Orientador: Arício Xavier Linhares / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-19T03:28:23Z (GMT). No. of bitstreams: 1 Stoppa_LailaSilvaLeoneldeAvila_M.pdf: 2902278 bytes, checksum: 52b99920b1d3a2f1d669e4cff58d53fe (MD5) Previous issue date: 2011 / Resumo: A entomologia forense utiliza principalmente dados de insetos necrófagos para estimar o intervalo pós-morte (IPM). A oviposição destes insetos pode ser influenciada por vários fatores, tais como se o corpo está envolto por algum material, se está enterrado, carbonizado, em ambiente aberto ou fechado, podendo assim alterar o cálculo do IPM. O propósito deste trabalho foi estudar as diferenças na exposição de carcaças abrigadas e desabrigadas, no processo de decomposição, na atratividade e oviposição pelos artrópodes e na sucessão entomológica. A influência do modo de exposição das carcaças em relação à abundância das espécies que as coloniza também foi reportado. Os experimentos foram realizados em duas estações do ano, quente e úmida e na seca e fria. Foram utilizadas três carcaças de suínos (Sus scrofa L.), fêmeas de aproximadamente 10kg, em cada estação. A fim de se evitar a predação por animais de grande porte, as carcaças foram colocadas em gaiolas de ferro e expostas em um arboreto, nas dependências da UNICAMP, situado entre os paralelos 22°48'57"S, 47°03'33"W, 640 metros de altitude. Uma carcaça ficou desabrigada (controle) e as outras duas (réplicas) dentro de barracas de camping de dimensões 2,05m x 1,45m x 1,00m, abrigadas, uma na sombra e outra no sol. Os insetos adultos alados foram coletados com puçá e os imaturos direto da bandeja de cada carcaça, todos os dias no mesmo horário. Amostras de solo também foram coletadas no local do experimento ao redor de cada carçaca. Foi obtido um total de 37.787 espécimes pertencentes ao Filo Arthropoda, distribuídos em 11 diferentes ordens, sendo que desse total 8.842 foram coletados e 28.945 foram criados, ou seja, os imaturos foram levados ao laboratório e mantidos em câmaras de criação (±26°C) até sua emergência. Dos coletados, 92,2% pertencem à ordem Diptera, onde se destacaram as famílias Calliphoridae (28,7%) e Muscidae (34,6%). Outras ordens coletadas foram Coleoptera (4,5%), Hymenoptera (1,9%), Lepidoptera (0,5%) e Hemiptera (0,3%). Foram coletados menos de 1% de artrópodes ocasionais pertencentes às ordens Blattodea, Mantodea e Neuroptera, além de Acari, Opiliones e Araneae. Dos criados, 99,8% pertenciam a ordem Diptera onde se destacaram as famílias Calliphoridae (93,2%) e Muscidae (5,4%). A ordem Coleoptera foi responsável por 0,2% restantes. Considerando-se diferentes tipos de exposição, observou-se diferença no tempo de decomposição, modo e duração de cada estágio entre as carcaças, sendo que na carcaça controle a decomposição foi muito mais rápida e obteve maior abundância de artrópodes que nas carcaças abrigadas, embora a composição de espécies ter sido similar em todas elas. Fatores abióticos exerceram influência retardando ou acelerando a atividade dos insetos necrófagos nas carcaças. Chrysomya albiceps foi à espécie mais abundante e juntamente com Hemilucilia segmentaria e Peckia (Euboetcheria) collusor podem ser considerados indicadores forenses para estimativa de IPM, enquanto Hemilucilia segmentaria e Hemilucilia semidiaphana podem ser usadas como indicador forense para região de mata, modo de exposição e estação fria e seca. Tais resultados demonstram a importância da utilização da entomologia forense como ferramenta nas investigações legais / Abstract: Forensic Entomology uses data, mainly from carrion insects, to estimate the Post Mortem Interval (PMI). The oviposition pattern of these insects can be influenced by several factors, such as the presence of materials in the vicinity of the body, whether the corpses are buried, burned or disposed in open or closed environments, etc. These factors can affect the PMI estimate. The purpose of this work was to study the differences in decomposition, attractiveness and oviposition patterns between carcasses exposed in sheltered and open environments. The influence of the mode of exposure in relation to the abundance of the species that colonized were also been studied. The experiments were conducted in two seasons: warm and wet (summer), and dry and cool (winter). Three carcasses of female pigs (Sus scrofa L.), weighing aproximatelly 10 kg were used per season. The carcasses were placed in iron cages in order to avoid predation by large animals, and exposed in an arboretum located on the State University of Campinas campus, (22 ° 48'57 "S 47 ° 03'33" W), at 640 m above sea level. One pig was placed in and unsheltered area (control) and the other two were exposed inside camping tents measuring 2,05 m x 1,45 m x 1,00 m. One tent was set in a shaded area and the other one was places under direct sunlight. Adult insects visiting the carcasses were collected with an insect net. The immatures leaving the carcasses to pupate were collected in trays placed beneath each carcass, taken to the laboratory, reared at 26 °C in growth chambers until adult emergence, and identified. Soil samples were also collected at the site of the experiment around each carcass. A total of 37.787 specimens of Arthropoda was obtanined, distributed in 11 different orders. Of this total, 8.842 were collected and 28.945 were reared from the collected larvae. Diptera totaled 92.2% of the adults collected (Calliphoridae-28.7% and Muscidae-34.6%). The other collected orders were: Coleoptera (4.5%), Hymenoptera (1.9%), Lepidoptera (0.5%) and Hemiptera (0.3%). Les than 1% of occasional arthropods belonging to the orders Blattodea, Mantodea and Neuroptera, and Acari, Opiliones and Araneae were also collected. More than 99% of the reared insects belonged to the order Diptera, Calliphoridae (93.2%) and Muscidae (5.4%) were the most abundant families. The order Coleoptera accounted for the remaining 0.2%. Considering the different types of exposure, there was a difference in the decomposition time, mode and duration of each decomposition stage of the carcasses. The decomposition process in the control carcasses was much faster and had greater abundance of arthropods in the sheltered carcasses, but species composition was similar. Abiotic factors exerted influence by delaying or accelerating the activity of insects in the carcasses. Chrysomya albiceps was the most abundant species and, together with Hemilucilia segmentaria and Peckia (Euboetcheria) collusor can be considered important indicators in the estimation of PMI, while Hemilucilia semidiaphana and Hemilucilia segmentaria can be also used as indicators for wooded areas, mode of exposure and season. These results evidence the importance of using forensic entomology as a tool in legal investigations / Mestrado / Parasitologia / Mestre em Parasitologia

This study was part of a larger study to develop an animal production model to obtain the optimum beef tenderness. There is a variety of different beef production systems being used in South Africa. The production systems investigated in this study were; animals reared on pasture until A age classification group (AP), animals reared on pasture until AB age classification group (ABP), animals reared on pasture until B age classification group (BP), animals reared at a feedlot until A age classification group (AF) and animals reared at a feedlot until AB age classification group (ABF). These production systems affect a wide range of components in the muscle that contribute to meat quality. Meat quality refers mainly to tenderness and colour. Scientists and industry role players are uncertain of which production system is the best to produce high quality meat. The aim of the study was to determine the effects of different production systems on the post slaughter muscle energy metabolism and the related effects on meat quality (tenderness and colour). In the experiment 180 steers of the following breed crosses were used: Nguni, Simmental and Brahman. These animals were reared until they reached the A, AB or B age classification, either on pasture or in a feedlot (B only on pasture). All the carcasses were electrically stimulated for approximately 15 sec, before analyses were done. The following properties were measured on the animals after slaughter: tenderness (Warner-Bratzler shear force), carcass pH, carcass temperature, sarcomere lengths, myofibrillar fragmentation length, calpain activity, muscle lactic acid concentration, muscle glycogen concentration, muscle ATP concentration, muscle glucose-6- phosphate concentration, muscle creatine phosphate concentration, muscle glucose concentration, water holding capacity, drip loss and meat colour. These measurements and determinations were done according to standard laboratory procedures at the ARC (Agricultural Research Council) at Irene. Breed had no effect on the muscle energy status for muscle glycolytic potential, muscle lactic acid concentration, muscle glucose concentration, muscle glycogen concentration, muscle glucose-6-phosphate concentration, muscle ATP concentration and creatine phosphate concentration (p>0.05). Older animals from the pasture had lower muscle energy levels than younger animals from the feedlot. Animals from the BP production system had the darkest colour meat with the highest hue angle. AF production system animals had the lightest colour meat with the lowest hue angle and the highest chroma. ABF production system animals had the lowest chroma. Animals from the ABF production system had the lowest shear force value at 1, 7 and 14 days post mortem and animalsfrom the AP production system had the highest shear force value at 1, 7 and 14 days post mortem. This study showed that the energy status in the muscle post mortem does not influence the tenderness of the meat nor the colour (p>0.05). Shear force had a weak to medium positive correlation with muscle pH (between 0.186 and 0.410) and a weak to medium negative correlation with muscle temperature (between -0.157 and -0.268) (p<0.05). Muscle lactic acid concentrations (between -0.033 and -0.322), muscle glucose concentrations (between -0.066 and -0.155) and sarcomere length (between -0.276 and -0.326) had a weak to medium negative correlation with shear force (p<0.05). Muscle glycogen concentrations (between 0.026 and 0.166) and myofibrillar fragment length (between 0.248 and 0.447) had a weak to medium positive correlation with shear force (p<0.05). Shear force had a weak positive correlation with calpastatin activity (between 0.064 and 0.253) and a weak to medium negative correlation with calpain I activity (between -0.183 and -0.313) (p<0.05). The ratio of calpastatin: calpain I (between 0.323 and 0.348) and the ratio of calpastatin: calpain I + II (between 0.183 and 0.275) had a weak to medium positive correlation with shear force (p<0.05). Breed had no effect on the muscle energy status for muscle glycolytic potential, muscle lactic acid concentration, muscle glucose concentration, muscle glycogen concentration, muscle glucose-6-phosphate concentration, muscle ATP concentration and creatine phosphate concentration (p>0.05). If electrical stimulation was not used in this study the difference between the production systems in terms of muscle energy status and colour would have been more prominent. The conclusion is that if animals are slaughtered under “ideal” circumstances in terms of stress being kept to a minimum before slaughter and the carcasses are electrically stimulated in order to prevent cold shortening, the production system shows a small effect on the energy status of the animal and consequently also levels out the meat quality characteristics such as tenderness and colour. For more dramatic results and academic value it would have been more useful to include more variations of non-ideal slaughter conditions and non-electrical stimulation, as well as more breeds. A follow up study with no electrical stimulation can be helpful to explain some uncertainties. This follow up study will present its own challenges, for example higher frequency of DFD. A follow up study on the effects of a larger variety of breeds can help to determine the exact effect ofmuscle energy metabolites in the different breeds, on the tenderness and colour of the meat. Copyright / Dissertation (MSc(Agric))--University of Pretoria, 2012. / Animal and Wildlife Sciences / unrestricted

Orientador: Patricia Jacqueline Thyssen / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-27T11:10:49Z (GMT). No. of bitstreams: 1 Alonso_MarcelaAquiyama_M.pdf: 1564049 bytes, checksum: 75bec9069782a8cb90070bdbf5ad0c93 (MD5) Previous issue date: 2015 / Resumo: Os insetos constituem o grupo mais diversificado e abundante do Reino Animal, com ampla diversidade morfológica, fisiológica e de hábitos alimentares e por isso podem ser encontrados em vários habitats e ecossistemas. Insetos necrófagos podem ser vestígios e fontes de informações de interesse forense, como para a estimativa do intervalo pós-morte (IPM), baseada, por exemplo, na idade dos imaturos que se criam em corpos em decomposição. O desenvolvimento desses insetos pode ser afetado pela variação de temperatura e presença de substâncias tóxicas nos tecidos de um cadáver, entre outros fatores. Chrysomya megacephala (F.) e Chrysomya putoria (W.) (Diptera: Calliphoridae), introduzidas no Brasil, são consideradas de importância forense, médico e veterinária, devido aos seus comportamentos sinantrópicos e necrófagos. No presente estudo foi avaliado o tempo de desenvolvimento de imaturos na fase embrionária e pós-embrionária de C. megacephala e C. putoria sob diferentes temperaturas e / ou presença de cloridrato de fluoxetina, um antidepressivo, em fígado de coelho, o substrato alimentar. A relação entre temperatura e desenvolvimento, na fase embrionária, foi similar entre ambas as espécies. O tempo de desenvolvimento dos ovos para C. megacephala variou aproximadamente de 64-7h a 13 e 35 °C, respectivamente, e para C. putoria de 69-8h a 13 e 35 °C, respectivamente. Houve eclosão de larvas a 13 °C, mas as mesmas não completaram o desenvolvimento. A temperatura e o cloreto de fluoxetina afetaram o desenvolvimento dos imaturos, na fase na pós-embrionária. Para ambas as espécies, as larvas do grupo controle completaram seu desenvolvimento 24h mais rápido que o grupo com fluoxetina a 17 °C, mas apresentaram o desenvolvimento 12h mais lento a 35 °C. Estudos considerando tempo real de desenvolvimento dos ovos e avaliando como a combinação de duas ou mais variáveis podem influenciar o desenvolvimento de insetos de interesse forense são de grande valia para aumentar a acurácia da estimativa do IPM / Abstract: Insects are the most diverse and abundant group of the Animal Kingdom, with great diversity of morphological, physiological and feeding habits and are found in nearly all habitats and ecosystems. Scavengers species can provide important information of forensic interest, as the post-mortem interval (PMI) estimate, based on, e.g., the age of the larvae reared in decomposing bodies. The development of these insects can be affected by temperature oscillation and presence of toxic substances, among other factors, on the rearing media. Chrysomya megacephala (F.) and Chrysomya putoria (W.) (Diptera: Calliphoridae), introduced in Brazil, are considered of forensic, medical and veterinary importance, due to their necrophagous and synanthropic behaviour. This study evaluated the developmental time of C. megacephala and C. putoria under different temperatures and / or with fluoxetine hydrochloride, an antidepressant drug, in rabbit liver, the rearing substrate. The relationship between temperature and development, on the embryonic phase, was similar for both species. Egg developmental time for C. megacephala was approximately of 64-7h at 13 and 35 °C, respectively, and for C. putoria was 69-8h at 13 and 35 °C, respectively. The larval hatching occurred at 13 °C, but, at this temperature, the larval development was not completed. Both temperature and fluoxetine hydrochloride, when present, affected the development of the larvae. For both species, the larvae of control group completed their development 24h faster than the fluoxetine hydrochloride group at 17 °C, but the development was 12h slower at 35 °C. Studies considering real egg developmental time and evaluating, simultaneously, the insects' response for two or more variables that might influence their development are of great value to increase the accuracy of PMI estimate / Mestrado / Relações Antrópicas, Meio Ambiente e Parasitologia / Mestra em Biologia Animal

No description available.

Estima-se atualmente que mais de 5% da população mundial vem fazendo uso recreativo de algum tipo de substância psicoativa, sendo que o direito a esse uso é tema recorrente da sociedade contemporânea. Por apresentar riscos associados à saúde e a segurança das populações, o uso abusivo dessas substâncias tem instigado a toxicologia social na busca de respostas, com as quais se possa caracterizar, analisar e gerenciar esses riscos. Drogas de grande consumo no Brasil são a anfetamina, cocaína e Cannabis sativa. Esta tese desenvolveu uma nova metodologia para detectar e quantificar anfetamina, cocaína e tetrahidrocanabinol em sangue total, com uso de microextração em fase líquida via fibra de polipropileno (HF-LPME), seguida de cromatografia gasosa acoplada a espectrometria de massa (GC-MS). Trata-se de uma técnica que apresenta vantagens sobre as tradicionais, uma vez que demanda quantidades menores de solvente orgânico, diminuindo riscos e custos de processo. Também propôs um estudo com a aplicação dos métodos em 69 amostras de sangue de vivos e de post mortem, as quais foram obtidas por convênio com a superintendência da polícia técnica científica de São Paulo (SPTC/SP). Os métodos desenvolvidos foram validados de acordo com diretrizes internacionais de interesse forense. Como resultado da validação, os métodos desenvolvidos se mostraram precisos e exatos para anfetamina e cocaína. O limite de detecção da cocaína foi de 5 ng . mL-1 e o limite de quantificação de 10 ng . mL-1. Quanto a anfetamina, os limites de detecção e de quantificação foram de 5 ng . mL-1. A técnica de HF-LPME não foi aplicável ao tetraidrocanabinol (&#916;9-THC). Como resultado da análise das amostras, 40% delas apresentaram resultados positivos para cocaína. Desses positivos, 35% foram oriundos das matrizes de sangue de vivos e 64% oriundos de sangue post mortem. Nenhuma delas apresentou resultado quantificável para anfetamina. / It is currently estimated that more than 5% of the world\'s population has been doing recreational use of some kind of psychoactive substances and the legal right to such use is a recurring theme debated by contemporary society. Due to the risks associated with populations health and safety, the abusive use of these substances has been instigating by social toxicology to search for answers to characterize, analyze and manage these risks. Drugs of great consumption in Brazil are, amphetamine cocaine and marijuana. This thesis proposes to develop a new methodology to detect and quantify psychoactive drugs in whole blood with the use of liquid phase microextraction by polypropylene fiber (HFLPME), followed by gas chromatography coupled to mass spectrometry (GC-MS). It is a technique that presents advantages compared with traditional ones, because of the smaller amounts demands of organic solvent, reducing risks and process costs. It also proposes a study with 69 blood samples taken from living persons and post mortem blood samples, which were obtained by agreement with the Superintendency of São Paulo Scientific Technical Police (SPTC / SP). The methods developed were validated according to international guidelines of forensic interest. As a result of the validation, the methods developed were precise and accurate for amphetamine and cocaine. The limit of cocaine detection was 5 ng . mL-1 and the limit of quantification was 10 ng . mL-1. As for amphetamine, the limits of detection and quantification were 5 ng . mL-1. The HF-LPME technique was not applicable to tetrahydrocannabinol (&#916;9-THC). As a result of the sample analysis, 40% of them presented positive results for cocaine. Of these, 35% were from blood samples taken from living persons and 64% from the post mortem blood samples. None of the samples presented quantifiable results for amphetamine.

O objetivo deste trabalho foi investigar a incidência de carnes DFD (dark, firm, dry) em um frigorífico comercial de frango e determinar as suas propriedades funcionais comparando com as do PSE (pale, soft, exudative) e Normal em filés de peito, Pectoralis major. Para a avaliação da ocorrência desses tipos de cores, as aves das linhagens Cobb e Ross receberam banho calmante, com aspersão de água em temperatura ambiente por aproximadamente 10 minutos, associado à ventilação, imediatamente antes do seu abate. Em seguida, foram sacrificadas e desossadas e os filés foram analisados ao tempo de 90min e 24h post mortem, para coleta de valores de pH e L* e classificados em carnes PSE, Normal e DFD, com base nos resultados de 24h post mortem (n=329). O resultado da incidência foi de 24,92, 73,55 e 1,52% para amostras PSE, Normal e DFD, respectivamente. O valor de L* ao tempo de 90 min post mortem foi de 51,57 (&#177;1,91), 48,55 (&#177;1,94) e 41,67 (&#177;1,80), respectivamente, enquanto para o pH foi de 6,10 (&#177;0,48), 6,31 (&#177;0,59) e 7,15 (&#177;0,43) para carnes PSE, Normal e DFD, respectivamente. Após 24h post-mortem os resultados foram para L* 54,32 (&#177;1,09), 50,56 (&#177;1,64), 42,23 (&#177;1,80) e para o pH 5,69 (&#177;0,14), 5,78 (&#177;0,14), 6,05 (&#177;0,43) respectivamente para amostras PSE, Normal e DFD, significativamente diferente (P<0,05). Para o estudo das propriedades funcionais, amostras (n=374) foram selecionadas nas linhas de produção, classificadas em carnes PSE, Normal e DFD de acordo com o L* e pH 24h post mortem. Foram realizadas as medidas do exsudado formado, perda de umidade durante o cozimento e estabilidade da emulsão. Os resultados encontrados na medida do exsudado foram 4,73 (&#177;2,61)%, ,55 (&#177;2,16)%, 1,11 (&#177;1,01)% para PSE, Normal e DFD respectivamente, significativamente diferentes (P<0,05). A medida da perda de umidade durante o cozimento para as amostras PSE foi de 26,03 (&#177;2,93)%, para o grupo Normal a quebra foi de 25,38 (&#177;3,96)% e para o grupo DFD 22,89 (&#177;2,65)%. Entre as amostras dos grupos PSE e Normal não ocorreu diferença significativa (P<0,05), apesar da leve tendência para menor quebra de peso para grupo considerado Normal, enquanto que a quebra de peso para o grupo Normal e DFD apresentou resultados significativamente diferentes (P<0,05) com diferença significativa de 2,50% para DFD. Finalmente, para a medida de instabilidade da emulsão os resultados foram 1,04 (&#177;0,32)%, 0,78 (&#177;0,20)%, 0,70 (&#177;0,12)%, respectivamente para PSE, Normal e DFD, significativamente diferente (P<0,05). Concluindo, as matérias-primas que apresentam características DFD tendem a conferir melhor qualidade funcional com melhor capacidade de absorção, retenção de umidade, baixo exsudado e baixa perda durante o cozimento. A constatação da diferença nas cores do filé na linha de processamento pode padronizar os processos contribuindo para a distribuição adequada das amostras de carne PSE, Normal e DFD especificamente para diferentes tipos de processamento trazendo benefícios econômicos às industrias e qualidade sensorial aos consumidores. / The objective of this work was to investigate the incidence of DFD (dark, firm, dry) meat, in a commercial chicken plant and to determine its functional properties in comparison to PSE (pale, soft, exudative) and Normal breast fillet, Pectoralis major. In order to evaluate the occurrence of different color types, commercial Cobb and Ross chickens were submitted to water shower and ventilation treatments during 10 minutes immediately before slaughtering. Birds (n=329) were sacrificed, deboned and analyzed after 90 minutes and 24h post mortem for pH and L* values evaluation and submitted to classification as PSE, Normal, DFD meat based upon 24h post mortem samples. The occurrence was 24.92, 73.55 and 1,52% for samples PSE, Normal and DFD meats, respectively. L* values for 90 minutes post mortem were 51.57 (&#177;1.91), 48.55 (&#177;1.94) and 41.67 (&#177;1.80), respectively, while for pH values were 6.10 (&#177;0.48), 6.31 (&#177;0.59) and 7.15 (&#177;0.43) for PSE, Normal e DFD meats, respectively. After 24h post-mortem, L* values results were 54.32 (&#177;1.09), 50.56 (&#177;1.64), 42.23 (&#177;1.80) for PSE, Normal and DFD meats, respectively, and pH values were 5.69 (&#177;0.14), 5.78 (&#177;0.14), 6.05 (&#177;0.43) respectively for PSE, Normal and DFD meats. All results were significantly different (P<0.05). For functional properties evaluation, samples (n=374) were selected from production lines and ranked as PSE, Normal e DFD meats according to L* and pH 24h post mortem. Measurement for formed exudate, cook drip loss and emulsion stability was carried out. Results for formed exudate were 4.73 (&#177;2.61), 2.55 (&#177;2.16), 1.11 (&#177;1.01)% for PSE, Normal e DFD meats, respectively and for cook drip loss for PSE 26.03 (&#177;2.93), for Normal 25.38 (&#177;3.96) and for DFD 22.89 (&#177;2.65)%, respectively. No significantly difference (P<0,05) was found between PSE and Normal samples although there was a tendency for lower weight loss for normal group while between normal and DFD samples a significantly difference (P<0.05) was found. Finally, for emulsion estability results were 1.04 (&#177;0.32), 0.78 (&#177;0.20), 0.70 (&#177;0.12)%, respectively for PSE, Normal and DFD samples, significantly different (P<0,05). In conclusion, broiler breast fillet meat which presents DFD characteristics tends to have better functional properties for water holding capacity, lower exudate formation and lower cook drip loss. The standardization of colour differences detection in broiler fillet meat in the processing line could direct these samples specifically to PSE, normal and DFD meats for processing meat products accordingly in order to have economical benefits to the poultry industries and particularly sensorial meat products improvement for consumers.

Estima-se atualmente que mais de 5% da população mundial vem fazendo uso recreativo de algum tipo de substância psicoativa, sendo que o direito a esse uso é tema recorrente da sociedade contemporânea. Por apresentar riscos associados à saúde e a segurança das populações, o uso abusivo dessas substâncias tem instigado a toxicologia social na busca de respostas, com as quais se possa caracterizar, analisar e gerenciar esses riscos. Drogas de grande consumo no Brasil são a anfetamina, cocaína e Cannabis sativa. Esta tese desenvolveu uma nova metodologia para detectar e quantificar anfetamina, cocaína e tetrahidrocanabinol em sangue total, com uso de microextração em fase líquida via fibra de polipropileno (HF-LPME), seguida de cromatografia gasosa acoplada a espectrometria de massa (GC-MS). Trata-se de uma técnica que apresenta vantagens sobre as tradicionais, uma vez que demanda quantidades menores de solvente orgânico, diminuindo riscos e custos de processo. Também propôs um estudo com a aplicação dos métodos em 69 amostras de sangue de vivos e de post mortem, as quais foram obtidas por convênio com a superintendência da polícia técnica científica de São Paulo (SPTC/SP). Os métodos desenvolvidos foram validados de acordo com diretrizes internacionais de interesse forense. Como resultado da validação, os métodos desenvolvidos se mostraram precisos e exatos para anfetamina e cocaína. O limite de detecção da cocaína foi de 5 ng . mL-1 e o limite de quantificação de 10 ng . mL-1. Quanto a anfetamina, os limites de detecção e de quantificação foram de 5 ng . mL-1. A técnica de HF-LPME não foi aplicável ao tetraidrocanabinol (&#916;9-THC). Como resultado da análise das amostras, 40% delas apresentaram resultados positivos para cocaína. Desses positivos, 35% foram oriundos das matrizes de sangue de vivos e 64% oriundos de sangue post mortem. Nenhuma delas apresentou resultado quantificável para anfetamina. / It is currently estimated that more than 5% of the world\'s population has been doing recreational use of some kind of psychoactive substances and the legal right to such use is a recurring theme debated by contemporary society. Due to the risks associated with populations health and safety, the abusive use of these substances has been instigating by social toxicology to search for answers to characterize, analyze and manage these risks. Drugs of great consumption in Brazil are, amphetamine cocaine and marijuana. This thesis proposes to develop a new methodology to detect and quantify psychoactive drugs in whole blood with the use of liquid phase microextraction by polypropylene fiber (HFLPME), followed by gas chromatography coupled to mass spectrometry (GC-MS). It is a technique that presents advantages compared with traditional ones, because of the smaller amounts demands of organic solvent, reducing risks and process costs. It also proposes a study with 69 blood samples taken from living persons and post mortem blood samples, which were obtained by agreement with the Superintendency of São Paulo Scientific Technical Police (SPTC / SP). The methods developed were validated according to international guidelines of forensic interest. As a result of the validation, the methods developed were precise and accurate for amphetamine and cocaine. The limit of cocaine detection was 5 ng . mL-1 and the limit of quantification was 10 ng . mL-1. As for amphetamine, the limits of detection and quantification were 5 ng . mL-1. The HF-LPME technique was not applicable to tetrahydrocannabinol (&#916;9-THC). As a result of the sample analysis, 40% of them presented positive results for cocaine. Of these, 35% were from blood samples taken from living persons and 64% from the post mortem blood samples. None of the samples presented quantifiable results for amphetamine.

L’estimation du délai post mortem est une étape fondamentale en anthropologie médico-légale. À ce jour, peu de méthodes précises et fiables existent. Les objectifs de notre travail étaient d’étudier le tissu osseux et son évolution dans les années et siècles suivant le décès en développant la Résonance Magnétique Nucléaire (RMN) des solides du carbone13C et du proton1H. Nous avons analysé une centaine d’os humains et animaux pour lesquels nous connaissions l’âge au décès, le sexe, la date de décès et les conditions de conservation. Nous avons caractérisé les os au niveau moléculaire en identifiant le collagène, les lipides et l’hydroxyapatite constitutifs du tissu osseux. Nous avons développé une méthode RMN permettant de distinguer des altérations de certains échantillons attestant de la présence d’adipocire au sein du tissu osseux, ou des dégradations sur des échantillons très anciens. L’étude de l’âge au décès et du sexe des sujets n’a pas mis en évidence une grande influence de ces facteurs sur les données RMN même si, pour des délais post mortem de 0 ou 1 an, les sujets féminins présentent quantitativement plus de lipides que les sujets masculins. L’analyse des conditions de conservation des individus montre un développement plus important d’adipocire pour les os laissés à l’air libre comparés aux os enterrés. Enfin, nous rapportons une décroissance quantitative du collagène et des lipides présents au sein du tissu osseux lorsque l’intervalle post mortem augmente. Cette décroissance est beaucoup plus rapide pour les lipides (quelques années) que pour le collagène (plusieurs millénaires) alors que l’hydroxyapatite présente une relative stabilité dans les premiers siècles suivant le décès. / The post mortem interval estimation is a fundamental step in forensic anthropology and up to now there are little accurate and reliable methods to do so. The objectives of our study were to investigate the bone composition and its evolution over years and centuries following the death by developing carbon 13C and proton 1H solid-state nuclear magnetic resonance (NMR). We analyzed about one hundred human and animal bones for which the age at death, sex, date of death and the storage conditions were known. Bones were characterized at the molecular level by identification of collagen, lipids and hydroxyapatite embedded in the bone matrix. We have designed a NMR-based method that allows determining alterations on some samples, evidencing the presence of adipocere (bone wax) within the bone, or finding bone tissue deterioration on some very old samples. Subject age at death and sex did not reveal significant changes on NMR data, except for post mortem interval ranging between 0 to 1 year, where female subjects had quantitatively more lipids in their bones than males. Storage conditions may promote a greater development of adipocere especially for bones left in the open air compared to those buried. Finally, we report a quantitative decrease of collagen and lipids present in the bone tissue when the post mortem interval increases. This decrease is much faster for lipids than for collagen where as the hydroxyapatite has a relative stability in the first centuries after the death. Decreases occur with very different time constants, ranging from years to millennia.

In this thesis we have addressed three different although related topics. First, we studied the post-mortem mutation damage rate of contaminated DNA sequences in ancient human remains focusing on the development of strategies to avoid pre-laboratory derived contaminations. We proposed a guideline to control them consisting in typing every single person involved on the manipulation of the remains, especially when they have not been excavated and washed under controlled conditions. Second, we successfully develop a non-invasive technique to sequence ancient remains but preserving it from the destruction. And third, we sequenced ancient human remains from different evolutionary times (from Paleolithic to post-Neolithic) to make inferences about the peopling of Western Europe focusing mainly in the Iberia peninsula. We found that there is a long term genetic continuity at least since the Neolithic. The only clear genetic discontinuity found is that involving two different human species, H. sapiens and H. neanderthalensis. / En la presente tesis hemos tratado tres temas diferentes aunque muy relacionados. Primero, hemos estudiado la tasa de mutación post-mortem de secuencias de ADN contaminante en restos humanos antiguos centrándonos en el desarrollo de estrategias para evitar que las muestras se contaminen antes de llegar al laboratorio. Proponemos una guía que consiste en el tipado genético de cada persona implicada en la manipulación de los restos, especialmente cuando estos han sido excavados y lavados bajo condiciones no controladas. Segundo, hemos desarrollado una técnica no invasiva para secuenciar DNA de restos humanos antiguos pero sin destruirlos. Y por ultimo, hemos secuenciado restos humanos antiguos pertenecientes a diferentes periodos evolutivos (desde el Paleolitico hasta el post-Neolitico) que nos han permitido hacer inferencias sobre el poblamiento Europeo centrándonos básicamente en la Península Ibérica. Hemos encontrado que ha habido una continuidad genética desde el Neolítico. La única clara discontinuidad genética encontrada es entre dos especies distintas: H. Sapiens y H.neanderthalensis.

O objetivo deste trabalho foi investigar a incidência de carnes DFD (dark, firm, dry) em um frigorífico comercial de frango e determinar as suas propriedades funcionais comparando com as do PSE (pale, soft, exudative) e Normal em filés de peito, Pectoralis major. Para a avaliação da ocorrência desses tipos de cores, as aves das linhagens Cobb e Ross receberam banho calmante, com aspersão de água em temperatura ambiente por aproximadamente 10 minutos, associado à ventilação, imediatamente antes do seu abate. Em seguida, foram sacrificadas e desossadas e os filés foram analisados ao tempo de 90min e 24h post mortem, para coleta de valores de pH e L* e classificados em carnes PSE, Normal e DFD, com base nos resultados de 24h post mortem (n=329). O resultado da incidência foi de 24,92, 73,55 e 1,52% para amostras PSE, Normal e DFD, respectivamente. O valor de L* ao tempo de 90 min post mortem foi de 51,57 (&#177;1,91), 48,55 (&#177;1,94) e 41,67 (&#177;1,80), respectivamente, enquanto para o pH foi de 6,10 (&#177;0,48), 6,31 (&#177;0,59) e 7,15 (&#177;0,43) para carnes PSE, Normal e DFD, respectivamente. Após 24h post-mortem os resultados foram para L* 54,32 (&#177;1,09), 50,56 (&#177;1,64), 42,23 (&#177;1,80) e para o pH 5,69 (&#177;0,14), 5,78 (&#177;0,14), 6,05 (&#177;0,43) respectivamente para amostras PSE, Normal e DFD, significativamente diferente (P<0,05). Para o estudo das propriedades funcionais, amostras (n=374) foram selecionadas nas linhas de produção, classificadas em carnes PSE, Normal e DFD de acordo com o L* e pH 24h post mortem. Foram realizadas as medidas do exsudado formado, perda de umidade durante o cozimento e estabilidade da emulsão. Os resultados encontrados na medida do exsudado foram 4,73 (&#177;2,61)%, ,55 (&#177;2,16)%, 1,11 (&#177;1,01)% para PSE, Normal e DFD respectivamente, significativamente diferentes (P<0,05). A medida da perda de umidade durante o cozimento para as amostras PSE foi de 26,03 (&#177;2,93)%, para o grupo Normal a quebra foi de 25,38 (&#177;3,96)% e para o grupo DFD 22,89 (&#177;2,65)%. Entre as amostras dos grupos PSE e Normal não ocorreu diferença significativa (P<0,05), apesar da leve tendência para menor quebra de peso para grupo considerado Normal, enquanto que a quebra de peso para o grupo Normal e DFD apresentou resultados significativamente diferentes (P<0,05) com diferença significativa de 2,50% para DFD. Finalmente, para a medida de instabilidade da emulsão os resultados foram 1,04 (&#177;0,32)%, 0,78 (&#177;0,20)%, 0,70 (&#177;0,12)%, respectivamente para PSE, Normal e DFD, significativamente diferente (P<0,05). Concluindo, as matérias-primas que apresentam características DFD tendem a conferir melhor qualidade funcional com melhor capacidade de absorção, retenção de umidade, baixo exsudado e baixa perda durante o cozimento. A constatação da diferença nas cores do filé na linha de processamento pode padronizar os processos contribuindo para a distribuição adequada das amostras de carne PSE, Normal e DFD especificamente para diferentes tipos de processamento trazendo benefícios econômicos às industrias e qualidade sensorial aos consumidores. / The objective of this work was to investigate the incidence of DFD (dark, firm, dry) meat, in a commercial chicken plant and to determine its functional properties in comparison to PSE (pale, soft, exudative) and Normal breast fillet, Pectoralis major. In order to evaluate the occurrence of different color types, commercial Cobb and Ross chickens were submitted to water shower and ventilation treatments during 10 minutes immediately before slaughtering. Birds (n=329) were sacrificed, deboned and analyzed after 90 minutes and 24h post mortem for pH and L* values evaluation and submitted to classification as PSE, Normal, DFD meat based upon 24h post mortem samples. The occurrence was 24.92, 73.55 and 1,52% for samples PSE, Normal and DFD meats, respectively. L* values for 90 minutes post mortem were 51.57 (&#177;1.91), 48.55 (&#177;1.94) and 41.67 (&#177;1.80), respectively, while for pH values were 6.10 (&#177;0.48), 6.31 (&#177;0.59) and 7.15 (&#177;0.43) for PSE, Normal e DFD meats, respectively. After 24h post-mortem, L* values results were 54.32 (&#177;1.09), 50.56 (&#177;1.64), 42.23 (&#177;1.80) for PSE, Normal and DFD meats, respectively, and pH values were 5.69 (&#177;0.14), 5.78 (&#177;0.14), 6.05 (&#177;0.43) respectively for PSE, Normal and DFD meats. All results were significantly different (P<0.05). For functional properties evaluation, samples (n=374) were selected from production lines and ranked as PSE, Normal e DFD meats according to L* and pH 24h post mortem. Measurement for formed exudate, cook drip loss and emulsion stability was carried out. Results for formed exudate were 4.73 (&#177;2.61), 2.55 (&#177;2.16), 1.11 (&#177;1.01)% for PSE, Normal e DFD meats, respectively and for cook drip loss for PSE 26.03 (&#177;2.93), for Normal 25.38 (&#177;3.96) and for DFD 22.89 (&#177;2.65)%, respectively. No significantly difference (P<0,05) was found between PSE and Normal samples although there was a tendency for lower weight loss for normal group while between normal and DFD samples a significantly difference (P<0.05) was found. Finally, for emulsion estability results were 1.04 (&#177;0.32), 0.78 (&#177;0.20), 0.70 (&#177;0.12)%, respectively for PSE, Normal and DFD samples, significantly different (P<0,05). In conclusion, broiler breast fillet meat which presents DFD characteristics tends to have better functional properties for water holding capacity, lower exudate formation and lower cook drip loss. The standardization of colour differences detection in broiler fillet meat in the processing line could direct these samples specifically to PSE, normal and DFD meats for processing meat products accordingly in order to have economical benefits to the poultry industries and particularly sensorial meat products improvement for consumers.