O Estado de São Paulo apresenta uma das mais altas taxas de infecções por vírus dengue no Mundo, mas apesar dessa situação, poucos são os estudos dirigidos às populações do mosquito Aedes aegypti. O objetivo deste trabalho foi caracterizar geneticamente e morfologicamente populações de Ae. aegypti localizadas em seis municípios (Santos, S.P., Campinas, São Carlos, Catanduva, S.J.R.P.) do Estado de São Paulo durante 2011 e 2012. Todos os marcadores biológicos indicaram estruturação populacional. Os oito loci microssatélites apontaram diferenciação genética moderada entre as populações (Fst= 0.04; p < 0,05) e os níveis de diversidade nucleotídica do gene COI (&pi; =0,0062) e do gene ND4 (&pi;=0,017) foram moderadamente altos. Duas linhagens geneticamente distintas foram encontradas no Estado. Ao longo dos meses que compreenderam o estudo, foram encontradas diferenças morfo-genéticas temporais entre as seis populações analisadas, possivelmente indicativas de microevolução. Os resultados obtidos podem ser úteis para compreendermos a dispersão deste mosquito vetor. / The State of São Paulo displays one of the highest rates of dengue infection in the world, but despite this fact, a few populational studies of Ae. aegypti have been undertaken. The aim of this study was to genetically and morphologically characterize Ae. aegypti populations from six locations in the São Paulo State (Santos, S.P., Campinas, São Carlos, Catanduva, S.J.R.P.) during 2011 and 2012. The phenetic and genetic analyses revealed that populations of Ae. aegypti are structured. Eight microsatellites loci were polymorphic and genetic differentiation among samples was moderate (Fst= 0.04; p < 0.05). Nucleotide diversities of COI (&pi; = 0.0062) and ND4 gene (&pi; = 0.017) were moderately high. Two lineages distinct genetically were found in the State. Over the months comprised by the study, we found the temporal genetics and morphologics differences among the six populations, a possibly indicative of microevolution of mosquitoes. The results of this study may be useful for understand the spread of this vector mosquitoes in the State of São Paulo.

Species conservation is typically founded upon a range of management strategies, which integrate both biological and socioeconomic data. In this thesis, population genetics, acoustic telemetry, spawning aggregation surveys and stakeholder assessments were used to address key knowledge gaps limiting effective conservation management for critically endangered Nassau grouper (Epinephelus striatus) stocks in The Bahamas. A panel of polymorphic microsatellite markers was optimised to assess the genetic population dynamics of more than 400 Nassau grouper sampled throughout the country. Microsatellite data indicate that contemporary Nassau grouper populations in The Bahamas are predominantly genetically diverse and weakly differentiated, but lack geographic population structure. Assessments of changes in effective population size (Ne) show substantive reductions in Ne within The Bahamas compared to historic values that are likely due to natural disturbances. Evidence for recent bottlenecks occurring in three islands as well as an active spawning site, along with higher inbreeding coefficients in two islands were also found, and can be attributed to more recent anthropogenic activities. Collapse of a historically important Nassau grouper fish spawning aggregation (FSA) was supported by both acoustic telemetry and spawning aggregation survey dives. Restriction-site-associated DNA sequencing (RAD-seq) of 94 Nassau grouper was used to explore intraspecific population dynamics, loci under selection and patterns of gene flow in The Bahamas. Genomic assessments of diversity were in accord with microsatellite data and examinations of gene flow support higher levels of connectivity in The Bahamas than was previously suggested. The increased resolution gained from assessments of genomic data support intraspecific population structuring that may be driven by differences in gene flow and putative loci under divergent selection. Telemetry data were successfully used to identify the origins of spawning adults, and support demographic connectivity through migrations between an active FSA in the central Bahamas and home reef habitats within the Exumas and a no-take marine protected area. Stakeholder assessments highlight the complexities of fisheries management within The Bahamas, with key stakeholders often exhibiting conflicting opinions regarding the status of Nassau grouper and the efficacy of management options. However, these groups mutually agree upon the need to better manage remaining Nassau grouper stocks within The Bahamas through science-grounded policies. Synthesis of these studies along with a review of fisheries governance in The Bahamas was used to develop a comprehensive national management plan for Nassau grouper to facilitate better conservation for remaining populations of this ecologically important marine species.

Species identifications have been historically difficult in the economically important spruce budworm (Choristoneura fumiferana) pest complex. Morphological, ecological, behavioural, and genetic characters have been studied to try to understand the taxonomy of this group, but diagnostic character states differ in frequency rather than being complete replacements between each species. I developed a morphology-based character system that focuses on forewing colour components (Chapter 2), as well as eight simple sequence repeats (SSRs, also referred to as microsatellite markers) (Chapter 3). I tested these along with a 470 bp region of COI mitochondrial DNA (mtDNA) (Chapter 2, 4) to determine their congruence with putative species that were identified by adaptive traits (larval host plant, length of larval diapause, larval and adult morphology, pheromone attraction, distribution). The morphometrics system was effective for identification of the five species tested, with only slight overlap between C. fumiferana and C. biennis. MtDNA distinguished C. fumiferana and C. pinus pinus, but the remaining species shared haplotypes. SSRs distinguished four species (C. fumiferana, C. pinus pinus, C. retiniana, C. lambertiana) but the remaining four species that were included in this survey (Chapter 4) remained mixed within two populations. There was evidence for hybridization between several species pairs. I also conducted a detailed study (Chapter 5) in Cypress Hills, an isolated remnant coniferous forest in western Canada, where identifying individuals from the Choristoneura fumiferana complex has been impossible due to the unusual ecogeographic characteristics of the area. I integrated data on behaviour, ecology, morphology, mtDNA, and SSRs, comparing Cypress Hills populations to those from other regions of North America to determine which species they resembled most. I delimited at least three populations, resembling C. fumiferana, C. occidentalis and C. lambertiana. Adult flight phenology, along with pheromone attraction, were identified as major isolating mechanisms between these populations. My studies highlighted the importance of integrative taxonomy for understanding species boundaries. Their patterns of differentiation suggest that spruce budworm species have recently diverged via natural selection in spite of some gene flow. Overall, this work is intended to contribute to more accurate identification of specimens and a better understanding of the evolutionary processes that drive speciation. / Systematics and Evolution

<p>In many forensic cases, such as mass disasters reconstruction cases, the recovered DNA is highly degraded. In such incidences, typing of STR loci has become one of the most powerful tools for retrieving information from the degraded DNA. However, as DNA degradation proceeds, three phenomena occur consecutively: loci imbalance, allele dropout and no amplification. To solve the problem of degraded DNA, redesigned primer sets have been developed in which the primers were positioned as close as possible to the STR repeat region. These reduced primer sets were called Miniplexes. Unfortunately, a few of the CODIS STR loci cannot be made into smaller amplicons. For this reason non-CODIS miniSTRs have been developed. The present study was undertaken for the population genetic analysis of microsatellite variation in four South African populations / Afrikaner, Xhosa, Mixed Ancestry and Asian Indian using eight non-CODIS miniSTR loci. These miniSTRs loci were characterized within the populations by estimating the levels of diversity of the markers, estimating the population genetic parameters, and studying the inter-population relationships. All of the miniSTRs were amplified successfully and the genetic variability parameters across all loci in Afrikaner, Mixed Ancestry, Asian Indian and Xhosa were estimated to be in the range of 3 (D4S2364) to 12 (D9S2157) alleles, the total number of alleles over all loci ranged from 100 to 204, the allelic richness ranged from 3.612 to 10.307 and the heterozygosity ranged from 0.4360 to 0.8073. Genetic distance was least between Afrikaner and Asian Indian and highest between Xhosa and Mixed Ancestry. Deviations from Hardy-Weinberg equilibrium were not observed for most of the loci. The low mean FIS (-0.027) and FIT (-0.010) and FST (0.017) values across the populations indicated low level of inbreeding within (FIS) and among (FST) the populations. The Asian Indian population showed higher levels of the inbreeding coefficient, indicating less gene exchange between it and other populations. These 8 markers can be used for genetic investigations and assessing population structure. The study contributed to the knowledge and genetic characterization of four South African populations. In addition, these MiniSTRs prove to be useful in cases where more genetic information is needed.</p>

S'ha estudiat la variabilitat genètica de les poblacions del nord d'Àfrica a partir de l'anàlisi de diverses regions genòmiques per tal d'entendre les poblacions analitzades d'una banda, i comprendre la dinàmica del genoma per l'altra. Els resultats obtinguts ens han permès verificar diferents hipòtesis sobre la història de les poblacions d'aquesta regió com són l'efecte paral·lel i independent de l'onada de difusió del neolític des de l'Orient Mitjà al llarg d'ambdues ribes de la Mediterrànea; i l'efecte de l'arabització. S'ha pogut estimar també la contribució genètica masculina nord africana a la península ibèrica i detectat certa contribució genètica del pobles sub-saharians a les poblacions nordafricanes. Per altra banda, el tipatge de marcadors genètics que evolucionen a velocitats diferents al cromosoma Y ha permès mostrar que el background genètic predomina sobre el background poblacional en l'estructura de la variació genètica dels microsatèl·lits en la regió no recombinant del cromosoma Y humà. / The genetic variability of the North African populations has been studied through the analysis of different genomic regions in order to understand both the analysed populations and the dynamics of the genome. The obtained results allow us to verify different hypotheses about the population history of this region including the parallel and independent effect of the Neolithic wave of advance from the Middle East and along both Mediterranean coasts; and the effect of Arabization phenomena. We also tried to estimate the North African male genetic contribution to the Iberian peninsula and detected Sub-Saharian genetic influences to the North African peoples. Moreover, the typing of genetic markers with different evolutionary rates on the Y chromosome allowed us to demonstrate that variation in microsatellites is deeply structured by genetic background on the non-recombining region of the human Y chromosome.

Η μεσογειακή μύγα αποτελεί το κύριο παράσιτο πολλών καλλιεργούμενων φρούτων προκαλώντας ετησίως μεγάλες καταστροφές σε γεωργικές καλλιέργειες. Η μελέτη του εντόμου τόσο σε γενετικό όσο και σε πληθυσμιακό επίπεδο μπορεί να συμβάλλει σημαντικά στην ανάπτυξη ή και τη βελτίωση αποτελεσματικών και φιλικών προς το περιβάλλον μεθόδων ελέγχου. Οι μικροδορυφόροι είναι απλές επαναλήψεις ενός νουκλεοτιδικού μοτίβου που αποτελείται 1-6 ζεύγη βάσεων. Αποτελούν πολύ χρήσιμους γενετικούς δείκτες διότι είναι άφθονοι και διάσπαρτοι στο γονιδίωμα των ευκαρυωτκών οργανισμών. Επιπλέον είναι υψηλά πολυμορφικοί, κληρονομούνται ως συνυπερέχοντες Μεντελικοί δείκτες και αναλύονται εύκολα μέσω PCR, χαρακτηριστικά που τους καθιστούν πολύτιμα εργαλεία για πληθυσμιακές και εξελικτικές μελέτες. Από τους μικροδορυφορικούς δείκτες που αναπτύχθηκαν στο εργαστήριό μας, επιλέχθηκαν 10 με βάση το βαθμό πολυμορφισμού που έδειξαν σε εργαστηριακά στελέχη. Οι δείκτες αυτοί χρησιμοποιήθηκαν στην ανάλυση 481 ενηλίκων ατόμων που προέρχονταν από 19 διαφορετικά δείγματα φρούτων που συλλέχθηκαν από εννέα διαφορετικές περιοχές της Δυτικής Ελλάδας και της Βόρειας Πελλοπονήσου. Η γενοτυπική ανάλυση πραγματοποιήθηκε με ηλεκτροφόρηση των προϊόντων της PCR για κάθε γενετικό δείκτη σε πήκτωμα πολυακρυλαμιδίου και επακόλουθη αυτοραδιογραφία. Η στατιστική επεξεργασία των αποτελεσμάτων, με τη βοήθεια υπολογιστικών προγραμμάτων αποκάλυψε σημαντική γενετική διαφοροποίηση μεταξύ των δειγμάτων, που αποδίδεται εκτός των άλλων παραγόντων (κλιματολογικές συνθήκες, γεωγραφική προέλευση) στο είδος του ξενιστή. / C. capitata, is the main pest of many cultivable fruits and responsible for a significant loss in annual products, resulting in great economic damage. Studies on the genetic and population analysis will make a contribution towards the development or the improvement of environmental friendly control methods. Microsatellites are tandem simple sequence repeats of short (1-6) nucleotide motifs. They are very important genetic markers because they are dispersed and abundant in most eukaryotic genomes. They are highly polymorphic, inherited as co-dominant Mendelian markers and easily scored by PCR. Consequently, they have become one of the most popular molecular markers with application in many genetic studies, including genetic analysis of natural populations and evolutionary studies. From the available microsatellites in our laboratory, were selected ten (10), based on their polymorphism in laboratory strains. They were used for the screening of 481 adult flies in the medfly, collected from nineteen (19) different samples of fruits from nine (9) different areas in west Greece and north Peloponnesus. Analysis of genotype composition in the samples was achieved by polyacrylamide electrophoresis of the PCR products, for every genetic marker and then autoradiography. The statistic analysis of our results using software programs, revealed an important genetic differentiation in samples. Except for many factors (climatic conditions, geographic origin), the host origin will be responsible for this genetic differentiation.

No description available.

No description available.

No description available.

<p>In many forensic cases, such as mass disasters reconstruction cases, the recovered DNA is highly degraded. In such incidences, typing of STR loci has become one of the most powerful tools for retrieving information from the degraded DNA. However, as DNA degradation proceeds, three phenomena occur consecutively: loci imbalance, allele dropout and no amplification. To solve the problem of degraded DNA, redesigned primer sets have been developed in which the primers were positioned as close as possible to the STR repeat region. These reduced primer sets were called Miniplexes. Unfortunately, a few of the CODIS STR loci cannot be made into smaller amplicons. For this reason non-CODIS miniSTRs have been developed. The present study was undertaken for the population genetic analysis of microsatellite variation in four South African populations / Afrikaner, Xhosa, Mixed Ancestry and Asian Indian using eight non-CODIS miniSTR loci. These miniSTRs loci were characterized within the populations by estimating the levels of diversity of the markers, estimating the population genetic parameters, and studying the inter-population relationships. All of the miniSTRs were amplified successfully and the genetic variability parameters across all loci in Afrikaner, Mixed Ancestry, Asian Indian and Xhosa were estimated to be in the range of 3 (D4S2364) to 12 (D9S2157) alleles, the total number of alleles over all loci ranged from 100 to 204, the allelic richness ranged from 3.612 to 10.307 and the heterozygosity ranged from 0.4360 to 0.8073. Genetic distance was least between Afrikaner and Asian Indian and highest between Xhosa and Mixed Ancestry. Deviations from Hardy-Weinberg equilibrium were not observed for most of the loci. The low mean FIS (-0.027) and FIT (-0.010) and FST (0.017) values across the populations indicated low level of inbreeding within (FIS) and among (FST) the populations. The Asian Indian population showed higher levels of the inbreeding coefficient, indicating less gene exchange between it and other populations. These 8 markers can be used for genetic investigations and assessing population structure. The study contributed to the knowledge and genetic characterization of four South African populations. In addition, these MiniSTRs prove to be useful in cases where more genetic information is needed.</p>