Post mortem interval (PMI – the time elapsed since death and discovery) is important to medicolegal investigations. It helps to construct crucial time lines and assists with the identification of unknown persons by inclusion or exclusion of a suspect’s known movements. Accurate methodologies for establishing PMI are limited to about 48-hours. Such methods involve use of increasing levels of potassium in vitreous humour, and algor mortis. This study is two-fold. Firstly, it explores the biomolecular changes in degrading porcine cartilage buried in soil environments and its potential to determine PMI in the crucial two days to two months period. Trotters were interred in a number of graves at two distinct locations exhibiting dissimilar soil environments. Weekly disinterments (for 6 weeks) resulted in dissection for cartilage samples which were processed for protein immunoblot analyses and cell vitality assays. Results demonstrate that aggrecan, a major structural proteoglycan, produces high (230kDa) and low (38kDa) molecular weight cross-reactive polypeptides (CRPs) within cartilage extracellular matrix. The 230kDa CRP degrades in a reproducible manner irrespective of the different soil environments utilised. As PMI increases, aggrecan diminishes and degrades forming heterogeneous subpopulations with time. Immunodetection of aggrecan ceases when joint exposure to the soil environment occurs. At this time, aggrecan is metabolised by soil microbes. The molecular breakdown of cartilage proteoglycans has potential for use as a reliable indicator of PMI, irrespective of differing soil environments, beyond the 48-hours period. Likewise, vitality assays also demonstrated viable chondrocytes for as long as 35 PM days. The second component of this study examined the fungal activity associated with trotters buried below ground. Results indicate that fungal growth was considerably influenced by soil chemistry and changes in the environment. Fungal colonisation did not demonstrate temporal patterns of succession. The results of this study indicate that cartilage has the potential to prolong PMI determination well beyond the current 48- and 100-hour limitations posed by various other soft tissue methods. Moreover, the long-term post mortem viability of chondrocytes presents an opportunity to explore DNA extraction from these cells for the purpose of establishing a positive identification for unidentified remains. On the contrary, the growth and colonisation patterns of post putrefactive fungi in relation to decomposing porcine trotters proved to be futile for estimating PMI. Therefore, fungi may not be a suitable candidate for evaluating PMI during the early phase fungal activity.

Providing a presumptive identification of badly decomposed or skeletonized remains is the responsibility of a forensic anthropologist. An important component of identification is the estimation of a post-mortem interval (PMI) for the deceased. This information can: provide a window period for death, reduce the number of potential victims, exclude possible assailants and substantiate witness testimony. Due to a large number of open and relatively desolate fields in South Africa, human remains are frequently discovered in an advanced stage of decomposition. The aim of this study was to evaluate the usability of the method of Megyesi and associates (2005) in which Total Body Score (TBS) and Accumulated Degree-Days (ADD) were retrospectively applied to estimate the post-mortem interval (PMI). To achieve this, a longitudinal examination of quantitative variables, TBS and ADD, was conducted over a period of 8 months. This period included both summer and winter seasons. Scatter plots between TBS and PMI, and TBS and ADD were used to illustrate patterns in decomposition. Patterns of decay differed in winter and summer, with winter exhibiting distinct inactivity. Using Loglinear Random-effects Maximum Likelihood Regression, the r2 values for ADD (0.6227) and PMI (0.5503) for combined seasons were less than r2 values for separated seasons (ADD 0.7652; PMI 0.7677). In contrast to other studies, seasonality influenced the ADD model and PMI. Linear regression formulae for ADD and PMI as well as 95% confidence interval charts for TBS for ADD were developed. These equations, along with data from a local weather station, can be used to estimate the PMI with relative accuracy. AFRIKAANS : Verskaffing van 'n vermoedelike identifikasie van erg ontbinde of skeletale oorskot is die verantwoordelikheid van ‘n forensiese antropoloog. ‘n Belangrike deel van identifikasie is die beraming van ‘n post-mortem interval (PMI) vir die oorledene. Hierdie inligting verskaf 'n venster tydperk van dood, verminder die aantal potensiële slagoffers, sluit moontlike aanvallers uit en ondersteun getuienis. As gevolg van 'n groot aantal relatief verlate en oop velde in Suid-Afrika, word menslike oorskot dikwels aangetref in ‘n gevorderde stadium van ontbinding. Die doel van hierdie studie was om die bruikbaarheid van die metode van Megyesi en medewerkers (2005) wat gebruik maak van Totale Liggaams Telling (TLT) en Opgehoopte Graad-Dae (OGD) om die postmortem interval (PMI) te skat, terugwerkend te evalueer. Hiervoor was 'n longitudinale studie van kwantitatiewe veranderlikes, TBS en ADD, oor ‘n tydperk van 8 maande gedoen. Hierdie tydperk sluit beide somer en winter in. Verspreidingsgrafieke tussen TBS en PMI, en TBS en ADD is gebruik om patrone in ontbinding te illustreer. Ontbindingspatrone het verskil tussen winter en somer met duidelike onaktiwiteit in die winter. Logliniêre Tweekansige-effek Maksimum Waarskynlikheid Regressie was gebruik om die r2 waardes van die gekombineerde en geskeide seisoene te bepaal. The r2 waardes vir die OGD (0.6227) en PMI (0.5503) vir gekombineer seisoene was minder as die r2 waardes vir seisoene apart (OGD 0.7652; PMI 0.7677). In teenstelling met ander studies, het seisoenaliteit die OGD model en PMI beinvloed. Lineêre regressie formules vir OGD en PMI sowel as 95% vertrouensinterval kaarte vir TLT vir OGD was saamgestel. Hierdie formules saam met data vanaf ‘n plaaslike weerstasie kan gebruik word om die PMI met relatiewe akkuraatheid te skat. / Dissertation (MSc)--University of Pretoria, 2010. / Anatomy / unrestricted

Submitted by (edna.saturno@ufrpe.br) on 2016-10-11T13:34:12Z No. of bitstreams: 1 Geraldo Martins.pdf: 2758105 bytes, checksum: 4e586fcda70973bcd0b330ad69b0e52c (MD5) / Made available in DSpace on 2016-10-11T13:34:12Z (GMT). No. of bitstreams: 1 Geraldo Martins.pdf: 2758105 bytes, checksum: 4e586fcda70973bcd0b330ad69b0e52c (MD5) Previous issue date: 2013-02-21 / The objective of this study was to identify and analyze the abundance of dipteran family Calliphoridae present along the decomposing carcass of pigs in the Cariri, Paraiba State, Brazil, in two periods: rainy and dry. The study was conducted in Reserva Particular do Patrimônio Natural (RPPN) Fazenda Almas, in São José dos Cordeiros-PB. We used two pig carcasses (Sus scrofa) by period, protected from scavengers by a metal cage inside trap like "Shannon". Samples were collected daily, twice a day, for eleven days in both periods. The meteorological data of precipitation were obtained at the Centro de Previsão e Estudos Climáticos do INMET and temperature and relative humidity were obtained in situ using a digital hygrometer. The insects collected were placed in the laboratory of entomology of UFPB and identified as well. We collected 8.811 individuals of the family Calliphoridae, belonging to six species (Chrysomya albiceps (Wiedemann, 1819), Cochliomyia macellaria (Fabricius, 1775), Chrysomya putoria (Wiedemann, 1818), Chloroprocta idioidea (Robineau-Devoidy, 1830), Chrysomya megacephala (Fabricius, 1794) e Lucilia eximia (Wiedemann, 1818)), which were presented the in both the periods, although a relative abundance of species was higher in the rainy period (4.706). We measured the abundance of species per period, where C. albiceps was the most abundant in the dry period (3.244/36, 81%) and C. idioidea in the rainy period (2.860/32, 45%). There were five stages of decomposition: fresh, chromatic, emphysematous, coliquative and skeletonization stage. The emphysematous was the stage where we most collected Calliphoridae (3.923/44, 52%), followed by skeletonization (3.458/39, 25%), chromatic (726 / 8.24%), coliquative (627/7, 12%) and fresh (77/0, 87%). It was also found that the highest abundance of C. idioidea occurred on the highest elevation of relative humidity and rainfall. This study is the first survey of the region of Caatinga in Paraiba state using active collecting, these results will serve to enrich the data to local forensic entomology and will contribute to the understanding the fauna of the region carrion flies. / Objetivou-se com este estudo inventariar e analisar a abundancia de dípteros da família Calliphoridae presentes ao longo da decomposição da carcaça de suínos na região do Cariri, estado da Paraíba, Brasil, em dois períodos: chuvoso e seco. O estudo foi desenvolvido na Reserva Particular do Patrimônio Natural (RPPN) na Fazenda Almas, no município de São José dos Cordeiros-PB. Foram utilizadas duas carcaças de suíno (Sus scrofa) por período, protegidas de animais carniceiros por uma gaiola metálica no interior de armadilha do tipo “Shannon”. As coletas foram realizadas diariamente, duas vezes ao dia, durante onze dias em ambos os períodos. Os dados meteorológicos de precipitação foram obtidos no Centro de Previsão de Tempo e de Estudos Climáticos do INMET e a temperatura e umidade relativa do ar foram obtidas in loco utilizando-se um termohigrômetro digital. Os insetos coletados foram encaminhados mediante autorização ao laboratório de entomologia da UFPB e posteriormente identificados e parte deles depositada na coleção de referencia. Foram coletados 8.811 indivíduos da família Calliphoridae, pertencentes a seis espécies: Chrysomya albiceps (Wiedemann, 1819), Cochliomyia macellaria (Fabricius, 1775), Chrysomya putoria (Wiedemann, 1818), Chloroprocta idioidea (Robineau-Devoidy, 1830), Chrysomya megacephala (Fabricius, 1794) e Lucilia eximia (Wiedemann, 1818), as quais estiveram presentes em ambos os períodos, porém a abundância relativa de espécies foi maior no período chuvoso (4.706/53,4%). Verificou-se a abundância de algumas espécies por período, onde a C. albiceps foi a espécie mais abundante no período seco (3.244/36,81%) e C. idioidea no período chuvoso (2.860/32,45%). Observaram-se cinco fases de decomposição: fresca, cromática, enfisematosa, coliquativa e esqueletização. A enfisematosa foi a fase onde mais se coletou Calliphoridae (3.923/44,52%), seguida pela esqueletização (3.458/39,25%), cromática (726/8,24%), coliquativa (627/7,12%) e fresca (77/0,87%). Constatou-se também que a maior abundancia de C. idioidea ocorreu no dia de maior elevação da umidade relativa e precipitação. Este é o primeiro estudo de levantamento realizado com coleta ativa em região de Caatinga paraibana, cujos resultados permitirão enriquecer os dados da entomologia forense local e contribuirão para o entendimento da fauna de dípteros necrófagos da região.

Orientador: Arício Xavier Linhares / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T21:53:49Z (GMT). No. of bitstreams: 1 Proencio_VanessaMinhoto_M.pdf: 2983676 bytes, checksum: c8bd0965eae3ffccfd30f8656cf54e04 (MD5) Previous issue date: 2011 / Resumo: Para estimar o intervalo pós-morte (IPM) e determinar a provável causa mortis, a ciência forense pode utilizar várias técnicas, sendo um exemplo a avaliação das fases de decomposição de um corpo. Estas fases podem ser influenciadas por fatores extrínsecos como o clima (temperatura e umidade), injúrias no corpo e por fatores intrínsecos, isto é, condições fisiológicas e metabólicas do próprio organismo e também pelo modo como ocorreu a morte. Além desta técnica a ciência forense pode utilizar a Entomologia Forense, através do estudo da fauna cadavérica e sua possível sucessão, para auxiliar nas estimativas de IPM e determinar a causa mortis. O objetivo deste trabalho foi avaliar a influência da carbonização na evolução das fases de decomposição, atratividade e possível padrão sucessional dos insetos. Foram utilizadas quatro carcaças de suínos (Sus scrofa L.), sendo duas no período mais quente e úmido do ano e duas no período mais frio e seco. Para cada período uma carcaça foi queimada com gasolina até o nível 2 da escala de carbonização de Crow-Glassman, sem que fosse totalmente destruída, e a outra, utilizada como controle. As carcaças foram expostas em área natural de mata e protegidas por gaiolas para evitar a ação de predadores. Imagens das fases de decomposição foram obtidas diariamente. Insetos imaturos foram coletados das carcaças e levados ao laboratório para posterior emergência dos adultos, contagem e identificação. Insetos adultos foram coletados utilizando uma rede entomológica e identificados no laboratório. Embora em alguns momentos ambas as carcaças estivessem na mesma fase de decomposição, houve um retardo na decomposição da carcaça carbonizada de 3 dias a partir do 5º estágio de decomposição em relação à carcaça controle na estação quente. Na estação fria este fato foi observado a partir do 3º estágio de decomposição, com uma média de atraso de 9 dias, sugerindo que a carbonização influencia no tempo de decomposição. Foi observada uma sucessão entomológica em ambas as carcaças (carbonizada e controle), apresentando um padrão diferente em cada estação do ano e relacionado ao modo de morte. A colonização por artrópodes mostrou-se diferente em ambas as carcaças demonstrando a influência da carbonização na atratividade e frequência de insetos, sugerindo que a espécie Hemilucilia semidiaphana pode ser considerada indicador forense para modo de morte, para região de mata e na estimativa de IPM, enquanto que a espécie Hemilucilia segmentaria pode ser usada como indicador forense para região de mata e na estimativa de IPM. Foi observado também que Chrysomya albiceps pode ser usada para a estimativa de IPM. Chrysomya putoria e Lucilia eximia podem ser utilizadas como indicadores forenses para modo de morte e estimativa de IPM. Espécies da família Sarcophagidae podem ser usadas como indicadores forenses para modo de morte e para a estimativa de IPM. Temperatura e umidade relativa do ar exerceram influência na atividade dos insetos e sobre a decomposição, retardando ou acelerando o processo. Este estudo mostra a importância da causa mortis, dos fatores abióticos, do tempo de decomposição e da entomofauna para o auxílio na estimativa do IPM / Abstract: To estimate the postmortem interval (PMI) and to determine the probable cause of death in forensic science several techniques can be used, such as the assessment of the stage of decomposition of a body. These stages may be affected by extrinsic factors such as meteorological conditions (temperature and humidity) and injuries on the body, and by intrinsic factors related to physiological and metabolic conditions of the body, and also by the circumstances of death. In addition, the study of the cadaveric arthropod fauna as well as its successional pattern may help to estimate the PMI and to determine the cause of death. The aim of this study was to assess: 1) the influence of carbonization in the carcass decomposition process; 2) the attractiveness exerted by the burned carcass to necrophagous insects, and 3) the successional pattern of necrophagous insects in the carcasses. Four pig (Sus scrofa L.), carcasses were used in the experiments: two in the hot and humid period of the year (summer) and two during the cold and dry period of the year (winter). For each period, one carcass was burned with gasoline up to level 2 of Crow-Glassman scale (CGS) and another, was used as control. The carcasses were exposed in a natural wooded area, and placed in wire cages tp prevent the access of large scavengers. Pictures of the stages of decomposition were taken daily. Maggots were collected daily from the carcasses and brought to the laboratory and kept until adult emergence, counted and identified. Adults were collected by using an entomological net and identified in the laboratory. Although both, control and burned carcasses, were at the same decomposition stage, there was a delay in the decomposition of the burned carcass of 3 days from the 5th stage of decomposition in relation to the carcass control in the hot season. In this cold season was observed from the 3rd stage of decomposition, with an average delay of 9 days suggesting that the carbonization influence the time of decomposition. Insect sucession was observed in both carcasses (burned and control), showing a different pattern in each season and related to the mode of death.The colonization by arthropods was different in both carcasses, showing an influence of burning in the attractiveness and occurrence of some insects, suggesting that the blowfly Hemilucilia semidiaphana, may be used as forensic indicator for the mode of death, wooded areas and estimates of PMI, while Hemilucilia segmentaria may be used as forensic indicator for wooded areas and estimates of PMI. It was also observed that Chrysomya albiceps may be used as forensic indicator to estimates of PMI. Chrysomya putoria and Lucilia eximia may be used as forensic indicators for mode of death and estimates of IPM. Species of the family Sarcophagidae may be used as forensic indicators for mode of death and to estimate of PMI. Temperature and relative humidity also had an important influence on the decomposition process, slowing or accelerating the process on both substrates, as well as influencing the activity of insects. This study reported the important influence exerted by the cause of death and the abiotic factors, on the time of decay and on the insect fauna. Therefore, these factors have to be taken into consideration for the determination of the PMI / Mestrado / Parasitologia / Mestre em Parasitologia

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.

Die Geruchswahrnehmung von Insekten ist an die spezifischen Anforderungen der Ökosysteme angepasst, in denen sie leben. Von Insekten wahrgenommene Volatile können Informationen über den Ort eines geeigneten Ovipositionsplatzes oder den physiologischen Zustand einer Wirtspflanze geben. Da der Geruchssinn der Insekten sich seit Millionen von Jahren an die Wahrnehmung bestimmter Volatile angepasst hat, können die wahrgenommenen Volatile als Markerstoffe angesehen werden, die zuverlässig einen Ökosystemzustand angeben können. Die Identifikation dieser Markerzustände und die Evaluation ihrer Nützlichkeit für spurenanalytische Anwendungen ist das Ziel dieser Arbeit. Es wurden drei Themen ausgewählt um die Verwendungsmöglichkeit des Insektengeruchssinns für Sensoranwendungen zu überprüfen: Fleischfrische, Todeszeitbestimmung (post mortem Intervall, PMI) und frühzeitige Feuererkennung. Spurenanalytik (GC-MS), Elektrophysiologie (EAG, GC-MS/EAD), Verhaltensversuche und Feldtests wurden angewandt, um die chemoökologische Interaktion der Schmeißfliege Calliphora vicina und der „Feuerkäfer“ Melanophila cuspidata, Merimna atrata and Acanthocnemus nigricans mit ihrer natürlichen Umgebung zu untersuchen. Die Resultate aus diesen Methoden erlaubten die Selektion von Volatilen, die drei Kriteria erfüllen: hohe Quantität, zuverlässige Emission und die ausschliessliche Emission von der untersuchten und keiner anderen Quelle. Diese drei Auswahlkriterien wurden aufgestellt um zu gewährleisten, dass die ausgewählten Markervolatile nicht nur zuverlässig mit dem physiologischen Status der Substrate (Alter von verderbendem Fleisch, PMI, Temperatur von erhitzten Spänen) korrelieren, sondern auch den Ansprüchen technischen Sensorsysteme genügen. Im Falle des alternden Fleisches unter warmen und trockenen Bedingungen ist Nonanal ein Zeiger für die korrekte Reifung des Fleisches zu Schinken. Dimethyl Trisulfid, Phenol und Indol zeigen das Verderben des Fleisches unter warmen und feuchten Bedingungen an. Bei niedrigen Temperaturen sind zunehmende 2,3 Butandiol Emission und abnehmende Nonanal Emission ein Zeiger für zunehmendes Verderben des Fleisches, sowohl unter trockenen als auch feuchten Bedingungen. Allerdings wurde 2,3 Butandiol nicht von C. vicina wahrgenommen, da die Fliege während der Vegetationszeit aktiv ist und unter diesen Bedingungen Dimethyl Trisulfid, Phenol und Indol emittiert werden. Nonanal, Hexanal, Dimethyl Disulfid, Dimethyl Trisulfid, Butan-1-ol und Phenol wurden als nützliche Volatile zur Eingrenzung der Leichenliegezeit ausgewählt. Die genannten Aldehyde sind typisch für die frühen Stadien der Verwesung, gefolgt von Butan-1-ol und den Schwefelsulfiden. Phenol wird hauptsächlich in den späteren Verwesungsstadien emittiert. Allerdings werden Phenol und 1-Butanol nicht von der Fliege wahrgenommen, da diese ein Generalist ist und nicht zwischen dem geblähten und aktivem Stadium der Verwesung unterscheiden muss. Daher wird zusätzlich die Geruchswahrnehmung des Speckkäfers Dermestes maculans betrachtet. Die Untersuchung anderer necrophager Insekten können weitere Einblicke in die Zeitabhängigkeit der Volatilemissionen während der Vertebratenverwesung geben. Die im Rahmen dieser Arbeit gewonnenen Ergebnisse erlauben eine Korrelation zwischen Emissionen und Leichenliegezeit und können eine Grundlage für eine volatilenbasierte post mortem Zeitbestimmung für Polizeikräfte sein. Terpene, aliphatische Aldehyde, Furfural und Methoxyphenole werden von erhitztem Holz emittiert. Komponenten aus all diesen Stoffklassen werden von M. cuspidata und M. atrata wahrgenommen, da diese Käfer den Erhitzungszustand von Baumstämmen nach einem Feuer erkennen können, um einen geeigneten Ovipositionsplatz zu finden. A. nigricans nimmt vor allem Methoxyphenole wahr, wohingegen M. atrata und M. acuminata Furfural zur Orientierung verwenden. Diese Unterschiede komplementieren das bekannte Verhalten dieser Käfer in ihrer natürlichen Umgebung, da A. nigricans auf verkohlten Stämmen, die große Mengen von Methoxyphenolen emittieren, seine Eier ablegt, und M. atrata und M. cuspidata auf Stämmen ihre Eier ablegen, die vom Feuer zwar äußerlich verkohlt, aber innerlich nur erhitzt wurden und daher Furfural emittieren. Halbleitergassensoren, die Furfural mit einer ausreichenden Selektivität vermessen können, können Holzbrände schon während ansteigender Temperatur und damit vor der Entzündung detektieren. Derartige Sensoren können sowohl als neuartige Frühwarnsysteme für die holzverarbeitende Industrie als auch für Waldbrandwarnsysteme oder den Haushalt dienen.