Efecto de los contenidos de C, Mn y Si en el comportamiento a fluencia en caliente de aceros de construcción al carbono y microaleados: aplicación a la obtención de grano ultrafino en productos largos laminados

Després d’aplicar alguns tractaments d’elaboració i conservació als aliments, queden bacteris lesionats. Aquests bacteris perden la capacitat de créixer en els medis de cultiu selectiu convencionals, de manera que se’n subestima el recompte. Malgrat això, poden recuperar-se als aliments i suposar un risc per la salut, ja que alguns encara poden mantenir activitat metabòlica i integritat estructural. En aquest projecte, es van optimitzar protocols de preparació de mostres per citometria de flux (CF) per avaluar l’estat fisiològic de patògens alimentaris (Escherichia coli O157:H7, Salmonella Enteritidis i Listeria monocytogenes) sotmesos a estrès. Es van estudiar principalment dos paràmetres fisiològics: la integritat de membrana, mitjançant iodur de propidi i fluorocroms de la família SYTO; i l’activitat respiratòria, per la reducció intracel•lular d’una sal de tetrazole, el CTC. En primer lloc, es van avaluar variables de protocol, com la concentració de colorant, la ràtio entre colorants, la solució de tinció i el temps d’incubació, en mostres control (cèl•lules sanes i mortes). A continuació, els protocols optimitzats es van aplicar a suspensions bacterianes en medi de cultiu que prèviament havien estat sotmeses a estressos físics i fisicoquímics. Durant l’etapa final del projecte, els coneixements adquirits sobre la preparació de mostres per CF es van aplicar a l’anàlisi de mostres de matriu complexa: amanides comercials inoculades amb E. coli O157:H7. Als assajos amb indicadors d’integritat de membrana en suspensions bacterianes sotmeses a estrès, es van poder quantificar cèl•lules amb la membrana parcialment danyada (presumptes cèl•lules lesionades). El recompte de cèl•lules que mantingueren l’activitat respiratòria després de ser sotmeses a estrès va ser superior al que es va obtenir mitjançant recompte en placa convencional, cosa que va evidenciar la presència de cèl•lules actives però no cultivables. La introducció d’estratègies per reduir les interferències provocades per les partícules alimentàries i l’ús d’un anticòs amb marcatge fluorescent va permetre detectar selectivament les cèl•lules d’E. coli O157:H7 i avaluar-ne la integritat de membrana simultàniament. L’anàlisi de cèl•lules bacterianes per CF requereix de la exhaustiva optimització dels protocols, que són específics per cada soca i matriu. Malgrat això, i a diferència del mètode convencional per recompte en placa, ofereix la possibilitat d’obtenir una gran quantitat d’informació sobre el sovint complex estat fisiològic d’una mostra.

Studies of genotoxicity and mutagenicity of nitroimidazoles: demystifying this critical relationship with the nitro group

Nitroimidazoles exhibit high microbicidal activity, but mutagenic, genotoxic and cytotoxic properties have been attributed to the presence of the nitro group. However, we synthesised nitroimidazoles with activity against the trypomastigotes of Trypanosoma cruzi, but that were not genotoxic. Herein, nitroimidazoles (11-19) bearing different substituent groups were investigated for their potential induction of genotoxicity (comet assay) and mutagenicity (Salmonella/Microsome assay) and the correlations of these effects with their trypanocidal effect and with megazol were investigated. The compounds were designed to analyse the role played by the position of the nitro group in the imidazole nucleus (C-4 or C-5) and the presence of oxidisable groups at N-1 as an anion receptor group and the role of a methyl group at C-2. Nitroimidazoles bearing NO2 at C-4 and CH3 at C-2 were not genotoxic compared to those bearing NO2 at C-5. However, when there was a CH3 at C-2, the position of the NO2 group had no influence on the genotoxic activity. Fluorinated compounds exhibited higher genotoxicity regardless of the presence of CH3 at C-2 or NO2 at C-4 or C-5. However, in compounds 11 (2-CH3; 4-NO2; N-CH2OHCH2Cl) and 12 (2-CH3; 4-NO2; N-CH2OHCH2F), the fluorine atom had no influence on genotoxicity. This study contributes to the future search for new and safer prototypes and provide.

Understanding the two-component sensing System of virulent bacteria

Report for the scientific sojourn carried out at the l’ Institute for Computational Molecular Science of the Temple University, United States, from 2010 to 2012. Two-component systems (TCS) are used by pathogenic bacteria to sense the environment within a host and activate mechanisms related to virulence and antimicrobial resistance. A prototypical example is the PhoQ/PhoP system, which is the major regulator of virulence in Salmonella. Hence, PhoQ is an attractive target for the design of new antibiotics against foodborne diseases. Inhibition of the PhoQ-mediated bacterial virulence does not result in growth inhibition, presenting less selective pressure for the generation of antibiotic resistance. Moreover, PhoQ is a histidine kinase (HK) and it is absent in animals. Nevertheless, the design of satisfactory HK inhibitors has been proven to be a challenge. To compete with the intracellular ATP concentrations, the affinity of a HK inhibidor must be in the micromolar-nanomolar range, whereas the current lead compounds have at best millimolar affinities. Moreover, the drug selectivity depends on the conformation of a highly variable loop, referred to as the “ATP-lid, which is difficult to study by X-Ray crystallography due to its flexibility. I have investigated the binding of different HK inhibitors to PhoQ. In particular, all-atom molecular dynamics simulations have been combined with enhanced sampling techniques in order to provide structural and dynamic information of the conformation of the ATP-lid. Transient interactions between these drugs and the ATP-lid have been identified and the free energy of the different binding modes has been estimated. The results obtained pinpoint the importance of protein flexibility in the HK-inhibitor binding, and constitute a first step in developing more potent and selective drugs. The computational resources of the hosting institution as well as the experience of the members of the group in drug binding and free energy methods have been crucial to carry out this work.

Hemoglobin enhances the biological activity of synthetic and natural bacterial (endotoxic) virulence factors: a general principle.

Although hemoglobin (Hb) is mainly present in the cytoplasm of erythrocytes (red blood cells), lower concentrations of pure, cell-free Hb are released permanently into the circulation due to an inherent intravascular hemolytic disruption of erythrocytes. Previously it was shown that the interaction of Hb with bacterial endotoxins (lipopolysaccharides, LPS) results in a significant increase of the biological activity of LPS. There is clear evidence that the enhancement of the biological activity of LPS by Hb is connected with a disaggregation of LPS. From these findings one questions whether the property to enhance the biological activity of endotoxin, in most cases proven by the ability to increase the cytokine (tumor-necrosis-factor-alpha, interleukins) production in human mononuclear cells, is restricted to bacterial endotoxin or is a more general principle in nature. To elucidate this question, we investigated the interaction of various synthetic and natural virulence (pathogenicity) factors with hemoglobin of human or sheep origin. In addition to enterobacterial R-type LPS a synthetic bacterial lipopeptide and synthetic phospholipid-like structures mimicking the lipid A portion of LPS were analysed. Furthermore, we also tested endotoxically inactive LPS and lipid A compounds such as those from Chlamydia trachomatis. We found that the observations made for endotoxically active form of LPS can be generalized for the other synthetic and natural virulence factors: In every case, the cytokine-production induced by them is increased by the addition of Hb. This biological property of Hb is connected with its physical property to convert the aggregate structures of the virulence factors into one with cubic symmetry, accompanied with a considerable reduction of the size and number of the original aggregates.

Small RNAs in bacterialcell-cell communication

When all three separate quorum-sensing signals act in concert in Vibrio harveyi, they maximize bioluminescence and fully repress type III secretion. V. harveyi has five qrr loci encoding small RNA regulatory molecules, each consisting of about 100 nucleotides; several of them are involved in repressing bioluminescence. Small RNAs also play roles in population density-dependent activities, including regulation of virulence factors, for bacterial pathogens such as Pseudomonas fluorescens, V. cholerae, Salmonella enterica, Pseudomonas aeruginosa, and Erwinia spp. Although some bacteria appear to carry redundant copies of small RNA genes with which to finely tune expression

Pesquisa de Contaminações Microbiológicas na Carne Bovina na Cidade da Praia

O presente trabalho relata as condições microbiológicas da carne bovina comercializada no talho do mercado do Plateau, bem como as condições higiénicas das superfícies de contacto das carnes. Para o estudo foi utilizado como parâmetros: presença / ausência de Salmonella, contagem de Staphilococcus coagulase positiva e Echerichia Coli β-glucuronidase positiva na carne bovina e contagem de Coliformes totais e Aeróbios mesofilos nas superfícies de contacto das carnes bovinas. Para a realização desse estudo foram seleccionadas três tipos de amostras da carne bovina (alcatra, costela e acém) e três tipos de superfícies de contacto da carne bovina (balanças, facas e mesas). As amostras foram recolhidas em oito semanas decorridas no período de Julho a Setembro de 2011 e analisadas no Laboratório de Controlo de Qualidades da Inpharma – InLab. Os resultados foram classificados de acordo com o Regulamento (CE) Nº 1441/2007 e APHA 2001. Os resultados evidenciam que as condições higiénicas das superfícies de contacto da carne bovina não são das melhores evidenciando números elevados de contagem de Coliformes totais e Aeróbios mesofilos que ultrapassam os limites da APHA. Em relação aos resultados das amostras das carnes bovinas, está muito contaminada evidenciando presenças de Salmonella e números elevados de Staphilococcus coagulase positiva e Echerichia Coli βglucuronidase positiva, pondo em causa a qualidade da carne bovina comercializadas neste estabelecimento público.

Qualidade Microbiológica e Físico – Química de Queijo Fresco de Leite de Cabra produzido em Cabo Verde

A Segurança Alimentar tem assumido uma importância cada vez maior a nível mundial, devido essencialmente ao aumento das trocas internacionais de alimentos e da consciencialização dos consumidores face ao direito de obter alimentos seguros. Tendo em conta este panorama, diversos países encaram a Segurança Alimentar como uma prioridade a nível de gestão governamental, onde se destaca os países em vias de desenvolvimento, como é o caso de Cabo Verde. No domínio do setor de produção de alimentos, têm sido implementadas ações que visam, principalmente, o aumento da qualidade. Em Cabo Verde existe uma grande tradição relacionada com a produção e consumo de queijo, sendo esta atividade possivelmente herdada dos antepassados coloniais. A maioria do queijo comercializado provém de fabricação artesanal e sua quantificação não consta em estatísticas oficiais, embora seja conhecida a existência de unidades de produção caseira e unidades semi-industriais. O presente trabalho, afeto ao curso de Mestrado em Segurança Alimentar da Faculdade de Farmácia – Universidade de Coimbra, teve como objetivo avaliar as condições sanitárias de produção e a qualidade microbiológica e físico-química de queijo fresco de leite de cabra produzidos em Cabo Verde. No total foram analisados 60 amostras de queijo, sob o ponto de vista microbiológico (30 amostras) e sob o ponto de vista físico-químico (30 amostras). Da avaliação sanitária conclui-se que ainda persistem algumas deficiências, como por exemplo, a realização de ordenha de forma inadequada e a produção de queijo sem tratamento térmico do leite. De acordo com os resultados microbiológicos, constataram-se elevadas contagens de microrganismos a 30ºC, na totalidade das amostras, e de E. coli, em 63% das amostras. Não se encontraram amostras positivas à presença de L. monocytogenes e Salmonella spp. As análises físico-químicas apresentaram teores médios, por 100g de produto, de 53% de humidade, 24% de matéria-gorda, 19% de proteína bruta, 3% de cinzas, 1,5% de açúcares (lactose) e 1,4% de sal, conduzindo a teores médios de 69% de humidade no queijo isento de matéria-gorda e de 51% de matéria gorda no resíduo-seco; o pH médio foi de 5,88 e o aw médio situou-se em 0,847. Foram identificadas variações inter-ilhas, a nível dos X parâmetros analisados, embora não muito marcantes, mas suficientes para conduzir a alguma diferenciação em função da proveniência. Recomendou-se o uso contínuo das boas práticas de higiene e fabrico na elaboração dos queijos, com o objetivo de melhorar a qualidade sanitária da produção e dos produtos finais.

Qualitat microbiològica dels productes vegetals frescos mínimament processats i desenvolupament de tecnologies de bioconservació mitjançant bacteris de l'àcid làctic

Food safety today is conditiones by the implementation of new Tecnologies for food preservation based on mild treatments and mínimum process, Novel foods, severe restrictions in the toxicològic profile of chemical preservatives, And the drastic limitation /prohibition of antibiotics, and as a consequence, by the New emergint pathogens or by the increase of classical food-borne pathogens. Biopreservatives appear within this context with strong expectations, because thei are safe microorganisms of ten isolated from foods, chemical compounds of natural origin -antimicrobial peptides and proteins, botanical extracts, enzymes- and sintetic compounds based on natural structures, but less toxic and more eficients, like the amtimicrobial peptides. Among the microbial biopreservatives, lactic acid bacteria have shown great possibilities in the preservation of cured meat products, ready to eat fresh fruit and vegetables, as well as to decrease microbial spoilage in food by products before processing for valorization. Our laboratory has performed an extense survey of the microbiological quality of fresh fruit and vegetables, and of ready-to-eat products, and have detect low levels, but significant of Salmonella sp., E. coli and Listeria spp., including L.monocytogenes, in retail markets and Supermarkets of Catalonia. Due to this reason, we started a project consisting of Developing application of lactic acid bacteria (LAB) obtained from these products, as biopreservatives. LAB were abundant in ready-to-eat fresh fruits and vegetables, specially in germinated seeds. From these products we obtained strains of Leuconostoc, Lactobacillus and Weissella, producing bacteriocins and with a Significant activity of control of L.monocytogenes in fresh apple and cut salad. Ather strains were efective in the inhibition of fungal rot during postharvest caused by Penicillium expansum

S. infantis -tartunnan hävittäminen seosrehua käyttävällä tilalla : tapausselostus

Mervi Yli-Hynnilä

Riskien hallitseminen rehualalla on salmonellatorjunnan perusedellytys

Pirjo Kortesniemi

Nautakarjan parantuminen salmonellasta

Summary: The time required for the elimination of bovine salmonella infection

Ongelmanselvittely pitkittyneellä salmonellatilalla

Summary: Prolonged bovine salmonella infection, what to do?

Valiolaisten meijereiden neuvonta salmonellatiloilla

Summary: Instruction of Valio Group dairies to Salmonella infected farms

Yhteislantanäytteet nautakarjojen salmonellatartuntojen diagnostiikassa

Summary: Comparison of pooled faeces samples with a method of individual faecel samples for salmonella investigation in cattle

Salmonellan esiintyvyys LSO Foods Oy:lle välitysvasikoita myyvillä tiloilla

Summary: Prevalence of salmonella in dairy herds producing feeder calves for LSO Foods Ltd