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Determination of the Leak Size Critical to Package Sterility MaintenanceKeller, Scott Wayne 09 April 1999 (has links)
This study was divided into four sections: the literature review; the mechanism by which a package defect becomes a leak; and the imposed pressures generated within a package during distribution; comparison of the threshold leak size to the critical leak size and their effect on loss of package sterility; and the relationships between microorganism characterisitics and the threshold leak size, and their effect on the critical leak size.
Section II. The mechanism by which a package defect converts to a leaker in an effort to develop a relationship between the threshold leak size and loss of package sterility was studied. The threshold leak size is the hole size at which the onset of leakage occurs. The threshold pressure is that which is required to initiate a leak. Leak initiation was studied in terms of the interaction between three components: liquid attributes of liquid food products, defect size, and pressures required to initiate liquid flow.
Liquid surface tension, viscosity, and density were obtained for sixteen liquids. The imposed pressures (Po) required to initiate flow through microtubes of IDs 0, 2, 5, 7, 10, 20 or 50 m, were measured using 63 test cells filled with safranin red dye, tryptic soy broth, and distilled water with surface tensions of 18.69 mN/m, 44.09 mN/m, and 64.67 mN/m, respectively. Significant differences were found between observed threshold pressures for safranin red dye, tryptic soy broth, and distilled water (p < 0.05). Liquids with small surface tensions such as safranin red dye required significantly lower threshold imposed pressures than liquids with large surface tensions such as distilled water (p < 0.05). An equation was developed to quantify the relationship between liquid surface tension, threshold imposed pressure, and defect size. Observed threshold pressures were not significantly different (p > 0.05) than those predicted by the equation.
Imposed pressures and vacuums generated within packages during random vibration and sweep resonance tests were measured for brick-style aseptic packages (250 ml), metal cans size 76.2-mm x 114.3-mm (425 ml), quart gable top packages (946 ml), one-half gallon gable top packages (1.89 L) and one-gallon milk jugs (4.25 L). Significant differences were found between packages for observed generated pressures during vibration testing (p < 0.05). An equation to calculate the threshold like size based on liquid surface tension and imposed pressure was established.
Section III. The onset of liquid flow through a defect as a result of imposed positive pressures or vacuum were linked to the sterility loss of a package. Five-hundred sixty-three test cells, each with microtubes of 0, 2, 5, 7, 10, 20 or 50 m, manufactured to simulate packages with defects, were biochallenged via an aerosol concentration of 106 cells/cm3 of Pseudomonas fragi Lacy-1052, under conditions of imposed positive pressure or vacuum of 20.7, 13.8, 6.9, 0, -6.9, -13.8, -20.7 kPa, respectively and temperatures of 4 , 25 and 37 C. A statistically significant relationship between loss of sterility due to microbial ingress in test cells and the initiation of liquid flow were found (p < 0.05). Microbial ingress was not found in test cells with microtube IDs of 2 m. Leak sizes critical to the sterility maintenance were found to be different based on the liquid surface tension, and imposed package pressures. The threshold leak size where the onset of liquid flow was initiated, and the critical leak size at which loss of sterility occured were not significantly different (p > 0.05).
Section IV. The effects of microorganism size and motility, and the imposed pressure required to initiate liquid flow, on the leak size critical to the sterility of a package were measured. Pseudomonas fragi Lacy-1052, Bacillus atrophaeus ATCC 49337, and Enterobacter aerogenes ATCC 29007 were employed to indicate loss of package sterility. One hundred twenty-six microtubes with interior diameters (I.D.s) of 5, 10, and 20 m and 7 mm in length were used as the manufactured defects. Forty-two solid microtubes were used as a control. An equation was used to calculate imposed pressures sufficient to initiate the flow of tryptic soy broth through all defects. No significant differences were found for loss of sterility as a result of microbial ingress into test cells with microtube ID sizes of 5, 10, and 20 m between the test organisms (p > 0.05). Interactions between the initiation of liquid flow as a result of imposed pressures, and the sterility loss of test cells were significant (p < 0.05). / Ph. D.
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Mapování genů ovlivňujících poddruhově specifické funkce meiotického genu Prdm9 / Maping of genes modifying the subspecies-specific roles of the meiotic gene Prdm9Škaloudová, Eliška January 2015 (has links)
The PRDM9 (PR domain containing 9) protein is an epigenetic factor that trimethylates lysine 4 of histone H3 and thereby determines the future meiotic double-strand breaks - sites important for proper segregation of homologous chromosomes. Males of the Mus musculus domesticus (Mmd) origin with homozygous deletion in Prdm9 (Prdm9-/- ) are sterile with a complete arrest in meiotic prophase I, in contrast to the same mutant males of the M. m. musculus (Mmm) subspecies. The aim of this diploma thesis was to identify the genomic loci responsible for the phenotypic difference of these Prdm9-/- males. The major research tool was a population of 182 Mmm x Mmd Prdm9-/- males. The mapping method of quantitative trait loci (QTLs) was based on relating the genotypes of single-nucleotide and microsatellite polymorphisms to the observed phenotypes. At least two QTLs on Chr X were identified. The Mmm alleles of these QTLs reduced fertility of Prdm9-/- males. Both QTLs were confirmed and narrowed down using two types of subconsomic strains. It was not possible to confirm other QTLs, particularly on autosomes. This QTL mapping is the first step towards the identification of genes that modify the resulting phenotype of Prdm9-/- animals. This identification should help designing studies of human infertility that...
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Vztah typu locus of control na výskyt depresivity u neplodných osob / Relation locus of control on prevalence of depresivity by infertil personBlašková, Ivana January 2016 (has links)
This master thesis deals with infertility and its treatment. Both in the theoretical and empirical part, it focuses on the female aspects of the issue. The theoretical part summarizes an overview of key information important to understand the presented problems. We define the terms, describe the psychological and physiological causes of infertility, the process of treatment of infertility and aspects thereof. We focus on the depression rate related to infertility and its treatment, furthermore, we deal with type locus of control related to the experienced rate of depression. Our aim in the empirical part is to describe the depression rate, type of locus of control within the two samples of women acquired by us and capture the possible connections. The sample is made of 58 women who are currently undergoing a treatment for infertility and of the control group, which is made of 58 healthy women. To collect the data, we used the Beck Depression Inventory BDI-II, the Rotter's Internal- External Scale and the anamnestic questionnaire. Based on acquired data, we found out some specifity in depression rate and type of locus of control within our samples of women.
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Obtenção e caracterização de linhagem transgênica de Aedes aegypti machos geneticamente estéreis. / Obtention and characterization of transgenic lines of Aedes aegypti for males genetically sterile.Carvalho, Danilo de Oliveira 08 August 2016 (has links)
Com o aumento progressivo do número de casos de infecção por diferentes arbovírus, por exemplo, dengue, zica e chikungunya, faz-se necessário o desenvolvimento de novas técnicas para o combate a esses arbovírus. A manipulação genética possibilitou a obtenção de mosquitos geneticamente modificados que sejam capazes de suprimir a população selvagem ou impedir a transmissão de agentes etiológicos gerando doenças. O estudo teve como objetivo o estabelecimento de linhagens para supressão populacional de Aedes aegypti. Esta construção apresenta esterilidade condicionada à presença ou ausência de antibiótico no meio em que esses mosquitos se desenvolvem durante a fase larval. Dessa forma, sem a necessidade de se utilizar radiação para obter insetos estéreis, é possível melhorar a qualidade dos machos adultos liberados e aumentar a competitividade dos mesmos em competir por fêmeas selvagens e adicionalmente gerar o desejado quadro de supressão populacional. / The increasing number of infection cases by arboviruses, for instance Dengue, Zika and Chikungunya, it is necessary to develop new techniques to fight against these arboviruses. Genetic manipulation allowed the production of genetically modified mosquitoes that are capable of suppressing the wild population or prevent pathogens transmission and avoid disease development. The project proposes to establish lines for population suppression of Aedes aegypti, one of the main vectors of those diseases. This construction presents the conditional sterility due to the presence or absence of antibiotics in the environment where these mosquitoes are developed during the larval stage. Thus, without the need to use radiation for sterile insects, it is possible to improve the quality of the released adult males and enhance the competitiveness thereof in competing with wild females and additionally generate population suppression.
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Estudo para validação de método rápido microbiológico aplicado a teste de esterilidade: técnica de bioluminescência de ATP / Validation of rapid microbiological method applied to sterility test: ATP bioluminescence techniquePicanço, Aline Marinho 25 August 2014 (has links)
Este estudo foi realizado com o objetivo de desenvolvimento e validação do método microbiológico rápido empregando a técnica de bioluminescência de Adenosina Trifosfato (ATP) como método alternativo para o teste de esterilidade. O ATP reage com o sistema enzimático luciferina/luciferase e gera um fóton de luz em presença de íons magnésio, reação que pode ser utilizada na detecção de microrganismos. A luz gerada na reação é medida por um dispositivo chamado luminômetro, que traduz o sinal em unidades relativas de luz (URL), metodologia altamente sensível que pode ser utilizada na análise de produtos estéreis com o objetivo de diminuição no tempo do ensaio. Enquanto a turbidez do meio de cultura só pode ser visualizada quando o contaminante chega à concentração de 106 UFC/ml, a tecnologia de bioluminescência de ATP pode detectar amostras com concentração em torno de 104 UFC. Foram empregados na validação dados obtidos a partir do método tradicional de esterilidade (técnica de filtração) realizado paralelamente ao método alternativo. As soluções parenterais utilizadas nos ensaios foram: solução fisiológica 0,9%; solução de dextrose 5%; ringer lactato; e solução de metronidazol 0,5%. As soluções-teste foram inoculadas intencionalmente com suspensões microbianas preparadas através da diluição de Bioballs®, com concentrações de 10 UFC/100 ml, 2 UFC/100 ml e 0,4 UFC/100 ml. Após a realização do teste convencional, as membranas resultantes do ensaio foram incubadas nos meios de cultura caldo caseína de soja e tioglicolato. Alíquotas destes meios foram retiradas após 96 horas de incubação para análise pelo método alternativo. Os seguintes microrganismos foram selecionados para o estudo: Staphylococcus aureus, Bacilus subtilis, Pseudomonas aeruginosa, Candida albicans, Clostridium sporogenes, Aspergillus brasiliensis, Kocuria rosea e Micrococcus luteus. A análise dos resultados obtidos mostrou que o método alternativo é capaz de detectar os microrganismos testados. Quanto à sensibilidade, o método alternativo apresentou vantagem na concentração 2 UFC/100 ml, e equivalência nas outras concentrações. A não interferência dos diferentes produtos e meios nos resultados encontrados permite vislumbrar evidência de robustez do método. Adicionalmente, em relação ao tempo de resposta, o método alternativo demonstrou ser equivalente ao convencional (p-valor=0,43). / This study is being conducted with a goal of validating and developing the fast microbiological method of ATP bioluminescence, as an alternative method to the sterility test. The ATP reacts with the enzymatic system luciferin-luciferase and produces light in the presence of magnesium ions, this reaction can be used for microorganism\'s detection. The light generated in this reaction can be measured by a device called luminometer that translates the signal in relative light units (RLU). This methodology has high sensibility and it can be used in the sterile products analysis with the objective of reducing the time of the sample. While the turbidity of the culture medium it can be visualized just when the sample reaches a concentration of 106 UCF per mL, the bioluminescence assay can detect samples at concentrations around 104 UCF per mL. The both methods, conventional (filtration technique) and alternative were done in parallel and the result data were used in the validation study. It was used in the assay the next parenteral solutions: physiological solution 0,9%, metronidazole solution 0,5%, dextrose solution 5% and Ringer lactate. The test solutions were inoculated with the microorganism suspensions, prepared by the dilution of the Bioballs® with result concentrations of 10 CFU/100 mL, 2 FU/100 mL and 0,4 CFU/ mL. After the conventional test was performed, the result membranes were incubated in thioglicollate and soybean casein broth. After 96 hours of incubation, aliquots from the broth were taken to perform the analysis by the alternative method. The following microorganisms were selected to perform the validation study: Staphylococcus aureus, Bacilus subtilis, Pseudomonas aeruginosa, Clostridium sporogenes, Candida albicans, Aspergillus brasiliensis, Kocuria rosea and Micrococcus luteus. The analysis of results shows that the alternative method can detect the test microorganisms. Regarding of the sensibility, the alternative method shows advantage in the 2 CFU/mL inoculum concentration and equivalence in the other two concentrations. The method shows evidence of robustness because the results were not affected by the products or culture media used in the assay. Additionally concerning the detection time, the alternative method was established equivalent to the conventional method (p-value=0,43).
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Esterilização de machos de Diabrotica speciosa (Coleoptera: Chrysomelidae) com irradiação gama visando controle em culturas de importância econômica / Sterilization of males Diabrotica speciosa (Coleoptera: Chrysomelidae) with gamma irradiation for control in cultures of economic importanceAraújo, Márcio Martins de 15 August 2017 (has links)
Diabrotica speciosa (Germar, 1824), causa danos diretos por alimentação e indiretos como vetor de vírus para diversos grupos de plantas. A principal forma de controle são os agroquímicos, portanto visando o manejo populacional sem impacto ambiental, o presente trabalho teve como objetivo determinar a dose de radiação gama que proporciona a esterilidade de machos, o seu consumo foliar e as alterações histológicas em suas gônadas. Os adultos foram submetidos à radiação gama (60Co) no terceiro dia após a emergência nas doses de 0, 25, 50, 75 e 100 Gy a uma taxa de 0,808 KGy/hora, totalizando 20 repetições/ dose. A dose esterilizante baseou-se na fertilidade de fêmeas sexualmente maduras acasaladas por machos irradiados. Os casais foram individualizados em \"arenas\" e alimentados com folíolos de feijão (Phaseolus vulgaris L.) com gaze preta umedecida para oviposição. Os ovos foram tratados e dispostos em recipientes plásticos forrados com papel de filtro. Após a eclosão, as larvas foram transferidas para um recipiente maior com tampa telada contendo vermiculita fina e plântulas de milho (Zea mays L.) que foram substituídas a cada 10 dias, até a emergência do adulto. Após o 4º dia de irradiação disponibilizou-se um disco foliar de 3,2 cm de diâmetro por 24 horas, para cada casal. Os discos foram digitalizados e analisados no software ImageJ. Para avaliação das gônadas foram utilizados 3 machos por dose com 8 dias de idade dissecados em PBS e através da técnica de Hematoxilina Eosina as laminas foram avaliadas em microscópio óptico. Verificou-se que a esterilidade dos machos ocorreu a partir de 75 Gy e sua longevidade média foi de 12,5 dias. O consumo da área foliar dos casais constituídos por um macho estéril foi de 42,9% e a analise histológica testicular demonstrou desorganização nos tecidos e lacunas entre as células germinativas nas maiores doses de 75 Gy e 100 Gy. / Diabrotica speciosa (Germar, 1824), causes direct and indirect damage on crops by feeding and acting as virus vector for diverse groups of plants. The main control form is the use of agrochemicals so, aiming to manage the population without environment impact, the present work had as objective to determine the dose of gamma radiation that provides male sterility, its foliar consumption and gonads histological changes. Adult males were submitted to gamma radiation (60Co) on the third day after emergence at doses of 0, 25, 50, 75 and 100 Gy at 0,808 KGy/ hour rate, totaling 20 replicates/ dose. The sterilizing dose was based on the fertility of sexually mature non-irradiated females mated by irradiated males. The couples were individualized in \"arenas\" and fed with leaflets Phaseolus vulgaris L. with black gauze moistened for oviposition. Eggs were treated and arranged in plastic containers lined with filter paper. After hatching, larvae were transferred to a larger container with a cover cap containing fine vermiculite and maize seedlings (Zea mays L.), which were replaced every 10 days, until adult emergence. After the 4th day of irradiation, a leaf disc with 3.2 cm in diameter was available for 24 hours for each couple. The discs were scanned and analyzed in ImageJ software. To evaluate the gonads, 3 males with 8 days of age were used per dose. They were dissected in PBS, throught the Hematoxylin-Eosin technique the laminas were evaluated under an optical microscope. We observed that male sterility occurred from 75 Gy and the average longevity of this group was of 12.5 days. Consumption of the leaf area by couples composed by a sterile male was 42.9% and the testicular histological analysis demonstrated tissue disorganization and gaps between germ cells at the highest doses of 75 Gy and 100 Gy.
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Proposta de um modelo de implementação da liberação paramétrica para produtos com esterilização terminal / Proposal of a model for implementation of parametric release for terminally sterilized productsRamos, Thais Mitleton Borges 04 August 2014 (has links)
A Liberação Paramétrica (L.P) é definida como a liberação de produtos submetidos à esterilização terminal, por meio do cumprimento de parâmetros críticos do processo de esterilização, sem a necessidade da realização do teste de esterilidade. O conceito da L.P para produtos com esterilização terminal surgiu como uma alternativa à deficiência do teste de esterilidade, pois o teste apresenta limitações, não sendo capaz de garantir que todas as unidades do lote se encontram estéreis. A L.P é um assunto recente no Brasil e que se encontra em processo de regulamentação. Em 2012, a Agência Nacional de Vigilância Sanitária - (ANVISA) iniciou a discussão sobre o tema, dada a importância que este apresenta para país e, recentemente, no inicio do ano de 2014, foi publicada Consulta pública sobre o tema. Desse modo, ainda existem poucas informações sobre o cenário regulatório brasileiro da L.P e como esse conceito deve ser implementado pelas indústrias brasileiras, o que torna relevante a discussão sobre o tema. Sendo assim, o presente trabalho tem como objetivo a proposição de um modelo conceitual para auxiliar a adoção da L.P para indústrias farmacêuticas na substituição ao teste de esterilidade terminal. Após a proposição do modelo, o trabalho busca ainda a aplicação deste em uma indústria de produtos com esterilização terminal, com intuito de avaliar o modelo e agregar novos conhecimentos sobre o conceito da L.P. A metodologia do trabalho que melhor se encaixa à proposta é a pesquisa exploratória, ou seja, pesquisa do tipo teórico-conceitual, como discussões conceituais e pesquisas bibliográficas sobre o tema. Esta pesquisa também é classificada como qualitativa, sem a pretensão de garantir a precisão dos dados, já que se trata da proposição e aplicação de um modelo de implementação da L.P. Por ter um caráter aplicado, a pesquisa também pretende avaliar o modelo proposto, segundo o método da pesquisa-ação em uma indústria de produtos com esterilização terminal, no qual o pesquisador interfere e participa da pesquisa. O trabalho desenvolvido conseguiu atingir resultados satisfatórios, mostrando que o modelo proposto traz benefícios à empresa que pretende utilizá-lo para realizar a adoção da LP, pois, através da aplicação do mesmo, a empresa pode se organizar melhor internamente para conduzir as atividades do projeto. O modelo também é vantajoso, pois trabalha com o conceito de equipes multidisciplinares. Isto favorece a integração entre os membros da organização, proporcionando a troca de conhecimento entre os profissionais e a criação de novos conhecimentos relacionados à LP. O modelo também permite que a empresa adquira melhor conhecimento sobre o processo de fabricação dos produtos estéreis no qual deseja adotar a LP. Pois, através da aplicação da ferramenta de análise de risco para LP, é possível visualizar os pontos críticos de controle para a LP, PCC para LP. E isto permite conhecer criticamente o processo e permite elaborar planos de ação para controlar os riscos existentes desses pontos, a fim de atenderem as exigências para a adoção da LP. / Parametric Release (PR) is defined as the release of products subject to terminal sterilization through fulfillment of critical parameters of the sterilization process without the necessity of completing the terminal sterility test. The concept of PR for terminal sterilization products has emerged as an alternative to the sterility test because of its inability to guarantee that all units in the batch are sterile. PR is a new issue in Brazil, undergoing the regulatory process. In 2012, the Brazilian Agency (Agência Nacional de Vigilância Sanitária - ANVISA) started a debate on the topic given the importance it has for the country and recently, at the beginning of the year 2014 a public survey on the subject was published. Thus, there is little information on PR in the Brazilian regulatory scenario and how this concept should be implemented by Brazilian industries, which becomes relevant to the discussion on the topic. Thus, the present work aims at proposing a conceptual framework to help the adoption of PR as a substitute for the terminal sterility test for pharmaceutical industries. After submitting the model, the present work will also seek to employ this product in a company using terminal sterilization in order to assess the model and add new know-how regarding the concept of PR. The methodology that best suits the proposal is exploratory research, i.e., a theoretical/conceptual-type study, such as conceptual discussions and bibliographic exploration on the topic. This kind of research is also classified as qualitative, without any intent to ensure the accuracy of the data, since it is a proposition and an application of a model for implementation of PR. Due to its character, applied to research, it also aims to evaluate the proposed model using the method of action research in a company using terminal sterilization of products, where the researcher interferes and participates in the study. The work has achieved satisfactory results showing that the proposed model is beneficial to the company that may want to use it to adopt PR, because through the same application, the company can better organize itself internally to conduct project activities. The model is also advantageous because it works with the concept of multidisciplinary teams. This improves the integration between members of the organization providing the knowledge exchange among professionals and generation new knowledge related to PR. The model also allows the company to acquire better knowledge about the process of manufacture of sterile products for which it may want to adopt the PR. For, through the application of risk analysis for the PR tool, one can view the critical control points for PR, (CCPs). And this allows to view the process critically and develop action plans to control the risks relative to these points to meet the requirements for the adoption of PR
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Validação de teste de esterilidade baseado em detecção de dióxido de carbono / Validation of sterility test based in carbon dioxide detectionLira, Rodolfo Santos de 09 April 2014 (has links)
De acordo com compêndios farmacêuticos, o teste de esterilidade, requer período de incubação de 14 dias para obtenção de resultado analítico, período em que produto farmacêutico é mantido em quarentena, o que demanda custos elevados. Os métodos alternativos com aplicação na avaliação da esterilidade são particularmente interessantes, quando possibilitam a redução do período de incubação do teste, o período de quarentena e, em consequência, os custos envolvidos, além de reduzir o tempo para preparação do material requerido, para execução do ensaio e para a capacitação técnica. O presente estudo avaliou o desempenho do método microbiológico rápido BacT/Alert 3D (Biomerieux®) aplicado ao teste de esterilidade em comparação ao teste de esterilidade por método indireto descrito nas farmacopeias (brasileira, americana, européia e japonesa). O Bact/Alert 3D se baseia na detecção do crescimento microbiano, que é feita por meio de detecção de alteração da cor de um sensor sensível à liberação de dióxido de carbono produzido por metabolismo microbiano. Os testes utilizaram três diferentes soluções parenterais de grande volume: solução salina 0,9%, concentrado polieletrolítico para diálise e solução de metronidazol. Micro-organismos desafio: Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538, Bacillus subtilis ATCC 6633, Clostridium sporogenes ATCC 19404, Candida albicans ATCC 10231 e Aspergillus brasiliensis ATCC 16404. Cargas de contaminação: 20, 2 e 0,2 UFC/membrana. Temperatura de incubação: 32,5 ± 0,5 °C. As amostras foram avaliadas por 14 dias. Amostras dos três produtos foram artificialmente contaminadas com BioBalls® SingleShot dos micro-organismos desafio de diferentes cargas de contaminação foram submetidas ao teste de esterilidade farmacopeico, com inoculação por método indireto. Após 18h de incubação, foram retiradas alíquotas de cada um dos meios de cultura, que foram inoculadas nos frascos de meio de cultura do BacT/Alert 3D. Bact/Alert 3D demonstrou equivalência ao método farmacopeico (p > 0,05) e tempo de detecção inferior. / Nowadays, the harmonized sterility test method described on pharmacopoeias requires an incubation period of 14 days. During this period the product stays on quarantine waiting the result for release, which demands warehousing and inventory costs. The application of Rapid microbiologial methods for sterility testing is particulary interesting considering the benefits of incubation time reduction, quarantine period reduction, time reduction for material preparation, reduction of time for test execution and technical training.This work evaluated comparetivelly the performance of Bact/Alert 3D system as a rapid microbiologial method applied to sterility testing and sterility testing according to described on Brazilian pharmacopeia and harmonized pharmacopeias (American pharmacopeia, European pharmacopeia and Japanese pharmacopeia). The Bact/Alert 3D is based on microrganisms gowth and has as principle the detection of pad color change which is sensible to carbon dioxide release by microorganism metabolism. To perform the evaluation it was considered Buffer saline solution, Metronidazol solution and Polieletrolitic dialysis concentrate solution. Challenge Microrganisms: Staphylococcus aureus (ATCC 6538), Bacilus subtilis (ATCC 6633), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 8739 ), Clostridium sporogenes (ATCC 19404), Candida albicans (ATCC 10231) and Aspergillus brasiliensis (ATCC 16404). Inoculum load: 20CFU/sample; 2 CFU/sample; 0,2CFU/sample. Incubation temperature: 32,5 ± 0,5 °C. The samples were evaluated during 14 days. Aseptic samples of the three products were intentionally contaminated with BioBalls® SingleShot of challenge microorganisms, according to inoculum concentration desired, and were tested by indirect inoculation. After 18h aliquotes of were taken from inoculated culture media and inoculated into BacT/Alert 3D culture media bottles. Bact/Alert showed equivalence to compendial method (p > 0,05) and showed results faster than compendial method.
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Esterilização de machos de Diabrotica speciosa (Coleoptera: Chrysomelidae) com irradiação gama visando controle em culturas de importância econômica / Sterilization of males Diabrotica speciosa (Coleoptera: Chrysomelidae) with gamma irradiation for control in cultures of economic importanceMárcio Martins de Araújo 15 August 2017 (has links)
Diabrotica speciosa (Germar, 1824), causa danos diretos por alimentação e indiretos como vetor de vírus para diversos grupos de plantas. A principal forma de controle são os agroquímicos, portanto visando o manejo populacional sem impacto ambiental, o presente trabalho teve como objetivo determinar a dose de radiação gama que proporciona a esterilidade de machos, o seu consumo foliar e as alterações histológicas em suas gônadas. Os adultos foram submetidos à radiação gama (60Co) no terceiro dia após a emergência nas doses de 0, 25, 50, 75 e 100 Gy a uma taxa de 0,808 KGy/hora, totalizando 20 repetições/ dose. A dose esterilizante baseou-se na fertilidade de fêmeas sexualmente maduras acasaladas por machos irradiados. Os casais foram individualizados em \"arenas\" e alimentados com folíolos de feijão (Phaseolus vulgaris L.) com gaze preta umedecida para oviposição. Os ovos foram tratados e dispostos em recipientes plásticos forrados com papel de filtro. Após a eclosão, as larvas foram transferidas para um recipiente maior com tampa telada contendo vermiculita fina e plântulas de milho (Zea mays L.) que foram substituídas a cada 10 dias, até a emergência do adulto. Após o 4º dia de irradiação disponibilizou-se um disco foliar de 3,2 cm de diâmetro por 24 horas, para cada casal. Os discos foram digitalizados e analisados no software ImageJ. Para avaliação das gônadas foram utilizados 3 machos por dose com 8 dias de idade dissecados em PBS e através da técnica de Hematoxilina Eosina as laminas foram avaliadas em microscópio óptico. Verificou-se que a esterilidade dos machos ocorreu a partir de 75 Gy e sua longevidade média foi de 12,5 dias. O consumo da área foliar dos casais constituídos por um macho estéril foi de 42,9% e a analise histológica testicular demonstrou desorganização nos tecidos e lacunas entre as células germinativas nas maiores doses de 75 Gy e 100 Gy. / Diabrotica speciosa (Germar, 1824), causes direct and indirect damage on crops by feeding and acting as virus vector for diverse groups of plants. The main control form is the use of agrochemicals so, aiming to manage the population without environment impact, the present work had as objective to determine the dose of gamma radiation that provides male sterility, its foliar consumption and gonads histological changes. Adult males were submitted to gamma radiation (60Co) on the third day after emergence at doses of 0, 25, 50, 75 and 100 Gy at 0,808 KGy/ hour rate, totaling 20 replicates/ dose. The sterilizing dose was based on the fertility of sexually mature non-irradiated females mated by irradiated males. The couples were individualized in \"arenas\" and fed with leaflets Phaseolus vulgaris L. with black gauze moistened for oviposition. Eggs were treated and arranged in plastic containers lined with filter paper. After hatching, larvae were transferred to a larger container with a cover cap containing fine vermiculite and maize seedlings (Zea mays L.), which were replaced every 10 days, until adult emergence. After the 4th day of irradiation, a leaf disc with 3.2 cm in diameter was available for 24 hours for each couple. The discs were scanned and analyzed in ImageJ software. To evaluate the gonads, 3 males with 8 days of age were used per dose. They were dissected in PBS, throught the Hematoxylin-Eosin technique the laminas were evaluated under an optical microscope. We observed that male sterility occurred from 75 Gy and the average longevity of this group was of 12.5 days. Consumption of the leaf area by couples composed by a sterile male was 42.9% and the testicular histological analysis demonstrated tissue disorganization and gaps between germ cells at the highest doses of 75 Gy and 100 Gy.
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Adiamento do projeto parental: um estudo psicol?gico com casais que enfrentam a esterilidade / The postponing of the parentting project: a psychological study with couples fighting sterilityNascimento, F?tima Regina Mibach do 03 February 2009 (has links)
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Previous issue date: 2009-02-03 / The aim of this study was to investigate the postponing of the parental project, emphasizing the psychological and social aspects that interfered, the sterility repercussion on the couple involvement and their experience with the aided reproductive treatments that were made. Participated in the research three couples, straight, over 30, no biological child and different social and economic life, which underwent treatment of assisted reproduction. The contents obtained through Opened Psychological Interview were analysed using the Content Analysis technique proposed by Mathieu as well as discussed according to the theoretical referential of Psychoanalysis and the Group Psychoanalysis. Results shows: specific social conditions (access to biotechnology resources, familiar, social, economic and religious influences), as well as unconscious factors (conflicts, resistances, ambivalence, fantasies and anguish) interfered on the parental project completion; damage on the wedlock (decrease in the spontaneity and sexual interest; difficulty on reorganizing the vital project and interference in daily life); lost in familiarity with relatives and friends; decreasing movement to the early stages of the psychological work, characterized by the persecuting and depressive anguish and the use of primitive ways to deal with it (refusing, illusion, projection, displacement and rationalization). The couple s identification with the research theme and the effectiveness prove to collect data, analysis and phenomenon interpretation to touch the couples were surpassed. / O objetivo deste estudo era investigar o adiamento do projeto parental, enfatizando aspectos psicol?gicos e sociais que o determinaram, as repercuss?es da esterilidade para o v?nculo conjugal e a viv?ncia dos tratamentos reprodutivos. Participaram 03 casais, heterossexuais, acima de 30 anos, sem filhos biol?gicos e de n?veis socioecon?micos variados, que realizaram tratamentos de reprodu??o assistida. Os conte?dos obtidos atrav?s da Entrevista Psicol?gica Aberta foram analisados de acordo com t?cnica de An?lise de Conte?do proposta por Mathieu e discutidos conforme o referencial te?rico da Psican?lise e da Grupalidade. Resultados revelam: condi??es sociais espec?ficas (acesso aos recursos biotecnol?gicos, influ?ncias familiares, sociais, econ?micas e religiosas) associadas ?s condi??es inconscientes (conflitos, resist?ncias, ambival?ncias, fantasias e ang?stias) interferiram na realiza??o do projeto parental; comprometimento do v?nculo conjugal (diminui??o da espontaneidade e interesse sexual; dificuldades para reformula??o do projeto vital e interfer?ncias no cotidiano); preju?zos ? conviv?ncia familiar e social; regress?o aos est?gios iniciais do funcionamento mental desencadeando ang?stias persecut?rias e depressivas e uso de defesas primitivas (nega??o, ilus?o, proje??o, deslocamento e racionaliza??o). Houve identifica??o dos casais com tema da pesquisa e comprova??o da efic?cia do dispositivo grupal para coleta, an?lise e interpreta??o dos fen?menos ps?quicos envolvidos.
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