent with these differences should collect epidemiologic information regarding animal exposures to determine if contact with animals resulted in the transmission of the outbreak.

Attempts to produce an effective vaccine against group B meningococcal disease have been unsuccessful due to poor immunogenicity of the capsular polysaccharide, and high variability in the currently identified outer membrane proteins. There is therefore a need to find other vaccine candidates. It is clear from studies of bacteria such as Salmonella typhimurium, that successful pathogens have the ability to specifically express genes in response to an in vivo environment. Many of these in vivo-expressed gene products will contribute to virulence of the organism and may therefore be target antigens for development of new vaccines. An in vivo promoter-probe strategy was developed using the Cre/lox recombination system to identify meningococcal promoters that are specifically expressed in vivo. The strategy employs the cre gene, which encodes a site-specific recombinase that targets two 34-base pair directly repeated lox sites. A vector, pUS1604, was constructed which contains two lox sites flanking a transcriptional terminator that separates the aph (encoding kanamycin resistance) coding sequence from its promoter. A promoterless ere gene was placed on a low copy number vector to limit the number of Cre molecules within each host bacterial cell. Expression of Cre is detected by its ability to recombine the two lox sites and reconstitute a functional kanamycin resistance gene. The system was initially tested in E. coli by cloning a known regulated promoter from the araBAD operon that is induced by L-arabinose and repressed by D-glucose. However, the promoter failed to tightly control the regulation of cre gene expression, resulting in partial loxP recombination, which produced a mixed population of streptomycin and kanamycin resistant cells within each colony. Also, attempts to place the system into the meningococcus were unsuccessful. Efforts were made to tighten the regulation of the cre gene by placing the system into a different E. coli strain and test the system using a library of meningococcal genomic DNA fragments, as a compromise to show the experiments intended to isolate promoters. A vector, pUS1623, was constructed that would place the cre gene under the control of external promoters. A group B meningococccal genomic library was cloned upstream of the promoterless cre gene, with the inserted DNA fragments ranging from 0.1-1kb in size. The library was introduced into the E.coli XLl-Blue strain carrying pUS1604, and the Cre/lox recombination event has been shown to behave as expected, yielding a proportion of kanamycin-resistant clones. This indicates that promoter-like sequences are driving Cre, which is seen through the kanamycin-resistant phenotype of the clone. Conversely, clones that remain streptomycin-resistant do not have promoterlike sequences driving the expression of Cre. This streptomycin-resistant pool of clones was used to inoculate 50% serum/50% PBS liquid medium to identify group B strain meningococcal promoter sequences that drive Cre expression in this environment. Several sequences found to induce Cre expression from the initial screen, and during the serum experiment were sequenced. The sequence data was next analysed for homology against the sequences on the group B meningococcal genome sequence database (http://www.tigr.org). to identify the function of the contig from which the isolated sequence is found within the meningococcal genome. Finally, the sequences were analysed for prokaryote -35 and -10 consensus regions and orientated to determine the direction of transcription relative to the cre gene and also to genomic open reading frames, to determine the likelihood of isolating actual promoters. Although the experiments carried out in E. coli do not give an accurate picture of the regulatory gene pathways in the meningococcus, the results give an insight into the suitability of the system as regards to its use in an IVET strategy.

Diploma work focuses mainly on the influence of vermicomposting process, which should reduce the incidence of bacteria of family Enterococcus and bacteria of family of Salmonella spp. in the product. As these are among the famous and important human and animal pathogens there was an experiment done to prove this statement. The experiment took 10 weeks. During the first 14 days (from 9th November 2015) there were samples (pomace of grape vine and apple pomace) taken in the research institute FAPPZ in Červený Újezd. The earthworms were taken form the materials and then taken to SZÚ in Prague with the material, where there was the preparation and analyses of the sample done. The procedure of sample work was as following. 50g of homogenised material was placed into 48 perforated dishes. The material for the lab experiment was prepared from vermicompost (75%) and from the raw material (25%) so that there was enough nourishment for the earthworms for the duration of the trial. There were certain numbers of microorganisms stated (CPM) in this material. Salmonella reached the values of < 1 CFU/g as the initial substrate. Enterococci in the pomace from the grape vine with earthworms reached the values of 1,1.10 CFU/g, at pomace from the grape vine without earthworms < 750 CFU/g and at apple pomace the value was always < 750 CFU/g. The eathworms were placed upto the half of the glass (2,5g) 6 dishes were chosen from each material, 2 of them served as a check, 2 were inoculated enterococci and 2 with salmonella. There was always one out of 2 dishes placed with earthworms. After the filling there was inoculated of the examined microorganisms. Inoculated variants with enterococci were inoculated of 2.0.107 CFU/g and variants of salmonella by 1.1.108 CFU/g. After inoculated there was sand placed to the dish. During 8 weeks in frequency of 14 days the chosen samples were taken for analyses. The first analyses was done 23rd November 2025, the 2nd analyses was done 7th December, 3rd analyses was done 21st December 2015 and the 4th analyses was done 4th January 2016. Before the analyses was done the eartworms were taken out of the dish, they were weighted then they vitality was stated. It was proved there there isn´t any influence of earthworms on the reduction of pathogens according to the experiments. It is impossible to state that there is the influence of eartworms on the process of making compost. The reduction of pathogens was the highest at the family of Salmonella spp. There was obvious reduction at the second week of analyses and in both materials. Bacteria of the family Enterococcus showed slower process of reduction because of higher resistance to temperatures, pH, chemical substances and preparations. The absolute reduction was obvious the during the last week of the experiment.

CARVALHO, Fátima Cristiane Teles de. Influências exôgenas na qualidade bacteriológica da água, solo e camarão (Litopenaeus vannamei), em quatro fazendas de camarão do Estado do Ceará. 2006. 87 f. Dissertação (Mestrado em Ciências Marinhas Tropicais) - Universidade Federal do Ceará, Instituto de Ciências do Mar, Fortaleza, 2006. / Submitted by Debora Oliveira (deby_borboletinha@hotmail.com) on 2011-11-25T15:28:55Z No. of bitstreams: 1 2006_dis_fctdcarvalho.pdf: 1074909 bytes, checksum: 70a95311bb90d6fa8060cd66892f0469 (MD5) / Approved for entry into archive by Nadsa Cid(nadsa@ufc.br) on 2011-12-02T19:19:04Z (GMT) No. of bitstreams: 1 2006_dis_fctdcarvalho.pdf: 1074909 bytes, checksum: 70a95311bb90d6fa8060cd66892f0469 (MD5) / Made available in DSpace on 2011-12-02T19:19:04Z (GMT). No. of bitstreams: 1 2006_dis_fctdcarvalho.pdf: 1074909 bytes, checksum: 70a95311bb90d6fa8060cd66892f0469 (MD5) Previous issue date: 2006 / The objective of the present study was to evaluate the influence of the environment upon marine shrimp farming with regard to bacteriological quality of the water, pond sediments and livestock (Litopenaeus vannamei). The study was carried out on four shrimp farms in Ceará (1, 2, 3 and 4). Of the eight samplings performed, 3 involved water (one outside the farm-PEX, one at the pumping site-PB, and one in the pond-PV); 3 involved sediment (at locations PEX, PB and PV), and two consisted of harvested and processed shrimp. Samplings Were carried out in different seasons (dry, intermediary and rainy) and totaled 288 individual samples (108 water; 108 sediment; 72 shrimp). The most probable numbers (MPN) of total coliforms (TC), fecal coliforms (FC) and Escherichia coli were determined and samples were investigated for Salmonella strains. The bacteriological contamination observed on the shrimp farms was predominantly caused by TC and FC and was most intense during the intermediary season. Farms 2 and 4 yielded the highest concentrations of FC in water and sediment, respectively, at sampling locations PEX and PB during the three seasons. No shrimp sample presented E. coli values above those permitted by the European Commission (EU). However, on account of the Salmonella values observed, Farms 2 and 4 would be barred from exporting shrimp to the EU. In addition, water sampled at Farm 3 contained Salmonella strains capable of infecting the livestock. Only Farm 1 presented an exogenous and endogenous environment free of fecal contamination. The presence of Salmonella and E. coli in water and shrimp samples is disquieting, considering the fact that theses pathogens are of fecal origin and that their natural habitat is the intestinal tract of warm-blooded animals. / O presente estudo teve por objetivo pesquisar a influência do meio exógeno à carcinicultura na qualidade bacteriológica da água, sedimento dos viveiros e no camarão de cultivo (Litopenaeus vannamei). Foram realizadas oito coletas em quatro Fazendas 1, 2, 3 e 4, sendo três de água (ponto externo-PEX, ponto de bombeamento-PB e viveiro-PV) e três de sedimento (PEX, PB e PV) e duas amostras de camarão (despescado e processado). As coletas foram realizadas em diferentes períodos sazonais (seco, intermediário e chuvoso), perfazendo um total de 288 amostras, sendo 108 de água, 108 de sedimento e 72 de camarão. Foi determinado o Número Mais Provável (NMP) de coliformes totais (CT), fecais (CF), e de Escherichia coli, além da pesquisa e identificação de Salmonella. Os resultados mostram que a contaminação microbiológica das fazendas é feita predominantemente por CT e CF, com maior intensidade na estação intermediária. As Fazendas que apresentaram valores mais elevados de CF nas três estações foram as Fazendas 2 (PEX e PB) e 4 (PEX e PB) nas amostras de água e sedimento, respectivamente. Nenhuma amostra de camarão, das fazendas, apresentou valores acima do permitido para E. coli pela Comissão Européia (CE). Porém se o parâmetro considerado for a presença de Salmonella, as Fazendas 4 e 2 não estariam habilitadas a exportar os crustáceos para países da União da Comunidade Européia. E ainda, das águas da fazenda 3 também foram isoladas Salmonella o que facilmente poderia contaminar os camarões cultivados. Dentre as quatro fazendas, somente a Fazenda 1, não apresentou ambiente endógeno e exógeno contaminado com material fecal. A detecção de Salmonella e a presença de E. coli nas amostras de água e camarão é preocupante, uma vez que estes patógenos são de origem fecal e seu habitat é o trato intestinal dos animais de sangue quente e pecilotérmicos.

Salmonella enterica subsp. enterica sorovar Derby (S. Derby) é um dos sorovares mais frequentemente detectados na cadeia produtiva de suínos na região sul do Brasil. Estudos prévios indicaram que isolados de S. Derby apresentando perfis similares de resistência a antimicrobianos estavam circulando ao longo dos anos nesta região. A sobrevivência de Salmonella no ambiente e a persistência no hospedeiro são facilitadas pela sua habilidade em formar biofilmes. Desta forma, o objetivo deste trabalho foi identificar a presença de grupos clonais entre 140 isolados de S. Derby coletados ao longo de um período de dez anos de diferentes origens da cadeia produtiva de suínos (linfonodos, conteúdo intestinal, ambiente e produtos suínos). Para tanto, o perfil de resistência a antimicrobianos, através de dois conceitos distintos de avaliação de resistência (valores ECOFF e valores de breakpoint), a habilidade em formar biofilmes e a caracterização genotípica por macro-restrição (PFGE) foram avaliados. A determinação do MIC foi realizada em todos os isolados, frente aos seguintes antimicrobianos: ácido nalidíxico, ampicilina, cefotaxima, ceftazidima, ciprofloxacina, colistina, florfenicol, gentamicina, estreptomicina e tetraciclina. Para avaliar a capacidade de S. Derby em formar biofilmes, três metodologias foram aplicadas: i. morfologia de colônia em ágar contendo Vermelho Congo, para determinar a detecção fenotípica de fímbria curli e celulose; ii. formação de biofilme na interface ar-líquido em caldo LB; iii. aderência em microplacas de poliestireno de 96 poços. No total, apenas 17 (12,1%) isolados puderam ser classificados como suscetíveis a todos os antimicrobianos testados. As maiores frequências de populações “non-Wild Type” (nWT) foram observadas frente a tetraciclina (75,7%), estreptomicina (70%) e colistina (11,4%), enquanto que não foram detectadas populações nWT para ciprofloxacina, ceftazidima e gentamicina. Multi-resistência (resistência a ≥ 3 antimicrobianos; de acordo com valores de breakpoint) foi detectada em 8,6% dos isolados. Em ágar contendo Vermelho Congo, três diferentes morfologias de colônia foram observadas. A morfologia Rdar (vermelha, seca e rugosa) foi a mais frequente, sendo detectada em 59 isolados (42,1%); enquanto que as morfologias Saw (branca e mucoide) e Pdar (rosa, seca e rugosa) foram observadas em 46 (32,9%) e em 35 (25%) dos isolados, respectivamente. Os morfotipos Rdar e Pdar, que em conjunto albergaram a maioria dos isolados, são considerados os mais usualmente observados em isolados de Salmonella capazes de formar biofilme. A formação de película rígida, sem dispersão por agitação, foi observada em 24 amostras (17,1%) em caldo LB convencional e em 78 amostras (55,7%) em caldo LB low salt, sendo todas pertencentes aos morfotipos Rdar ou Pdar. Em placas de poliestireno de 96 poços, um grande número (n=96; 68,6%) dos isolados foram fracamente ou moderadamente produtores de biofilme, incluindo todos os isolados que apresentaram os morfotipos Rdar e Pdar. Através dos perfis de formação de biofilme e de resistência a antimicrobianos observados, 36 isolados foram selecionados para macro-restrição por eletroforese em campo pulsado (PFGE). Destas, 27 foram agrupadas em 11 pulsotipos que apresentavam mais de uma cepa com 100% de similaridade. Desta forma, foi possível identificar clones e cepas intimamente relacionadas de S. Derby que circularam em granjas, matadouros-frigoríficos e foram encontrados em alimentos ao longo de uma década. / Salmonella enterica subsp. enterica serovar Derby (S. Derby) is one of the serovars most frequently detected in the swine production chain of southern Brazil. Previous studies indicated that isolates of S. Derby displaying similar antimicrobial resistance profiles circulated over the years in this region. Survival of Salmonella in the environment and persistence in the host are facilitated by the ability to form biofilms. Thus, the aim of this study was to identify the presence of clonal groups between 140 S. Derby isolates collected over a ten-year period from various porcine origins (lymph nodes, intestinal content, environment and pork). Therefore, the antimicrobial resistance profile, through two distinct concepts of resistance evaluation (ECOFF value and breakpoint value), the ability to form biofilm and the genotypic characterization by macro-restriction (PFGE) were evaluated. MIC determinations were performed on all isolates, against the following antimicrobials: nalidixic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, colistin, florfenicol, gentamicin, streptomycin and tetracycline. To evaluate the S. Derby ability to form biofilm, three methods were applied: i. colony morphology on Congo Red agar, to determine the phenotypic detection of curli fimbriae and cellulose; ii. biofilm formation in liquid-air interface of LB broth; iii. adherence on a 96-well polystyrene microtiter plate. In total, only 17 (12.1%) isolates were classified as susceptible to all antimicrobials tested. The highest frequencies of non-Wild Type (nWT) populations were observed against tetracycline (75.7%), streptomycin (70%) and colistin (11.4%), whereas there were no detectable nWT population to ciprofloxacin, ceftazidime and gentamicin. Multidrug resistance (resistance to ≥ 3 antimicrobials) was detected in 8.6% of isolates. On Congo Red agar, three different colony morphology types were observed. The Rdar type (read, dry and rough) was the most frequent, being detected in 59 isolates (42.1%); while Saw (white and smoth) and Pdar (pink, dry and rough) types were observed in 46 (32.9%) and 35 isolates (25%), respectively. The Rdar and Pdar types, which together encompassed the majority of the isolates, are considered the most often observed in Salmonella isolates able to form biofilm. The formation of a rigid pellicle, which could not be dispersed by agitation, was observed in 24 (17.1%) isolates in conventional LB broth and in 78 (55.7%) isolates in low salt LB broth, all belonging to morphotypes Rdar or Pdar. On 96-well polystyrene microtiter plate, a larger number (n=96; 68.6%) of isolates showed to be weak or moderate biofilm producers, including all isolates that showed the Rdar and Pdar type. Through the biofilm and resistance profiles observed, 36 isolates were selected for macro-restriction for pulsed-feld gel electrophoresis (PFGE). Of these, 27 isolates were clustered in 11 pulsotypes which presented more than one strain with 100% of similarity. Therefore, was possible to identify clones and strains closely related of S. Derby that circulated in pig farms, slaughter plants and encountered in foods along a decade.

A boa qualidade do ar é fator essencial para manutenção da saúde humana e ambiental. O material particulado atmosférico é um poluente do ar que consiste de uma variedade de substâncias orgânicas e inorgânicas, entre as quais, algumas apresentam características genotóxicas. O ramo petroquímico é um importante componente da atividade industrial do estado do Rio Grande do Sul, Brasil, com histórico de emissões prejudiciais à qualidade do ar. O presente estudo objetivou avaliar os efeitos biológicos associados à dispersão de material particulado atmosférico inalável em área sob influência de um complexo petroquímico, integrando os resultados de biomonitoramento ambiental e humano através de biomarcadores de genotoxicidade. Amostras de material particulado foram coletadas em área de característica mista rural, urbana e industrial, utilizando filtros de membrana de Teflon e amostradores de grandes volumes de ar, semanalmente, por períodos de 24hs. Partículas inaláveis finas (MP2,5) foram amostradas em dois locais– NO e NE - posicionados na primeira e segunda direção preferencial dos ventos na região de estudo, a 2,5Km de distância da principal fonte emissora da central de matérias-primas do complexo petroquímico. Em um terceiro local (NO II), afastado 35Km da fonte emissora e na primeira direção dos ventos foi analisado partículas inaláveis grossas (MP10). Extratos orgânicos foram obtidos dos filtros com solvente diclorometano e avaliados pelo ensaio Salmonella/microssoma, método de microssuspensão, com as linhagens TA98, YG1021 e YG1024. Para avaliar a mutagenicidade dos metabólitos utilizou-se a fração de metabolização de mamíferos (S9). Também foram empregados Ensaio Cometa (EC) e Teste de Micronúcleos em linhagem celular de pulmão de hamster chinês (V79), nos extratos dos locais NO e NE. Os 16 hidrocarbonetos políclicos aromáticos (HPAs) considerados preferenciais pela agência ambiental norte americana (USEPA) foram avaliados nos extratos orgânicos para caracterização dos três locais. Foi realizado o biomonitoramento humano em crianças de escolas públicas do Local NO e NO II, com coleta de sangue periférico e células esfoliadas da mucosa oral. Em sangue periférico foi utilizado o Ensaio do micronúcleo (MN) com bloqueio da citocinese- citoma (CBMN-cyt) para avaliar as frequências de células com MN e anomalias nucleares (brotos nucleares - NBUDs, pontes nucleoplásmicas -NPBs), e EC avaliando o parâmetro intensidade de cauda. Utilizou-se o Ensaio do MN em mucosa oral-citoma (BMCyt) para detectar MN e anomalias nucleares. Todos os extratos de MP2,5 mostraram mutagenicidade pelo ensaio Salmonella, sendo destacada a presença de compostos nitrogenados. Verificaram-se respostas genotóxicas no EC em 87% dos extratos testados e indução de micronúcleos em 74% dos ensaios realizados, além de citotoxicidade em V79, para todas as amostras avaliadas. Os resultados obtidos nos três biomarcadores in vitro, e o perfil de HPAs mais tóxicos no material particulado, apontaram pior qualidade do ar do Local NO, sendo compatível com a maior dispersão de poluentes na primeira direção dos ventos. O biomonitoramento contou com a participação de 54 crianças de 5 a 12 anos, com média de 8,3 ± 1,8 anos. Danos ao DNA pelo EC foram significativamente mais elevados no local NO, em relação ao local NO II. A frequência de MN no CBMN-cyt não diferiu entre os grupos, porém, foram significantemente maiores em relação a um local de referência externo. A ocorrência de NBUDs foi significativamente mais elevada no local NO II. Quanto ao ensaio BMCyt, não houve diferença entre os grupos para MNs e NBUDs. As frequências das alterações nucleares, cariorréxis e cariólise, foram significativamente mais elevadas no local mais afastado da fonte emissora (NOII). A avaliação ambiental associando o Ensaio Salmonella e análise de HPAs mostrou que mesmo amostras dentro dos padrões de controle da qualidade, apresentam potencial genotóxico. Estes resultados permitiram evidenciar que as crianças avaliadas estão expostas a uma mistura de contaminantes de diferentes fontes, sendo a proximidade da indústria petroquímica um contribuinte aos fatores de risco. Medidas são necessárias para identificar e reduzir emissões e efeitos perigosos, uma vez que os padrões de qualidade do ar não são suficientes para garantir a saúde das populações expostas. A complexa composição do MP2,5 pode provocar diversos efeitos genotóxicos, sendo a utilização de diferentes bioensaios fundamental para o entendimento dos efeitos dessa matriz. / Good air quality is essential key to human and environmental health maintenance. Atmospheric particulate matter is an air pollutant consisting of a variety of organic and inorganic substances, some of which presents genotoxic characteristics. The petrochemical industry is an important activity in Rio Grande do Sul state, Brazil, with a history of emissions that are detrimental to air quality. The present study aimed to evaluate the biological effects associated with the dispersion of inhalable particulate matter in an area under the influence of a petrochemical complex, integrating the results of environmental and human biomonitoring through genotoxicity biomarkers. Particulate matter samples were collected in mixed rural, urban and industrial area, using Teflon membrane filters and large air volume samplers, weekly, for periods of 24 hours. Fine inhalable particles (PM2.5) were sampled at two locations - NO and NE - positioned in the first and second preferential wind directions in the study region, 2.5 km away from the main emission source of industrial raw material center. In a third location (NO II), 35 km away from the emission source and in the first wind direction, coarse inhalable particles (MP10) were analyzed. Organic extracts were obtained with dichloromethane from sampled filters and evaluated by the Salmonella/microsome assay, microsuspension method, with the strains TA98, YG1021 and YG1024. The mammalian metabolization fraction (S9) was used to evaluate metabolite mutagenicity. Also, the Comet Assay (EC) and Micronucleus Test in Chinese hamster lung cell line (V79) was used to test the NO and NE samples. The 16 polycyclic aromatic hydrocarbons (PAHs) considered preferential by the US Environmental Agency (USEPA) were evaluated in the organic extracts for characterization of the three sites. Human biomonitoring was carried out in children of public schools from NO and NO II sites, through the collection of peripheral blood and buccal exfoliated cells. To peripheral blood samples, the cytokinesis-block MN cytome assay (CBMN-cyt) was used to evaluate the frequencies of cells with micronuclei (MN) and nuclear abnormalities (nuclear buds - NBUDs, nucleoplasmic bridges - NPBs), and EC evaluating the tail intensity. The buccal MN cytome assay (BMCyt) was used to detect MN and nuclear abnormalities. All PM2.5 extracts showed mutagenicity through the Salmonella assay, highlighting the nitrogenated compounds effects. Genotoxic responses in EC were observed in 87% of the tested extracts and MN induction in 74% of the tests, in addition all the samples showed cytotoxicity to V79 cells. The results of the three biomarkers in vitro and the more toxic PAHs profile presented by the particulate matter, showed a lower air quality of NO site. It is compatible with the greater pollutants dispersion in the first winds direction. Biomonitoring included 54 school children aged 5 to 12 years, with a mean of 8.3 ± 1.82 years. DNA damage evaluated by EC was significantly higher in children from NO compared to NO II. The MN frequency by CBMN-cyt did not differ between groups, however, it was significantly higher in relation to an external reference site. The occurrence of nuclear buds was higher at the NO II site. Regarding the BMCyt assay, there was no difference between groups for MNs and NBUDs. The frequencies of nuclear abnormalities, karyorrhexis and karyolysis were significantly higher at the site farthest from the emission source (NOII). The environmental evaluation with the Salmonella assay and PAH analysis showed genotoxic potential even to samples within the quality standards. These results showed that the schoolchildren are exposed to a mixture of contaminants from different sources, being the proximity of the petrochemical industry a risk factor. Actions are needed to identify and reduce emissions and adverse effects since air quality standards are not enough to ensure the health of exposed populations. The complex PM2.5 composition could cause several genotoxic effects, and the use of different bioassays is fundamental for understanding the effects of this environmental matrix.

Submitted by Fabio Sossai Possebon null (fabio.cid@fmvz.unesp.br) on 2016-02-24T18:24:37Z No. of bitstreams: 1 Dissertação Fábio S Possebon - Salmonella em suínos Final.pdf: 1036715 bytes, checksum: 11bc9e5b832c6262c21bfb938afd4b94 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-02-25T19:42:40Z (GMT) No. of bitstreams: 1 possebon_fs_me_bot.pdf: 1036715 bytes, checksum: 11bc9e5b832c6262c21bfb938afd4b94 (MD5) / Made available in DSpace on 2016-02-25T19:42:40Z (GMT). No. of bitstreams: 1 possebon_fs_me_bot.pdf: 1036715 bytes, checksum: 11bc9e5b832c6262c21bfb938afd4b94 (MD5) Previous issue date: 2016-02-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente trabalho avaliou 250 animais de 25 lotes distintos abatidos em 4 estabelecimentos do estado de São Paulo para a presença de Salmonella nos linfonodos mesentéricos, e caracterizou o perfil de resistência das cepas isoladas aos principais antibióticos. O patógeno estava presente em 36,4% das amostras e 72% dos lotes analisados. Dos 91 isolados, 67 foram sorotipados, e os principais sorovares encontrados foram S. Typhimurium (n=13), S. 1.4,5,12:i:- (n=12) S. Infantis (n=12) e S. Havana (n=11). Os compostos com menor eficácia frente aos isolados foram estreptomicina e tetraciclina (68,1% de resistência) ampicilina e sulfonamidas (62,6%), cloranfenicol (56,0%), trimetoprim-sulfametoxazol (41,8%) e ácido nalidíxico (40,7%). Os mais efetivos foram aztreonam e cefoxitina, (ambos com 3,3% de resistência) e cefepima e ceftriaxona, (ambos com 7,7%). Cepas multidrogas resistentes (MDR) corresponderam a 70,3% dos isolados. Oito cepas foram submetidas ao MLST: quatro S. Typhimurium e uma S. 1.4,5,12:i:- todas pertencentes ao ST 19, duas S. Infantis pertenceram ao ST 32 e uma S. Derby, pertencente ao ST 40. A grande prevalência do patógeno nos animais analisados, com altas taxas de resistência aos antibióticos e pertencentes a grupos genéticos frequentemente associados a surtos e doenças em humanos demonstram que a cadeia produtiva da carne suína é uma fonte de contaminação em potencial nos casos de salmonelose, sendo necessárias medidas preventivas eficazes para o controle do patógeno e diminuição do risco de veiculação de patógenos por alimentos. / This study evaluated 250 animals of 25 different batches processed in four slaughterhouses in São Paulo state - Brazil for the presence of Salmonella in the mesenteric lymph nodes, and characterized the resistance profile for the main antibiotics. The pathogen was present in 36.4% of samples and 72% of the analyzed batches. Of the 91 isolates, 67 were serotyped, and the main serovars found were S. Typhimurium (n = 13), S. 1.4,5,12:i- (n = 12) S. Infantis (n = 12) and S. Havana (n = 11). The compounds with less efficacy were streptomycin and tetracycline (68.1% resistant) ampicillin and sulphonamides (62.6%), chloramphenicol (56.0%), trimethoprim-sulfamethoxazole (41.8%), and nalidixic acid (40.7%). The most effective were cephalothin and aztreonam, (both with 3.3% resistant) and ceftriaxone and cefepime (both with 7.7%). Multidrug-resistant strains (MDR) accounted for 70.3% of the isolates. Eight strains were submitted to MLST: Four S. Typhimurium and one S.1.4,5,12:i:- all belonging to the ST 19, two S. Infantis, belonging to the ST 32 and one S. Derby, belonging to ST 40. The high prevalence of the pathogen in the analyzed animals, with high rates of resistance to antibiotics and belonging to genetic groups that are often associated with outbreaks and disease in humans, shows that the production chain of pork is a potential source of contamination in salmonellosis cases, with the necessity of effective preventive measures for pathogen control and lower the risk of foodborne diseases transmission.

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-02-17Bitstream added on 2014-06-13T19:04:11Z : No. of bitstreams: 1 oliveira_gh_dr_jabo.pdf: 314775 bytes, checksum: 0fd8bccb749e336852eff962616e3d04 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Foi desenvolvido um ensaio imunoenzimático do tipo ELISA indireto para a detecção de resposta sorológica de aves para Salmonella sorotipos Gallinarum, Pullorum, Enteritidis e Typhimurium. Utilizou-se antígeno solúvel obtido por meio de sonicação de cultura de Salmonella Gallinarum (AgSG), Salmonella Enteritidis cepa aflagelar (AgSE) e Salmonella Typhimuirum cepa aflagelar (AgTM), os conjugados peroxidase e fosfatase alcalina e amostras de soros positivos e negativos de vários sorotipos de salmonelas. Os resultados demonstraram que o AgSG pode ser utilizado diluído a 1:25.000 (peroxidase e fosfatase alcalina). Observou-se que o ELISA contendo S. Gallinarum como antígeno e fosfatase alcalina como enzima, propicia a separação de reações positivas para Gallinarum e Pullorum de Enteritidis. O AgSE pode ser utilizado diluído a 1:10.000 (peroxidase) ou 1:5.000 (fosfatase alcalina). Nestas condições, o ELISA/AgSE detectou resposta sorológica para os sorotipos Enteritidis, Gallinarum e Pullorum. O ELISA com o AgTM demonstrou que o antígeno pode ser diluído a 1:20.000 para ambos os conjugados. O ELISA/AgTM demonstrou reatividade entre salmonelas dos grupos B e D. Todas as amostras de soros testes devem ser analisadas diluídas a 1:1.000. Concluindo, o ELISA mostrou-se um teste útil para identificar aves com reação sorológica contra S. Gallinarum, S. Pullorum, S. Enteritidis e S. Typhimurium, podendo ainda identificar aves com sorologia positiva para S. Gallinarum, S. Pullorum sem que haja reação cruzada com amostras de soro de aves vacinadas ou infectada por S. Enteritidis. / This study was done to assess the enzyme-linked immunosorbent assays (ELISA) for detection chicken serologic response against Salmonella enterica sorotypes Gallinarum, Pullorum, Enteritidis and Typhimurium. The test was performed using soluble proteins from Salmonella Gallinarum strain 9 (AgSG), from non-flagellate Salmonella Enteritidis strain (AgSE) and from not flagellate Salmonella Typhimurium (AgTM) strain as detecting antigen and peroxidase and alkaline phosphatase enzymes, as conjugate. According to the results, the antigen has to be diluted at 1:25.000 (AgSG, peroxidase and alkaline phosphatase). In addition, using alkaline phosphatase enzyme, the assay was helpful to separate positive serological reaction to serotypes Gallinarum and Pullorum from Enteritidis. To the ELISA/AgSE, the antigen has to be diluted at 1:10.000 for peroxidase assay and at 1:5.000 for alkaline phosphatase assay. In this condition, the ELISA/AgSE can detect serological reaction to S. Enteritidis, S. Gallinarum and S. Pullorum. To the ELISA/AgTM the antigen has to be diluted at 1:20.000 to both enzymes. In this condition the ELISA/AgTM showed sensibility but was no possible to separate positive serological reaction to serotype concerning at the group B and group D. In all test, the sample of serum has to be diluted at 1:1.000. Therefore, the ELISA was able to identity reactors birds to Salmonella antigens and also to detect serological response to S. Gallinarum, S. Pullorum antigen with no cross-reaction with serum samples taken from birds either challenged or vaccinated against S. Enteritidis.