Dissertationen zum Thema „DNA fingerprints“

Tandem-repetitive hypervariable minisatellites detected in a DNA fingerprint provide highly informative genetic markers. To identify and localize specific loci represented in a DNA fingerprint, it is necessary to clone individual minisatellites. This thesis is concerned with the characterization of single locus minisatellite probes cloned from DNA fingerprints. Seven single locus human minisatellite probes have been cloned by screening ? libraries with DNA fingerprint probes 33.6 and 33.15. Each locus consists of a minisatellite, with repeat units ranging in length from 9 to 47 base pairs depending on the locus. These autosomal loci are amongst the most variable loci characterized to date. The heterozygosity values of D1S7, D1S8, D5S43, D7S21, D7S22 and D12S11 range from 85% to >99%. Clustering of minisatellites was initially detected at the D12S11 locus. This observation led to the subsequent discovery of minisatellites showing close physical linkage as well as a tendency for minisatellites to be localized in proterminal chromosomal regions. An association of a minisatellite with a dispersed repetitive element was identified when studying the organization of cloned D7S22. This phenomenon was later found to be common amongst minisatellites. Pedigree analysis revealed a high level of instability of the locus detected by D1S7. This manifestation of detectable mutant alleles demonstrated the feasibility of direct estimation of mutation rates at minisatellite loci. The hypervariability of loci detected by minisatellites and their sensitivity in blot hybridizations make minisatellites a powerful tool in genetic analysis. These probes have already proved instrumental in many genetic and clinical studies. The high degree of individual specificity and the relatively simple banding pattern generated make these probes invaluable in forensic medicine. D1S7 and D7S21 were used in the first example of DNA-based identification in a rape and murder enquiry. One minisatellite probe was found to detect two loci, DNF21S1 and DNF21S2, on chromosomes 6 and 16 respectively. The 39 base pair repeat unit of this minisatellite is itself repetitive. The heterozygosity values of DNF21S1 and DNF21S2 are 61% and 16% respectively. Genomic mapping and sequence analyses revealed close similarity between these loci. Human population and pedigree studies showed that some individuals carry two alleles at DNF21S2, some carry one allele, some carry a duplicated allele while some are devoid of this locus. A model of duplication of a large proterminal segment of chromosome 6 DNA containing a minisatellite and transposition into an interstitial region of chromosome 16 in some human individuals is suggested. This is, to my knowledge, the first report of a human DNA polymorphism arising via transposition of DNA. The duplication unit on chromosome 16 is large (>15 kb) and has inserted into a member of a target site family present in 5-10 copies per genome. This sequence family represents a novel class of human repetitive DNA.

The function of a DNA sequence is commonly predicted by measuring its nucleotide similarity to known functional sets. However, the use of structural properties to identify patterns within families is justified by the discovery that many very different sequences have similar structural properties. The aim of this thesis is to develop tools that detect any unusual structural characteristics of a particular sequence or that identify DNA structure-activity fingerprints common to a set. This work uses the Octamer Database to describe DNA. The database's contents are split into two categories: those parameters that describe minimum energy structure and those that measure flexibility. Information from both of these categories has been combined to describe structural tendencies, offering an alternative measure of sequence similarity. A structural DNA profile gives a graphical illustration of how a parameter from the Octamer Database varies across either a single sequence's length or across a set of sequences. Profile Manager is an application that has been developed to automate single sequence profile generation and is used to study the A-tract phenomenon. The use of profiles to explore patterns in flexibility across a set of pre-aligned promoters is then investigated with interesting transitions in decreasing twist flexibility discovered. Multiple sequence queries are harder to solve than those of single sequences, due to the inherent need for the sequences to be aligned. It is only under rare circumstances that sequences are pre-aligned by an experimentally determined position. More commonly a multiple alignment must be generated. An extended, structure-based, hidden Markov model technique that successfully generates structural alignment~ is presented. Its. application is tested on four DNA protein binding site datasets with comparisons made to the traditional sequence method. Structural alignments of two out of the four datasets were comparable in performance to sequence with useful insights into underlying structural mechanisms.

As part of an ongoing multidisciplinary effort at California Polytechnic State University, biologists and computer scientists have developed a new Library-Dependent Microbial Source Tracking method for identifying the host animals causing fecal contamination in local water sources. The Cal Poly Library of Pyroprints (CPLOP) is a database which stores E. coli representations of fecal samples from known hosts acquired from a novel method developed by the biologists called Pyroprinting. The research group considers E. coli samples whose Pyroprints match above a certain threshold to be part of the same bacterial strain. If an environmental sample from an unknown host animal matches one of the strains in CPLOP, then it is likely that the host of the unknown sample is the same species as one of the hosts that the strain was previously found in. The computer science technique for finding groups of related data (ie. strains) in a data set is called clustering. In this thesis, we evaluate the use of density-based clustering for identifying strains in CPLOP. Density-based clustering finds clusters of points which have a minimum number of other points within a given radius. We contribute a clustering algorithm based on the original DBSCAN algorithm which removes points from the search space after they have been seen once. We also present a new method for comparing Pyroprints which is algebraically related to the current method. The method has mathematical properties which make it possible to use Pyroprints in a spatial index we designed especially for Pyroprints, which can be utilized by the DBSCAN algorithm to speed up clustering.

Fingerprints are a worldwide well known tool for law enforcement agencies to reach the individualization of people convicted of a crime. Moreover, all major countries have huge fingerprint databases and efficient automated systems (AFIS) to perform electronic screening of fingerprints marks recovered by crime scene investigation. Fingerprints are permanent and even if, from a scientific point of view, they could not be considered unique, friction ridge is highly selective and allows a discrimination between different individuals with a very high proficiency. Up to now, the most common techniques for enhancing latent fingerprints from articles collected in the crime scene are based on chemical-physical processes, or optical detection techniques, based on absorption, photoluminescence, diffused reflection or ultraviolet imaging, with appropriate band-pass and/or narrow-band filtering. Chemical-physical processes have shown really good performances, but they are destructive with respect to the latent finger mark deposit and in most cases these methods partially affect subsequent DNA analysis. On the other side, optical detection processes have the advantage of being non-destructive of the fingerprint. As a result, these techniques allow later performing of DNA analysis and/or the further application of conventional fingerprint development procedures. The majority of the optical techniques, with the possible exception of the ultraviolet inspection, allow further biological analysis. And as the aforementioned methods have the advantage of being non alterative with the respect of the fingerprint deposit, subsequent application of chemical and/or physical methods is not precluded. Moreover, some recent studies are investigating the X-ray fluorescence of fingerprints, and some others are attempting to discriminate the IR spectrum of the finger mark deposit from the IR spectrum of the surface. It is easy to understand how crucial is to develop a robust technique of optical analysis, able to reach a high-resolution imaging of finger marks, requiring no chemical conventional or non-conventional pre-process and producing no modification either on the finger perspiration deposit or on the background surface. The proper image of the fingerprint, obtained from the item surface, could allow us to perform a complete fingerprint analysis, which potentially leads us to the individualization of the perpetrator. Moreover, fingerprint imaging could exactly point out the particular region of the whole surface where we can surely find the DNA of the donor, with a higher probability of successful analysis.

The present thesis has been developed considering three different livestock species such as chicken, cattle and sheep. The aim of the study was the evaluation of the application of molecular makers in order to assay the genetic population structure on seven local breeds of chicken, to evaluate the applicability of candidate genes as support of conventional breeding on Piedmontese cattle breed and to detect new SNPs on a sheep population. The first two researchs were carried out at Department of Animal Science of University of Padova while the last one at Reprogen (Faculty of Veterinary Science, University of Sydney, NSW, AUS).

[Truncated abstract] The collection of buccal cells is common practise in the epidemiological and forensic science. Unlike venipuncture collection of blood; it is a safer, non-invasive method for collection of biological material. The methods by which these cells are collected from the inner cheek of an individual and stored are the key elements in preserving DNA. Typically, forensic samples require long term storage. Samples are commonly collected on cotton swabs and stored moist at low to ultra-low temperatures (less than -20oC). Although this is the method of choice in most forensic facilities, there are drawbacks. The samples are inherently contaminated with microflora within the oral cavity and the moisture allows a plethora of microorganisms to grow. As the time frame that has elapsed from collection to storage increases, there is an exponential increase in bacterial cells. Storage of containers containing swabs coated with cells at temperatures below 20oC is also costly due to requirements for large freezers which are running and monitored over 24 hours. In the pass 10 to 15 years, researchers have focussed on alternative ways to store buccal cells. The FTA card system by Whatman is one such development. The FTA card is unique in that it provides a means for the collection of buccal cells for storage at room temperature. DNA profiling from samples stored in this way for 11 years has been successfully achieved. The filter paper matrix of the FTA card binds and subsequently lyses cells. ... (2) The second component of this thesis describes a study which subjected cells on buccal swabs to various conditions of increased temperature over periods of time to establish if DNA could be amplified. The aim was to mimic exposure to the vigours of field conditions, particularly in the extreme local environments that prevail in the United Arab Emirates. a. Initially, buccal cells stored at -20oC over 360 days were used to mimic standard archiving procedures. The cells were subsequently transferred to FTA cards, amplified and profiled by using ABI AmpFLSTR Identifiler PCR Amplification Kit (Applied Biosystems, Foster City, CA). Complete STR profiles were successfully recovered from the archived swabs. In most cases 100% of alleles were recovered, suggesting that it is feasible to transfer DNA from properly archived buccal swabs to FTA cards. b. The second phase involved the storage of fresh swabs that had been artificially aged by using incubation temperatures ranging from 40oC to 100oC. Partial profiles resulted from artificially aged samples, indicating that the prevailing conditions prior to low temperature storage of the swabs plays an important role in ensuring cellular integrity and thus, DNA quality. Results from this study suggest that it is possible for biological samples stored under correct conditions to be transferred from swabs to FTA card. In combination, the two chapters presented in this study show that it is feasible to transfer achieved forensic biology samples from swabs to the FTA card system. However, it is necessary to ensure that the samples are treated in the correct manner so as to minimise contamination from external sources and to maintain the correct environmental state to maintain intact cells and usable DNA.

Mestrado em Ciências Forenses<br>Master Degree Course in Forensic Sciences<br>A possibilidade de analisar amostras com quantidades exíguas de material genético (amostras Low Copy Number ou LCN), em que estão presentes apenas algumas células, tem alterado a forma de encarar a cena do crime. Alguns vestígios que até agora não eram considerados como susceptíveis de proporcionarem resultados, podem actualmente ser analisados com sucesso. As impressões digitais são um bom exemplo. Estes vestígios apresentam um baixo número de células, permitindo apenas recuperar quantidades de DNA inferiores a 100 pg. Assim, para a análise do DNA nuclear, é necessário implementar sistemas muito sensíveis que consistem, habitualmente, no aumento do número de ciclos da PCR. Contudo, alguns artefactos são produzidos, tornando difícil a interpretação dos electroforectogramas. Neste trabalho pretendeu-se comparar a aplicação do estudo de STR autossómicos, Y-STR e miniSTR na análise genética de impressões digitais, partindo do conceito do aumento do número de ciclos como estratégia para se obter maior sensibilidade. Procedeu-se também à amplificação total do genoma e nested-PCR, como métodos alternativos ao aumento do número de ciclos. Adicionalemente, neste estudo tentou-se perceber a influência dos principais métodos reveladores de impressões digitais (cianoacrilato, pó magnético e pó branco) na análise do DNA. Os resultados mostram que o aumento do número de ciclos é a melhor opção como método para aumentar a sensibilidade. Constata-se também que o DNA extraído de impressões digitais encontra-se parcialmente degradado, obtendo-se diferenças significativas entre loci com fragmentos de amplificação menores e maiores do que 200 pb. Dos diferentes marcadores caracterizados verifica-se que, em termos de percentagem de alelos detectados, os miniSTR proporcionam os melhores resultados. Por outro lado, os Y-STR parecem altamente sensíveis à degradação ou presença de inibidores, pelo que são menos robustos para este tipo de análises. Verifica-se também que os perfis LCN são drasticamente afectados por artefactos, principalmente os derivados de variação estocástica, como o allele dropout e o desequilíbrio heterozigótico. A determinação de perfis de consenso permite reduzir alguns destes artefactos. Dos métodos de revelação estudados, o cianoacrilato é o que apresenta menor influência na análise e, pelo contrário, o pó branco provoca os resultados mais negativos.<br>The possibility to perform low copy number DNA typing, when just a few cells are available, as changed the way how crime scene investigations is faced. Nowadays it is possible to successfully type some evidence that couldn t be considered until now. Fingerprints are a good example of those. Since that just a few cells are present in this evidence (enabling recovery of low quantities of DNA, fewer than 100pg) just very sensitive systems can detect nuclear DNA. The most used method is definitely increasing the number of PCR cycles. However, increased occurrence of stutters and artifacts that reduced the quality of the DNA profile is normally observed. The present work aimed to compare the application of autosomic STR, Y-STR and miniSTR markers, based on the concept of increased number of PCR cycles as a strategy to achieve more sensitivity. Some other methods, such as whole genome amplification and nested-PCR, were also evaluated as an alternative way to reach the desired sensitivity. Another goal was to determine the influence of several reagents for developing latent fingerprints (cyanoacrylate fuming, magnetic powder and white powder) in DNA typing. The results shows that increasing the number of PCR cycles still is the best way to attain the required sensitivity. Moreover we could realize that DNA was partially degraded, once there were observed significant differences between loci larger and smaller than 200bp. Among all markers miniSTR showed to perform the best results in terms of detected alleles percentage. On the other hand, Y-STR seemed to be highly affected in the presence of degraded DNA and PCR inhibitors, which makes them less robust for these analyses. LCN profiles are significantly affected by artifacts, like allele dropout and heterozygous imbalance, derived from stochastic fluctuation. Reporting consensus profiles reduces artifact inherent errors. Finally, cyanoacrylate proved to have a minimum negative effect on DNA profiling, while white powder was the worst reagent.

Mestrado em Ciências Forenses<br>Master Degree Course in Forensic Sciences<br>A possibilidade de analisar amostras com quantidades exíguas de material genético (amostras Low Copy Number ou LCN), em que estão presentes apenas algumas células, tem alterado a forma de encarar a cena do crime. Alguns vestígios que até agora não eram considerados como susceptíveis de proporcionarem resultados, podem actualmente ser analisados com sucesso. As impressões digitais são um bom exemplo. Estes vestígios apresentam um baixo número de células, permitindo apenas recuperar quantidades de DNA inferiores a 100 pg. Assim, para a análise do DNA nuclear, é necessário implementar sistemas muito sensíveis que consistem, habitualmente, no aumento do número de ciclos da PCR. Contudo, alguns artefactos são produzidos, tornando difícil a interpretação dos electroforectogramas. Neste trabalho pretendeu-se comparar a aplicação do estudo de STR autossómicos, Y-STR e miniSTR na análise genética de impressões digitais, partindo do conceito do aumento do número de ciclos como estratégia para se obter maior sensibilidade. Procedeu-se também à amplificação total do genoma e nested-PCR, como métodos alternativos ao aumento do número de ciclos. Adicionalemente, neste estudo tentou-se perceber a influência dos principais métodos reveladores de impressões digitais (cianoacrilato, pó magnético e pó branco) na análise do DNA. Os resultados mostram que o aumento do número de ciclos é a melhor opção como método para aumentar a sensibilidade. Constata-se também que o DNA extraído de impressões digitais encontra-se parcialmente degradado, obtendo-se diferenças significativas entre loci com fragmentos de amplificação menores e maiores do que 200 pb. Dos diferentes marcadores caracterizados verifica-se que, em termos de percentagem de alelos detectados, os miniSTR proporcionam os melhores resultados. Por outro lado, os Y-STR parecem altamente sensíveis à degradação ou presença de inibidores, pelo que são menos robustos para este tipo de análises. Verifica-se também que os perfis LCN são drasticamente afectados por artefactos, principalmente os derivados de variação estocástica, como o allele dropout e o desequilíbrio heterozigótico. A determinação de perfis de consenso permite reduzir alguns destes artefactos. Dos métodos de revelação estudados, o cianoacrilato é o que apresenta menor influência na análise e, pelo contrário, o pó branco provoca os resultados mais negativos.<br>The possibility to perform low copy number DNA typing, when just a few cells are available, as changed the way how crime scene investigations is faced. Nowadays it is possible to successfully type some evidence that couldn t be considered until now. Fingerprints are a good example of those. Since that just a few cells are present in this evidence (enabling recovery of low quantities of DNA, fewer than 100pg) just very sensitive systems can detect nuclear DNA. The most used method is definitely increasing the number of PCR cycles. However, increased occurrence of stutters and artifacts that reduced the quality of the DNA profile is normally observed. The present work aimed to compare the application of autosomic STR, Y-STR and miniSTR markers, based on the concept of increased number of PCR cycles as a strategy to achieve more sensitivity. Some other methods, such as whole genome amplification and nested-PCR, were also evaluated as an alternative way to reach the desired sensitivity. Another goal was to determine the influence of several reagents for developing latent fingerprints (cyanoacrylate fuming, magnetic powder and white powder) in DNA typing. The results shows that increasing the number of PCR cycles still is the best way to attain the required sensitivity. Moreover we could realize that DNA was partially degraded, once there were observed significant differences between loci larger and smaller than 200bp. Among all markers miniSTR showed to perform the best results in terms of detected alleles percentage. On the other hand, Y-STR seemed to be highly affected in the presence of degraded DNA and PCR inhibitors, which makes them less robust for these analyses. LCN profiles are significantly affected by artifacts, like allele dropout and heterozygous imbalance, derived from stochastic fluctuation. Reporting consensus profiles reduces artifact inherent errors. Finally, cyanoacrylate proved to have a minimum negative effect on DNA profiling, while white powder was the worst reagent.

The requirements for brewing beer from barley (Hordeum vulgare L.) malt are specific and unique for each brewer. Anheuser-Busch InBev and Miller Coors Brewing Company (MillerCoors) are two major brewers in the United States that target different malt quality profiles for six-rowed barley malt. Two closely related cultivars developed by the University of Minnesota, Robust and Stander, differ greatly in agronomic and malt quality performance. Robust malt fits the requirements of MillerCoors and Stander malt has many of the parameters desired by Anheuser-Busch InBev. The close relationship between these two cultivars increases the chance of recognizing chromosome regions with the genes controlling malt quality traits. A total of 53 doubled-haploid (DH) lines (original population) and the parents from the Robust x Stander cross were grown at eleven locations in North Dakota and one location in Idaho the past six years. An additional 138 Robust x Stander DH lines were generated in 2009 and were evaluated alongside the original DH population in the summer of 2011 at two North Dakota locations. Agronomic data were collected at all locations and cleaned grain samples of the original population from six of the locations were micro-malted at NDSU. Three linkage maps were developed using the original and 191 DH line (entire) populations. The first linkage map was constructed using the original DH population, along with a total of 102 SNP, SSR, and DArT markers. The second and third linkage maps were developed using only 67 SNP markers, with the original and entire Robust x Stander DH population, respectively. The first map was used to identify QTL controlling malt quality and wort carbohydrate traits on chromosomes 4H, 5H, and 6H. The SNP map constructed using the original DH population was used to identify QTL controlling agronomic traits on chromosome 6H. The third map was used to identify QTL controlling agronomic traits on chromosomes 4H and 6H. The ultimate goal for this research in years to come is to develop a genetic haplotype that helps distinguish six-rowed barley lines suitable for MillerCoors and Anheuser-Busch InBev.

Six-rowed barley (Hordeum vulgare L.) malt is an important raw material of beer. Each barley cultivar has its own unique malt profile for specific traits. The two biggest brewers in the United States, Anheuser-Busch InBev (ABI) and MillerCoors Brewing Company (MillerCoors), have different malt profiles for their ideal malt. MillerCoors wants moderate levels of protein and enzymatic activity while ABI wants higher levels of protein and enzymatic activity. Two cultivars that have the ideal malt profile for each company are Robust for MillerCoors and Stander for ABI. The pedigree of these two cultivars is very narrow; thus, understanding the genetic basis for the differences observed between Robust and Stander may help us in developing new cultivars that meet specific brewers? needs. The objectives of this investigation is to use the Robust x Stander doubled-haploid population to develop a genetic haplotype that helps distinguish six-rowed barley lines for ABI and MillerCoors.

Submitted by Geandra Rodrigues (geandrar@gmail.com) on 2018-03-27T12:00:11Z No. of bitstreams: 0<br>Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2018-03-27T13:49:39Z (GMT) No. of bitstreams: 0<br>Made available in DSpace on 2018-03-27T13:49:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-02<br>CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior<br>O queijo Minas Frescal (QMF) representa um dos queijos mais consumidos no Brasil. Diversos fatores do seu processamento influenciam suas características microbiológicas, e consequentemente, sua qualidade e propriedades organolépticas. Seu alto teor de umidade e os riscos de contaminação durante a cadeia produtiva favorecem a ocorrência de microrganismos contaminantes, muitas vezes apresentando resistência aos antimicrobianos. Dessa forma, do ponto de vista da segurança alimentar e frente ao crescente fenômeno da resistência bacteriana às drogas, torna-se importante a investigação sobre a estrutura da comunidade bacteriana em QMF, bem como a avaliação da ocorrência de marcadores genéticos microbianos relacionados à resistência a drogas e seu potencial de mobilização. Neste estudo foram obtidas 5 amostras de um mesmo lote de 7 marcas de QMF identificadas de A a G, totalizando 35 amostras. Após a extração de DNA total microbiano das amostras, foram utilizadas abordagens de DNA fingerprint, pela amplificação de sequências palindrômicas extragênicas repetitivas (rep-PCR) para avaliação comparativa da similaridade da estrutura global da comunidade bacteriana. Posteriormente, PCR-DGGE foi utilizada para avaliar o perfil e a riqueza das amostras com relação a grupos de bactérias láticas. Matrizes de similaridade foram obtidas utilizando o método de agrupamento UPGMA. Os resultados obtidos pela técnica de rep-PCR revelaram que as amostras de queijos foram claramente agrupadas de acordo com as suas respectivas marcas. Além disso, perfis semelhantes entre amostras de marcas diferentes foram observados, indicando a presença de um núcleo microbiano comum. As amostras avaliadas também foram agrupadas de acordo com suas respectivas marcas de fabricação de acordo com os padrões de DGGE obtidos para bactérias láticas. A elevada similaridade entre a maioria das amostras do mesmo lote obtida nas técnicas de fingerprint sugere a reprodutibilidade e aplicabilidade das técnicas, e controle no processamento dos queijos ao longo da cadeia produtiva. Para a avaliação do resistoma clínico, a presença de 40 marcadores de resistência a diferentes classes de antibióticos foi avaliada por reação de PCR. Um núcleo comum de marcadores genéticos em todas as marcas foi detectado, associado à resistência aos beta-lactâmicos, tetraciclinas, quinolonas e sulfonamidas. Outros marcadores, incluindo aqueles relacionados a bombas de efluxo e resistência aos aminoglicosídeos, também foram observados. Integrons de classes 1 e 2 foram detectados, respectivamente, em 77% e 97% das amostras. As diferentes amostras de QMF puderam ser agrupadas de acordo com seu perfil de marcadores genéticos de resistência aos antimicrobianos, o que sugere epidemiologia peculiar que pode estar relacionada a qualidade e aos níveis de contaminação dos queijos ao longo da cadeia produtiva. Em conjunto, os dados sugerem que embora a cadeia produtiva do QMF seja controlada na indústria, riscos sanitários são inerentes pela contaminação dos queijos por bactérias putativas resistentes a antimicrobianos. Como um todo, os dados apontam para a necessidade de discussão dos parâmetros de qualidade microbiológica na produção, armazenamento e distribuição de QMF. Além disso, a detecção de integrons de classe 1 e 2 levanta questões a respeito do potencial de transferência horizontal de genes de resistência para a microbiota humana através do consumo destes alimentos.<br>Minas Frescal cheese (QMF) represents one of the most consumed cheeses in the country. Several factors of its processing influence its microbiological characteristics, and, consequently, its quality and properties. Its high moisture content and the risks of contamination during the production chain favor the occurrence of contaminating microorganisms, often presenting antimicrobial resistance. Thus, from the point of view of food safety and the growing phenomenon of bacterial resistance to drugs, it is important to investigate the structure of the bacterial community in Minas Frescal cheese, as well as the evaluation of the occurrence of microbial genetic markers related to drug resistance and its potential for mobilization. In this study 5 samples from the same batch of 7 brands of Minas Frescal cheeses were identified from A to G, totaling 35 samples. After the extraction of total DNA from the samples, DNA fingerprint approaches were used, by the amplification of repetitive extragenic palindromic sequences (rep-PCR) to evaluate the similarity of the global structure of the bacterial community. Afterwards, PCR-DGGE was used to evaluate the profile and richness of the samples in relation to groups of lactic bacteria. Similarity matrices were obtained using the UPGMA clustering method. The results obtained by the rep-PCR technique revealed that the cheese samples were clearly brand-clustered. In addition, similar profiles among samples of different brands were observed, indicating the presence of a common microbial nucleus. The evaluated samples were also separated according to their respective manufacturing brands by the DGGE for lactic acid bacteria. The high similarity among the majority of the samples from the same batch obtained in the fingeprint techniques suggests the reproducibility and applicability of the techniques, and control in the cheese processing along the production chain. For the evaluation of clinical resistance, the presence of resistance markers to different classes of antibiotics was evaluated by PCR reaction. A common core of genetic markers was detected, associated with resistance to beta-lactams, tetracyclines, quinolones and sulfonamides. Other markers, including those related to efflux pumps and aminoglycoside resistance, have also been observed, but not in all brands. Integrons of classes 1 and 2 were detected, respectively, in 77% and 97% of the samples. The different QMF samples could be grouped according to their profile of genetic markers of antimicrobial resistance, which suggests peculiar epidemiology that may be related to the quality and levels of contamination of the cheeses along the production chain. Taken together, the data suggest that although the productive chain of QMF is controlled in the industry, health risks are inherent in the contamination of cheeses by putative antimicrobial resistant bacteria. As a whole, the data point to the need to discuss the parameters of microbiological quality in the production, storage and distribution of QMF. In addition, the detection of class 1 and 2 integrons raises questions about the potential for horizontal transfer of resistance genes to the human microbiota through the consumption of these foods.

Made available in DSpace on 2016-08-10T10:38:43Z (GMT). No. of bitstreams: 1 RAQUEL VAZ RESENDE.pdf: 8228076 bytes, checksum: 2434f1bf0bafb200c6e01978c59427a6 (MD5) Previous issue date: 2013-02-08<br>The importance of scientific proof for the current Brazilian justice system is notorious. Article 158 of the CPC provides that when the offense is a trace essential examination of the corpus delicti. But many fingerprints arriving in section showdown Police Technician - Scientific Goiás, do not present conditions for analysis are blurred or incomplete, and thus unusable. The possibility of extracting DNA of these appears as an option in criminal investigations. The present study detected by light microscopy, scaly epidermal cells in 98% of the fifty sheets containing fingerprints subjected to Leishman stain, and the amount varied from fifteen to seven hundred and seventy cells per slide. After DNA extraction sixty-nine samples, deposited on five different media (aluminum, wood, paper, plastic and glass) were obtained concentrations ranging from 0.3 ng / uL to 25.4 ng / uL. Analyzing the concentrations of each surface separately observed that wood was the one with the highest average concentration of DNA (10.67 ng / uL), while paper and plastic had equal means and the lowest (5.92 ng / uL) . Comparing the media by student t test, we found three statistically significant analysis, the largest difference was observed between the surfaces of wood and paper (p = 0.001). When extracting DNA prints developed with ninhydrin or impregnated by black powder, concentration obtained in 70% of samples with ninhydrin and 60% of samples with dust. This study corroborates several studies have shown that it is possible to extract DNA from surfaces that have been touched by the hands of just one person. Our experiments also showed obtaining a higher concentration in the porous surfaces in relation to smooth surfaces and that using ninhydrin and black powder also allow the extraction of said genetic material.<br>A importância da prova científica para o atual sistema de justiça brasileiro é notória. O artigo 158 do CPP determina que quando a infração deixar vestígios será indispensável o exame do corpo de delito. Porém, muitas impressões digitais que chegam à seção de confronto da Polícia Técnico - Científica de Goiás, não apresentam condições de análises por estarem borradas ou incompletas, sendo assim, inutilizadas. A possibilidade de extrair DNA destas surge como uma opção nas investigações criminais. O presente estudo detectou, à microscopia óptica, células descamativas da epiderme em 98% das cinquenta lâminas contendo impressões digitais submetidas à coloração de Leishman, sendo que a quantidade variou de quinze a setecentos e setenta células por lâmina. Após a extração de DNA de sessenta e nove amostras, depositadas em cinco suportes diferentes (alumínio, madeira, papel, plástico e vidro) foram obtidas concentrações que variaram entre 0,3 ng/µL a 25,4 ng/µL. Analisando as concentrações de cada superfície separadamente observamos que a madeira foi a que apresentou a maior concentração média de DNA (10,67 ng/µL), enquanto que o papel e plástico apresentaram médias iguais e as menores (5,92 ng/µL). Na comparação entre os suportes pelo teste t student, encontramos três análises estatisticamente significativas, sendo a maior diferença foi observada entre as superfícies de madeira e papel (p = 0,001). Ao extrair DNA de impressões reveladas com ninidrina ou impregnadas pelo pó preto, obtivemos concentração em 70% das amostras com ninidrina e 60% das amostras analisadas com pó. O presente trabalho corrobora com vários estudos que já demonstraram ser possível extrair DNA de superfícies que foram simplesmente tocadas pelas mãos de uma pessoa. Nossos experimentos demonstraram, ainda, a obtenção de uma maior concentração nas superfícies porosas em relação às superfícies lisas e que o uso de ninidrina e pó de cor preta também permitem a extração do referido material genético.

The empirical research discussed in this thesis is aimed at investigating the establishment and implementation of the Italian DNA databank arranged on the base of the Prüm treaty and set up with the law n.85 of 30th June 2009. This archive of criminal DNA profiles operates as part of a broader set of tools for the regulation and the government of phenomena and situations that at some point have been presented as a problem and defined as a threat. The implementation of this “instrument” has been taken into account as an exemplary moment of the production of a larger dispositif of securitization and has been contextualised within a broader process of constitution, development and integration of national databases in the European framework of police and judicial cross-border cooperation. At a more general level the inquiry deals with classifications that define certain "kinds of people" (the criminal, the recidivist, the suspect). Special attention has been devoted to the scientific knowledge that support and justifies them, to the political discourses that makes theme effective and to the technical instruments they use, particularly that peculiar kind of tools represented by genetic and biometrics databases used for personal identification in criminal investigations, and in the management of public security and borders in EU.

In der vorliegenden Arbeit wurden 2460 Viertelgemelksproben aus 16 hessischen Milcherzeugerbetrieben untersucht. 115 S. canis-Isolate konnten gefunden und auf ihre morphologischen, biochemischen und bei molekularbiologischen Eigenschaften untersucht werden. Die Isolate stammten von Viertelgemelksproben bzw. Tankproben, die zu einem oder mehreren Zeitpunkten in den Betrieben genommen wurden. Die Untersuchung der biochemischen Eigenschaften erbrachte 24 verschiedene Reaktionsmuster. Der Vergleich dieser 24 Biotypen mit einem S. canis-Referenzstamm mittels tDNA-PCR und 16S-RNA-PCR ergab eine völlige Übereinstimmung (100%) und damit eine sichere Spezies-Identifizierung. Zur Aufklärung epidemiologischer Zusammenhänge und zur Intra-Spezies-Identifizierung wurde von allen 115 Isolaten mittels PFGE nach Makrorestriktionsverdau mit SmaI ein DNA-Fingerprint erstellt. Dabei ergaben sich 21 verschiedene Restriktionsmuster. Von den 21 nach Makrorestriktion mit Sma I und anschließender PFGE unterscheidbaren Restriktionsmustern wurde je ein Isolat zur Bestimmung der Differenzierungsfähigkeit der Restriktionsenzyme Cla I und Apa I sowie der RAPD-PCR weitergehend untersucht. Für die Beurteilung epidemiologischer Zusammenhänge bei S. canis erwies sich die PFGE nach Makrorestriktion mittels Sma I als die differenzierteste Variante. Die mittels PFGE nach Makrorestriktionsverdau mit Sma I durchgeführten Untersuchungen der 115 Isolate zeigten, dass zu einem Probennahme-Termin gewonnene Isolate identisch waren; vom gleichen Betrieb zu unterschiedlichen Zeiten entnommene Proben zeigten z.T. deutliche Unterschiede, und bei Isolaten von verschiedenen Betrieben konnten keine Verwandtschaftsbeziehungen nachgewiesen werden. Aufgrund dieser genotypischen Eigenschaften der Kulturen konnte gezeigt werden, dass es sich bei durch S. canis verursachte Mastitiden um ein infektiöses Bestandsproblem handelt, bei dem der Erreger von Viertel zu Viertel und von Kuh zu Kuh übertragen wird.

Cette thèse précise les conditions pour introduire une sélection sur apparentés de caractères de qualité assistée par empreintes génétiques dans un programme de sélection massale sur la croissance chez la truite arc-enciel. Ce travail confirme l’intérêt à maîtriser l’effet maternel avec une doublement de l’héritabilité du poids à 70 g (0,36 vs 0,16), des estimations d’héritabilités des caractères mesurés de valeurs intermédiaires (0,37-0,54), l’intérêt d’utiliser la résiduelle des parties mesurées régressées linéairement au poids vif pour une sélection indépendante du poids pour les rendements, l’intérêt à remplacer la mesure du rendement au filetage par celle du rendement en carcasse éviscérée-étêtée plus héritable et très corrélé au rendement au filetage (0,97),la corrélation génétique élevée entre les rendements avec le ratio échographique e8/e23 (0,72-0,85) permettant une sélection sur candidats plus efficace qu’une sélection sur apparenté sur le rendement lui-même, des précisions élevées des valeurs génétiques (0.63–0.82) malgré très peu d’individus par famille (3,5-4) et une efficacité surestimée d’une sélection sur apparentés pré-sélectionnés (de 14 % à 62 %). Les conclusions sont qu’il est possible d’introduire une sélection sur apparentés dans un programme de sélection massale avec des gains génétiques au moins équivalents à ceux attendus en sélection familiale classique avec familles élevées initialement de façon séparées<br>The thesis aims at describing conditions to improve mass selection on growth by sib-selection on quality traits assisted by DNA parentage assignment in rainbow trout. Main results are that the control and management of differences in egg size between dams doubles heritability of body weight at 70 g (0.36 vs 0.16), heritabilities of quality traits are intermediates (0.37-0.54), the opportunity to use the residual of the body part (e.g. fillet weight) regressed to the whole body weight to select independently of body weight for ratios, the higher efficiency to replace fillet yield by deheaded and gutted carcass weight more heritable and highly genetically correlated to fillet yield (0.97), the possibility to use e8/e23ultrasound measures to predict genetically yields (0.72-0.85) allowing a direct selection on breeding candidates more efficient than in using sib information, a surprising intermediate to high accuracy of EBV even with a very low mean number of sib per full-sib family (3.5-4) and the medium to high overestimation of EBV when using pre- selected sibs (14 % to 62 %). The general conclusion are that sib-selection on quality traits can be introduced in mass selection with at least similar expected genetic gains than in traditional breeding design based on families reared initially separated

This by-practice project is the first to provide an extensive investigation of the marginalisation of era appropriate science (EAS) and professional investigators by detective and historical detective fiction authors. The purpose of the thesis is to analyse specific detective fiction authors from the earliest formats of the nineteenth century through to the 1990s and contemporary, selected historical detective fiction authors. Its aim is to examine the creation, development and perpetuation of the marginalisation tradition. This generic trend can be read as the authors privileging their detective’s innate skillset, metonymic connectivity and deductive abilities, while underplaying and belittling EAS and professional investigators. Chapter One establishes the project’s critique of the generic trend by considering parental authors, E. T. A Hoffmann, Edgar Allan Poe, Émile Gaboriau and Wilkie Collins. Reading how these authors instigated and purposed the downplaying demonstrates its founding within detective fiction at the earliest point. By comparing how the authors sidelined and omitted specific EAS and professional investigators, alongside science available at the time, this thesis provides a framework for examining how it continued in detective fiction. In following chapters, the framework established in Chapter One and the theoretical views of Charles Rzepka, Lee Horsley, Stephen Knight and Martin Priestman, are used to discuss how minimising EAS and professional investigators developed into a tradition; and became a generic trend in the recognised detective fiction formula that was used by Sir Arthur Conan Doyle, Freeman Wills Crofts, H. C. Bailey, R. Austin Freeman, Agatha Christie, Ruth Rendell and P. D. James. I then examine how the device transferred to historical detective fiction, using the framework to consider Ellis Peters, Umberto Eco and other selected contemporary authors of historical detective fiction. Throughout, the critical aspect considers how the trivialisation developed and perpetuated through a generic trend. The research concludes that there is a trend embedded within detective and historical detective fiction. One that was created, developed and perpetuated by authors to augment their fictional detective’s innate skillset and to help produce narratives using it is a creative process. It further concludes that the trend can be reimagined to plausibly use EAS and professional investigators in detective and historical detective fiction. The aim of the creative aspect of the project is to employ the research and demonstrate how the tradition can be successfully reinterpreted. To do so, the historical detective fiction novel A Hidden Life uses traditional features of the detective fiction formula to support and strengthen plausible EAS and professional investigators within the narrative. The end result is a historical detective fiction novel. One that proves the thesis conclusion and is fundamentally crafted by the critical research.

碩士<br>國立中興大學<br>畜牧學系<br>86<br>Minisatellite, RAPD and RAMPO techniques were employed for studying of fingerprints in ostriches, chickens, mice, goats and pigs. They were then used for investigation the difference of various species and inividual variations. The minisatellite fingerprints of ostrich and chicken DNA digested with HaeIII and hybridized with probe (TG)6 showed a better resolution as compared to that by HinfI digestion. But it did not have a good resolution in mouse and goat. The band pattern of minisatellite fingerprints of pig DNA digested with either HaeIII or HinfI restriction enzyme was shapeless. The 94％(51/54) of random primers in OPAA, OPAO, OPAV, OPC and OPE serials can generated polymorphic RAPD fingerprints in ostrich, chicken, mouse, goat and pig. Numbers(％) of primers which can generated polymorphic RAMPO DNA fingerprints in oftrich, chicken, mouse, goat and pig was 26 (40.1％), 11(23.7％), 46(85.2％), 38(70.4％) and 43(79.6％), respectively. The RAPD fingerprints amplified with different primers had distinct results, and the RAMPO fingerprint after hybridization from RAPD products by probe (TG)6 exhibited differences among individuals in the same species. The nucleotide sequence of the bands at same position in the RAPD fingerprint coule be different and the visual bands on the RAMPO fingerprint did not observed in the RAPD fingerprint. Among species, the minisatellite DNA fingerprints of ostrich, chicken, mouse, goat and pig with HaeIII and HinfI restriction digestion and probe (TG)6 hybridization were different extinctly. Numbers or lengths of bands in RAPD and RAMPO fingerprints of various species were different distinguishably when fingerprints were revealed with the 51 primers. Most of DNA products of ostrich and chicken amplified with different primers by RAPD-PCR did not hybridize with probe (TG)6 . Comparison on fingerprints with methods of minisatellite, RAPD and RAMPO, the minisatellite methods was repeatable but more time-, labor-, cost- and DNA-consuming. Based upon the fact that a less requirement os DNA amounts in RAPD and RAMPO fingerprinting and generation of extinct banding among species, the RAPD fingerprint was applicable for the difference of various species. When both RAPD fingerprint and RAMPO fingerprint methods were used conjunctly, it could avoid misjudging of specified bands.

碩士<br>國立中興大學<br>畜牧學系<br>82<br>DNA指紋具孟德爾遺傳特性，在鑑定生物之遺傳變異上為一極新而有力之 工具。 本試驗之目的以重複序列寡核酸做為探針，進行荷蘭種乳牛、 台灣水牛及豬之DNA指紋分析，供個體鑑別、系譜分析、品種鑑定、性別 鑑定或與經濟性狀有關連性等之探討。牛與豬之基因組DNA經限制HinfI 或HaeⅢ切割後， 進行瓊脂糖 之將膠體乾燥，與放射線標定之重複序列 探針(TG)~bs1;6~bs0;進行膠體原位雜交，最後膠片覆以X-光片，使之自 動放射顯影，產生清晰的DNA指紋。試驗結果證明重複序列探針可用於牛 與豬之DNA指紋分析。 限制不同、電泳條件不同，產生的DNA指紋有極 大的差異。與HinfI比較，限制HaeⅢ切割後，需較長的電泳時間以分開 DNA片段，產生的環帶數亦較多。分別計算牛與豬之共有環帶頻率， 可供 分析DNA指紋資料之用。結果顯示，不同品種動物間之遺傳變異大於同種 動物，同種動物之DNA指紋有較高之相似性。探針(TG)~bs1;6~bs0;偵測則 缺乏品種特異性。以HaeⅢ切割不同性別荷蘭種乳牛之基因組 DNA，雜交 後之DNA指紋中，有一長度約15Kb之公牛特有環帶存在。 此DNA片段回收 後，用限制HindⅢ或EcoRI割切後， 接入載體 pUC18以篩選重組質體， 得到9個重組質體，長度在0.3-1Kb間。為了解其序列組成，以雙去氧法進 行序列分析。其中7個重組質體之部分序列巳定序清楚。 這些重組質體是 否有性別特異序列或重複序列存在，供性別鑑定探針之用，有待進一步探 討。 Genomic DNA was digested with restriction endonuclease HinfI or HaeⅢ and hybridized with probe (TG)~bs1;6~bs0;. fingerprints were obtained in these traits. The results showed that the simple tadem repeat probe may be suitable for the DNA fingerprinting in cattle and pig. The band- sharing probability were also estimated for related and unrelated animals. Results showed that individuals within breeds tented to be more similar to each other with regard to DNA fingerprint pattern than to individuals of other breeds. The DNA fingerprints of Holstein cattle showed a 15 kb sex-specific band presented in the male only when genomic DNA was digested with HaeⅢ and hybridized with~bs0;. The male- specific DNA fragement was recovered from an agarose gel and digested with HindⅢ or EcoRI,then ligated into pUC18.Nine clones had been screened,these inserted fragments were 0.3-1 Sequencing of these clones have been done excepted 2 There are two clones showing highly homology.These clones can be used as probe for sex determination or not need to be investigated in the future.

碩士<br>國立中興大學<br>獸醫學系<br>84<br>We established DNA fingerprints and assayed the correlation of seven dog breeds which included Formosan dog, German Shepherd, Shiba Inu, Pit bullterrier, Akita, Cocker Spaniel, and mixed-breed dog by random amplified DNA polymerase chain reaction (RAPD PCR). Twenty primers were selected from breed- specific DNA pools by screening 180 different ten-base oligonucleic primers. Analysis of the specific products was obtained. 43.8% of the specific products were polymorphic. The correlation index assayed by distance matrix methods between six breeds and mixed-breed dog was from 0.704 to 0.847. In addition, applicationof the selected primers to analyzed the individual DNA of 40 dogs within the six breeds and mixed-breed dogs by RAPD PCR revealed that some of the amplified products between those breeds were different. These results suggested that DNA fingerprints established by RAPD PCR can be served a potential reference to identify dog breed.

博士<br>中國醫藥學院<br>中國藥學研究所<br>89<br>DNA Fingerprints of Lycopodiaceae Revealed by RAPD Analysis Chiang-Tsuan Liao Graduate Institute of Chinese Pharmaceutical Science China Medical College* Abstract There are about 23 species of Lycopodiaceae in Taiwan. Up to now, we have collected and identified 19 species based on morphological study. They are listed as follows :（1）Lycopodium annotinum Linnaeus （2） L. carinatm Desvaux （3）L. casuarinoides Spring （4）L. cernuum Linnaeusu（5）L. pseudoclavatum Ching （6）L. japonicum Thunberg ex Murray（7）L. fargesii Herter（8）L. fordii Baker（9）L. multispicatum Wilce （10） L. yueshanense Kuo （11） L. obscurum Linnaeus （12）L. phlegmaria Linnaeus （13） L. salvinioides （Herter） Tagawa （14）L. quasipolytrchoides Hayata （15）L. taiwanense Kuo（16） L. serratum Thunberg var. longipetiolatum Spring （17） L. serratum Thunberg myriophyllifolium Hayata （18）L. somae Hayata（19）L. veitchii D. Christ。 The following studies have been performed , through RAPD approach : 1. to discover the polymorphism of Lycopodiaceae and to provide a rapid way to differentiate the species of Lycopodiaceae by molecular markers, e.g. RAPD. 2. to distinguish the difference between the varieties of Lycopodium serratum L. in Taiwan . 3. to use molecular marker as a new approach for the identification in Chinese Crude Drug “Jing-bu-huan”. 4. to study if the difference between two species“Shisong”and“Diswa gi”in Taiwan could be revealed by RAPD markers. Key words: Lycopodiaceae、 Macromorphology、 Key、 Qiancenta （Lycopodium serratum Thunb.） Lycopodium serratum Thunb. var. longipetiolatum Spring、L. pseudoclavatum Ching、L. japonicum Thunberg ex Murray、L. multispicatum Wilce、L. yueshanense Kuo、 PCR （ polymerase chain reaction）、RAPD（randomly amplified polymorphic DNA）. * No. 91 Hsueh-Shih Road,Taichung 404, Taiwan, R.O.C.

碩士<br>國立中興大學<br>獸醫學系<br>82<br>1990年 Williams等人提出了一個聚合鎖反應的新方法。這個由傳統PCR 為基礎所衍生出來的技術，只使用一個由隨意序列所組成、長為十個鹼基 的引子，而隨機增殖任何基因組不同基因位置之DNA片段，因而被稱為隨 機增殖聚合鏈鎖反應。　　我們利用這最新發展出來的方法，來試驗 RAPD技術是否適用於球蟲品種鑑定及不同族群的區分。同時也分析一些可 能由DNA、鎂離子、引子、去氧核糖三磷酸及Taq聚合等PCR反應物所造 成的人為變異。兩種未芽胞化的球蟲， Eimeria tenella 及 Isospora mikei，經 RAPD-PCR反應後，分析得到46及39個RAPD markers 而成功地 區分這兩種球蟲。此外，由11株Eimeria tenella 以12個引子經增殖作用 總共產生 78個特定的DNA片段。不同株之Eimeria tenella 以 binary data 紀錄特定片段之有或無，並以Sorrensen's similarity coefficient 計算其相似指數。其結果顯示，RAPD-PCR技術的確適用於 Eimeria tenella 之鑑定。 A polymerase chain reaction PCR-based method, described first by Williams et al.(19900, employs a single 10 base-long primers of arbitrary DNA sequence and amplifies DNA fragments of differ- ent loci from any genome know as random amplified polymorphic DNA(RAPD). We applied the lately developed technique to deter- mine if RAPD of coccidium DNA could be used for the identifi- cation of unknown specimens, and differentiation of population. The artificial variation of the origns of the DNA extract, mag- nesium, primer, dNTPs, and Taq DNA polymerase was analyzed. Two unsporulated coccidium coccidium, Eimeria tenella and Isospora mikei, were differentiated successfully using RAPD markers, which wrer 46 and 39 polymorhpic fragments respec- tively, obtained by the polymerase chain reaction. On the other hand, a total of 78 reproducibility, distinct fragments of elevent Eimeria tenella amplified by 12 ten-base primers of arbitrary sequence in the polymerase chain reaction was pro- duced. Different strains of Eimeria tenella were scored for presence or absence of RAPD fragments by binary data method and calculated the similarity index of the pairwise strains by Sorenson's similarity coefficient. The results indicate the RAPD-PCR technique will be useful in study of E. tenella identification.

MESHWR<br>Department of Ecology and Resources Management<br>The presence of harmful algal blooms (HABs) and cyanobacteria toxins in drinking water sources are known to pose a great threat to humans. The main aim of this study was to use molecular technique to determine the origins of the cyanobacteria species at Musina raw water abstraction point by identifying and comparing the non-toxic and toxic cyanobacteria species in the Limpopo River and some of its tributaries based on the phylogenetic analyses of 16S rRNA gene. The Musina water treatment plant is located downstream of a weir and the Beit bridge on the Limpopo River and the raw water supply is abstracted from 22 boreholes of which 14 are along the Limpopo River and 8 boreholes are inside the Limpopo River channel. The bottom sediments samples were collected from these rivers: Limpopo, Crocodile, Mokolo, Mogalakwena, Nzhelele, Lephalale, Sand rivers (South Africa); Notwane (Botswana), Shashe River and Mzingwane River (Zimbabwe). The physical-chemical analysis of the bottom sediments showed the availability of nutrients, nitrates and phosphates, in excess of 0.5 mg/l for most the of rivers, alkaline pH and salinity in excess of 500 mg/l. Total genomic DNA were extracted from cyanobacteria species on the bottom sediments and Polymerase Chain Reaction (PCR) method was used to detect the genetic profile of the cyanobacteria species. Molecular identification of cyanobacteria was based on PCR amplification and sequencing of the 16S rRNA gene. The 16S rRNA gene was absent from sediments of the Mogalakwena and Lephalale rivers but present in all other selected rivers. The cyanotoxins detection was also based on PCR by amplification of microcystin/nodularin and cylindrospermopsin polyketide synthetase genes. Most of the samples showed no amplification of the toxin genes. While two samples showed the amplification of cylindrospermopsin polyketide synthetase gene (Sand River and Nzhelele River Next to Tshipise) and two samples showed amplification for microcystin/nodularin synthetase gene, Crocodile River and Mzingwane River. The first was the confirmation of similarity of samples from Crocodile River downstream of hartbeespoort Dam and Shashe River to Leptolyngbya boryana with 99% bootstrap confidence. The similarity of sample from Musina borehole to Sand River upstream to Alkalinema pantanalense with 98% bootstrap. Thus, the presence of toxic genes may imply the presence of toxic cyanobacteria species in the river sediments and may be hazardous to humans because rural communities and commercial farmers abstract water from Limpopo River catchment for human consumption, livestock and irrigation. The waters of the Limpopo River basin also provide drinking water to wildlife and a habitant for aquatic organisms/animals.

碩士<br>國防管理學院<br>國防資訊研究所<br>87<br>In this thesis, we described the design of a DNA fingerprint recognition system, which is based on digital image analysis theories. Technologies including noise removal, feature extraction, run-length encoding, and correlation matching, are used and discussed. By using MATLAB tool, a database of DNA fingerprints is created. The system can recognize differences among DNA fingerprints and reduce searching time. This approach provides further manipulation and process in many applications.

abstract: Deoxyribonucleic Acid (DNA) evidence has been shown to have a strong effect on juror decision-making when presented in court. While DNA evidence has been shown to be extremely reliable, fingerprint evidence, and the way it is presented in court, has come under much scrutiny. Forensic fingerprint experts have been working on a uniformed way to present fingerprint evidence in court. The most promising has been the Probabilistic Based Fingerprint Evidence (PBFE) created by Forensic Science Services (FSS) (G. Langenburg, personal communication, April 16, 2011). The current study examined how the presence and strength of DNA evidence influenced jurors' interpretation of probabilistic fingerprint evidence. Mock jurors read a summary of a murder case that included fingerprint evidence and testimony from a fingerprint expert and, in some conditions, DNA evidence and testimony from a DNA expert. Results showed that when DNA evidence was found at the crime scene and matched the defendant other evidence and the overall case was rated as stronger than when no DNA was present. Fingerprint evidence did not cause a stronger rating of other evidence and the overall case. Fingerprint evidence was underrated in some cases, and jurors generally weighed all the different strengths of fingerprint testimony to the same degree.<br>Dissertation/Thesis<br>M.S. Psychology 2012

Forensic science has a significant historical and contemporary relationship with the criminal justice system. It is a relationship between two disciplines whose origins stem from different backgrounds. It is trite that effective communication assist in resolving underlying problems in any given context. However, a lack of communication continues to characterise the intersection between law and science. As recently as 2019, a six-part symposium on the use of forensic science in the criminal justice system again posed the question on how the justice system could ensure the reliability of forensic science evidence presented during trials. As the law demands finality, science is always evolving and can never be considered finite or final. Legal systems do not always adapt to the nature of scientific knowledge, and are not willing to abandon finality when that scientific knowledge shifts. Advocacy plays an important role in the promotion of forensic science, particularly advocacy to the broader scientific community for financial support, much needed research and more testing. However, despite its important function, advocacy should not be conflated with science. The foundation of advocacy is a cause; whereas the foundation of science is fact. The objective of this research was to conduct a qualitative literature review of the field of forensic science; to identify gaps in the knowledge of forensic science and its integration in the criminal justice system. The literature review will provide researchers within the field of forensic science with suggested research topics requiring further examination and research. To achieve its objective, the study critically analysed the historical development of, and evaluated the use of forensic science evidence in legal systems generally, including its role regarding the admissibility or inadmissibility of the evidence in the courtroom. In conclusion, it was determined that the breadth of forensic scientific knowledge is comprehensive but scattered. The foundational underpinning of the four disciplines, discussed in this dissertation, has been put to the legal test on countless occasions. Some gaps still remain that require further research in order to strengthen the foundation of the disciplines. Human influence will always be present in examinations and interpretations and will lean towards subjective decision making.<br>Jurisprudence<br>D. Phil.

Organy procesowe coraz chętniej korzystają z opinii kompleksowych z zakresu badań daktyloskopijnych i genetycznych, co wynika z coraz większych możliwości badawczych obu dyscyplin kryminalistycznych. Wykonanie ekspertyzy kompleksowej z zakresu badań daktyloskopijnych i genetycznych wymaga od biegłych ścisłego i dobrego współdziałania, nie pozostawiającego miejsca na „rywalizację” obu tych specjalności. To współdziałanie sprowadza się między innymi do tego, że biegli każdorazowo decydują o tym, które badania w poszczególnych przypadkach są skuteczniejsze, kiedy się one uzupełniają, a kiedy wykluczają, albo też kiedy wystarczy wykonać badania przy użyciu tylko jednej z technik. W każdym przypadku biegli muszą kierować się dobrem postępowania, czego przejawem niekiedy powinna być decyzja o odstąpieniu od badań z zakresu tej specjalności, co do której wiadomo z bardzo dużym prawdopodobieństwem, że badania zakończą się niepowodzeniem. Czasem trudno jest przewidzieć wynik badań bez ich przeprowadzenia, dlatego nie do przecenienia jest doświadczenie biegłych, ich wiedza oraz zdolność do wzajemnej komunikacji i chęć zawierania kompromisu, co jest niezbędne w kompleksowym podejściu do ekspertyzy. Głównym celem rozprawy, było jak najszersze ujęcie zagadnienie opiniowania kompleksowego z zakresu dwóch wiodących metod identyfikacji człowieka, jakimi są daktyloskopia i genetyka, uwypuklenie korzyści płynące z tego typu opiniowania oraz zwrócenie uwagi na pojawiające się problemy. Rozprawa składa się sześciu rozdziałów. W rozdziale 1 opisano drogę, jaką przemierzyła kryminalistyka, będąca stosunkowo młodą dziedziną nauki, aż do momentu pojawienia się genetyki sądowej. Przedstawiono pojęcie identyfikacji osobniczej, omówiono rodzaje identyfikacji, pokazano jak ewoluowała definicja śladu kryminalistycznego, a także wyjaśniono czym jest tzw. ślad kontaktowy, stanowiący podstawę badań kompleksowych z zakresu daktyloskopii i genetyki sądowej. W rozdziale 2 przedstawiono historię badań daktyloskopijnych oraz medycyny sądowej i biologii kryminalistycznej. W rozdziale 3 omówiono procesowe i kryminalistyczne aspekty opinii kompleksowej. Rozdział 4 poświęcony jest w całości daktyloskopii. Pokazano w nim jednak, że ta dziedzina kryminalistyki ma swoje korzenie w naukach przyrodniczych. Rozdział 5 jest dopełnieniem poprzedzającego go rozdziału i skupia się wokół zasad, na jakich jest oparta identyfikacja daktyloskopijna i genetyczna, oraz omawia wzajemne zależności, podobieństwa i różnice miedzy nimi. W rozdziale 6 przedstawiono wyniki analizy opinii kompleksowych z zakresu badań daktyloskopijnych i genetycznych, której celem było między innymi ustalenie czy opinie kompleksowe z zakresu badań daktyloskopijnych i genetycznych opracowywane są wspólnie w formie jednego dokumentu, czy jako opinie odrębne, w przypadku jakich przestępstw są zlecane opinie kompleksowe z zakresu badań daktyloskopijnych i genetycznych, jaka jest efektywność obu rodzajów badań oraz jakie dowody rzeczowe są badane w przypadku opiniowania kompleksowego z zakresu badań daktyloskopijnych i genetycznych, a także na czym polega kompleksowe podejście do badań? W celu weryfikacji założeń teoretycznych dotyczących kompleksowego opiniowania na podstawie śladów biologicznych i daktyloskopijnych, przeprowadzono analizę 122 opinii kompleksowych z zakresu badań daktyloskopijnych i genetycznych, wydanych przez pięć polskich laboratoriów policyjnych w latach 2010-2013. Analizowane ekspertyzy przeprowadzono w Centralnym Laboratorium Kryminalistycznym Policji (wydanych 67 opinii) oraz w 4 innych laboratoriach policyjnych zlokalizowanych w dużych ośrodkach miejskich (Laboratorium Kryminalistyczne Komendy Wojewódzkiej Policji w Łodzi - 23 opinie; Laboratorium Kryminalistyczne Komendy Wojewódzkiej Policji w Krakowie - 11 opinii; Laboratorium Kryminalistyczne Komendy Stołecznej Policji w Warszawie - 11 opinii; Laboratorium Kryminalistyczne Komendy Wojewódzkiej Policji w Olsztynie - 10 opinii). The judicial bodies are increasingly requesting comprehensive fingerprint and DNA casework examination. This is due to the growing examination possibilities of these two forensic disciplines. Performing comprehensive fingerprint and DNA casework examination requires a tight and good cooperation between forensic experts, which does not leave any space for „competition” between these disciplines. Among others, this interaction comes down to deciding by experts on case-to-case basis what types of examination would be effective, how they complement each other, in which situations they eliminate the possibility of applying the other technique or when the application of one method only is sufficient. In such scenarios, experts must always take action in the interest of proceedings, which sometimes means they abandon examination in the area which is quite likely to yield unproductive results. Although sometimes it is difficult to predict the result of examination a priori, nevertheless the experience of forensic experts cannot be overestimated along with their knowledge and ability to communicate and compromise, which is deemed essential in a comprehensive approach towards casework examination. The doctoral dissertation consists of six chapters and it is dedicated to widely understood comprehensive casework examination problems in scope of two most important forensic fields: fingerprint and DNA examination, emphasize benefits come from this tape of examination and point to inconveniences that can arise. Chapter I describes the distance, that forensic science has crossed – as a relatively young discipline – up to the point of emergence of forensic genetics. The concept of human identification has been discussed in addition to the types of human identification and evolution of definition of forensic trace. Finally the so-called contact traces (trace DNA, touch DNA) have been addressed. In Chapter II the history of fingerprint examination, forensic medicine and forensic biology was presented. The procedural and forensic aspects of a comprehensive opinions remain in the focus of Chapter III. Chapter IV is devoted to dactyloscopy but there is pointed, that this forensics science originated from biological science. Chapter V complements the previous chapter and focus on fingerprinting and DNA examination rules, as well as describes their mutual relationships, similarities and differences. Chapter VI contains the results of fingerprint and DNA casework examination analysis which aimed at answering the following questions: wheather comprehensive opinion are drawn up in the form of a single document or as an separate opinion? What types of criminal offences where comprehensive fingerprint/DNA examination opinion are requested? What is the effectiveness of both types of examination and what evidence is being investigated in the case of comprehensive evaluation? And finally what is the comprehensive approach to examination by the experts? For the purpose of verification of theoretical assumptions concerning comprehensive DNA/fingerprint examination opinions, the analysis of 122 comprehensive opinions from DNA/fingerprint casework examinations was carried out in relation to five police forensic laboratories in Poland in the period between 2010-2013. The study was conducted in the Central Forensic Laboratory of the Police (67 forensic opinions issued) as well as in 4 other chief police forensic laboratories (Voivodeship Police Forensic Laboratory in Łódź- 23 forensic opinions; Voivodeship Police Forensic Laboratory in Cracow - 11 forensic opinions; Metropolitan Police Forensic Laboratory in Warsaw - 11 forensic opinions; Voivodeship Police Forensic Laboratory in Olsztyn - 10 forensic opinions).

Increasing use is being made of various types of scientific evidence in court. The general requirement for the admissibility of such evidence is relevance. Although expert evidence is considered to be opinion evidence, it is admissible if it can assist the court to decide a fact in issue; provided that it is also reliable. In South Africa, the initial wide judicial discretion to either admit or exclude unconstitutionally obtained evidence, has developed into a more narrowly defined discretion under the final Constitution. Examples of scientific evidence, namely, DNA evidence, fingerprints, psychiatric evidence, bite-mark evidence and polygraph evidence are considered and problems inherent in the presentation of such evidence in courts in various jurisdictions are highlighted. An investigation of the presentation and evaluation of evidence in both the accusatorial and inquisitorial systems seems to indicate that the adversarial procedure has a marked influence on the evaluation of evidence<br>Criminal & Procedural Law<br>LL.M. (Law)