Studies were conducted on the effects of alcohol and opioids on the development of forensically important flies. In addition different methods of degree-day calculations and development thresholds were used to determine the effects on PMI estimates. The first study determined the effects of ethanol on the development of Phormia regina in vitro. Ground pork loin was treated with a 1, 5, or 10% ethanol solution to give an equivalent Blood Alcohol Concentration (BAC) of 0.01, 0.04, and 0.8 % w/v. A significant difference in the time for second instars to complete the stage was seen between the 1% treated and control. Significant differences were also found among pupal and adult weights between all treatments and the control. A significant difference was shown between growth curves of the 5% treated and control for third instar larvae using the Kolmogorov-Smirnov test. The ethanol content of Phormia regina in migrating third instar larvae that fed on treated meat was examined using headspace-gas chromatography (HSGC). All larvae had a content of 0.01% w/v, including the control. The effects of ante-mortem injection of oxycodone in pigs were examined with respect to insect succession patterns and the development rates of Phormia regina. Pigs were given a subcutaneous injection of oxycodone hydrochloride (3 mg/kg by weight) and antemortem blood samples were collected prior to and following drug injection. Shortly after death the carcasses were placed at an open field site and allowed to decompose in a field cage. Insect samples were collected from carcasses for seven days post-mortem and the collected data were used to develop occurrence matrices. The Simple Matching Coefficient showed that successional patterns were similar between treated and untreated animals. Loin and liver from the carcasses were used as rearing media for in vitro development studies of Phormia regina. Kolmogorov-Smirnov test showed that third instar P. regina maggots from treated loin tissue were significantly longer in length than maggots feeding on untreated loin tissue. A significant difference in time was found among larvae on loin for the time from eclosion to completion of the second instar. Significant differences were seen in the weight of adults reared on liver and loin. A chi-square for homogeneity showed that adults were biased towards males (2:1) from untreated loin tissue. A final study compared weather data sources, Accumulated Degree-Day (ADD) methods, and postmortem interval (PMI) estimations based on threshold and developmental data source. Four pigs were used for statistical comparisons. Pigs were taken to a test site and allowed to decompose in an experimental cage. Probes recorded ambient temperatures and body temperatures. Maggot sampling was completed every day for each pig. A three way factorial linear fit model was used to test for statistical differences. Significant differences were seen in the calculated ADD based on probe location and the development threshold used. The ADD calculated from local weather station locations: Kentland Farm, Moore Farm, and Blacksburg Airport were also compared. A significant difference in ADD was found in the main effects among locations (Airport 44.1 ADD, Kentland 37.5 ADD, Moore 48.6 ADD), as well as among the thresholds used (10,12.2, and 14°C). Different PMI estimations also resulted when using development data from different development studies on Phormia regina. / Ph. D.

The liver is one of the most frequently injured abdominal organs in motor vehicle collisions. Although previous studies have quantified the tensile failure properties of human liver parenchyma at 48hrs postmortem, it is currently unknown how the material properties change between time of death and 48hrs postmortem. Therefore, the purpose of this study was to quantify the effects of postmortem degradation on the tensile material properties of bovine liver parenchyma when stored in DMEM or saline. Fourteen fresh bovine livers were obtained from a local slaughter house and stored in either DMEM or saline as large blocks, small blocks, or slices of tissue. Multiple parenchyma dog-bone samples from each liver were tested once to failure at three time points: ~6hrs, ~24hrs, and ~48hrs postmortem. The data were then analyzed to determine if there were significant changes in the material properties with respect to postmortem time. The results showed that the failure strain decreased significantly between 6hrs and 48hrs after death when stored as large blocks in saline. Conversely, neither the failure stress nor failure strain changed significantly with respect to postmortem time when stored as large blocks in DMEM. The modulus did not significantly change for tissue stored as large blocks in either fluid. Preliminary results indicated that reducing the tissue storage size had a negative effect on the material properties and cellular architecture. Overall, this study illustrated that the effects of postmortem liver degradation varied with respect to the preservation fluid, storage time, and storage block size. / MS

Fresh turkey meat color is influenced by a myriad of biological factors which include muscle fiber type composition and heme protein concentrations. These factors either contribute to or are subject to the biochemical events involved in the conversion of muscle to meat. Subtle deviations in the processing environment can also result in aberrant fresh meat quality development and may ultimately alter the quality characteristics of cooked product. Our objective was to describe the underlying cause and significance of two-toning in fresh turkey breast. In the first experiment, pectoralis muscles were collected and subjected to image processing software to describe color of fresh turkey. In the second experiment, shackling time was tested as an aggravator of fresh turkey color. Results showed turkey breast possess two-lobes that differ in pH, drip loss, energy metabolism and muscle fiber type composition. Results also showed fresh turkey color was enhanced during the time from stun to exsanguination (P < 0.05). These results suggest inherent differences in breast muscle are responsible for variations in fresh turkey color. / Master of Science

Os barbitúricos são fármacos com atividade depressora do sistema nervoso central e estão relacionados com elevados números de casos de intoxicações e uso não-médico em vários países. No Brasil, a droga antiepiléptica mais encontrada em casos de intoxicação é o fenobarbital, pois os pacientes relatam que \"essa é uma substância com ação forte no cérebro\". De fato, os barbitúricos estão altamente relacionados com tentativa de suicídio e homicídio. Nesses casos existe a necessidade da quantificação dessas substâncias para correlacionar com a causa mortis. No entanto, as análises toxicológicas postmortem são de difícil execução e interpretação, pois a concentração de agentes tóxicos encontrados é bastante complexa e afetada não só pela condição de deterioração do corpo, mas também por um processo conhecido como redistribuição postmortem. Em geral, concentrações mais elevadas são encontradas no sangue situado nos sítios centrais (como o sangue coletado da cavidade cardíaca) em comparação aos níveis verificados nos vasos periféricos (como a veia femoral). Em outros casos, o tempo entre a morte e o exame postmortem é suficiente para que algumas substâncias que normalmente estariam presentes no sangue não estejam mais disponíveis neste fluido biológico. Há ainda um agravante, pois não existem valores de referências para a maioria das amostras biológicas não-convencionais, dificultando assim a interpretação dos resultados. Os exames toxicológicos devem ser realizados em amostras biológicas e tem como objetivo a avaliação da intoxicação como circunstância qualificadora do delito, como causa de periculosidade ou imputabilidade. O objetivo deste trabalho foi o desenvolvimento e aplicação de métodos de identificação de barbitúricos (butalbital, secobarbital, pentobarbital e fenobarbital) em amostras postmortem (sangue cardíaco, sangue femoral e fígado). Os analitos foram extraídos das amostras utilizando a micro extração em fase líquida (LPME), identificados e quantificados por cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). Após o desenvolvimento e validação, os métodos analíticos foram aplicados em amostras postmortem de onze cadáveres necropsiados pelo Serviço de Verificação de Óbito da Cidade de São Paulo (SVO (SVO-USP), com suspeita de envolvimento de barbitúricos. Nove casos apresentaram resultado positivo para fenobarbital. A média da razão sangue femoral/sangue cardíaco foi de 0,91 com o desvio padrão de 0,23. Para a correlação fígado/sangue femoral a média foi de 1,17 com desvio padrão de 1,29. Os barbitúricos foram escolhidos como modelo de estudo devido à grande incidência de casos de intoxicação aguda com estes fármacos no Brasil. / Barbiturates are a class of drugs that act as central nervous system depressant and are associated with high numbers of poisoning cases and non-medical use in several countries. In Brazil, phenobarbital is the most related antiepileptic drug involved in intoxication cases. Patients report that \"this drug is a substance with strong action in the brain.\" In fact, barbiturates are highly related to attempted suicide and homicide cases, in which quantification of these substances to correlate with the possible cause of death is necessary. However, postmortem toxicological analyses are difficult to perform and interpret, because the concentration of toxic agents found is quite complex and affected not only by deterioration condition of the body but also by a process known as postmortem redistribution. In general, higher concentrations are found in the blood located in central sites (e.g. heart cavity) compared with the levels found in peripheral vessels (such as the femoral vein). In other cases, the time between death and postmortem examination is enough for some substances that would normally be present in the blood are no longer available in this biological fluid. Besides, there are few reference values for most non-conventional biological samples, making it difficult to interpret the results. The objective of this work was the development and application of methods for identification of barbiturates (butalbital, secobarbital, pentobarbital and phenobarbital) in postmortem samples (heart blood, femoral blood and liver). The analytes were extracted by using liquid-phase micro extraction (LPME) and quantified by gas chromatography-mass spectrometry (GC-MS). After the development and validation, analytical methods were applied in real cases of eleven corpses autopsied by Death Verification Service of São Paulo City (USP-SVO), with suspected of barbiturates involvement. Nine cases were positive for phenobarbital. The mean ratio of blood femoral / cardiac blood was 0.91 with a standard deviation of 0.23. For the correlation liver / femoral blood the average was 1.17 with a standard deviation of 1.29. Barbiturates were chosen as model for this study because the high incidence of cases of acute poisoning with these drugs in Brazil.

Os barbitúricos são fármacos com atividade depressora do sistema nervoso central e estão relacionados com elevados números de casos de intoxicações e uso não-médico em vários países. No Brasil, a droga antiepiléptica mais encontrada em casos de intoxicação é o fenobarbital, pois os pacientes relatam que \"essa é uma substância com ação forte no cérebro\". De fato, os barbitúricos estão altamente relacionados com tentativa de suicídio e homicídio. Nesses casos existe a necessidade da quantificação dessas substâncias para correlacionar com a causa mortis. No entanto, as análises toxicológicas postmortem são de difícil execução e interpretação, pois a concentração de agentes tóxicos encontrados é bastante complexa e afetada não só pela condição de deterioração do corpo, mas também por um processo conhecido como redistribuição postmortem. Em geral, concentrações mais elevadas são encontradas no sangue situado nos sítios centrais (como o sangue coletado da cavidade cardíaca) em comparação aos níveis verificados nos vasos periféricos (como a veia femoral). Em outros casos, o tempo entre a morte e o exame postmortem é suficiente para que algumas substâncias que normalmente estariam presentes no sangue não estejam mais disponíveis neste fluido biológico. Há ainda um agravante, pois não existem valores de referências para a maioria das amostras biológicas não-convencionais, dificultando assim a interpretação dos resultados. Os exames toxicológicos devem ser realizados em amostras biológicas e tem como objetivo a avaliação da intoxicação como circunstância qualificadora do delito, como causa de periculosidade ou imputabilidade. O objetivo deste trabalho foi o desenvolvimento e aplicação de métodos de identificação de barbitúricos (butalbital, secobarbital, pentobarbital e fenobarbital) em amostras postmortem (sangue cardíaco, sangue femoral e fígado). Os analitos foram extraídos das amostras utilizando a micro extração em fase líquida (LPME), identificados e quantificados por cromatografia em fase gasosa acoplada à espectrometria de massas (GC-MS). Após o desenvolvimento e validação, os métodos analíticos foram aplicados em amostras postmortem de onze cadáveres necropsiados pelo Serviço de Verificação de Óbito da Cidade de São Paulo (SVO (SVO-USP), com suspeita de envolvimento de barbitúricos. Nove casos apresentaram resultado positivo para fenobarbital. A média da razão sangue femoral/sangue cardíaco foi de 0,91 com o desvio padrão de 0,23. Para a correlação fígado/sangue femoral a média foi de 1,17 com desvio padrão de 1,29. Os barbitúricos foram escolhidos como modelo de estudo devido à grande incidência de casos de intoxicação aguda com estes fármacos no Brasil. / Barbiturates are a class of drugs that act as central nervous system depressant and are associated with high numbers of poisoning cases and non-medical use in several countries. In Brazil, phenobarbital is the most related antiepileptic drug involved in intoxication cases. Patients report that \"this drug is a substance with strong action in the brain.\" In fact, barbiturates are highly related to attempted suicide and homicide cases, in which quantification of these substances to correlate with the possible cause of death is necessary. However, postmortem toxicological analyses are difficult to perform and interpret, because the concentration of toxic agents found is quite complex and affected not only by deterioration condition of the body but also by a process known as postmortem redistribution. In general, higher concentrations are found in the blood located in central sites (e.g. heart cavity) compared with the levels found in peripheral vessels (such as the femoral vein). In other cases, the time between death and postmortem examination is enough for some substances that would normally be present in the blood are no longer available in this biological fluid. Besides, there are few reference values for most non-conventional biological samples, making it difficult to interpret the results. The objective of this work was the development and application of methods for identification of barbiturates (butalbital, secobarbital, pentobarbital and phenobarbital) in postmortem samples (heart blood, femoral blood and liver). The analytes were extracted by using liquid-phase micro extraction (LPME) and quantified by gas chromatography-mass spectrometry (GC-MS). After the development and validation, analytical methods were applied in real cases of eleven corpses autopsied by Death Verification Service of São Paulo City (USP-SVO), with suspected of barbiturates involvement. Nine cases were positive for phenobarbital. The mean ratio of blood femoral / cardiac blood was 0.91 with a standard deviation of 0.23. For the correlation liver / femoral blood the average was 1.17 with a standard deviation of 1.29. Barbiturates were chosen as model for this study because the high incidence of cases of acute poisoning with these drugs in Brazil.

[Truncated abstract] A common application of forensic entomology is the estimation of post-mortem interval (PMI). This is most frequently estimated from the age of calliphorid specimens collected from a corpse, and in many cases it is the immature stages that are encountered. A critical step in the estimation of PMI is the accurate identification of insects to species level, with misidentification potentially resulting in the application of unsuitable developmental data and therefore inaccuracy in the resulting estimate. Identification has long been attempted on a morphological basis, but complicated by the lack of larval keys to the Calliphoridae, limited diagnostic features in immature stages and the poor preservation of specimens. Standard practice in forensic entomology is the rearing of immatures collected from the corpse through to the more distinctive adult stages, however this process is time-consuming and may be hindered where specimens die during rearing. Furthermore, many cases are presented for forensic entomologist as an afterthought and specimens are already preserved. Consequently, a new approach to the identification of calliphorids is sought which will overcome the problems of the morphological and rearing methods. ... The culmination of this study is the consideration of applications of molecular data to forensic entomology. A sequence-specific priming (SSP) technique is presented for the identification of the forensically significant calliphorids of Australia and New Zealand, along with a new method for the extraction and storage of calliphorid DNA samples using Whatman FTA cards. These techniques will potentially improve the efficiency and accuracy of identification in the estimation of PMI using calliphorids. The use of calliphorid DNA is not limited to PMI estimation, but may also be applied to museum studies. DNA was extracted from pupal casings from 300 year old mummified corpses, however difficulty was encountered in amplifying the DNA reproducibly. This illustrates however, the wide-ranging implications of the calliphorid sequence data gathered in this study. This thesis makes a significant contribution to the consideration of the status of some global calliphorid species. The new technique presented for identification of Australian and New Zealand species is the culmination of an important body of data that will ultimately contribute to the strong foundation of forensic entomology and our future accuracy, efficiency and utility as a routine investigative tool.

Thesis (M.A.)--Texas State University--San Marcos, 2009. / Vita. Appendices: leaves 40-61. Includes bibliographical references (leaves 62-66). Also available on microfilm.

Thesis (M.A.) -- University of Tennessee, Knoxville, 2005. / Title from title page screen (viewed on Feb. 2, 2006). Thesis advisor: Lee Meadows Jantz. Vita. Includes bibliographical references.

Thesis (M.S.)--Oregon State University, 2006. / Printout. Includes bibliographical references. Also available via the World Wide Web.

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.