<p>Forensic science has gained popularity in the last few decades. Many new techniques are being studied and implemented. It includes a wide range of scientific disciplines, such as entomology, physics and biology. An important improvement to forensic science is the development of different DNA techniques which are implemented during an investigation, making DNA a gold standard for forensics. Most of the time DNA is mentioned it is in terms of human DNA, but there are microscopic organisms with useful DNA. In the last decade, with the development of next-generation sequencing (NGS), studies focusing on bacterial communities have been published, but fungal communities have not been extensively studied.</p> For this project, the potential of fungi in forensic science was investigated through three different studies. Human flora was looked at by NGS from thirty-seven human bodies with differing post-mortem intervals (PMIs). The communities were analyzed statistically and quantitatively, resulting in unique operational taxonomic units (OTUs) and genera which were only present in certain PMIs, and in some which were present through the entire PMI time scale. These attributes can help, not only to give a better view on human mycoflora during decomposition, but they can also help in determining fungal signatures during decomposition. These signatures can help in a PMI determination. Moreover, swine carcasses – the model animals for human forensic studies – were investigated as well to create a checklist of fungal flora after five months of winter decomposition in the West Lafayette, Indiana area. Furthermore, due to the increased importance of wildlife forensics, a wildlife study was also conducted using four wildlife species (mute swan, red tailed hawk, river otter, bobcat). The fungal flora from these species were compared within species at the beginning of the study and at skeletonization stage to look at any indicator fungal species and to create a general checklist for wildlife studies in the West Lafayette, Indiana area for future studies. Additionally, the fungal communities were compared across species as well.

Several studies were designed to investigate effect of cold winter transportation on broiler meat quality and physiological parameters. The first study assessed the effect of microclimate environment during winter transportation (-27 to +11ºC) from farm to the processing plant (3-4 h) on core body temperature (CBT) and breast meat quality parameters of 522 broiler chickens using an actively ventilated transport vehicle. Temperatures below freezing during transportation resulted in significantly (P < 0.05) higher ultimate pH (pHu) and darker color breast meat, which led to 8% incidence of dark, firm and dry (DFD) breast meat (pH > 6.1; L*< 46).<p> The effects of extreme cold conditions (-18 to -4ºC) along with age (5 and 6 wk), gender and lairage (0 and 2 h) on physiology parameters, breast muscle metabolites and meat quality of 360 birds using a simulated transport system were investigated. Five and 6 wk birds exposed to temperatures below -8 and -14ºC, respectively, showed significantly (P < 0.05) lower CBT and blood glucose and higher live shrink with breast meat that was darker in color, higher in pHu, water holding and water binding capacity compared to breast meat of birds exposed to warmer temperatures. A high incidence of DFD breast meat (> 50%) was observed for 5 and 6 wk birds exposed to temperatures below -8 and -14ºC respectively. Lairage following acute cold exposure caused an increase of up to 20% in DFD breast meat following exposure of the birds to temperatures below -8ºC.<p> Characteristics of cold-induced DFD breast meat were investigated in broiler chickens. DFD breast meat was significantly darker, redder, and less yellow in color and had higher pH, water binding capacity, processing cook yield and lower thaw and cook loss compared to normal meat. However, no significant difference in initial energy reserves was observed between DFD and normal breast meat.<p> Effect of extreme cold-stress (-15 to -9ºC) was further assessed on thigh meat quality and incidence of DFD from 160 male broilers. A greater impact of cold stress was observed on muscle metabolites and pH of thigh compared to breast meat, with 85% DFD incidence in thigh compared to 42% in breast meat of cold-stressed birds.<p> Biochemical basis of cold-induced DFD breast and thigh meat was investigated by measuring muscle metabolites at different times post-mortem. Lack of energy reserves at slaughter clearly explained the incidence of DFD thigh meat, but other factors might contribute to DFD breast meat aside from glycogen reserve at slaughter. However, the factors leading to DFD breast meat still remain unresolved and association of post-mortem enzyme activity and development of DFD breast meat merits further investigation.

Several studies were designed to investigate effect of cold winter transportation on broiler meat quality and physiological parameters. The first study assessed the effect of microclimate environment during winter transportation (-27 to +11ºC) from farm to the processing plant (3-4 h) on core body temperature (CBT) and breast meat quality parameters of 522 broiler chickens using an actively ventilated transport vehicle. Temperatures below freezing during transportation resulted in significantly (P < 0.05) higher ultimate pH (pHu) and darker color breast meat, which led to 8% incidence of dark, firm and dry (DFD) breast meat (pH > 6.1; L*< 46).<p> The effects of extreme cold conditions (-18 to -4ºC) along with age (5 and 6 wk), gender and lairage (0 and 2 h) on physiology parameters, breast muscle metabolites and meat quality of 360 birds using a simulated transport system were investigated. Five and 6 wk birds exposed to temperatures below -8 and -14ºC, respectively, showed significantly (P < 0.05) lower CBT and blood glucose and higher live shrink with breast meat that was darker in color, higher in pHu, water holding and water binding capacity compared to breast meat of birds exposed to warmer temperatures. A high incidence of DFD breast meat (> 50%) was observed for 5 and 6 wk birds exposed to temperatures below -8 and -14ºC respectively. Lairage following acute cold exposure caused an increase of up to 20% in DFD breast meat following exposure of the birds to temperatures below -8ºC.<p> Characteristics of cold-induced DFD breast meat were investigated in broiler chickens. DFD breast meat was significantly darker, redder, and less yellow in color and had higher pH, water binding capacity, processing cook yield and lower thaw and cook loss compared to normal meat. However, no significant difference in initial energy reserves was observed between DFD and normal breast meat.<p> Effect of extreme cold-stress (-15 to -9ºC) was further assessed on thigh meat quality and incidence of DFD from 160 male broilers. A greater impact of cold stress was observed on muscle metabolites and pH of thigh compared to breast meat, with 85% DFD incidence in thigh compared to 42% in breast meat of cold-stressed birds.<p> Biochemical basis of cold-induced DFD breast and thigh meat was investigated by measuring muscle metabolites at different times post-mortem. Lack of energy reserves at slaughter clearly explained the incidence of DFD thigh meat, but other factors might contribute to DFD breast meat aside from glycogen reserve at slaughter. However, the factors leading to DFD breast meat still remain unresolved and association of post-mortem enzyme activity and development of DFD breast meat merits further investigation.

Water and lipid-soluble meat flavour precursors are gradually formed post-mortem via biochemical reactions. Storage time and temperature can affect final flavour precursor concentrations which in turn will affect the sensory quality of cooked meat. Selected key flavour precursors were monitored in Bison bison longissimus dorsi muscles from six animals stored at 2, 4, 8, 15 and 21 days at 4°C, in order to evaluate the effect of post-mortem conditioning on the formation of flavour precursors. Results were correlated with sensory data obtained using quantitative descriptive analysis with 8 trained panelists. While lipid-soluble flavour precursors remained mostly unchanged, significant increases (P<0.05) in concentrations of water-soluble flavour precursors including reducing sugars (eg. ribose, xylose), free amino acids (eg. valine, leucine) and adenosine-5’-triphosphate (ATP) degradation products (eg. inosine and hypoxanthine) were obtained with chilled storage conditioning post-mortem. The overall balance and correlations of water-soluble flavour precursors with storage day 15 and 21 were reported and can potentially impact the eating quality of cooked bison meat.

Water and lipid-soluble meat flavour precursors are gradually formed post-mortem via biochemical reactions. Storage time and temperature can affect final flavour precursor concentrations which in turn will affect the sensory quality of cooked meat. Selected key flavour precursors were monitored in Bison bison longissimus dorsi muscles from six animals stored at 2, 4, 8, 15 and 21 days at 4°C, in order to evaluate the effect of post-mortem conditioning on the formation of flavour precursors. Results were correlated with sensory data obtained using quantitative descriptive analysis with 8 trained panelists. While lipid-soluble flavour precursors remained mostly unchanged, significant increases (P<0.05) in concentrations of water-soluble flavour precursors including reducing sugars (eg. ribose, xylose), free amino acids (eg. valine, leucine) and adenosine-5’-triphosphate (ATP) degradation products (eg. inosine and hypoxanthine) were obtained with chilled storage conditioning post-mortem. The overall balance and correlations of water-soluble flavour precursors with storage day 15 and 21 were reported and can potentially impact the eating quality of cooked bison meat.

TCC (graduação) - Universidade Federal de Santa Catarina. Centro de Ciências Jurídicas. Direito. / Submitted by Ana Sofia de Souza Fabre null (ana.sofia.fabre@grad.ufsc.br) on 2014-12-02T04:29:45Z No. of bitstreams: 1 Ana_Sofia_Fabre_tcc.pdf: 859923 bytes, checksum: 049a1c81e6ad155c0453ba1337a31baa (MD5) / Approved for entry into archive by Josiane Rose Petry null (josiane.rp@ufsc.br) on 2014-12-02T11:59:26Z (GMT) No. of bitstreams: 1 Ana_Sofia_Fabre_tcc.pdf: 859923 bytes, checksum: 049a1c81e6ad155c0453ba1337a31baa (MD5) / Made available in DSpace on 2014-12-02T11:59:26Z (GMT). No. of bitstreams: 1 Ana_Sofia_Fabre_tcc.pdf: 859923 bytes, checksum: 049a1c81e6ad155c0453ba1337a31baa (MD5) / A Medicina e a tecnologia avançam de tal forma que muitas vezes vão além do alcance do Direito. Prova disso são as técnicas de reprodução assistida, que hoje permitem a concretização da inseminação artificial homóloga post mortem, uma forma de manter vivos os planos familiares iniciais de um casal cujo cônjuge veio a falecer. Numa análise de todos os direitos dos envolvidos, sejam eles o direito da viúva ao material genético do de cujus, o direito de filiação da criança que será concebida e a própria abertura de sucessão para essa criança, entende-se que, quando o Direito apresenta lacunas, é preciso encontrar formas diferentes, porém justas, de se lidar com os casos concretos, mesmo que excepcionais. Sopesando-se os princípios constitucionais da dignidade da pessoa humana, da legalidade, da liberdade, da isonomia de tratamento entre os filhos e da proteção ao melhor interesse da criança, juntamente com o direito fundamental à herança e suas normas de funcionamento, e a presunção de paternidade, presentes no Código Civil brasileiro, conclui-se que, não só há o direito de essa criança ser concebida, como ainda ela possuirá tudo aquilo a que tem direito, como se nascida por qualquer outra forma de reprodução fosse. Afinal, de todos os entes envolvidos na questão, é ela, a criança, o que merece a maior proteção e os menores ônus possíveis.

Made available in DSpace on 2014-06-12T15:03:00Z (GMT). No. of bitstreams: 2 arquivo1599_1.pdf: 10160665 bytes, checksum: 18093a04cd5f95193e064e8b75b921af (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010 / Universidade Federal de Pernambuco / O resultado de óbitos mal-definidos torna indispensável à necropsia para que seja elucidada a causa morte. Dessa forma, este estudo visou verificar infecções fúngicas isoladas ou em associação com outras doenças relacionadas ao óbito, bem como dados epidemiológicos que possam ter contribuído. Durante a necropsia, foram realizados exames micológicos e histopatológicos. Os órgãos acometidos por fungos foram pulmões, cérebro, rim, meninges, fígado e coração, sendo diagnosticadas post mortem aspergilose, candidíase, fusariose e trichosporonose. Os indivíduos eram de ambos os sexos, com idade variando de 29 a 91 anos e desempenhavam variadas ocupações. Diabetes mellitus, tuberculose, esquistossomose, cardiopatia, hipertensão intracraniana, neoplasia cerebral, insuficiência renal foram doenças de base associadas às infecções fúngicas. Os dados refletem a dificuldade diagnóstica e terapêutica dessas infecções e confirmam a importância da necropsia

La consommation excessive d’alcool présente des risques élevés pour l’individu et pour la société ; elle est fréquemment associée à une augmentation du risque d’accidents, d’actes de violence, et peut également conduire à court et/ou à long terme à de graves maladies et à des problèmes sociaux. Dès lors, l’utilisation de marqueurs fiables permettant de détecter une consommation excessive d’alcool, ponctuelle ou chronique, s’avère nécessaire pour prévenir des conséquences néfastes de l’abus d’alcool. L’éthylglucuronide (EtG) est un marqueur d’alcoolisation utilisé en toxicologie clinique (alcoologie) et médicolégale. Par rapport aux marqueurs indirects d’alcoolisation (CDT, &#947;-GT), ce métabolite mineur de l’éthanol est très spécifique et est quantifiable dans diverses matrices biologiques. La production d’EtG est catalysée par des enzymes de la famille des UDP-glucuronosyl-transférases (UGT). Cependant, les UGT impliquées dans la glucuronoconjugaison de l'éthanol, ainsi que les sources potentielles de variabilité interindividuelle de la production d'EtG, sont encore mal connues. Nos travaux ont ainsi consisté à (1) développer et valider une méthode de dosage de l’EtG dans différentes matrices biologiques par chromatographie en phase gazeuse couplée à la spectrométrie de masse en tandem, (2) identifier les UGT humaines impliquées dans la glucuronoconjugaison de l’éthanol et étudier la contribution relative de chaque isoforme active au niveau hépatique, (3) étudier l’impact de substances fréquemment utilisées par les consommateurs d’alcool sur la production d’EtG in vitro, (4) étudier l’impact de polymorphismes génétiques fonctionnels des UGT sur la production hépatique d’EtG, et enfin (5) évaluer l’impact de la consommation de cannabis et d’autres drogues sur la production d’EtG à l’aide de prélèvements post-mortem. Ces travaux ont notamment permis de montrer que (1) l'éthanol est glucuronoconjugué principalement par le foie, puis dans une moindre mesure par les reins et par l'intestin, (2) les UGT1A9 et 2B7 sont les deux enzymes majoritairement impliquées dans la glucuronoconjugaison de l’éthanol, quel que soit l’organe considéré, (3) la morphine, la codéine, la nicotine et la cotinine n’entraînent aucune modification des taux de production d’EtG in vitro ; le lorazépam et l'oxazépam augmentent légèrement cette production (p = 0,2 et 0,065, respectivement) ; le cannabidiol inhibe la glucuronoconjugaison de l’éthanol par un mécanisme non-compétitif (CI50 = 1,17 mg/L; Ki = 3,1 mg/L), alors que le cannabinol augmente cette glucuroconjugaison de manière concentration-dépendante (p <0,05), (4) les SNP c.-900G>A affectant l’UGT2B7 et IVS1+399T>C affectant l’UGT1A9 augmentent légèrement la production d’EtG in vitro. Enfin (5) le rapport des concentrations sanguines EtG/éthanol apparaît significativement plus élevé chez des co-consommateurs de cannabis et/ou d’autres drogues que chez des consommateurs d’alcool seul. L’ensemble de ces résultats démontre l’existence de plusieurs facteurs pouvant potentiellement influencer la production d’EtG et devraient donc être pris en considération lors de l’interprétation de sa concentration in vivo. / Alcohol abuse is frequently associated with an increased risk of road accidents and violence, and can also lead to serious social and health problems. Therefore, the use of reliable markers to detect excessive punctual and/or chronic consumption of alcohol is necessary to prevent the harmful consequences of alcohol abuse. Ethylglucuronide (EtG) has been proposed as a marker of alcohol consumption in a variety of clinical and forensic contexts. Compared with the indirect markers (e.g. CDT, &#947;-GT), this minor metabolite of ethanol is very sensitive and specific, and is quantifiable in various biological matrices. It is formed by conjugation of ethanol with uridine 5’-diphosphate glucuronic acid (UDP-GA) via the action of UDP-glucuronosyltransferase (UGT) enzymes. However, the knowledge of the UGTs involved in the glucuronidation of ethanol, and the potential sources of the interindividual variability of EtG production are still not clearly established. The aims of our work were (1) to develop and validate a method for the determination of EtG in different biological matrices by gas chromatography coupled with tandem mass spectrometry, (2) to identify the human UGT isoforms involved in the glucuronidation of ethanol, and then to evaluate qualitatively and quantitatively their specific contribution in the formation of EtG, (3) to study the impact of the co-administration of drugs frequently used by ethanol consumers on the in vitro production of EtG, (4) to study the impact of functional genetic polymorphisms of two UGTs on the hepatic production of EtG, and finally (5) to study the impact of the consumption of cannabis and other drugs on the production of EtG using post-mortem samples. The main results of our study showed that (1) ethanol is primarily glucuronidated by the liver and, to a lesser extent, by kidneys, (2) UGT1A9 and 2B7 were identified as the main human UGTs involved in ethanol glucuronidation, (3) morphine, codeine, nicotine, and cotinine did not modify EtG in vitro formation rate; lorazepam and oxazepam produced a minor, but not significant, increase of EtG formation. Only cannabinol and cannabidiol significantly affected ethanol glucuronidation; cannabinol significantly increased the glucuronidation of ethanol in a concentration-dependent manner, whereas cannabidiol inhibited the glucuronoconjugaison of ethanol by a non-competitive mechanism (CI50 = 1.17 mg / L; Ki = 3.1 mg / L), (4) the SNP c.-900G>A and IVS1+399T>C affecting UGT2B7 and UGT1A9, respectively, seem to increase the in vitro production of EtG, and (5) cannabis and/or drugs consumption (mainly opioids, benzodiazepines, and paracetamol) seem to be associated with ratios of blood concentrations of EtG/ethanol significantly higher than those observed among only alcohol consumers. Taken together, these results show the existence of several factors that could potentially influence the production of EtG, and that should be taken into account when interpreting its concentration in vivo.

This study was conducted to investigate the causes and the effects of breed, gender and season on carcass condemnations post-mortem in a large South African abattoir. Condemnation of carcasses as a results of animal diseases and conditions has been identified as a problem in the South African beef industry, but the causes and extent of carcass condemnations have not been studied in detail. The importance of cattle management during transportation and pre-slaughter is generally appreciated, but a better understanding of the influence of breed, gender and season on the prevalence of carcass condemnations can help to improve slaughter management at large abattoirs. Diseases and conditions evaluated during this study were parafilaria, bruising, soiling, fever, peritonitis and pleuritis, abscesses, measles, oedema and intramuscular haemorrhage. Breeds of cattle slaughtered were Friesland, Bonsmara, Brahman, Angus, Afrikander, Hereford and Nguni cattle, grouped as males and females, and the seasonal effects investigated were confined to Autumn, Winter and Summer combined with Spring as it was not easy to distiquish between the two. Data was collected in a Grade A abattoir in South Africa in 2010. The experimental design involved 42 combinations based on their effects and interactions namely 7 breeds x 2 genders x 3 seasons. The model used in this procedure was based on the presence of conditions = (Intercept) x i x j x k where i~breed, j~gender and k~season. The binary response variable was the presence (value = 1) or absence (value = 2) of the respective condition. Overall, the significant order and extent of causes of bovine carcass condemnations evaluated in this particular abattoir during the period of study were peritonitis and pleuritis at 2.49%, soiling 2.33%, bruising 2.10%, parafilaria 0.54%, while other conditions had a negligible effect. Almost half of these carcass condemnations were due to soiling and bruising, which can be addressed by implementing better abattoir management and better management during transportation. Peritonitis and pleuritis were prevalent to a greater degree in Autumn, Soiling in Summer, bruising in winter and parafilaria more prevalent in Summer as well. Bonsmara bulls were more affected by peritonitis and pleuritis, soiling, bruising and parafilari compared to other breeds.. Overall breed, gender and season influenced carcass condemnations in large South African abattoir It is suggested that proper dehorning should be done at an early stage, especially in bulls of breeds like Bonsmara, Nguni, Friesland and Hereford, which predispose cattle to bruising which increases the number of carcass condemnations. Steps should be taken (i.e. proper handling techniques should be applied) to avoid injuries which may lead to internal bleeding, and eventually bruises (most probably due to fighting during transportation or in lairage), that can be avoided by using improved and upgraded facilities which include enough space per animal. In case of soiling, well-trained slaughter personnel, cattle handlers and cleaners should be hired to prevent bile and soil contamination during evisceration and to avoid mud or manure contamination durinmg lairage. It is recommended that employees be trained to help reduce the high rate of carcass condemnations. Breeds like Bonsmara and Brahman were more susceptible to peritonitis, pleuritis and parafilaria, so treatrment programmes are advisable for these breeds, especially those from regions where these conditions or diseases are more prevalent. / Dissertation (MSc Agric)--University of Pretoria, 2013. / gm2014 / Animal and Wildlife Sciences / unrestricted

The question of post-mortem interval (PMI) or time since death is often the most sought after piece of information associated with a medical death investigation. Based on the observation that DNA degradation disproportionately affects the analysis of larger genetic loci, it was proposed that DNA degradation, as a result of autolysis or putrefaction, could prove suitable as a potential rate-of-change indicator of PMI. Nine randomly amplified polymorphic DNA (RAPD) analysis primers and three sets of directed amplification primers were evaluated to determine their suitability for use in assessing the degree of DNA fragmentation in tissue samples. They were assessed for amplicon specificity, total DNA target sensitivity, allele monomorphism and the observance of degradation-based profile changes. Markers meeting the requisite criteria were then used to assess a range samples degraded under controlled and uncontrolled conditions. Tissue samples collected from seven domestic pigs (Sus scrofa) were incubated under controlled laboratory or uncontrolled field conditions to produce samples simulating those potentially collected in a forensic case. DNA samples isolated from these specimens were then analyzed at those loci which had been determined to meet the requisite criteria. Collectively, data generated from these analyses indicate that genetic profiles generated by this approach can provide information useful for estimating the post-mortem interval, with the locus and amplicons used being most useful during the first 72 hours after death.