Microrganismos geneticamente modificados (OGMs), obtidos através das técnicas de biologia molecular são amplamente utilizados para as mais diversas finalidades, atingindo o ambiente, solo, águas, animais e até mesmo o Homem. Contudo, o comportamento e o destino destes microrganismos no ambiente, depois que sua função tenha sido completada, e onde estes não podem ser controlados, tem sido objeto de preocupação. Para diminuir os riscos potenciais associados à liberação (acidental ou intencional) dos OGMs no ambiente pesquisadores desenvolveram sistemas de contenção de microrganismos geneticamente engenheirados, baseados na clonagem de genes que codificam proteínas tóxicas, sob regulação de promotores indutíveis (para revisão ver Molin et aI., 1993). Estes sistemas permitem o controle da morte celular em tempo pré-determinado, com a indução do promotor que regula o gene que codifica a proteína tóxica. Contudo, embora as leveduras sejam tão importantes e úteis quanto as bactérias, não existem sistemas suicidas usados para contenção destes microrganismos. No presente trabalho, construímos um sistema suicida para a levedura Saccharomyces cerevisiae o qual apresenta o gene da nuclease de Serratia marcescens clonado sob controle do promotor ADH2GAPDH, que é reprimido por glicose. Desde que a nuclease é capaz de destruir DNA e RNA, tal sistema não permitirá que o material genético do OGM em questão, seja transferido para outros microrganismos, após a morte celular. A atividade da nuclease foi detectada in vivo, através da diminuição da viabilidade dos transformantes de levedura, após o consumo da glicose. Quando leveduras mutantes rad52 foram usadas como células hospedeiras, o efeito letal do plasmídeo foi dramaticamente aumentado. Por outro lado, a atividade da nuclease produzida nos tranformantes de levedura foi analisada in vitro, através da incubação de extratos celulares com DNA plasmidiano mostrando a completa degradação deste em gel de agarose. Além disso, em ensaio de microcosmos, a viabilidade das leveduras portadoras do plasmídeo suicida no ambiente diminuiu drasticamente quando comparada com células controles. Estes resultados mostram que a nuclease de Serratía marcescens é expressa de forma ativa em S. cerevísíae sendo capaz de evitar a permanência dos transformantes de leveduras no ambiente, em laboratório, ou em condições simulando o ambiente, sem a necessidade de um indutor adicional. / Genetic Engineered Microorganisms (GEMs), obtained by molecular biology techniques, are becoming of widespread use for different purposes thus reaching the environment, including soil, water, plants, animais and Man. However, the behavior and fate of these microorganisms after their function has been completed in environments where they cannot be physically contained is a matter of concern. To minimize the potential risks associated with the release (deliberate or accidental) of GEMs in the environment, researchers have developed biological containment systems for bacteria which are based on the cloning of genes coding for toxic proteins, under the regulation of regulatable promoters (for review, see Molin et al., 1993). Such suicide systems allow the predetermined control of cell death from the moment the promoter of the killer gene is switched on. However, even if yeasts are as useful and important in biotechnological processes as bacteria, there is no suicide system described so far for the containment of these microorganisms. In the present work, we constructed a suicide system for the yeast Saccharomyces cerevisiae, consisting of the Serratia marcescens nuclease gene, cloned under the control of the yeast AOH2GAPOH promoter, which is repressed by glucose. Since this nuclease is able to destroy DNA and RNA, such suicide system does not allow the genetic material of the GEM to be transferred to other microorganisms upon cell death. The nuclease activity in S. cerevisiae was detected in vivo, through the decrease in cell viability of the yeast transformants, upon glucose consumption. When yeast rad52 mutants were used as the host cells, the lethal effect of the suicide plasmid was dramatically increased. On the other hand, the activity of the nuclease produced in yeast transformants was analyzed in vitro, by incubation of cell-free extracts with plasmid DNA, showing complete degradation on agarose gels. Furthermore, in microcosmos assays, the viability of the yeast cells carrying the suicide plasmid decreased drastically when compared to control cells. These results showed that the S. marcescens nuclease is expressed in an active form in S. cerevisiae, being able to avoid the maintenance of the yeast transformants, either in the laboratory, or in conditions simulating the environment, without the need of adding any inducer.

8 pp. / By making several simple horse care changes, you can significantly decrease your horse’s risk of exposure to disease. Using the tips below, you may prevent your horse from being exposed to sick horses while away at a show. If your horse were to bring a virus home, proper Biosecurity practices could result in just one sick horse, as opposed to a whole barn full. The intent of this article is to inject a little humor while providing key common sense tips on Biosecurity. Each tip is accompanied by an easy to remember theme and cartoon. These changes seem small but they can make a big difference in protecting the health of your horse. .

O aumento de casos de doenças infecto-contagiosas, como hepatite e AIDS, despertou para a necessidade de elaboração de regras mais rígidas para controle de infecções em ambiente odontológico. As práticas de esterilização e desinfecção, entre as medidas de biossegurança, tornaram-se imprescindíveis. Em virtude disto, o objetivo desse estudo foi verificar a qualidade desse processo em 31 consultórios odontológicos da rede pública do município de Porto Velho (Rondônia, BR). Para isso os cirurgiões-dentistas ou auxiliares foram questionados sobre os procedimentos relacionados ao processo de esterilização. Também foi realizado o monitoramento biológico dos equipamentos, utilizando esporos de Bacillus subtilis para a estufa e Geobacillus stearothermophilus para a autoclave. Em 31 dos consultórios investigados foram detectados problemas nos processos de esterilização, como: utilização das relações tempo X temperatura incorretos (100% para autoclaves, 70,4% para estufas), falta de termômetros nos equipamentos (59,3%), entre outros. Embora fossem pouco utilizadas, todas as autoclaves apresentaram resultado negativo na avaliação do monitoramento biológico. Já no monitoramento das estufas, 20% apresentaram resultados positivos, indicando falha no processo de esterilização. Portanto os resultados apontam para existência de problemas na eficácia e nas condições dos processos de esterilização realizados na rede pública do município de Porto Velho, o que coloca em risco a saúde dos pacientes atendidos nessas unidades. / The increase in the number of cases of infectious-contagious diseases, like hepatitis and HIV, has brought to attention the need to elaborate stricter infection controls in the dental environment. The practices of sterilization and disinfection, among the measures of bio-security, became indispensable. For this reason, the objective of this study was to verify the quality of these procedures in 31 dental offices of the public system of the municipality of Porto Velho (Rondônia, BR). For this survey, dentists or their assistants have been questioned on the subject of the procedures related to the sterilization. Biological monitoring of the equipment was also carried out by using Bacillus subtilis for the oven and Geobacillus stearothermophilus for the autoclave. Failures in sterilization were detected in 31 investigated offices, such as: the use of incorrect ratio between time X temperature (100% for autoclaves, 70.4% for ovens), the absence of thermometers on equipment (59.3%), among other factors. Even though infrequently used, all the autoclaves showed negative results in the biological monitoring. In the monitoring of the ovens, 20% showed positive results, indicating failure in the sterilization process. Therefore, the results point to the existence of problems in the effectiveness and in the conditions of the sterilization procedures of the public system of the municipality of Porto Velho, endangering the health of the patients treated in those units.

Objetivo: Comparar el nivel de conocimiento y aplicación de medidas de bioseguridad en estudiantes de pregrado y egresados de la carrera de odontología del periodo 2013-2018 de una universidad privada peruana. Materiales y Métodos: Estudio piloto descriptivo de tipo transversal observacional, siendo la población los estudiantes de pregrado y egresados de la carrera de odontología de una universidad privada peruana. Se tomó una muestra de 32 estudiantes de pregrado y 32 egresados de odontología de la misma institución, evaluándose el nivel de conocimiento mediante una encuesta autoaplicable y el nivel de aplicación de las medidas de bioseguridad por medio de una ficha observacional. El análisis univariado se obtuvo mediante frecuencias absolutas y relativas; para el análisis bivariado se utilizó la prueba de Chi-cuadrado y prueba exacta de Fisher. (p≤0.05) Resultados: El conocimiento sobre medidas de bioseguridad de los alumnos de pregrado y egresados es de nivel regular, con valores de 40.6% y 46.8% respectivamente, el 34.37% de alumnos de pregrado y el 25.56% del grupo de egresados obtuvieron un buen nivel de aplicación de las medidas de bioseguridad. Se encontró diferencias estadísticamente significativas entre el nivel de conocimiento de los alumnos de pregrado y egresados. (p= 0.001). Conclusión: Los grupos de pregrado y egresados en referencia al conocimiento en medidas de bioseguridad, obtuvieron el nivel de regular, y lo que respecta a la aplicación de medidas de bioseguridad, ambos grupos lograron el nivel de bueno. / Objective: Compare the level of knowledge and application of biosafety measures in undergraduate and graduate students of the 2013-2018 dental career of a Peruvian private university. Materials and Methods: A descriptive, observational cross-sectional pilot study, with the population being undergraduate and graduate students of the dental career of a Peruvian private university. A sample of 32 undergraduate students and 32 graduates of dentistry from the same institution were taken, assessing the level of knowledge through a self-applying survey and the level of application of biosecurity measures through an observational record. Univariate analysis was obtained by absolute and relative frequencies; The Chi-square test and Fisher's exact test were used for the bivariate analysis. (p≤0.05) Results: The knowledge on biosecurity measures of undergraduate and graduate students is of a regular level, with values ​​of 40.6% and 46.8% respectively, 34.37% of undergraduate students and 25.56% of the group of graduates obtained a good level of application of biosecurity measures. Statistically significant differences were found between the level of knowledge of undergraduate and graduate students. (p = 0.001). Conclusion: Undergraduate and graduate groups in reference to knowledge in biosecurity measures, obtained the level of regular, and with regard to the application of biosecurity measures, both groups achieved the level of good. / Tesis

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CHDS State/Local / Approved for public release; distribution is unlimited / The concept of biosecurity as it pertains to Biological Select Agents and Toxins in American biomedical research institutions is explored in some depth. Posing the research question "How can specific public biomedical research universities securely use and store biological select agents?" the thesis outlines the dynamics of the select agent and toxin list, the relevant history of the control of biological agents both in the international and domestic settings, including federal regulations pertaining to biosecurity (42CFR73). Two specific case studies are presented in the thesis. The biosecurity strategies and tactics at these two distinct biomedical research are compared. An answer to the research question is proposed and additional areas for research are outlined. / Captain, the University of Texas at Houston Police Department

Doctor of Philosophy / Department of Diagnostic Medicine and Pathobiology / Michael W. Sanderson / The last outbreak for Foot and Mouth Disease (FMD) in the United States (U.S.) was in 1929. Since that time the U.S. has not had any exposure to the disease or vaccination, creating a very susceptible livestock population. The central U.S. has a large susceptible livestock population including cattle, swine, sheep, and goats. The impact of FMD in the U.S. would be devastating. Simulation modeling is the only avenue available to study the potential impacts of an introduction in the U.S. Simulation models are dependent on accurate estimates of the frequency and distance distribution of contacts between livestock operations to provide valid model results for planning and decision making including the relative importance of different control strategies. Due to limited data on livestock movement rates and distance distribution for contacts a survey was conducted of livestock producers in Colorado and Kansas. These data fill a need for region specific contact rates to provide parameters for modeling a foreign animal disease. FMD outbreaks often require quarantine, depopulation and disposal of whole herds in order to prevent the continued spread of the disease. Experts were included in a Delphi survey and round table discussion to critically evaluate the feasibility of depopulating a large feedlot. No clearly acceptable method of rapidly depopulating a large feedlot was identified. Participants agreed that regardless of the method used for depopulation of cattle in a large feedlot, it would be very difficult to complete the task quickly, humanely, and be able to dispose of the carcasses in a timely fashion. Simulation models were developed to assess the impact of livestock herd types and vaccination on FMD outbreaks in the central U.S. using the North American Animal Disease Spread Model (NAADSM), a spatially explicit, stochastic state-transition simulation model. Simulation scenarios with large vaccination zones had decreased outbreak length and number of herds destroyed. Vaccination did not provide additional benefit to control compared to depopulation alone when biosecurity and movement controls were high, however the ability to achieve high levels of biosecurity and movement control may be limited by labor and animal welfare concerns.

É reconhecida a importância da avicultura para o agronegócio e para o desenvolvimento do Brasil. Mas o País tem enfrentado grande concorrência internacional, que se traduz em barreiras sanitárias e exigências cada vez maiores de controle de seu rebanho por parte dos importadores. Neste sentido, o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) publicou uma série de atos legais para viabilizar a organização dos programas de saúde animal, entre eles as Instruções Normativas n° 56/2007, n° 59/2009, n°36/2012 e n°10/2013, para estabelecer os procedimentos para registro e fiscalização de estabelecimentos avícolas comerciais e de reprodução, voltados à biosseguridade do sistema avícola. Como todos os setores produtivos, a avicultura é fortemente influenciada pela sua estrutura de custos, de modo que há um certo sentimento de que a adequação às medidas de biosseguridade, preconizadas pelas normativas, pode impactar a atividade de produção de ovos comerciais de modo a levar, eventualmente, alguns avicultores ao abandono da mesma. O objetivo deste trabalho foi analisar o impacto socioeconômico das políticas sanitárias para estabelecimentos avícolas comerciais de postura da regional agropecuária de Limeira, Estado de São Paulo, analisando o perfil socioeconômico da propriedade e do produtor e, estimando o custo de produção e seu impacto em atendimento às medidas de biosseguridade preconizadas pelas normativas. Para a análise do perfil das propriedades foram utilizados os dados dos 28 estabelecimentos avícolas, levantados por meio de documentação para registro. Para a análise do perfil do produtor e para estimar os custos de produção foram considerados dez estabelecimentos, voluntários à pesquisa, visitados entre os meses de junho e julho de 2013. O resultado do estudo sugere que as adequações à biosseguridade podem ser factíveis de serem realizadas economicamente, sendo o custo relativamente pequeno frente aos possíveis riscos de enfermidades, representando entre 1,61% e 2,09% do custo total de produção. No entanto, as sucessivas alterações nas legislações podem fazer com que o programa de sanidade avícola perca a credibilidade diante da sociedade e dos produtores que demonstram resistência às mudanças nos paradigmas zoosanitários / The importance of the poultry industry for agribusiness and development is recognized in Brazil, but the country has faced strong international competition, translated into sanitary barriers and increasing requests for the control of Brazilian flock by importers. In this regard, the Ministry of Agriculture, Livestock and Supply (MAPA) has published a series of legal acts to facilitate the organization of animal health programs, including Regulatory Instructions n°56/2007, n°59/2009, n°36/2012 & n°10/2013, to establish procedures for the registration and inspection of boilers, layer hens and commercial poultry breeding, focused on biosecurity systems. Like all productive sectors, the poultry one is strongly influenced by its cost structure, thus some people believe that the adequacy of biosecurity measures could affect the activity of commercial egg production and, eventually, lead some producers to the abandonment of the activity. The aim of this study was to analyze the socioeconomic impact of health policies on farm eggs of Limeira regional, Sao Paulo State, by analyzing the socioeconomic profile of the property and the producer, estimating the cost of production and its impact on compliance with biosecurity measures recommended by the Regulatory Instructions. For the analysis of the profile of farms we used data of twenty-eight poultry establishments, raised through documentation register. For the analysis of the profile of the producer and to estimate the costs of production, data was raised in ten farm eggs, volunteers of this research, which were visited between June and July 2013. The result of the study suggests that adaptations to biosecurity are economically feasible and costs are relatively small compared to the potential risks of disease, representing between 1.61% and 2.09% of the total production cost, thus not justifying the abandonment of activity. Nevertheless, successive changes in legislation may lead poultry health programs to lose credibility in society and with producers who demonstrate resistance to changes in zoo-sanitary paradigms

A presente dissertação tem por objetivo demonstrar que a maneira como vem ocorrendo o comércio transfronteiriço de alimentos geneticamente modificados gera dúvidas sobre a aptidão da legislação internacional de regulamentá-lo. Os alimentos transgênicos são resultado do avanço da biotecnologia moderna e interferem diretamente na saúde, na alimentação, na economia, nas relações de consumo e no equilíbrio ambiental. Assim, é direito indispensável de todo cidadão o poder optar e decidir a respeito do alimento que irá ingerir. Embora a biotecnologia moderna tenha trazido grandes avanços para a humanidade, ainda não é possível afirmar que os alimentos transgênicos são totalmente isentos de riscos para a saúde humana, animal e para o meio ambiente. Dessa forma, notase que existem posicionamentos opostos sobre a liberação comercial de OGMs no mundo: de um lado, o entendimento de que há inúmeros benefícios que podem advir da disseminação dos cultivos transgênicos para a agricultura e para o comércio; de outro, o entendimento de que há inúmeras incertezas quantos aos potenciais impactos negativos para a humanidade e para a natureza. Apesar das discussões permanecerem, a comercialização e o consumo de alimentos geneticamente modificados vem crescendo de maneira muito rápida e a nota-se que há uma grande preocupação internacional em se regulamentar o tema, porém, a pluralidade de posicionamentos pode gerar carência de coerência e efetividade na regulamentação existente. O avanço da Ciência é inegavelmente mais rápido que o do Direito. A complexidade do tema é enorme, vez que, ainda que a legislação acompanhe o progresso científico, existem divergências entre compromissos globais comerciais e ambientais, que, devido aos diferentes mecanismos de responsabilização e solução de conflitos do Protocolo de Cartagena e dos Acordos da Organização Mundial do Comércio, tendem à prevalência do interesse econômico em detrimento da proteção ambiental. O desafio está em se compatibilizar o avanço tecnológico com a preservação ambiental da maneira mais consciente possível, evitando situações de irreversibilidade de danos ambientais, o que, inevitavelmente traria impactos sobre o comércio internacional, daí a grande dificuldade. / This dissertation aims to demonstrate that the way that has been occurring cross-border trade of genetically modified foods raises doubts about the ability of international law to regulate it. Transgenic food is a result of the advancement of modern biotechnology and directly interferes in the health, in nutrition, in economy, in consumer relations and in environmental balance. It is therefore essential right of every citizen to be able to choose and decide about the food that will ingest. Although modern biotechnology has brought great advances for humanity, it is not yet possible to affirm that transgenic food are completely free of risks to human, animal health and the environment. Thus, we note that there are opposing positions on the commercial release of GMOs in the world: on one hand, the understanding that there are numerous benefits that can accrue the spread of GM crops for agriculture and trade, on the other, the understanding that there are many uncertainties to numerous potential negative impacts for humanity and nature. Despite discussions remains, marketing and consumption of genetically modified foods has been growing very quickly and we note that there is a great concern in the international regulatory issue, however, the plurality of positions can generate a lack of coherence and effectiveness in existing regulating. The advancement of science is undeniably faster than the law. The complexity of the issue is huge, since, although the legislation accompanies scientific progress, there are differences between commercial and global environmental commitments, which, due to the different mechanisms of accountability and conflict resolution of the Cartagena Protocol and the agreements of the World Trade Organization, tend to the prevalence of economic interests over environmental protection. The challenge is to reconcile the technological advancement with environmental preservation in the more conscientious way possible, avoiding situations of irreversibility of environmental damage, which , would inevitably impacts on international trade, hence the great difficulty.