Orientador: Renato Peixoto Dagnino / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Geociências / Made available in DSpace on 2018-08-15T15:16:15Z (GMT). No. of bitstreams: 1 Lima_MarciaMariaTait_M.pdf: 1555790 bytes, checksum: 9231ac8c84dd06895dd5ddd500eb7017 (MD5) Previous issue date: 2009 / Resumo: O enlace atual entre ciência, tecnologia e mercado - referido pela palavra tecnociência - tem como exemplo paradigmático as novas biotecnologias. A engenharia genética e o desenvolvimento de organismos geneticamente modificados (OGMs) constituem temas-chave de diversas controvérsias que se estabelecem entre as dimensões tecnocientífica, econômica e social. A pesquisa utilizou o discurso de cientistas brasileiros para mostrar como se constroem e difundem suas concepções de tecnociência. Esses discursos se referem à Comissão Técnica Nacional de Biossegurança (CTNBio), responsável pelos pareceres técnicos sobre OGMs e por assessorar o governo na definição da política de biossegurança. A pesquisa apontou que a política de biossegurança brasileira e a forma como a CTNBio se constituiu e atua não foram determinadas por opções de caráter estritamente "científico". Também permitiu entender como as concepções de tecnociência estão presentes neste contexto. No capítulo conclusivo foi incluída uma proposta de abordagem construtivista engajada para o tratamento das novas biotecnologias. / Abstract: The current link between science, technology and the market - referred to as technoscience - has as an emblematic example the new biotechnologies, among which those related to genetics and the development of genetically modified organisms may be considered key issues for scientific and environmental controversies. Speech fragments from the Brazilian research community are extracted from the current debate to show how those conceptions of technoscience on which they are based are constructed, propagated and legitimized. They refer to the National Biosafety Technical Commission (CTNBio) presented as electronic news by Science Journal (Jornal da Ciência) and as letters, lectures and other public documents. At the end of this dissertation, a relationship between the views of the research community, technoscientific practices and biosafety policies are established. The research points out that the Brazilian biosafety policy and the way that CTNBio was constituted and acts were not determined strictly scientific choices or by the end of uncertainty and clear and consensual boundaries regarding technological risks. Additionally, a normative constructivist approach is suggested as a form of understanding the new biotechnologies. / Mestrado / Mestre em Política Científica e Tecnológica

Trata-se de um estudo descritivo, exploratório, com abordagem de análise quantitativa, cujo objetivo foi verificar os acidentes com material biológico potencialmente contaminado, ocorridos entre acadêmicos de enfermagem de três instituições de ensino de um município do interior paulista. Para a coleta de dados aplicou-se um questionário com questões abertas e fechadas, após a observação dos aspectos ético-legais. Participaram do estudo 355 alunos que estavam presentes em sala de aula por ocasião da coleta de dados e que já realizavam atividades de ensino-aprendizagem. Os dados foram analisados em Epi Info versão 3.4.3. Verificou-se que, 44 (12,4%) estudantes referiram ter sofrido exposição acidental a material biológico. A maioria era do sexo feminino (93,1%), tinha entre 22 a 25 anos (56,8%), cursava o quarto ano do curso de enfermagem (68,1%), em período integral (75%). O número de acidentes mencionados pelos alunos totalizou 55. Quanto ao tipo de exposição, a de pele íntegra foi a mais freqüente (70,9%), seguida de acidente percutâneo (25,5%), exposição em mucosa (1,8%) e pele lesada (1,8%). As topografias mais afetadas nesses acidentes foram os dedos e as mãos (74,6%). As agulhas foram citadas como o objeto causador de 64,3% dos acidentes percutâneos e o sangue foi o material biológico envolvido em 72,7% do total de acidentes. O maior número de acidentes ocorreu na retirada da punção venosa/soro (18,2%), ao puncionar ou coletar sangue (16,4%) e ao aspirar paciente (12,7%). Os alunos não utilizavam Equipamento de Proteção Individual em 49,1 % dos acidentes, embora em mais de 80% das atividades seria recomendado ao menos o uso das luvas de procedimento. Os alunos lavaram o local do acidente em 92,7% dos casos e dois alunos espremeram a área lesada após acidente percutâneo. Em 40% dos acidentes, os estudantes não notificaram a ocorrência aos docentes supervisores; em 38,2% das exposições os alunos não tomaram nenhuma conduta, não notificaram e não procuraram atendimento médico. A maioria dos docentes, quando notificados, orientou seus alunos quanto às condutas corretas e os encaminharam para avaliação médica; um docente, apesar de notificado, ignorou o fato. As situações mais apontadas pelos alunos como as que favoreceram a exposição acidental foram não utilizar Equipamento de Proteção Individual (49,1%) e a desatenção (45,5%). Como sugestões para prevenir e controlar acidentes com material biológico entre alunos, 21,4% referiu a educação permanente/continuada e 19,7% a utilização de Equipamento de Proteção Individual e a conscientização quanto ao uso. Para minimização de acidentes com material biológico potencialmente contaminado, entre alunos de enfermagem, nota-se a necessidade de envolver não apenas os alunos, mas também docentes e profissionais dos campos de ensino-aprendizagem, num programa efetivo de prevenção e controle dos acidentes. / This is a descriptive and exploratory study with a quantitative analytic approach. The aim was to verify the occurrence of accidents involving potentially contaminated biological material among nursing students from three teaching institutions in an interior city of São Paulo State, Brazil. After observing ethical-legal aspects, a questionnaire with open and closed questions was applied for data collection. Study participants were 355 students present in the classroom at the moment of data collection and who had already been involved in practical teachinglearning activities. Data were analyzed in Epi Info version 3.4.3. Forty-four (12.4%) students indicated accidental exposure to biological material. Most of them were female (93.1%), between 22 and 25 years old (56.8%), taking the forth year of the nursing course (68.1%) and studying full-time (75%). The students mentioned 55 accidents. As to the type of exposure, intact skin exposure was the most frequent (70.9%), followed by percutaneous accident (25.5%), mucous membrane exposure (1.8%) and injured skin (1.8%). The most affected topographic areas during these accidents were fingers and hands (74.6%). Needles were mentioned as the causal object in 64.3% of percutaneous accidents and blood was the biological material involved in 72.7% of all accidents. The largest number of accidents occurred when removing venipuncture/serum (18.2%), while puncturing or collecting blood (16.4%) and while aspirating a patient (12.7%). The students were not using Individual Protection Equipment in 49.1% of the accidents, although at least the use of procedure gloves would be recommended in more than 80% of activities. The students washed the accident site in 92.7% of cases and two students squeezed the injured area after a percutaneous accident. In 40% of the accidents, the students did not notify the occurrence to the supervising faculty members; in 38.2% of the exposures, the students did not take any action and neither notified nor looked for medical help. When notified, most faculty members advised their students about correct conducts and referred them for medical evaluation; despite the notification, one faculty member ignored the fact. The situations students most indicated as favoring accidental exposure were not using Individual Protection Equipment (49.1%) and lack of attention (45.5%). As suggestions to prevent and control accidents involving biological material among students, 21.4% mentioned permanent/continuing education and 19.7% the use of Individual Protection Equipment and creating awareness about the use. To minimize accidents with potentially contaminated biological material among nursing students, the need to involve not only students is observed, but also faculty members and professionals from teaching-learning areas, as part of an effective accident prevention and control program.

Trata-se de um estudo descritivo exploratório com abordagem de análise quantitativa, cujo objetivo foi estudar a ocorrência de acidentes com material biológico potencialmente contaminado e os fatores de risco envolvidos entre os alunos do Curso de Odontologia do interior do Estado do Paraná, Brasil. Para a coleta de dados, aplicou-se um questionário contendo perguntas abertas e fechadas. Este instrumento foi aplicado aos alunos após a observação dos aspectos ético-legais. Participaram do estudo 172 alunos da 3ª, 4ª e 5ª série do curso. Destes, 122 (70,9%) alegaram ter sofrido exposição acidental com material potencialmente contaminado. A maioria dos alunos que sofreram exposição acidental é do sexo feminino (61,0%) e estão cursando a 5ª série do curso, a idade predominante ficou entre 20 e 26 anos (86,1%). O número de acidentes mencionados pelos 122 alunos totalizou 448. Quanto ao tipo de exposição, a de pele íntegra foi a mais freqüente, seguida da exposição em mucosa e em 3º o acidente percutâneo perfazendo uma média total de 2,6 acidentes por aluno sendo a média de 0,23 para acidentes percutâneos. As topografias mais afetadas nestas exposições foram os olhos, mãos e dedos, enquanto que as brocas e agulha de anestesia foram citadas como os principais objetos causadores destes acidentes. A situação de ocorrência mais encontrada foi o ato de se utilizar normalmente um instrumental e o momento de realizar a limpeza dos instrumentais. Foi nas disciplinas de Clínica Integrada, Periodontia e Dentística que ocorreram a maioria das exposições. Quanto à severidade, 41,8% das exposições foram consideradas superficiais e 22,1% como moderadas. Apenas 10,7% dos alunos que sofreram exposição procuraram atendimento para avaliação e conduta em relação à exposição, 88,6% dos sujeitos receberam as três doses da vacina contra hepatite B. Observou-se ainda que nem sempre os alunos faziam uso dos Equipamentos de Proteção Individual, sendo que o uso dos óculos de proteção e a utilização de luvas de borracha grossa para realizar a limpeza dos materiais foram os mais negligenciados. Este estudo permitiu concluir que o risco de exposição a material biológico é iminente para esta categoria e que é necessário estabelecer estratégias de intervenção urgentes tanto em nível institucional quanto acadêmico a fim de reduzir estes riscos. / This is an exploratory descriptive study with a quantitative analysis approach, which objective was to study the occurrence of accidents with potentially biologically contaminated material and the risk factors involving the students from the Course of Odontology in the interior of the State of Paraná, Brazil. For the data collection, a questionnaire was applied containing open and closed questions. This instrument was applied to the students after the observation of the ethical-legal aspects. One hundred and seventy two students from the 3rd, 4th and 5th years of the course participated in the study. From these students, 122 (70.9%) alleged to have suffered accidental exposition with potentially contaminated material. Most of the students that suffered accidental exposition were female (61.0%) and they were at the 5th year of the course, the predominant age was between 20 and 26 years old (86.1%). The number of accidents mentioned by the 122 students summed 448. The most frequent kind of accident occurrences was the intact skin, followed by the exposition of mucous membrane and the percutaneous accident which made a total average of 2.6 accidents for each student and the average of 0.23 for percutaneous accidents alone. The most affected areas in these expositions were the eyes, hands and fingers, and the drills and the anesthesia needles were mentioned as the main objects that causes these accidents. The most found occurrence situation was the common action of using an instrument and at the moment of cleaning them. It was in the courses of Integrated Clinic, Periodontics and Dentistics that happened most of the expositions. As for the severity of the cases, 41.8% of the expositions were considered superficial and 22.1% were moderate. Only 10.7% of the students exposed to some kind of injury, sought for a service of checking and possible treatment; 88.6% of the persons received the three doses of the hepatitis B vaccine. It was observed though, that students didn’t usually make use of the Equipments of Individual Protection. The use of the protection glasses and gloves, made of thick rubber at the time of cleaning the materials were the most neglectful. This study allowed us to conclude that the exposition risk to biological material is imminent for this work category and that it is necessary to establish urgent intervention strategies on an institutional or academic level in order to reduce these risks.

No

<p>Bacillus anthracis is a risk class III organism and needs to be handled inside a biosafety level 3-laboratory. A major problem when working with airborne, spore-forming bacteria like B. anthracis are the hazardous aerosols created when using an automated DNA-extraction method to prepare samples suspected to contain the organism. This study has therefore evaluated the possibility of enclosing a DNA-extraction-robot inside an air tight container (glove box). A prototype of a class III safety cabinet (also known as a glove box) was designed and built to enclose a BioRobot EZ1 from Qiagen. The purpose of this prototype was to evaluate the measurements needed to and also the feasibility of working with the robot inside the cabinet. During the manual DNA-extractions, there was some contamination found on the glove box gloves, probably due to the significantly lowered dexterity that was seen with the thick gloves. The enclosing of the robot revealed no obstacles as the machine was very easy to operate. In addition, protocols have been created for the operation of a transportable class III safety cabinet from Germfree available at SVA. The protocols include the different pressure tests that needed before every experiment take place and also decontamination steps before and after each run. Bacillus cereus was used as a model organism for different DNA-extractions, i.e. automated and manual extractions. The extracted DNA was analysed by real-time polymerase chain reaction (PCR). DNA was also extracted and analysed from B. cereus-spores. When using a manual DNA-extraction kit, B. cereus-DNA was detected at the femtogram level, i.e. 10-15 g DNA / PCR. When using the automated BioRobot EZ1, detection level was found to be at 10-16 g DNA / PCR. The PCR-efficiency for the manual kit was 89-90 % for all samples, whereas with the EZ1, efficiency was 99 %, showing the strengths of the magnetic bead separation used by the machine. A novel PCR-machine, the AlphaHelix QuanTyper™, was evaluated and compared to an ABI 7500 with regards to efficiency, speed and consistency. The QuanTyper™ was found to be superior in ramping speeds, performing a 40-cycle real-time PCR-run with melting point analysis in only 14 minutes. The fastest run accomplished on the ABI 7500 took 1 h 40 min. A ready made master mix for PCR was used for most tests (Platinum® SYBR® Green qPCR SuperMix-UDG), but faster and more robust enzymes are available and further studies need to be performed on the QuanTyper™ to fully evaluate the platform. Three target genes in Bacillus anthracis-DNA were analysed in only 38 minutes with efficiencies between 96-104 % for the virulence plasmids and detection at femtogram amount of DNA. This master thesis has addressed rapid pathogen-detection with automated DNA-extraction and novel PCR-technology, coupled with a strong biosafety aspect. The thesis will hopefully contribute to the surprisingly small area of biosafety and safety cabinet research.</p>

Laboratórios de biossegurança são classificados em quatro níveis de risco (1 a 4), sendo constituídos de combinações de práticas e técnicas de laboratório, equipamento de segurança e instalações do laboratório. Cada combinação é especificamente adequada para as operações realizadas, vias de transmissões documentadas ou suspeitas de agentes infecciosos e em monitoramento das atividades de laboratórios. O objetivo deste trabalho foi o de tornar possível a construção de laboratórios de Biossegurança Nível 3 (NB3) com baixo custo, mantendo-se as condições mínimas de biossegurança no aspecto da engenharia. Como resultado seis laboratórios NB3 foram construídos para o projeto da FAPESP, denominado Rede de Diversidade Genética Viral (VGDN), com finalidade de diagnóstico de vírus respiratórios e mais uma laboratório para o Ministério da Agricultura, situado no LANAGRO de Pernambuco na cidade de Recife. / Biosafety laboratories are classified into four levels of risk (1 to 4), and consist of combinations of laboratory practices and techniques, safety equipment and facilities of the laboratory. Each combination is particularly suitable for the operations; routes of transmission documented or suspected infectious agents in tracking the activities of laboratories. The objective of this work was to make possible the construction of laboratories for Biosafety Level 3 (BSL3) with low cost, remaining the minimum conditions of biosecurity in the aspect of engineering. As results were built six laboratories BSL3 of FAPESP for the project, called Viral Genetics Diversity Network (VGDN), with the purpose of diagnosis of respiratory viruses and another laboratory for the Ministério da Agricultura, located in LANAGRO of Pernambuco in the city of Recife.

Foram selecionados, aleatoriamente, 12 equipos em 7 clínicas da FOB/USP, dois foram preenchidos com água destilada (Ortodontia e Urgência), um com água de torneira (UBAs), em um o reservatório ficou seco por 30 dias (Odontopediatria) em outro por 60 dias (Pós-graduação), um equipo com a tecnologia B-Safe® (Multidisciplinar) e, em três, a limpeza com o detergente enzimático foi avaliado (Dentística). Amostras de 10 mL de água dos reservatórios e tubulações das canetas de alta rotação foram obtidas e diluições feitas até 10-4, semeadas nos meios de cultura R2A, Peptona Diluída - PD, Plate Count Agar - PCA e Sabouraund Dextrose Agar SDA. Alíquotas de 100 L das amostras foram semeadas nos meios de cultura R2A, PD, PCA e SDA pela técnica de esgotamento, alíquotas de 25 L foram semeadas (R2A, PD, PCA e SDA) pela técnica da gota e alíquotas de 100 L das amostras foram semeadas no meio PCA pela técnica de pour plate. As placas de R2A, PD, PCA foram incubadas por 72 horas a 24°C e as placas de SDA por 4 a 7 dias a 24°C. Foi feita a identificação bacteriana através do kit Bactray I, II ou III e a fúngica através do microcultivo. A média bacteriana obtida foi de 128.151 UFC/mL na Odontopediatria, 1.834.807 UFC/mL na Ortodontia, 60.422 UFC/mL na Pós-Graduação, 615,68 UFC/mL na UBAs, 899,64 UFC/mL na Urgência, 97.632 UFC/mL na Multidisciplinar e 417.619 UFC/mL na Dentística sem limpeza e 135.924 UFC/mL após a limpeza. A média dos fungos foram 205 UFC/mL na Odontopediatria, 702,50 UFC/mL na Ortodontia, 12,50 UFC/mL na Pós-Graduação, 41.475 UFC/mL UBAs, 117.500 UFC/mL Urgência, 4.469 UFC/mL na Multidisciplinar e 64.642 UFC/mL e de 23.627 UFC/mL, antes e após a limpeza na Dentística. A presença de micro-organismos foi detectada nos reservatórios e tubulações de água em todas as 7 clínicas; para quantificar as bactérias, o meio R2A seguido do PD foram melhores que o PCA e, para detectar diferentes espécies o meio PCA foi superior ao R2A e PD; a técnica da gota foi melhor do que a de esgotamento e pour plate para as bactérias, enquanto a de esgotamento foi superior para os fungos. O detergente enzimático foi eficaz em desestruturar o biofilme, atuando mais sobre os fungos do que para as bactérias, que não foram eliminadas após as limpezas. Foram identificadas 22 espécies de bactérias: Acinetobacter baumanni/calcoaceticus, Aeromonas hydrophila, Alcaligenes xylosoxidans denitrificans, Brevundimonas vesicularis, Burkholderia cepacia, Burkholderia pseudomallei, Chromobacterium violaceum, Hafnia alvei, Hafnia alvei (Biogrupo 1), Ochrobactrum anthropi, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, Pseudomonas luteola, Pseudomonas oryzihabitans, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Serratia liquefaciens, Serratia plymuthica, Sphingobacterium multivorum, Tatumella ptyseos. Foram identificados 12 gêneros de fungos: Acremonium sp, Alternaria sp, Aspergillus sp, Cladosporium sp, Curvularia sp, Exophiala sp, Fonsecaea sp, Fusarium sp, Paecylomices sp, Penicillium sp, Rhinocladiella sp, Verticillium sp. / Twelve dental units of 7 clinics of FOB/USP were randomly selected, two were filled with distilled water (Orthodontics and Urgency), one with tap water (UBAs), one reservoir was dry for 30 days (Odontopediatry) and another for 60 days (Postgraduate Clinic), one dental unit was filled with the B - Safe ® technology (Multidisciplinary Clinic). Three dental clinics were evaluated concerning the cleaning procedure with enzymatic detergent. Samples of 10 mL of water reservoirs and high-speed handpieces were collected and made up to 10-4 dilutions, plated on R2A media, Peptone Diluted culture - PD, Plate Count Agar - PCA and Sabouraund Dextrose Agar - SDA. Aliquots of 100 L of the samples were plated on R2A media, PD, PCA and SDA culture technique for spreading. Aliquots of 25 L were seeded (R2A, PD, PCA and SDA) in drops and aliquots of 100 L samples were seeded in PCA by the pour plate technique. R2A plates, PD, PCA were incubated for 72 hours at 24°C and SDA plates for 4 to 7 days at 24°C. Bacterial identification was performed using Bactray I, II or III kit and fungal identification by microculture. Bacterial average was 128.151 CFU/mL in Odontopediatry, 1.834.807 CFU/mL in Orthodontics, 60.422 CFU/mL in Postgraduate Clinic, 615.68 CFU/mL in UBAs, 899.64 CFU/mL in Urgency, 97.632 CFU/mL in Multidisciplinary Clinic and 417.619 CFU/mL in Dentistry without the cleaning procedure and 135.924 CFU/mL after it. The average of fungi was 205 CFU/mL in Odontopediatry, 702.50 CFU/mL in Orthodontics, 12.50 CFU/mL in Postgraduate Clinic, 41.475 CFU/mL in UBAs, 117.500 CFU/mL in Urgency, 4.469 CFU/mL in Multidisciplinary and 64.642 CFU/mL and 23.627 CFU/mL before and after cleaning procedure in Dentistry. The presence of micro - organisms occurred in reservoirs and waterlines in all of the 7 clinics evaluated. To quantify bacteria, R2A and PD medium provided more accurate results than PCA. To detect different species, PCA medium was better than R2A and PD. The drop technique was better than the spreading technique and pour plate for bacteria; however, the spreading technique provided better results for the yeasts. The enzymatic detergent was effective on disrupting the biofilm eliminating more yeasts than residual bacteria. Twenty-two species of bacteria were identified: Acinetobacter baumanni/calcoaceticus, Aeromonas hydrophila, Alcaligenes xylosoxidans denitrificans, Brevundimonas vesicularis, Burkholderia cepacia, Burkholderia pseudomallei, Chromobacterium violaceum, Hafnia alvei, Hafnia alvei (Biogroup 1), Ochrobactrum anthropi, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, Pseudomonas luteola, Pseudomonas oryzihabitans, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Serratia liquefaciens, Serratia plymuthica, Sphingobacterium multivorum, Tatumella ptyseos. Twelve genus of fungi were identified: Acremonium sp, Alternaria sp, Aspergillus sp, Cladosporium sp, Curvularia sp, Exophiala sp, Fonsecaea sp, Fusarium sp, Paecylomices sp, Penicillium sp, Rhinocladiella sp, Verticillium sp.

Made available in DSpace on 2016-04-27T18:15:45Z (GMT). No. of bitstreams: 1 Edson Cinaqui Filho.pdf: 2131297 bytes, checksum: aef7f7cc013ddbf281ee30bc79baa5a2 (MD5) Previous issue date: 2015-03-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Transgenic organisms have inserted into its genome one or more DNA sequences of a species of interest, to ensure the inserted genic expression. Of these genes. These sequences (genes) may come from any living organismo (bactéria, fungus, plants, animals, protozoan) or nonliving (vírus). The permission for planting and commercialization of these organisms which are modified to resist various environmental pressusres raised the debate about its biosafety to the environment and to human and animal health. In this research were analyzed and discussed the decision-making processes involved in the approval of three varieties of transgenic corn in Brazil: the Liberty Link herbicide tolerant, insect resistant MON810 and Bt11 also resistant to insects. The choice of analyzing corn soybeans, is due the fact that Brazil is one of the centers of diversity of this plant and also for its importance to indigenous tribes and traditional communities. The studies submitted by the companies planning release the transgenic seeds were inadequate perfomed at short notice and grounded in statistical methods poorly documented. The samples were not representative and the number of repetitions insuficiente. Studies on the possible effects of there transgenic varieties on populations of non-target organisms were performed improperly, disregarding its ecological importante. The releases of these three varieties of transgenic com in Brazil contradicted Article 225 of the Federal Constitution which states that everyone has the right to a balanced environment which is essential to a healthy quality of life, and the Government should protect and preserve it for presente and future generations.The permission for planting and sala of transgenics occurred throught a comission in which the majority of members were interested in a transgenic research funded by multinationals. In addition to the political nature of problems, the release of transgenic corn wounded three basic environmental principles: precaution, sustainability and responsibility, for damage. Currently several studies attest problems in human and animal health and in the environment arising from transgenic organisms. Thus, the adoption of the Precautionary Principle should be essential before the release of transgenic corn in Brazil, to ensure that with so many scientific uncertainties the environment small farmers and the health human beings and other animals would be sabe. , to ensure that with so many scientific uncertainties, the environment, small farmers and the health of human beings and other animals would be safe / Organismos transgênicos possuem inseridos em seu genoma uma ou mais sequências de DNA de uma espécie de interesse, de modo a garantir a expressão gênica destes genes inseridos. Essas sequências (genes) podem provir de qualquer organismo vivo (bactérias, fungos, plantas, animais, protozoários) ou não vivo (vírus). A liberação para plantio e comercialização destes organismos modificados e resistentes a diversas pressões ambientais levantaram o debate sobre a biossegurança destes ao meio ambiente, saúde humana e animal. Neste trabalho foram analisados e discutidos os processos decisórios envolvidos na aprovação de três variedades de milhos transgênicos no Brasil: o Liberty Link tolerante a herbicida, o MON810 resistente a insetos e o Bt11 também resistente a insetos. A escolha de analisar o milho e não a soja, se deve ao fato do Brasil ser um dos centros de diversidade dessa planta, e também por sua importância para os povos indígenas e para as comunidades tradicionais. Os estudos apresentados pelas empresas que pleiteavam liberar suas sementes transgênicas foram inadequados, realizados à curto prazo e embasados em métodos estatísticos mal documentados. As amostragens não foram representativas e os números de repetições insuficientes. Estudos sobre os possíveis efeitos destas variedades transgênicas em populações de organismos não-alvo foram realizados de forma inadequada, desprezando a importância ecológica destes. As liberações destas três variedades de milho transgênico no Brasil contrariaram o Artigo 225 da Constituição Federal que afirma que todos têm direito ao meio ambiente em equilíbrio e essencial à sadia qualidade de vida, e que caberia ao Poder Público defende-lo e preservá-lo para as gerações presentes e futuras. As liberações para plantio e comercialização de transgênicos ocorreram por meio de uma comissão cuja maioria dos membros estavam interessados em pesquisas de transgenia com financiamento de multinacionais. Além dos problemas de cunho político, a liberação do milho transgênico feriu três princípios ambientais básicos: da precaução, da sustentabilidade e da responsabilidade por danos. Atualmente diversos estudos atestam problemas na saúde humana, animal e ao meio ambiente decorrentes de organismos transgênicos. Desta forma, a adoção do Princípio da Precaução se fazia fundamental antes da liberação do milho transgênico no Brasil, para garantir que diante de tantas incertezas científicas o meio ambiente, o pequeno agricultor e a saúde do ser humano e demais animais estariam seguros

Submitted by Erika Demachki (erikademachki@gmail.com) on 2014-09-05T20:46:26Z No. of bitstreams: 2 (LEILA ABOU SALHA DISSERTAÇÃO MEPES FINAL28022014 DIFINITIVA).pdf: 1146310 bytes, checksum: ceea6748760a294b7f289fc3f04e6ee5 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-09-05T20:46:26Z (GMT). No. of bitstreams: 2 (LEILA ABOU SALHA DISSERTAÇÃO MEPES FINAL28022014 DIFINITIVA).pdf: 1146310 bytes, checksum: ceea6748760a294b7f289fc3f04e6ee5 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-02-28 / Professional practice is also guided by the teaching- learning and knowledge in biosafety training of pharmacists , is of paramount importance for the safe and conscientious professional practice . The study aims to investigate the knowledge of undergraduate students about concepts of biosafety aspects in the use of personal protective equipment , standard precautions and immunization . A questionnaire was applied to interview students of pharmacy of a public institution to collect data on biosafety , during the period November 2012 to May 2013. Among the 300 participants , 132 ( 64.4 %) use lab coat and glove 240 ( 80 % ) have knowledge of waste disposal , 246 ( 82 % ) know about measures to protect the safety and health of workers in health services , 289 ( 96 % ) know the correct handwashing and 71 ( 26 % ) reported knowledge of occupational immunization. It was observed that approximately 65 % of the students showed biosafety knowledge and this knowledge did not show the expected periods over the course of the addition. There is need for changes in teaching strategies to be effective in practice biosecurity as essential to professional practice safely content, reaffirming content as transversal and interdisciplinary . / A prática profissional se orienta também pelo processo de ensino-aprendizagem e o conhecimento em biossegurança na formação dos farmacêuticos, é de suma importância para o exercício profissional seguro e consciente. O estudo tem por objetivo investigar o conhecimento de graduandos acerca de conceitos de biossegurança nos aspectos da utilização de equipamentos de proteção individual, precauções-padrão e imunização. Aplicou-se questionário para entrevistar alunos do curso de farmácia de uma instituição pública para coleta de dados sobre biossegurança, durante o período de novembro de 2012 a maio de 2013. Dos 300 participantes, 132 (64,4%) utilizam jaleco e luva, 240 (80%) tem conhecimento sobre descarte de resíduos, 246 (82%) sabem sobre medidas de proteção à segurança e a saúde de trabalhadores dos serviços de saúde, 289 (96%) conhecem a higienização correta das mãos e 71 (26%) relataram conhecimento acerca de imunização ocupacional. Observou-se que aproximadamente 65% dos graduandos mostraram conhecimento em biossegurança e esse conhecimento não apresentou o acréscimo esperado ao longo dos períodos do curso. Há necessidade de mudanças nas estratégias de ensino para se efetivar na prática a biossegurança como conteúdo imprescindível ao exercício profissional com segurança, reafirmando o conteúdo como transversal e interdisciplinar.

Bacillus anthracis is a risk class III organism and needs to be handled inside a biosafety level 3-laboratory. A major problem when working with airborne, spore-forming bacteria like B. anthracis are the hazardous aerosols created when using an automated DNA-extraction method to prepare samples suspected to contain the organism. This study has therefore evaluated the possibility of enclosing a DNA-extraction-robot inside an air tight container (glove box). A prototype of a class III safety cabinet (also known as a glove box) was designed and built to enclose a BioRobot EZ1 from Qiagen. The purpose of this prototype was to evaluate the measurements needed to and also the feasibility of working with the robot inside the cabinet. During the manual DNA-extractions, there was some contamination found on the glove box gloves, probably due to the significantly lowered dexterity that was seen with the thick gloves. The enclosing of the robot revealed no obstacles as the machine was very easy to operate. In addition, protocols have been created for the operation of a transportable class III safety cabinet from Germfree available at SVA. The protocols include the different pressure tests that needed before every experiment take place and also decontamination steps before and after each run. Bacillus cereus was used as a model organism for different DNA-extractions, i.e. automated and manual extractions. The extracted DNA was analysed by real-time polymerase chain reaction (PCR). DNA was also extracted and analysed from B. cereus-spores. When using a manual DNA-extraction kit, B. cereus-DNA was detected at the femtogram level, i.e. 10-15 g DNA / PCR. When using the automated BioRobot EZ1, detection level was found to be at 10-16 g DNA / PCR. The PCR-efficiency for the manual kit was 89-90 % for all samples, whereas with the EZ1, efficiency was 99 %, showing the strengths of the magnetic bead separation used by the machine. A novel PCR-machine, the AlphaHelix QuanTyper™, was evaluated and compared to an ABI 7500 with regards to efficiency, speed and consistency. The QuanTyper™ was found to be superior in ramping speeds, performing a 40-cycle real-time PCR-run with melting point analysis in only 14 minutes. The fastest run accomplished on the ABI 7500 took 1 h 40 min. A ready made master mix for PCR was used for most tests (Platinum® SYBR® Green qPCR SuperMix-UDG), but faster and more robust enzymes are available and further studies need to be performed on the QuanTyper™ to fully evaluate the platform. Three target genes in Bacillus anthracis-DNA were analysed in only 38 minutes with efficiencies between 96-104 % for the virulence plasmids and detection at femtogram amount of DNA. This master thesis has addressed rapid pathogen-detection with automated DNA-extraction and novel PCR-technology, coupled with a strong biosafety aspect. The thesis will hopefully contribute to the surprisingly small area of biosafety and safety cabinet research.