No description available.

Thesis (PhD (Conservation Ecology and Entomology))--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Codling moth, Cydia pomonella, is a worldwide pest and of major economic importance to the South African pome fruit industry. Sterile insect release is applied as a component of area-wide integrated pest management and includes the mass-rearing, sterilization and the release of the sterile insects. For sterile insect release, the improvements of rearing methods in terms of the quality of the diet ingredients and the economical aspect of the rearing method are examined. The effect of genetically modified maize meal, containing the Bacillus thuringiensis gene, in an artificial medium for codling moth rearing, is determined. The use of even a small amount of Bacillus thuringiensis resulted in larval mortality and prolonged development. These results are detrimental to a mass-rearing facility and must be considered by any rearing facility that uses genetically modified maize meal if the insect is sensitive to the gene. An alternative to maize meal in the artificial medium was tested and whole wheat flour was considered to be a suitable replacement. Agar agar is an expensive gelling agent used in the artificial medium. An alternative for agar agar (Kelcogel, Elastigel and carrageenen) is tested and the biological effect on codling moth is determined. Factors such as mortality, pupal and moth weight, longevity, fecundity and development time were used as quality parameters. Results showed that Elastigel was a suitable replacement for agar agar, with bigger pupae and moths, higher fecundity and increased longevity. The economical advantage of the replacement is a 40.91% reduction of the diet cost. The other gelling agents tested also gave acceptable results and can be considered if shortages of agar agar or Elastigel occur. A new method of mass-rearing codling moth larvae in a closed rearing system using large trays placed in a ventilated box is designed. This method is more cost and space effective as a smaller area is needed to rear a large number of moths. The risk of diet contamination is less because of the closed environment and more economical and effective air handling. This is the first report of its kind to describe the mass-rearing of codling moth in a closed environment and the risks involved in using genetically modified maize meal in an artificial diet for the codling moth. These results should be incorporated into existing mass-rearing facilities or taking into consideration when designing new mass-rearing facilities. / AFRIKAANSE OPSOMMING: Die kodlingmot, Cydia pomonella, is van ekonomiese belang vir die Suid-Afrikaanse kernvrugte bedryf. Die steriele insek tegniek word gebruik as ‘n komponent in area-wye geïntegreerde plaagbeheer en sluit in die massa-aanteel, sterilisering en vrylaat van steriele insekte. Vir die steriele insek tegniek is die verbetering van die massa-aanteel van die kodlingmot in terme van kwaliteit van die dieet en die ekonomiese aspek van die aanteel metode ondersoek. Die effek van genetiese gemanipuleerde mieliemeel wat die Bacillus thuringiensis geen bevat, in ‘n kunsmatige voedselmedium vir die aanteel van kodlingmot, is bepaal. Daar is gevind dat die gebruik van selfs ‘n klein persentasie Bacillus thuringiensis in die mieliemeel, mortaliteit en ‘n verlengde lewenssiklus in kodlingmot veroorsaak. Die gevolge is nadelig vir ‘n massa-aanteel fasiliteit en behoort in ag geneem te word vir enige insek wat op ‘n kunsmatige medium, wat mieliemeel bevat, geteel word, mits die insek sensitief is vir Bacillus thuringiensis. ‘n Alternatiewe bestanddeel vir mieliemeel, volkoringmeel, word aanbeveel. Agar agar is ‘n duur verdikkingsagent wat in kunsmatige mediums gebruik word. ‘n Alternatief vir agar agar (Kelcogel, Elastigel en carrageenen) is getoets en die biologiese effek op die kodlingmot is bepaal. Faktore soos mortaliteit, papie en mot gewig, langlewendheid, vrugbaarheid en lengte van lewenssiklus was gebruik as kwaliteit parameters. Resultate het getoon dat Elastigel ‘n geskikte plaasvevanger is van agar agar, met groter papies en motte, groter vrugbaarheid en langlewendheid. Die ekonomiese gevolg van die plaasvervanger, is ‘n vermindering van 40.91% van die dieetkoste. Die ander verdikkingagente wat is getoets is, het aanvaarbare resultate gelewer wat noodsaaklik is indien daar ‘n tekort van Elastigel of agar agar ontwikkel. ‘n Nuwe metode van massa-aanteel van kodlingmot larwes is bepaal. Die metode behels ‘n geslote sisteem, waar groter aanteel bakke in ‘n geslote, geventileerde boks geplaas word. Die metode is koste en spasie effektief en ‘n kleiner area word benodig om ‘n groter aantal motte te lewer. Die risiko van kontaminasie van die dieet word verminder as gevolg van die geslote sisteem wat gebruik word en meer ekonomiese en effektiewe lugversorging word gebruik. Hierdie is die eerste verslag van sy soort wat die massa-aanteel van kodlingmot in ‘n geslote sisteem beskryf en wat die risiko aandui van geneties gemanipuleerde mieliemeel in ‘n kunsmatige medium vir die kodlingmot. Hierdie resultate behoort in ag geneem te word vir reeds bestaande massa-aanteel fasiliteite of met die ontwerp van nuwe massa-aanteel fasiliteite.

In this thesis I test the modified Newtonian dynamics as an alternative to the cold dark matter hypothesis. In the Milky Way, I show that the dynamics of the dwarf galaxies are well described by the paradigm and I confirm its distant low surface brightness globular clusters provide a strong test, for which I make predictions. Through analysis of a sample of 26 X-ray bright galaxy groups and clusters I demonstrate that the three active neutrinos and their anti-particles are insufficient to reconcile modified Newtonian dynamics with the observed temperatures of the X-ray emitting gas, nor with weak-lensing measurements, in particular for the bullet cluster. To this end, I propose an 11eV sterile neutrino to serendipitously resolve the residual mass problem in X-ray bright groups and clusters, as well as matching the angular power spectrum of the Cosmic Microwave Background. With this in mind, I show that the large collision velocity of the bullet cluster and the high number of colliding clusters is more naturally reproduced in MOND than in standard dynamics.

Genetic control strategies are a novel method for reducing populations of pest insects such as the yellow fever mosquito Aedes aegypti, a major vector of several important arboviral diseases. This thesis describes efforts to develop new tools to engineer the Ae. aegypti genome and to better understand existing tools, and furthermore to use these to engineer a gene drive system in Ae. aegypti. The piggyBac transposon was found to be extremely stable in the germline of Ae. aegypti, and transposons engineered into the germline could not be remobilized with either an endogenous or exogenous source of piggyBac transposase. Conversely, somatic remobilization of piggyBac transposons was found to be readily detectable in the presence of a source of active transposase, the first report of such remobilization in Ae. aegypti. Toward new tools for genome engineering, the site-specific integrase from the phage φC31 was successfully used to promote exchange between a transgene cassette inserted into the genome of Ae. aegypti and a cassette in a plasmid vector, in the first demonstration of recombinase mediated cassette exchange technology in a pest insect species. The integrases from phages φRV1 and Bxb1 were not found to be active in the germline of the mosquito. Finally, development of a gene drive system in Ae. aegypti using an RNAi-mediated killer-rescue mechanism was attempted. Tissue-specific expression of tTAV-regulated-toxic effectors genes, using the promoter regions of the blood meal induced genes Carboxypeptidase A-1, 30Kb and Vitellogenin A, was possible, but sex-specificity was not achieved. A blood meal inducible lethal phenotype was not possible using the chosen promoters, with expression of the effectors either leading to death in early development or to a sublethal phenotype. RNAi against tTAV fused to the Mnp fragment of the dengue virus’ genome was tissue specific, but was found to be highly effective in the fat body suggesting that the Vitellogenin A was the best candidate for the engineering of killer-rescue systems in the mosquito.

The cosmic neutrino background (CNB) can be composed of both active and hypothetical sterileneutrinos. At approximately one second after big bang, neutrinos decoupled from radiationand matter at a temperature of approximately one MeV. Neutrinos played an important role inthe origin and evolution of our universe and have been indirectly verified by cosmological dataon the BBN (Big Bang nucleosynthesis) of the Big Bang.It was Steven Weinberg in 1962 that first theorized on the direct detection of relic neutrinos.The signal of the relic neutrino capture on a tritium target can be observed by studying theendpoint of the electrons kinetic energy that are above the endpoint energy of the beta decayspectrum. The PTOLEMY project aims to archive direct detection of the relic neutrinobackground with a large tritium target of 100 gram, MAC-E-Filter, RF-tracking, Time of flighttracking and a cryogenic calorimetry.In this thesis the MAC-E-Filter have been simulated in two filter configurations. In the firstconfiguration, the electron were simulated five times in the filter. Two in the opposite sideof the detector, one in the middle, and two at the detector. In the second configuration theelectrons was simulated in the entrance solenoid at a fixed position of y = -0.19634954 m fromthe center of the filter and in random positions. Both multiple electrons and single electronswere simulated in the second configuration.In the single electron configuration the electron had a starting position of y = -0.19634954 mfrom the center of the filter, and an initial kinetic energy of 18.6 KeV. The first filter configurationsuccessfully accomplished to simulate the electron track, as the electron was reflectedback and forth between the entry and detector solenoid. The electric and magnetic field profilediered at the entry and detector solenoid. The second filter configuration successfully showedthat the electron will reach the end solenoid, when the filter length was 0.5 m. When the filterlength was increased to 0.7 m, then the electron was reflected in the middle of the filter. Thesimulation showed that the electron energy dropped below 1 eV from 18.6 KeV as the electronpropagated through the filter. The magnetic and electric fields decreased exponentially in thedirection of the detector solenoid. The Simulation of multiple electrons showed mixed resultsand would need more modifications in order to come to a final conclusion.

Le mécanisme de génération de masses des neutrinos, la nature de la matière noire et l’origine de l’asymétrie baryonique de l’Univers sont les trois questions les plus pressantes dans la physique moderne des astroparticules, qui exigent l’introduction d’une nouvelle physique au-delà du Modèle Standard. Dans cette thèse, nous nous concentrons sur ces trois questions en fournissant une solution possible en termes d'une extension minimale du Modèle Standard, constituée par l’ajout d'un ensemble de fermions stériles au contenu des champs de la théorie. Les fermions stériles sont des champs qui sont singlets de jauge et qui peuvent interagir avec les neutrinos actifs à travers des termes de mélange. Nous nous concentrons sur le mécanisme dit de l’Inverse Seesaw (ISS), qui est caractérisé par une faible échelle de la nouvelle physique (de l’ordre TeV ou inférieure) et qui peut être testé dans les installations expérimentales actuelles et futures.
Nous présentons l'analyse qui permet d’identifier les réalisations minimales de ce mécanisme et l'étude phénoménologique pour prendre en compte la masses des neutrinos légers et pour imposer toutes les contraintes expérimentales pertinentes au modèle, ainsi que les signatures expérimentales attendues. Nous montrons la viabilité de l’hypothèse que les neutrinos stériles constituent la matière noire, et les caractéristiques de cette solution dans le mécanisme minimale de l’ISS. La possibilité d’expliquer avec succès l'asymétrie baryonique à travers un processus de leptogenèse dans une réalisation testable du mécanisme est aussi adressée.
Il est important de chercher des manifestations des fermions stériles dans les expériences de laboratoire. Nous abordons ce point en faisant des prévisions sur les rapports des branchement attendus pour les désintégrations des bosons vectoriels qui violent le saveur leptonique, qui peuvent être véhiculés par les fermions stériles. Nous étudions aussi l'impact des fermions stériles sur les fits globaux des données de précision électrofaible. / The neutrino mass generation mechanism, the nature of dark matter and the origin of the baryon asymmetry of the Universe are three compelling questions that cannot be accounted for in the Standard Model of particle physics. In this thesis we focus on all these issues by providing a possible solution in terms of a minimal extension of the Standard Model, consisting in the addition of a set of sterile fermions to the field content of the theory. Sterile fermions are gauge singlet fields, that can interact via mixing with the active neutrinos. We focus on the Inverse Seesaw mechanism, which is characterised by a low (TeV or lower) new physics scale and that can be tested in current and future experimental facilities. We present the model building analysis that points towards the minimal realisations of the mechanism, and the phenomenological study in order to accommodate light neutrino masses and to impose all the relevant experimental constraints in the model, as well as the expected experimental signatures. We show the viability of the sterile neutrino hypothesis as dark matter component, together with the characteristic features of this scenario in the minimal Inverse Seesaw mechanism. The possibility of successfully accounting for the baryon asymmetry in a testable realisation of the leptogenesis mechanism is also addressed.On the other side it is important to look for manifestations of sterile fermions in laboratory experiments. We address this point by making predictions for the expected rates of rare lepton number violating decays of vector bosons, that can be mediated by sterile fermions, as well as by studying the impact of sterile fermions on global fit of electroweak precision data.

En Afrique sub-saharienne, près de 10 millions de km2 de terres, les plus fertiles en productions fourragères et agricoles, sont infestées de mouches tsé-tsé limitant ainsi les initiatives de développement d’une agriculture durable. Les tsé-tsé transmettent des trypanosomes qui sont responsables des trypanosomoses animales et humaines africaines. En 2000, les Chefs d’Etats et de Gouvernements africains ont décidé de redoubler d’efforts pour lutter contre les mouches tsé-tsé et les trypanosomoses en créant la Pan African Tsetse and Trypanosomosis Eradication Campaign (PATTEC). Dans ce contexte, le gouvernement sénégalais a initié un programme d’éradication des glossines dans la zone des Niayes en utilisant une souche de Glossina palpalis gambiensis originaire du Burkina Faso. La présente thèse visait à optimiser l’utilisation de la technique de l’insecte stérile (TIS) en Afrique de l’Ouest pour lutter contre les glossines. Un dispositif de transport sur de longues distances de pupes matures a été développé et validé à partir de pupes mâles de G. p. gambiensis produites et irradiées à Bobo-Dioulasso, Burkina Faso et à Bratislava, Slovaquie (irradiation faite à Seibersdorf, Autriche) et transportées par voie aérienne jusqu’à Dakar, Sénégal. Le dispositif constitué d’une boîte isotherme et des packs S8 a permis de maintenir les pupes à une température de 10 ± 3°C et de les transporter pendant 2-3 jours jusqu’au centre d’émergence de l’ISRA, pour produire des mâles stériles utilisables pour la technique de l’insecte stérile. Un contrôle qualité a été réalisé sur un échantillon de 50 pupes prélevé dans chaque lot de pupes (minimum 2 lots par envoi) pour déterminer l’aptitude d’envol des mâles stériles et leur survie sous stress (à jeun). Le reste des pupes utilisé pour les lâchers sur le terrain a été considéré comme témoin. Le protocole qualité décrit permettra un suivi précis de la qualité des mâles stériles utilisés dans les programmes opérationnels d’éradication dans le cadre de la PATTEC. Un outil moléculaire de discrimination de mâles stériles lâchés et sauvages a également été développé à partir du gène mitochondrial COI (cytochrome oxydase) et a montré que les séquences COI des mouches lâchées (produites en insectarium) sont 100% identiques entre elles et différentes de celles des mouches sauvages. Par ailleurs, afin de déterminer les conditions optimales d’élevage de souches de G. p. gambiensis et de déterminer la souche qui sera la plus adaptée à tel ou tel environnement ou pays dans le cadre d’une lutte avec une composante lâcher de mâles stériles, les traits de vie (survie et fécondité) de trois souches de G. p. gambiensis (souches originaires du Burkina Faso (BKF), Sénégal (SEN) et souche introgressée (SENbkf) ont été évalués dans différentes conditions de températures et d’humidités relatives. La température optimale d’élevage en masse a été de 25 ± 1°C, 24,6 ± 1°C et 23,9 ± 1°C pour BKF, SENbkf et SEN respectivement. La variation de l’humidité relative (entre 40 et 75%) a eu très peu d’influence sur la survie et la fécondité. La souche BKF a mieux résisté à de fortes températures que les souches SEN et SENbkf, mais la température limite de survie a été de 32°C pour les trois souches. / In sub-Saharan Africa, nearly 10 million km² of land, the most fertile for animal and agricultural productions, are infested with tsetse flies limiting all development initiatives for sustainable agriculture. Tsetse flies transmit trypanosomes that cause human and animal african trypanosomosis, a debilitating disease of humans (sleeping sickness) and livestock (nagana). In 2000, the African Heads of State and Government decided to increase efforts to address the tsetse and trypanosomosis problem on the African continent and created the Pan-African Tsetse and Trypanosomosis Eradication Campaign (PATTEC). In this context, the Government of Senegal initiated a tsetse eradication program in the Niayes area using a Glossina palpalis gambiensis strain originating from Burkina Faso. The objective of this thesis was to optimize the use of the sterile insect technique (SIT) in West Africa in order to control the tsetse flies. A system to transport mature pupae over long distances has been developed and validated for male G. p. gambiensis pupae produced and irradiated either in Bobo-Dioulasso, Burkina Faso or in Bratislava, Slovakia (irradiation done in Seibersdorf, Austria) and then transported by air to Dakar, Senegal. The system, constituted of an insulated box and S8 packs, allowed the maintenance of pupae at a temperature of 10 ± 3°C and their transport during 2-3 days to the emergence center of ISRA, where they molted into sterile males which were used for the sterile insect technique.A quality control was carried out on a sample of 50 pupae from each batch (at least 2 batches per shipment) to determine the flight ability of sterile males and their survival under stress conditions (without feeding). The remaining emerging pupae were released in the target area of the eradication programme and were considered as control group. The described protocol for quality control will allow accurate monitoring of the quality of sterile males used in operational eradication programs organized in the context of PATTEC.A molecular tool to distinguish between sterile and wild males was also developed using the mitochondrial gene COI (cytochrome oxidase). We showed that COI sequences of released flies (reared in insectary) are 100% identical and different from those of wild flies.Furthermore, in order to determine the optimal rearing conditions for G. p. gambiensis strains and to identify the strain that would be the best adapted to a particular environment or country in the context of a control with an SIT component, life history (survival and fecundity) of three G. p. gambiensis strains (strains originating from Burkina Faso (BKF), Senegal (SEN), and an introgressed strain (SENbkf)) were investigated at different temperatures and relative humidity conditions. The optimal temperature for the mass-rearing was 25 ± 1°C, 24.6 ± 1°C and 23.9 ± 1°C for BKF, SENbkf and SEN respectively. The relative humidity ranging from 40 to 75% had very little influence on the survival and fecundity. The BKF strain resisted better at higher temperatures than the SENbkf and SEN strains but the temperature limit for survival was about 32°C for all three strains.

Diversos países vêm adotando diretrizes que relacionam prazo de validade de esterilidade de materiais críticos utilizados na assistência à saúde com a ocorrência de eventos relacionados. Pesquisas têm validado a manutenção da esterilidade do material durante o transporte e armazenamento por longos intervalos de tempo. O racional teórico já refutava o antigo paradigma de prazo de validade de esterilidade, baseado no tempo relógio/calendário justificado pela eficiência das embalagens com características de barreira microbiana, selagem hermética e a teoria da ausência de geração espontânea dos micro-organismos. No entanto, no Brasil, pode-se afirmar que os Centros de Material e Esterilização ainda praticam o controle de materiais esterilizados, baseado em data arbitrariamente atribuída, sendo esta prática incentivada por recomendações de documentos elaborados por órgãos oficiais normatizadores e fiscalizadores nacionais. Na crença de que realizar uma investigação experimental in vitro, produzindo evidências científicas robustas contribuiria no fortalecimento da quebra do antigo paradigma, propôs-se avaliar a manutenção da esterilidade de materiais armazenados por até 6 meses, após as embalagens diversas sofrerem contaminação intencional em suas superfícies externas. O experimento consistiu em embalar corpos de prova que simulassem instrumental cirúrgico (cilindros de porcelana) em quatro diferentes tipos de invólucros: algodão tecido, papel crepado, tecido nãotecido (SMS) e papel grau-cirúrgico. Todos os pacotes foram previamente autoclavados em um mesmo ciclo de esterilização. Posteriormente, as superfícies externas dos pacotes foram intencionalmente manipuladas com mãos contaminadas com Serratia marcescens (106 U.F.C./mL). Após intervalos predeterminados de armazenamento (0, 7, 14, 28, 90 e 180 dias), uma quantidade determinada de pacotes de cada tipo de embalagem foi aberta e os corpos de prova foram semeados diretamente em meio de cultura caseína-soja. Os pacotes com corpos de prova com tempo zero de armazenamento foram considerados como grupo controle negativo. Com a finalidade de garantir a viabilidade do micro-organismo-teste na superfície externa das embalagens foram realizados semanalmente, até 180 dias, cultivos das amostras de cada uma das embalagens contaminadas com o micro-organismo-teste, constituindo-se o grupo controle positivo. Para cada intervalo de tempo de armazenamento, foram analisados 600 corpos de prova (150 para cada tipo de embalagem). Considerando o intervalo de confiança exato para distribuição binomial, o tamanho de amostra calculado na pesquisa garantiu um intervalo de confiança de 95% com probabilidade de 0 a 0,006 de se encontrar uma amostra contaminada. Como resultado, não foram recuperados micro-organismos-teste Serratia marcescens em nenhum dos intervalos de tempo de armazenamento. Todos os testes realizados no grupo controle positivo apresentaram recuperação do micro-organismo-teste. Com base nos resultados deste estudo, observou-se que pacotes embalados em invólucros com propriedades de barreiras microbianas e selados hermeticamente são capazes de proteger Resumo Giovana Abrahão de Araújo Moriya o conteúdo esterilizado por até 6 meses. Este estudo contribui para a prática em Centros de Material e Esterilização, impedindo o reprocessamento desnecessário dos materiais, ao mesmo tempo, alerta para que os eventos relacionados sejam controlados e que cada material, antes da sua utilização, seja inspecionado quanto à integridade da embalagem e selagem. / Many countries have adopted guidelines that relate sterility validity period of health care critical materials with the occurrence of related events. Research has validated the maintenance of sterility of the material during transportation and storage for long periods of time. The theoretical rationale has already refuted the old paradigm of sterility validity period based on the time clock/calendar (time-related) due to the efficiency of packaging with microbial barrier characteristics, hermetic seal and the theory of absence of spontaneous generation of micro-organisms. However, in Brazil, it can be stated that the Central Supply and Sterilization Departments still practice the control of sterilized materials based on arbitrarily assigned deadlines, this practice is encouraged by recommendation documents drawn up by official national entities of standardizing and supervising. In the belief that performing an experimental investigation in vitro, producing robust scientific evidence, would help in strengthening the breaking of the old paradigm, we proposed to evaluate the maintenance of sterility of materials stored for up to 6 months after the packages suffer intentional contamination of their external surfaces. The experiment consisted of packing the specimens that simulate surgical instrument (porcelain cylinders) in four different types of wrappers: cotton, crepe paper, nonwoven fabric (SMS) and paper-plastic pouches. All packages have previously been autoclaved at the same sterilization cycle. Subsequently, the external surfaces of the packages were intentionally manipulated with hands contaminated with Serratia marcescens (106 C.F.U./mL). After storage at predetermined intervals (0, 7, 14, 28, 90 and 180 days), a specific quantity of packages of each type of packaging has been opened and the samples were seeded directly in soybean-casein culture medium. The packages with samples with zero time of storage were considered as negative controls. In order to ensure the viability of the test micro-organism on the outer surface of the packages, cultivation of samples of each package contaminated with the test microorganism were made weekly up to 180 days, constituting the positive control group. For each interval of time of storage, 600 samples were analyzed (150 for each type of package). Considering the confidence interval for the exact binomial distribution, the sample size calculated in the study ensured a confidence interval of 95% with 0 to 0.006 probability of finding a contaminated sample. As result, there were no recovered test micro-organism Serratia marcescens in any of the time intervals of storage. Based on the results of this study, it was observed that packages wrapped in wrappers with microbial barrier properties and hermetically sealed are capable of protecting the sterile contents up to 6 months. This study contributes to the clinical practice in Central Supply and Sterilization Department, preventing unnecessary reprocessing of the material, while warns that the related events should be monitored and each material, before use, must be inspected for its integrity of the packaging and sealing.

Várias espécies de moscas-das-frutas têm sido alvo de investigações visando sua criação massal, incluindo Ceratitis capitata (Wiedemann) (mosca-do-Mediterrâneo ou medfly) e, mais recentemente, Anastrepha fraterculus (Wiedemann) (mosca-das-frutas sul americana), sendo as espécies de maior importância econômica e quarentenária em várias áreas produtoras de frutas do Brasil, Uruguai, Argentina e Peru. Um dos fatores determinantes para que a Técnica do Inseto Estéril (TIE) seja aplicada contra uma determinada praga é a existência de métodos de criação massal da espécie. Para se alcançar a criação em larga escala dessas espécies de moscas-das-frutas, é necessário, inicialmente, o preparo de uma dieta larval adequada. Como a dieta de criação pode oferecer uma quantidade enorme de substâncias nutritivas, as larvas criadas artificialmente amadurecem mais cedo do que as selvagens, o que acelera a sucessão de gerações. Essas mudanças detectadas durante o estágio larval podem formar adultos com maiores reservas nutritivas, atingindo a maturidade sexual mais cedo. Existem várias biofábricas de mosca-das-frutas no mundo capazes de produzir machos estéreis que são utilizados para a erradicação ou supressão populacional em área-ampla. Visando-se diminuir os custos de produção dos insetos, pesquisas voltadas para a otimização e desenvolvimento de novas dietas não podem parar. Portanto, os objetivos deste trabalho foram determinar a adequação e densidade larval ideal das moscas C. capitata e A. fraterculus em novas dietas à base de farinha de milho (FM), bagaço de cana-de-açúcar (BC), pó de cenoura liofilizada (PC) e carragenina (MC), escolhendo a dieta mais eficiente e viável economicamente para produção de cada uma dessas espécies com base na quantidade e qualidade de insetos produzidos e no custo de produção. As densidades larvais ideais para a criação de C. capitata nas dietas larvais FM, BC e PC foram, respectivamente, 0,5; 2,0; e 1,0 mL de ovos/kg de dieta. A dieta larval com maior produtividade foi a dieta BC, produzindo uma média de 6.514 pupas/kg de dieta, seguida das dietas FM e PC, que produziram, respectivamente, uma média de 5.018 e 4.135 pupas por kg de dieta. A ordem decrescente de viabilidade econômica entre as três dietas larvais para C. capitata foi a mesma (BC>FM>PC), sendo necessário um investimento médio de R$ 471,29; R$ 482,26; e R$ 1.085,85, respectivamente, para a produção de 1 milhão de pupas. Os resultados com A. fraterculus mostraram que as densidades larvais ideais nas dietas FM e MC foram de 2,0/kg de dieta e 1,5 mL de ovos/L de dieta, respectivamente. A dieta larval PC foi considerada inviável devido à baixa produtividade e peso de pupas. Em termos de produtividade, a dieta larval MC (produção média de 22.451 pupas/L de dieta), foi mais eficiente do que a FM (6.688 pupas/kg de dieta). Quanto à viabilidade econômica, a dieta larval MC foi mais vantajosa, demandando um investimento médio de R$ 312,67 para a produção de 1 milhão de pupas, enquanto que a dieta FM custou R$ 1.266,36 para produzir a mesma quantidade de pupas / Several species of fruit flies have been the subject of research for their mass rearing, including Ceratitis capitata (Wiedemann) (Mediterranean fruit fly or medfly) and, more recently, Anastrepha fraterculus (Wiedemann) (South American fruit fly), being these species the ones with most economic and quarantine importance in several fruit producing areas of Brazil, Uruguay, Argentina and Peru. One of the requirements for the Sterile Insect Technique (SIT) is the existence of mass rearing methods of the target species. In order to achieve the large-scale rearing of these species of fruit flies, it is necessary initially to develop a suitable larval diet. As the rearing diet can offer a huge amount of nutrients, artificially reared larvae mature earlier than wild ones, what accelerates the succession of generations. These changes during the larval stage may form adults with higher nutritional reserves and they can reach sexual maturity earlier. There are several mass-rearing facilities for fruit flies in the world capable of producing sterile males that are used for population eradication and suppression in area-wide. In order to reduce insect production costs, the optimization or development of new diets cannot stop. Therefore, the objectives of this study were to determine the adequacy and optimal larval density of C. capitata and A. fraterculus flies in new diets based on corn flour (FM), sugarcane bagasse (BC), lyophilized carrot powder (PC) and carrageenan (MC), choosing the most efficient and less costly diet for the production of each of the species, based on the quantity and quality of insects reared and production costs. The optimal larval densities for C. capitata in larval diets FM, BC and PC were, respectively, 0.5; 2.0 and 1.0 mL of eggs/kg of diet. The larval diet that reached the highest productivity was the BC diet, giving an average of 6,514 pupae/ kg of diet, followed by FM and PC diets, which produced an average of 5,018 and 4,115 pupae per kg of diet, respectively. The decreasing order of economic viability among the three larval diets used was the same (BC>FM>PC), requiring a mean investment of R$ 471.29; R$ 482.26; and R$ 1,085.85, respectively, for the production of 1 million pupae. The results with A. fraterculus showed that the ideal larval densities in FM and MC diets were 2.0 eggs/kg of diet and 1.5 mL of eggs/L of diet, respectively. The PC larval diet was considered infeasible based on the low production and weight of pupae. In terms of productivity, the MC larval diet, which resulted in an average of 22,451 pupae produced per L of diet, was more efficient than FM, which produced a mean of 6,688 pupae per kg of diet. Considering the economic costs, the MC larval diet was also superior, requiring R$ 312,67.00 for the production of 1 million pupae, while the FM diet required R$ 1,266.36 to produce the same amount of pupae

O controle microbiológico durante a produção de preparações farmacêuticas é de grande importância para garantir a qualidade do produto final, quanto às propriedades terapêuticas e de segurança ao paciente. O monitoramento ambiental é uma valiosa ferramenta como forma de mensurar a efetividade das medidas que integram a estratégia de controle de contaminação microbiana. Neste contexto, pouco destaque tem sido dado à manufatura de produtos farmacêuticos não-estéreis, por representarem as classes cujos riscos de contaminação microbiana são menores, quando comparados aos produtos parenterais. Dessa maneira, este estudo teve como objetivo caracterizar os isolados microbianos de amostras de ar ativo e passivo e de superfícies de áreas produtivas não-estéreis. Ainda, visou-se avaliar estatisticamente os dados de monitoramento ambiental, como base para o desenvolvimento de uma abordagem para determinação de limites de alerta e ação. Os resultados obtidos revelaram que a maioria dos microrganismos encontrados são de origem humana, seguidos por bactérias e fungos provenientes do solo. As diferenças sazonais foram observadas, principalmente, para a ocorrência de fungos, mais prevalentes no período seco. Foi desenvolvida uma abordagem estatística baseada em (1) determinação de subgrupos racionais, (2) avaliação da distribuição estatística e (3) determinação de limites, utilizando, como critério, o índice de capacidade do processo (Cpk). Um melhor entendimento do perfil microbiano das áreas produtivas e a determinação de limites de acordo com a distribuição real dos dados levará à destinação dos recursos necessários a ações que visem a qualidade do produto e a segurança do paciente. / The microbiological control during the production of pharmaceutical preparations is of great importance for quality assurance of the final product regarding to therapeutic properties and patient safety. Environmental monitoring is a valuable tool to measure the effectiveness of the actions that integrate the microbial contamination control strategy. In this context, little attention has been given to the manufacture of non-sterile pharmaceutical products, because they represent classes whose microbial contamination risks are lower when compared to parenteral products. Considering this scenario, this study aimed to characterize microbial isolates from surfaces, active and passive air sampling of non-sterile manufacturing areas. Furthermore, it was expected to statistically evaluate the environmental monitoring data, as a basis for the development of an approach for determining alert and action limits. The results showed that most of the microorganisms found are from human source, followed by bacteria and fungi typically found in the soil. The seasonal differences were mainly observed for fungi recovery, which were more prevalent in the dry period. A statistical approach was developed based on (1) the determination of rational subgroups, (2) evaluation of the statistical distribution and (3) limit determination, using the process capacity index (Cpk) as criteria. A better understanding of the typical manufacturing areas microbial profile and the determination of limits according to the actual data distribution will lead to the allocation of the necessary resources to actions focusing on product quality and patient safety.