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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
81

Revealing the past : the potential of a novel small nucleolar RNA (snoRNA) marker system for studying plant evolution

Hochschartner, Gerald January 2011 (has links)
Despite the existence of various molecular marker systems there are still limitations in distinguishing between closely related species based on molecular divergence, especially when hybridization events have occurred in the past. The characterisation of plant small nucleolar RNA (snoRNA) genes and their organisation into multigene clusters provides a potential nuclear marker system which could help in resolving the phylogenetic history of plants and might be applicable in DNA barcoding. Using closely and distantly related Senecio species, I investigated a combination of fragment length and sequence variation of snoRNA genes/snoRNA gene clusters to assess the utility of this marker system for barcoding and resolving species relationships. SnoRNA gene and gene cluster sequences identified in Arabidopsis thaliana were used to find homologues in other species and subsequently used for the design of universal primers. Most of the universal primer pairs designed were successful in amplifying snoRNA fragments in most Senecio species and fragment length variation between and within species could be detected. Furthermore, the combination of some fragment length datasets produced by different primer pairs enabled the separation of species and the detection of reticulate evolution indicating a high potential of snoRNA gene/gene cluster fragment length polymorphisms (SRFLPs) for phylogenetic reconstructions in Senecio and other plant genera. Most of the examined gene clusters showed a similar gene order in Senecio and Arabidopsis. However, the majority of these clusters appeared to exhibit more copies in Senecio, some of which were distinguishable by a combined sequencing/fragment profiling approach, and shown to be putative single copy regions with the potential to be used as co-dominant markers. However, a high number of paralogues and possible differences in copy number between species excludes these regions from being used in DNA barcoding. This is because specific primers would have to be developed for specific copies which would preclude development of a universal application for barcoding. None of the regions showed enough sequence variation to delimit distinctly closely related Senecio species and were therefore also considered to be unsuitable for DNA barcoding. Although most snoRNA genes and gene clusters might be inapplicable for DNA barcoding, they are likely to be valuable for phylogenetic studies of species groups, genera and families. On this scale, specific primers might act universally and the number of paralogous copies is likely to be equal across the species group of interest.
82

Caracterização molecular de isolados de Sarcocystis spp. obtidos de marsupiais do gênero Didelphis spp. pela análise de gene mitocondrial, gene de apicoplasto, espaçador interno transcrito (ITS-1) e genes codificadores de antígenos de superfície (SAGs) / Molecular characterization of Sarcocystis spp. isolates from marsupials of the genus Didelphis spp. through the analysis of mitochondrial and apicoplast genes, internal transcribed spacer region (ITS-1) and surface antigen genes (SAGs)

Valadas, Samantha Yuri Oshiro Branco 08 May 2015 (has links)
Em um trabalho anterior, foi avaliada a variabilidade de Sarcocystis spp. isolados de gambás oriundos do estado do Rio Grande do Sul pesquisando sequências gênicas codificadoras de antígenos de superfície (SAGs). Os resultados indicaram haver linhagens de isolados de Sarcocystis (relacionadas geneticamente a S. falcatula) que trocam genes em prováveis processos de recombinação sexual. A proposta deste estudo foi de conhecer as variações gênicas e relações filogenéticas em Sarcocystis spp. isolados de gambás através da análise de loci gênicos de genoma mitocondrial (CytB), de genoma de apicoplasto (ClpC) e das regiões espaçadoras transcritas internas (ITS-1), comparando a diversidade encontrada nestes grupos com a observada em integrantes da sub-familia Toxoplasmatinae. E também de conhecer a diversidade de genes codificadores de SAG-2, SAG-3 e SAG-4. Neste estudo foram obtidos esporocistos de Sarcocystis spp. através de raspado intestinal e fezes provenientes de gambás do gênero Didelphis, oriundos do Estado de São Paulo, Rio Grande do Sul e Rio Grande do Norte. Os marcadores moleculares CytB, ClpC e ITS-1 revelaram uma ampla variabilidade genotípica e alelos inéditos entre os isolados do Brasil. Os resultados levam a crer que é possível a existência de espécies “híbridas” de Sarcocystis no Brasil, com mistura de genes de ambas as espécies. Em relação aos genes codificadores de SAGs, foi encontrada uma diversidade ainda maior do que trabalho similar realizado. Os resultados sugerem uma possível ocorrência de reassortment e recombinação de alelos em sequências SAGs, contribuindo ainda mais para a geração de variabilidade e consequentemente na configuração antigênica do parasito. Estes resultados demonstram que isolados brasileiros possuem uma composição genética diferente do reportado em demais localidade, o que pode ter reflexos importantes no conhecimento da diversidade das espécies de Sarcocystis que infectam gambás em nosso meio e em toda a epidemiologia das infecções produzidas pelos protozoários deste grupo / A great variability in surface antigens genes (SAGs) of Sarcocystis spp. sporocysts was found in a previous study, through the intestinal scraping technique of opossums from the south region of Brazil. The results indicated probable sexual recombination between lineages of Sarcocystis isolates (genetically related to Sarcocystis falcatula). The aim of this research is the study of genetic variability and phylogenetic relationships among Sarcocystis spp. isolates from captured opossums in Brazil, by the analysis of molecular markers less subjected to selective pressure, such as DNA barcode, for this purpose, genetic loci from mitochondrial (CytB) and apicoplast genome (ClpC), and internal transcribed spacer regions (ITS-1) were applied and compare the results with the diversity found within members of the Toxoplasmatinae sub-family. The diversity among the SAG-2, SAG-3 and SAG-4 genes was also studied. The Sarcocystis sporocysts were obtained from opposum’s intestinal scraping and faeces from São Paulo State, Rio Grande do Sul State and Rio Grande do Norte State, Brazil. The molecular markers CytB, ClpC and ITS-1 revealed a vast genotypic variability and novel alleles among the Brazilian isolates. The results indicate possible existence of hybrid species of Sarcocystis in Brazil, with a gene mixture from both species. Regarding the SAG genes, an even greater variability was found than previously reported. The results also suggest the occurrence of reassortment and allele recombination of SAG sequences, contributing even more to the variability and thus in the parasites antigenic configuration. In this study, the results demonstrate that isolates from Brazil have a different genetic composition than reported in other locations, which may have important impacts in the knowledge of the diversity of Sarcocystis species shed by opossums in our environment and in all of the epidemiology of infections of protozoa of this particular group
83

Integração dos estudos cromossômicos e DNA barcoding em Rhamphichthys (Pisces: Gymnotiformes)

SILVA, Patrícia Corrêa da 16 May 2016 (has links)
Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-08-30T12:00:23Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_IntegracaoEstudosCromossomicos.pdf: 1817366 bytes, checksum: 3c1634ef74508886d0d4b52b6d5a125d (MD5) / Approved for entry into archive by Irvana Coutinho (irvana@ufpa.br) on 2017-08-31T12:39:11Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_IntegracaoEstudosCromossomicos.pdf: 1817366 bytes, checksum: 3c1634ef74508886d0d4b52b6d5a125d (MD5) / Made available in DSpace on 2017-08-31T12:39:12Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese_IntegracaoEstudosCromossomicos.pdf: 1817366 bytes, checksum: 3c1634ef74508886d0d4b52b6d5a125d (MD5) Previous issue date: 2016-05-16 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / FAPESPA - Fundação Amazônia de Amparo a Estudos e Pesquisas / A Ordem Gymnotiformes é composta por 219 espécies válidas, que estão distribuídas em cinco famílias. Os gêneros mais investigados são Eigenmannia e Gymnotus. Nosso trabalho concentrou-se na família Rhamphichthyidae, gênero Rhamphichthys que, assim como os demais Gymnotiformes, apresentam maior abundância e diversidade na região Amazônica. Foram realizadas coletas nos municípios de Abaetetuba, Barcarena e Belém (Pará) e em Tefé, Reserva Ecológica de Mamirauá (Amazonas), com objetivo de melhor definir as espécies, através da integração de dados citogenéticos clássicos, citogenômicos (através das sondas de sequências repetitivas de DNA) e do DNA Barcoding e assim compreender a evolução deste gênero de peixes na Amazônia. Foram identificados um novo citótipo para o gênero R. rostratus com a presença de cromossomos B e fórmula cariotípica FC=48m/sm+2st/a+(5-10)B, um novo citótipo da região do Amazonas, Rhamphichthys sp. FC = 44m/sm +6st/a, e também de R. marmoratus, FC = 46+4st/a no estado do Pará. A análise das sequências repetitivas nos novos citótipos demonstrou que as sondas de 18S são coincidentes com as regiões de constrição secundárias que são marcadas com nitrato de prata na técnica de coloração clássica da NOR. As sondas de DNA 5S marcam sítios múltiplos, deixando evidente que a evolução da família de genes ribossomais ocorre de maneira independente, pelo menos no gênero Rhamphichthys. Os retroelementos REX1 e REX3 marcaram de forma dispersa pelo genoma, como já foi descrito na literatura para outros peixes. O elemento REX1 marca ainda a região de constrição secundária em R. rostratus, o que também já foi descrito em outras espécies de peixes que habitam ambientes poluídos, expostos a estresses ambientais e também em indivíduos híbridos. A análise de DNA barcoding permitiu a construção de uma árvore bayesiana, que está de acordo com os dados de citogenética. Assim, as populações de R. rostratus com e sem cromossomos B constituem taxa distintos. Por sua vez, a amostra de Mamirauá, aqui denominada Rhamphichthys sp. por não haver sido descrita formalmente, é mais similar tanto nos dados cariotípicos como na análise de barcoding a R. hanni do Sudeste brasileiro. Nossos dados apontam para um número subestimado de espécies em Rhamphichthys, o que reforça a necessidade de uma revisão taxonômica para o gênero. / The Order Gymnotiformes is composed by 219 valid species, which are distributed in five families. The most investigated genera are Eigenmannia and Gymnotus. Our work focused on family Rhamphichthyidae, genus Rhamphichthys that, like other Gymnotiformes, present greater abundance and diversity in the Amazon region. Sampling was carried out in the municipalities of Abaetetuba, Barcarena and Belém (Pará) and Tefé, Ecological Reserve Mamirauá (Amazonas), in order to better define the species, through the integration of classical cytogenetic data, cytogenomic analysis (probes for repetitive DNA sequences) and DNA Barcoding and thus understand the evolution of this fish in the Amazon. A new karyotype was identified for R. rostratus with the presence of B chromosomes and karyotype formula FC = 48m / sm + 2st / a + (5-10) B, as well as a new cytotype from the Amazon region, in Rhamphichthys sp. FC = 44m / sm + 6st / a, and also in R. marmoratus, FC = 46 + 4st / a in the state of Pará. The analysis of repetitive sequences in the new cytotypes demonstrated that probes 18S coincided with the regions of constriction secondary that are marked with silver nitrate in the classical NOR staining technique. The DNA probes 5S mark multiple sites, letting clear that the evolution of the ribosomal gene family is independent, at least in the genus Rhamphichthys. Retroelements REX1 and REX3 marked in a dispersed fashion throughout the genome, as already described in literature for other fishes. The REX1 element also marks the secondary constriction in R. rostratus, which has also been described in other species of fishes that inhabit polluted environments, exposed to environmental stresses and also in hybrid individuals. The barcoding DNA analysis allowed the construction of a Bayesian tree, which is in agreement with the cytogenetic data. Thus, populations of R. rostratus with and without B chromosomes are separate taxa. In turn, the sample from Mamirauá, herein called Rhamphichthys sp., since it was not been formally described, it is more similar in both karyotypic data as the barcoding analysis with R. hanni from southeastern Brazil. Our data let clear that the number of species in Rhamphichthys is underestimated, which reinforces the need for a taxonomic revision of the genus.
84

Etude du régime alimentaire des carnivores par des techniques moléculaires

Shehzad, Wasim 14 December 2011 (has links) (PDF)
La caractérisation des réseaux trophiques est nécessaire pour comprendre le fonctionnement des écosystèmes et les mécanismes impliqués dans leur stabilité. Il est parfois difficile de déterminer les régimes alimentaires notamment pour des espèces discrètes et difficiles à observer comme les grands carnivores. Cependant, ces espèces jouent un rôle clé dans les écosystèmes dont elles influencent le fonctionnement et la biodiversité. Ainsi, connaitre le régime alimentaire des grands prédateurs avec précision est essentiel pour établir des stratégies de conservation. Diverses méthodes basées sur le monitoring, l'analyse d'échantillons invasifs ou non ont été utilisées pour étudier les régimes alimentaires. Elles sont généralement biaisées ou peu résolutives. Les méthodes basées sur l'identification des fragments d'ADN dans les fèces ont le potentiel de fournir une meilleure information, notamment dans le cadre d'une approche métabarcoding. Il s'agit de caractériser simultanément l'ensemble des espèces dont l'ADN est présent dans un échantillon environnemental, en utilisant les Nouvelles Techniques de Séquençage. Dans ce cas, les amorces universelles nécessaires pour amplifier toutes les proies potentielles amplifient également l'ADN du prédateur s'il y a proximité taxonomique (par exemple mammifères). Ainsi les produits PCR obtenus à partir des fèces sont essentiellement composés d'ADN du prédateur et ne reflètent pas l'ensemble du régime alimentaire. L'utilisation d'un oligonucléotide de blocage limitant spécifiquement l'amplification de l'ADN du prédateur peut résoudre ce problème. Nous avons développé une méthode de ce type basée sur l'utilisation d'amorces universelles pour les vertébrés (amplifiant la région 12SV5) et d'oligonucléotides de blocage. Bien que non quantitative, cette méthode s'est montrée robuste, adaptée à l'étude de prédateurs à très large spectre de proies, et très résolutive pour identifier les proies au niveau du genre et de l'espèce. Nous l'avons appliquée à l'étude du régime alimentaire du chat léopard (Prionailurus bengalensis) qui s'est avéré très diversifié (mammifères, oiseaux, amphibiens et poissons) dans les deux populations du Pakistan étudiées. Avec la même approche, nous avons démontré la réalité du conflit entre l'homme et le léopard commun (Panthera pardus) dont le régime est presque exclusivement composé d'animaux domestiques. Enfin, nous avons pu proposer des actions de conservations pertinentes après avoir montré que le régime de la très menacée panthère des neiges (Panthera uncia) est principalement composé d'ongulés sauvages.
85

COI Barcoding of the Shorebirds: Rates of Evolution and the Identification of Species

Elbourne, Rebecca 07 December 2011 (has links)
This study assembles COI barcodes from 1814 specimens from the shorebird order, Charadriiformes and examines variation relative to time, rate of evolution and taxonomic level. In the suborder Scolopaci, 95% of sampled species were identified correctly. COI barcode variation within monotypic species was low (0-1% maximum distance) but showed a wide range within polytypic species (0-5%). Preliminary Charadrii results suggest similar trends but success is reduced in the third suborder, Lari. Rates of COI evolution are found to be lowest in the Lari and this leads to reduced species identification in recently radiated families: just 49% of the Laridae and 57% of the Stercoraridae are identified but 100% of the older Alcidae. In the faster Scolopaci, subspecies are at the limit of resolution with some well differentiated subspecies not distinguished by barcodes. The interplay of evolutionary rates, divergence dates and gene flow appears to determine COI barcode differentiation between taxa.
86

COI Barcoding of the Shorebirds: Rates of Evolution and the Identification of Species

Elbourne, Rebecca 07 December 2011 (has links)
This study assembles COI barcodes from 1814 specimens from the shorebird order, Charadriiformes and examines variation relative to time, rate of evolution and taxonomic level. In the suborder Scolopaci, 95% of sampled species were identified correctly. COI barcode variation within monotypic species was low (0-1% maximum distance) but showed a wide range within polytypic species (0-5%). Preliminary Charadrii results suggest similar trends but success is reduced in the third suborder, Lari. Rates of COI evolution are found to be lowest in the Lari and this leads to reduced species identification in recently radiated families: just 49% of the Laridae and 57% of the Stercoraridae are identified but 100% of the older Alcidae. In the faster Scolopaci, subspecies are at the limit of resolution with some well differentiated subspecies not distinguished by barcodes. The interplay of evolutionary rates, divergence dates and gene flow appears to determine COI barcode differentiation between taxa.
87

Caracterização molecular de isolados de Sarcocystis spp. obtidos de marsupiais do gênero Didelphis spp. pela análise de gene mitocondrial, gene de apicoplasto, espaçador interno transcrito (ITS-1) e genes codificadores de antígenos de superfície (SAGs) / Molecular characterization of Sarcocystis spp. isolates from marsupials of the genus Didelphis spp. through the analysis of mitochondrial and apicoplast genes, internal transcribed spacer region (ITS-1) and surface antigen genes (SAGs)

Samantha Yuri Oshiro Branco Valadas 08 May 2015 (has links)
Em um trabalho anterior, foi avaliada a variabilidade de Sarcocystis spp. isolados de gambás oriundos do estado do Rio Grande do Sul pesquisando sequências gênicas codificadoras de antígenos de superfície (SAGs). Os resultados indicaram haver linhagens de isolados de Sarcocystis (relacionadas geneticamente a S. falcatula) que trocam genes em prováveis processos de recombinação sexual. A proposta deste estudo foi de conhecer as variações gênicas e relações filogenéticas em Sarcocystis spp. isolados de gambás através da análise de loci gênicos de genoma mitocondrial (CytB), de genoma de apicoplasto (ClpC) e das regiões espaçadoras transcritas internas (ITS-1), comparando a diversidade encontrada nestes grupos com a observada em integrantes da sub-familia Toxoplasmatinae. E também de conhecer a diversidade de genes codificadores de SAG-2, SAG-3 e SAG-4. Neste estudo foram obtidos esporocistos de Sarcocystis spp. através de raspado intestinal e fezes provenientes de gambás do gênero Didelphis, oriundos do Estado de São Paulo, Rio Grande do Sul e Rio Grande do Norte. Os marcadores moleculares CytB, ClpC e ITS-1 revelaram uma ampla variabilidade genotípica e alelos inéditos entre os isolados do Brasil. Os resultados levam a crer que é possível a existência de espécies “híbridas” de Sarcocystis no Brasil, com mistura de genes de ambas as espécies. Em relação aos genes codificadores de SAGs, foi encontrada uma diversidade ainda maior do que trabalho similar realizado. Os resultados sugerem uma possível ocorrência de reassortment e recombinação de alelos em sequências SAGs, contribuindo ainda mais para a geração de variabilidade e consequentemente na configuração antigênica do parasito. Estes resultados demonstram que isolados brasileiros possuem uma composição genética diferente do reportado em demais localidade, o que pode ter reflexos importantes no conhecimento da diversidade das espécies de Sarcocystis que infectam gambás em nosso meio e em toda a epidemiologia das infecções produzidas pelos protozoários deste grupo / A great variability in surface antigens genes (SAGs) of Sarcocystis spp. sporocysts was found in a previous study, through the intestinal scraping technique of opossums from the south region of Brazil. The results indicated probable sexual recombination between lineages of Sarcocystis isolates (genetically related to Sarcocystis falcatula). The aim of this research is the study of genetic variability and phylogenetic relationships among Sarcocystis spp. isolates from captured opossums in Brazil, by the analysis of molecular markers less subjected to selective pressure, such as DNA barcode, for this purpose, genetic loci from mitochondrial (CytB) and apicoplast genome (ClpC), and internal transcribed spacer regions (ITS-1) were applied and compare the results with the diversity found within members of the Toxoplasmatinae sub-family. The diversity among the SAG-2, SAG-3 and SAG-4 genes was also studied. The Sarcocystis sporocysts were obtained from opposum’s intestinal scraping and faeces from São Paulo State, Rio Grande do Sul State and Rio Grande do Norte State, Brazil. The molecular markers CytB, ClpC and ITS-1 revealed a vast genotypic variability and novel alleles among the Brazilian isolates. The results indicate possible existence of hybrid species of Sarcocystis in Brazil, with a gene mixture from both species. Regarding the SAG genes, an even greater variability was found than previously reported. The results also suggest the occurrence of reassortment and allele recombination of SAG sequences, contributing even more to the variability and thus in the parasites antigenic configuration. In this study, the results demonstrate that isolates from Brazil have a different genetic composition than reported in other locations, which may have important impacts in the knowledge of the diversity of Sarcocystis species shed by opossums in our environment and in all of the epidemiology of infections of protozoa of this particular group
88

CHARACTERIZING BILLBUG (SPHENOPHORUS SPP.) SEASONAL BIOLOGY USING DNA BARCODES AND A SIMPLE MORPHOMETRIC ANALYSIS

Marian M Rodriguez-Soto (10726101) 30 April 2021 (has links)
Insect species complexes challenge entomologists in a variety of ways ranging from quarantine protection to pest management. Billbugs (Coleoptera: Curculionidae: <i>Sphenophorus</i> spp. Schönherr) represent one such species complex that has been problematic from a pest management perspective. These grass-feeding weevils reduce the aesthetic and functional qualities of turfgrass. Sixty-four species of billbugs are native to North America, and at least ten are associated with damage to turfgrass. Billbug species are sympatric in distribution and their species composition and seasonal biology varies regionally. Since their management relies heavily on proper choice of insecticide active ingredients and timing of insecticide applications that target specific life stages, understanding billbug seasonal biology underpins the development of efficient management programs. However, billbug seasonal biology investigations are currently hindered by our inability to identify the damaging larval stage to species level. DNA barcoding, which involves the use of short DNA sequences that are unique for each species, represents one potential tool that can aid these efforts. By combining DNA-based species identification with morphometric measures capable of serving as a proxy of larval development, it may be possible to gain a more holistic understanding of billbug seasonal biology. In this study, we developed a DNA barcoding reference library using cytochrome oxidase subunit 1 (COI) sequences from morphologically identified adult billbugs collected across Indiana, Missouri, Arizona, and Utah. Next, we applied our reference library for comparison and identification of unknown larval specimens collected across the growing season in Utah and Indiana. We then used a combination of DNA barcoding and larval head capsule diameters acquired from samples collected across a short span of the growing season to produce larval phenology maps. Adult billbug COI sequences varied within species, but the variation was not shaped by geography, indicating that this locus itself could resolve larval species identity. Overlaid with head capsule diameter data from specimens collected across the growing season, a better understanding of billbug species composition and seasonal biology emerged. This knowledge will provide researchers with the tools necessary to fill critical gaps in our understanding of billbug biology thereby improving turfgrass pest management. Using this approach researchers will be able to support efforts to provide growers with the information necessary to develop more prescriptive, location-based management programs and reduce the ecological footprint of turfgrass pest management.
89

Identification of Species in Ground Meat Products Sold on the U.S. Commercial Market Using DNA-Based Methods

Kane, Dawn 01 May 2015 (has links)
Mislabeling of ground meat products is a form of food fraud that can lead to economic deception and interfere with dietary restrictions related to allergens or religious beliefs. In various parts of the world, including Ireland, Mexico and Turkey, high levels of meat mislabeling have been reported between 2000-2015. However, there is currently a lack of information regarding this practice in the United States. Therefore, the objective of this study was to test a variety of ground meat products sold on the U.S. commercial market for the presence of potential mislabeling. Forty-eight ground meat samples were purchased from online and local retail sources, including both supermarkets and specialty meat retailers. DNA was extracted from each sample in duplicate and tested using DNA barcoding of the cytochrome c oxidase subunit I (COI) gene. The resulting sequences were identified at the species level using the Barcode of Life Database (BOLD). Any samples that failed DNA barcoding went through repeat extraction and sequencing. Due to the possibility of a species mixture, these samples were also tested with real-time polymerase chain reaction (PCR) targeting beef, chicken, lamb, turkey, pork and horse. Of the 48 products analyzed in this study, 10 were found to be mislabeled, with nine containing multiple meat species. Meat samples purchased from online specialty meat distributors had a higher rate of being mislabeled (35%) compared to samples purchased from a local butcher (18%) and samples purchased at local vii supermarkets (5.8%). Horsemeat, which is illegal to sell on the U.S. commercial market, was detected in two of the samples acquired from online specialty meat distributors. Overall, the mislabeling detected in this study appears to be due to reasons such as intentional mixing of lower-cost meat species into higher cost products or unintentional mixing of meat species due to cross-contamination during processing.
90

Identifica??o de esp?cies de carn?voros (mammalia, carn?vora) utilizando sequ?ncias de DNA e sua aplica??o em amostras n?o-invasivas

Chaves, Paulo Bomfim 20 March 2008 (has links)
Submitted by PPG Zoologia (zoologia-pg@pucrs.br) on 2018-05-18T17:22:44Z No. of bitstreams: 1 dissertacao_mestrado_final_paulochaves.pdf: 4426171 bytes, checksum: 8be6ef944f497d1a9518754ebfbc27c1 (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-05-28T12:19:27Z (GMT) No. of bitstreams: 1 dissertacao_mestrado_final_paulochaves.pdf: 4426171 bytes, checksum: 8be6ef944f497d1a9518754ebfbc27c1 (MD5) / Made available in DSpace on 2018-05-28T12:35:54Z (GMT). No. of bitstreams: 1 dissertacao_mestrado_final_paulochaves.pdf: 4426171 bytes, checksum: 8be6ef944f497d1a9518754ebfbc27c1 (MD5) Previous issue date: 2008-03-20 / Sequ?ncias de DNA usadas na identifica??o de material biol?gico t?m alcan?ado consider?vel popularidade nos ?ltimos anos, especialmente no contexto dos c?digos de barras de DNA. Aferir a esp?cie de origem em amostras de pelos, penas, peles e particularmente fezes ? um passo fundamental para quem estuda a ecologia e evolu??o de diversos animais com este tipo de amostra. Este ? o caso em carn?voros, cujos h?bitos furtivos e baixas densidades populacionais de algumas esp?cies evidenciam a import?ncia de estudos baseados em amostras n?o-invasivas. Entretanto a atual escassez de ensaios padronizados de identifica??o de carn?voros freq?entemente dificulta a aplica??o dessas amostras em larga escala e compara??es de resultados entre diferentes localidades. No presente estudo n?s avaliamos dois segmentos curtos (<250 pb) de DNA mitochondrial (mtDNA) localizados nos genes ATP sintase 6 e citocromo oxidase I com potencial de servirem como marcadores-padr?o para identifica??o de carn?voros. Entre um e 11 indiv?duos de 66 esp?cies de carn?voros foram seq?enciados para um ou ambos os segmentos do mtDNA e analisados usando tr?s diferentes m?todos (?rvore de dist?ncia, dist?ncia gen?tica e an?lise de caracteres). Em geral, indiv?duos conspec?ficos apresentaram menor dist?ncia gen?tica entre si do que em rela??o a outras esp?cies, formando agrupamentos monofil?ticos. Exce??es foram algumas esp?cies que divergiram recentemente, algumas das quais ainda puderam ser identificadas pelo m?todo de caracteres, hapl?tipos esp?cie-espec?ficos, ou reduzindo a abrang?ncia geogr?fica das compara??es (restringindo a an?lise a uma regi?o zoogeogr?fica). An?lises in silico, usando um segmento curto do citocromo b freq?entemente empregado em carn?voros, tamb?m foram realizadas para comparar o desempenho deste segmento em rela??o aos outros dois propostos. N?s ent?o testamos o desempenho destes segmentos na identifica??o de fezes de carn?voros por meio de tr?s estudos de caso: (i) fezes de felinos de zool?gico, objetivando-se verificar o potencial de contamina??o das seq?encias com DNA da presa (coelho); (ii) fezes coletadas no Cerrado brasileiro contendo restos de presas (p?los, ossos, penas), supostamente proveniente de lobo-guar?, objetivando-se investigar a efici?ncia de identifica??o do predador e ocorr?ncia de interfer?ncia do DNA da presa na identifica??o; e (iii) fezes coletadas em uma reserva na Mata Atl?ntica, tamb?m com o objetivo de avaliar a efici?ncia de identifica??o. Apesar de diferen?as em alguns aspectos de sua performance, nossos resultados indicam que os dois segmentos propostos t?m um bom potencial de servir como marcadores moleculares eficientes para identifica??o acurada de amostras de carn?voros ao n?vel de esp?cie. / DNA sequences for species-level identification of biological materials have achieved considerable popularity in the last few years, especially in the context of the DNA barcoding initiative. Species assignment of biological samples such as hairs, feathers, pelts and particularly faeces is a crucial step for those interested in studying ecology and evolution of many species with these samples. This is especially the case for carnivores, whose elusive habits and low densities highlight the importance of studies based on noninvasive samples. However, the current lack of standardized assays for carnivore identification often poses challenges to the large-scale application of this approach, as well as the cross-comparison of results among sites. Here we evaluate the potential of two short (<250 pb) mitochondrial DNA (mtDNA) segments located within the genes ATP synthase 6 and cytochrome oxidase I as standardized markers for carnivore identification. Between one and eleven individuals of 66 carnivore species were sequenced for one or both of these mtDNA segments and analyzed using three different approaches (tree-based, distance-based and character-based), in conjunction with sequences retrieved from public databases. In most cases, conspecific individuals had lower genetic distances from each other relative to other species, resulting in diagnosable monophyletic clusters. Notable exceptions were the more recently diverged species, some of which could still be identified using diagnostic character attributes, species-specific haplotypes, or by reducing the geographic scope of the comparison (restricting the analysis to a single zoogeographic region). Additional in silico analyses using a short cytochrome b segment frequently employed in carnivore identification were also performed aiming to compare performance to that of our two focal markers. We then tested the performance of these segments in the identification of carnivore faeces via three case studies: (i) felid faeces collected in a controlled zoo experiment, aimed at assessing whether DNA from rabbit prey would contaminate the resulting sequences; (ii) field-collected faeces from the Brazilian Cerrado presumed to be from maned wolves and containing prey remains (hairs, bones, feathers), aimed at investigating the efficiency of predator identification and occurrence of prey DNA interference; and (iii) field-collected scats from an Atlantic Forest study site, also addressing the issue of PCR success rate and identification efficiency. In spite of some relevant differences in some aspects of their performance, our results indicate that both of our focal segments have a good potential to serve as efficient molecular markers for accurate species-level identification of carnivore samples.

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