Development of molecular-based techniques for the detection, identification and quantification of food-borne pathogens

La presencia de microorganismos patógenos en alimentos es uno de los problemas esenciales en salud pública, y las enfermedades producidas por los mismos es una de las causas más importantes de enfermedad. Por tanto, la aplicación de controles microbiológicos dentro de los programas de aseguramiento de la calidad es una premisa para minimizar el riesgo de infección de los consumidores. Los métodos microbiológicos clásicos requieren, en general, el uso de pre-enriquecimientos no-selectivos, enriquecimientos selectivos, aislamiento en medios selectivos y la confirmación posterior usando pruebas basadas en la morfología, bioquímica y serología propias de cada uno de los microorganismos objeto de estudio. Por lo tanto, estos métodos son laboriosos, requieren un largo proceso para obtener resultados definitivos y, además, no siempre pueden realizarse. Para solucionar estos inconvenientes se han desarrollado diversas metodologías alternativas para la detección identificación y cuantificación de microorganismos patógenos de origen alimentario, entre las que destacan los métodos inmunológicos y moleculares. En esta última categoría, la técnica basada en la reacción en cadena de la polimerasa (PCR) se ha convertido en la técnica diagnóstica más popular en microbiología, y recientemente, la introducción de una mejora de ésta, la PCR a tiempo real, ha producido una segunda revolución en la metodología diagnóstica molecular, como pude observarse por el número creciente de publicaciones científicas y la aparición continua de nuevos kits comerciales. La PCR a tiempo real es una técnica altamente sensible -detección de hasta una molécula- que permite la cuantificación exacta de secuencias de ADN específicas de microorganismos patógenos de origen alimentario. Además, otras ventajas que favorecen su implantación potencial en laboratorios de análisis de alimentos son su rapidez, sencillez y el formato en tubo cerrado que puede evitar contaminaciones post-PCR y favorece la automatización y un alto rendimiento. En este trabajo se han desarrollado técnicas moleculares (PCR y NASBA) sensibles y fiables para la detección, identificación y cuantificación de bacterias patogénicas de origen alimentario (Listeria spp., Mycobacterium avium subsp. paratuberculosis y Salmonella spp.). En concreto, se han diseñado y optimizado métodos basados en la técnica de PCR a tiempo real para cada uno de estos agentes: L. monocytogenes, L. innocua, Listeria spp. M. avium subsp. paratuberculosis, y también se ha optimizado y evaluado en diferentes centros un método previamente desarrollado para Salmonella spp. Además, se ha diseñado y optimizado un método basado en la técnica NASBA para la detección específica de M. avium subsp. paratuberculosis. También se evaluó la aplicación potencial de la técnica NASBA para la detección específica de formas viables de este microorganismo. Todos los métodos presentaron una especificidad del 100 % con una sensibilidad adecuada para su aplicación potencial a muestras reales de alimentos. Además, se han desarrollado y evaluado procedimientos de preparación de las muestras en productos cárnicos, productos pesqueros, leche y agua. De esta manera se han desarrollado métodos basados en la PCR a tiempo real totalmente específicos y altamente sensibles para la determinación cuantitativa de L. monocytogenes en productos cárnicos y en salmón y productos derivados como el salmón ahumado y de M. avium subsp. paratuberculosis en muestras de agua y leche. Además este último método ha sido también aplicado para evaluar la presencia de este microorganismo en el intestino de pacientes con la enfermedad de Crohn's, a partir de biopsias obtenidas de colonoscopia de voluntarios afectados. En conclusión, este estudio presenta ensayos moleculares selectivos y sensibles para la detección de patógenos en alimentos (Listeria spp., Mycobacterium avium subsp. paratuberculosis) y para una rápida e inambigua identificación de Salmonella spp. La exactitud relativa de los ensayos ha sido excelente, si se comparan con los métodos microbiológicos de referencia y pueden serusados para la cuantificación de tanto ADN genómico como de suspensiones celulares. Por otro lado, la combinación con tratamientos de preamplificación ha resultado ser de gran eficiencia para el análisis de las bacterias objeto de estudio. Por tanto, pueden constituir una estrategia útil para la detección rápida y sensible de patógenos en alimentos y deberían ser una herramienta adicional al rango de herramientas diagnósticas disponibles para el estudio de patógenos de origen alimentario.

Eye tracking data representation and visualization : on information and communication studies at CETAC.MEDIA

Eye tracking has become a preponderant technique in the evaluation of user interaction and behaviour with study objects in defined contexts. Common eye tracking related data representation techniques offer valuable input regarding user interaction and eye gaze behaviour, namely through fixations and saccades measurement. However, these and other techniques may be insufficient for the representation of acquired data in specific studies, namely because of the complexity of the study object being analysed. This paper intends to contribute with a summary of data representation and information visualization techniques used in data analysis within different contexts (advertising, websites, television news and video games). Additionally, several methodological approaches are presented in this paper, which resulted from several studies developed and under development at CETAC.MEDIA - Communication Sciences and Technologies Research Centre. In the studies described, traditional data representation techniques were insufficient. As a result, new approaches were necessary and therefore, new forms of representing data, based on common techniques were developed with the objective of improving communication and information strategies. In each of these studies, a brief summary of the contribution to their respective area will be presented, as well as the data representation techniques used and some of the acquired results.

Can genetically-modified cotton contribute to sustainable development in Africa?

Genetically modified (GM) crops and sustainable development remain the foci of much media attention, especially given current concerns about a global food crisis. However, whilst the latter is embraced with enthusiasm by almost all groups, GM crops generate very mixed views. Some countries have welcomed GM, but others, notably those in Europe, adopt a cautious stance. This article aims to review the contribution that GM crops can make to agricultural sustainability in the developing world. Following brief reviews of both issues and their linkages, notably the pros and cons of GM cotton as a contributory factor in sustainability, a number of case studies from resourcepoor cotton farmers in Makhathini Flats, South Africa, is presented for a six-year period. Data on expenditure, productivity and income indicate that Bacillus thuringiensis (Bt) cotton is advantageous because it reduces costs, for example, of pesticides, and increases income, and the indications are that those benefits continued over at least the six years covered by the studies. There are repercussions of the additional income in the households; debts are reduced and money is invested in children's education and in the farms. However, in the general GM debate, the results show that GM crops are not miracle products which alleviate poverty at a stroke, but nor is there evidence that they will cause the scale of environmental damage associated with indiscriminate pesticide use. Indeed, for some GM antagonists, perhaps even the majority, such debates are irrelevant – the transfer of genes between species is unnatural and unethical. For them, GM crops will never be acceptable despite the evidence and pressure to increase world food production.

The development and impact of the Livestock Guru: meeting the knowledge needs of poor livestock keepers in Tamil Nadu, India

Livestock keepers comprise 2/3rds of the 2.8 billion households living on less than two dollars per day. However, as a group they tend to be marginalised and excluded from formal service provision, particularly in relation to animal health. Therefore, the following paper describes the development of the Livestock Guru, a multi-media learning programme created to meet the knowledge needs of poor livestock keepers in Tamil Nadu, India. The findings from the study illustrate the importance of both appropriate visuals, voice-overs but also the need for addressing issues in the environment in which learning will take place.

Planning for development or decline? Education policy for Chinese language in Singapore

This article examines how political discourse, language ideologies, recent Chinese curriculum reforms, and their representations in the media are inextricably related. Using the Speak Mandarin Campaign as background for the inquiry, I focus on textual features of the various media sources, TV advertisements, campaign slogans, official speeches, and newspaper excerpts to illuminate the status and changing role of the Chinese language in Singapore’s sociocultural, economic, and political development. Using critical discourse analysis as an analytical framework, I examine the contradictory ideologies that underpin the government’s language policies and planning activities. On the one hand, the government emphasizes the cultural and economic values of the Chinese language; on the other hand, government schools teach Chinese as a subject. In particular, the recent reforms in Chinese language curriculum have arguably further diluted the content of teaching. In addition I point out how conflicting ideologies behind language policies can lead to cultural confusion and educational uncertainty. These mixed messages make it difficult for schools to offer a consistent language education curriculum that will help students appreciate the value, be it economic, cultural or educational, of the Chinese language.

Culex pipiens development is greatly influenced by native bacteria and exogenous yeast

Culex pipiens is the most cosmopolitan mosquito of the Pipiens Assemblage. By studying the nature of interactions between this species and microorganisms common to its breeding environment we can unravel important pitfalls encountered during development. We tested the survival rate of larval stages, pupae and adults of a Cx. pipiens colony exposed to a variety of microorganisms in laboratory conditions and assessed the transmission to offspring (F1) by those organisms that secured development up to adulthood. Three complementary experiments were designed to: 1) explore the nutritional value of yeasts and other microorganisms during Cx. pipiens development; 2) elucidate the transstadial transmission of yeast to the host offspring; and 3) to examine the relevance of all these microorganisms in female choice for oviposition-substratum. The yeast Saccharomyces cerevisiae proved to be the most nutritional diet, but despite showing the highest survival rates, vertical transmission to F1 was never confirmed. In addition, during the oviposition trials, none of the gravid females was attracted to the yeast substratum. Notably, the two native bacterial strains, Klebsiella sp. and Aeromonas sp., were the preferred oviposition media, the same two bacteria that managed to feed neonates until molting into 2nd instar larvae. Our results not only suggest that Klebsiella sp. or Aeromonas sp. serve as attractants for oviposition habitat selection, but also nurture the most fragile instar, L1, to assure molting into a more resilient stage, L2, while yeast proves to be the most supportive diet for completing development. These experiments unearthed survival traits that might be considered in the future development of strategies of Cx. pipiens control. These studies can be extended to other members of the Pipiens Assemblage

Characterization of the products of aniline peroxydisulfate oligo/polymerization in media with different pH by resonance Raman spectroscopy at 413.1 and 1064 nm excitation wavelengths

The pH-structure correlation of the products of aniline peroxydisulfate reaction was mainly investigated by resonance Raman spectroscopy. The reactions of aniline and ammonium peroxydisulfate were carried out in aqueous solutions of initial pH ranging from 4.9 to 13.2 and monomer/oxidant molar ratio of 4/1. For an initial pH of 4.9, the spectroscopic techniques showed that the emeraldine salt form of polyaniline (PANI-ES) is the main product, corroborating that the usual head-to-tail coupling mechanism is taking place. The resonance Raman spectra at 1064 nm exciting wavelength were useful to detect the emeraldine salt as a minor product for reactions at an initial pH of 5.3-11.5. The Raman spectra of the main product of the reaction at initial pH of 13.2 excited at 1064 and 413.1 nm showed new spectral features consistent with 1,4-Michael-type adducts of aniline monomers and 1,4-benzoquinone-monoimine unit. These compounds and their products of hydrolysis/oxidation are the predominant species for the reaction media of initial pH from 5.3 to 13.2. In order to get PANI with different nanoscale morphologies, a pH value of more than 0 or 1 was used in the aniline polymerization. The spectroscopic data obtained in this work reveal that head-to-tail coupling does not occur when aniline reacts at media pH higher than about 5. It is suggested that chemical structures of the products of aniline oxidation by an unusual mechanism are the driving force for the development of assorted morphologies. Copyright (C) 2011 John Wiley & Sons, Ltd.

Development of a spectrophotometric Sequential Injection Analysis (SIA) procedure for determination of ammonium: A Response Surface Methodology (RSM) approach

This paper describes the optimization and use of a Sequential Injection Analysis (SIA) procedure for ammonium determination in waters. Response Surface Methodology (RSM) was used as a tool for optimization of a procedure based on the modified Berthelot reaction. The SIA system was designed to (i) prepare the reaction media by injecting an air-segmented zone containing the reagents in a mixing chamber, (ii) to aspirate the mixture back to the holding coil after homogenization, (iii) drive it to a thermostated reaction coil, where the flow is stopped for a previously established time, and (iv) to pump the mixture toward the detector flow cell for the spectrophotometric measurements. Using a 100 mu mol L(-1) ammonium solution, the following factors were considered for optimization: reaction temperature (25 - 45 degrees C), reaction time (30 - 90 s), hypochlorite concentration (20 - 40 mmol L(-1)) nitroprusside concentration (10 - 40 mmol L(-1)) and salicylate concentration (0.1 - 0.3 mol L(-1)). The proposed system fed the statistical program with absorbance data for fast construction of response surface plots. After optimization of the method, figures of merit were evaluated, as well as the ammonium concentration in some water samples. No evidence of statistical difference was observed in the results obtained by the proposed method in comparison to those obtained by a reference method based on the phenol reaction. (C) 2010 Elsevier B.V. All rights reserved.

Historical Consciousness, Historical Media, and History Education

This thesis by publication contains an introductory summary chapter and three papers. The first paper presents a study of how the concept of historical consciousness has been defined, applied, and justified in Swedish history didactical research. It finds that there is consensus regarding the definition of what a historical consciousness is, but that there is variation in how the concept is applied. It is suggested that this variation makes historical con­sciousness a complex and vague concept.      The second paper uses the results presented in the first paper as a point of departure and from thence argues for a broadened understanding of the concept of historical consciousness that incorporates its definition, applica­tion, development, and significance. The study includes research about his­torical consciousness primarily from Sweden, the UK, the USA and Canada. The paper presents a typology of historical consciousness and argues that level of contextualisation is what distinguishes different types of historical consciousnesses and that an ability to contextualise is also what makes his­torical consciousness an important concept for identity constitution and morality.      The third paper proposes a methodological framework of historical con­sciousness based on the theory of historical consciosusness presented in the second paper. It presents arguments for why the framework of historical consciousness proposed can be useful for the analysis of historical media and it discusses how aspects of the framework can be applied in analysis. It then presents a textbook analysis that has been performed according to the stipulated framework and discusses its results regarding how textbooks can be used to analyse historical consciousness and its development.

Media piracy in emerging economies

Media Piracy in Emerging Economies is the first independent, large-scale study of music, film and software piracy in the developing world, with a focus on Brazil, India, Russia, South Africa, Mexico and Bolivia. Based on three years of work by some thirty-five researchers, the study tells two overarching stories: one tracing the explosive growth of piracy as digital technologies became cheap and ubiquitous around the world, and another following the growth of industry lobbies that have reshaped laws and law enforcement around copyright protection. The report argues that enforcement efforts have largely failed, and that the problem of piracy is better addressed as a failure of affordable access to media in legal markets.

Practices in social media in the travel education business

Praticas na midia social se refere a vista ou a percepcao de uma marca nao so com base em suas ofertas e servicos, mas tambem sobre seus valores e cultura como percebido principalmente pelos consumidores. As organizacoes podem utilizer a imagem que é interessante e aceitavel para os clients. Nos ultimos anos, uma das formas que as organizacoes de forma consistente comercializar ou publicitar a sua marca envolve midias sociais. O ojetivo deste estudo é explorer quais sao as praticas de midia social no negocio de educacao de viagens. Para obter os dados, a pesquisadora entrevistou pessoas de seis empresas de educacao de viagens e cotejadas informacoes sobre o tema a partir de fonts primarias e secundarias existentes. O pesquisador conduziu a pesquisa para determiner praticas de marketing de midia social em que se aplicam nos sectores do turismo e educacao. Com base nos dados obtidos pelo pesquisador, as praticas de marketing de midia social incluem a utilizacao de plataformas de midia social com um alcance internacional generalizada tipo Facebook, Twitter, Instagram e YouTube. Outra estratégia é para postar constantemente as atualizações que não são apenas interessante, mas também informativo sobre os produtos e serviços oferecidos por uma marca ou organização. As empresas ou organizações devem também interagir com os clientes ou clientes on-line, a fim de manter os interesses deste último em ofertas e serviços da ex. O envolvimento do cliente é uma das razões por que os clientes optam por seguir as empresas on-line através da mídia social. A pesquisa também revela outras vantagens e benefícios da mídia social que constituem as melhores práticas, tais como a conversão de não-clientes para os clientes, o aumento da presença da mídia para aumentar a popularidade, a comunicação eficaz das metas e objetivos organizacionais, bem como a formação de um bom relacionamento com os clientes. As organizações também podem usar turístico criado conteúdo (TCC) e outros tipos de conteúdo para orientar a tomada de decisão no desenvolvimento do produto gerado pelo usuário.

Social media crises in the organization: exploring management strategies through cases from France and Brazil

The aim of this Master’s thesis has been to shed light on the response strategies that organizations are implementing when facing a crisis created on or amplified by social media. Since the development of social media in the late 1990s, the interplay between the online and the offline spheres has become more complex, and characterized by dynamics of a new magnitude, as exemplified by the wave of “Twitter” Revolutions or the Wikileaks scandal in the mid 2000s, where online behaviors deeply affected an offline reality. The corporate world does not escape to this worldwide phenomenon, and there are more and more examples of organizational reputations destroyed by social media “fireballs”. As such, this research aims to investigate, through the analysis of six recent cases of corporate crises (2013-2015) from France and Brazil, different strategies currently in use in order to identify examples of good and bad practices for companies to adopt or avoid when facing a social media crisis. The first part of this research is dedicated to a review of the literature on crisis management and social media. From that review, we were able to design a matrix model, the Social Media Crisis Management Matrix, with which we analyzed the response strategies of the six companies we selected. This model allows the conceptualization of social media crises in a multidimensional matrix built to allow the choice, according to four parameters, of the most efficient (that is: which will limit the reputational damage) response strategy. Attribution of responsibility for the crisis to the company by stakeholders, the origin of the crisis (internal or external), the degree of reputational threat, and the emotions conveyed online by stakeholders help companies determining whether to adopt a defensive response, or an accommodative response. The results of the analysis suggest that social media crises are rather manichean objects for they are, unlike their traditional offline counterparts, characterized by emotional involvement and irrationality, and cannot be dealt with traditionally. Thus analyzing the emotions of stakeholders proved to be, in these cases, an accurate thermometer of the seriousness of the crisis, and as such, a better rudder to follow when selecting a response strategy. Consequently, in the cases, companies minimized their reputational damage when responding to their stakeholders in an accommodative way, regardless of the “objective” situation, which might be a change of paradigm in crisis management.

Larval development of Notolopas brasiliensis Miers, 1886 (Brachyura: Majoidea: Pisidae) described from laboratory reared material and a reappraisal of the characters of Pisidae

O completo desenvolvimento larval de Notolopas brasiliensis é descrito, a partir de material criado em laboratório, com ênfase na morfologia externa de Majoidea e comparado aos demais gêneros de Pisidae. O desenvolvimento larval de N. brasiliensis consiste em dois estágios de zoea e um de megalopa. A duração media de cada estágio foi de 4.2 ± 1.0 dias para a Zoea I e 3.8 ± 0.7 dias para a Zoea II, a megalopa aparece entre 8.1 ± 0.4 dias após a eclosão. Os caracteres previamente utilizados para definir as formas larvais de Pisidae ou são simplesiomórficos ou altamente homoplásticos. Foi observado que não existe um conjunto de caracteres capazes de definir Pisidae até o presente.Contudo foi mostrado que uma combinação de caracteres pode ser utilizada para diferenciar Notolopas dos demais gêneros da família.