No description available.

No description available.

The main thrust of this thesis was to characterize P.falciparum genetic diversity in northern Ghana. To do this, I used simple techniques to purify P. falciparum DNA from clinical samples across a rural setting for whole-genome sequencing. The goal was to provide a framework for exploring host-parasite genetic interactions. Utilizing Illumina deep sequencing data for 277 isolates I analyzed P. falciparum genetic diversity and described within-host diversity across this area. I observed random mating (ie no population structure) in the local parasite population, and a high genetic diversity indicative of high out-crossing. Moreover, when I aggregated my data with similar published data from Burkina Faso and Mali (sites ≈500km apart), no population structure was evident. In contrast, sites sampled in Cambodia and Thailand (≈ 800km apart) were found to have greater population structure and high potential for inbreeding. This may be driven by differences in transmission intensity between the sites sampled in West Africa and southeast Asia. To demonstrate the utility of deep sequencing data, I focused on the genomic regions of pfdhfr, pfdhps and pfcrt, known to be under antimalarial drug selection. I surveyed the full diversity of point mutations already characterized in these genes and discovered previously unknown variants. However, in order to provide a means to follow up on new variants or interesting candidate regions in large clinical samples with limited parasite DNA, I assessed the Sequenom iPLEX platform for high-throughput genotyping of P. falciparum polymorphisms. This necessitated developing a method appropriate for assigning genotypes in haploid genome mixtures common in natural infections. Finally, I used this method to type host and parasite markers in a case-control sample set from this region for exploring host-parasite genetic interactions. I found that children who have the sickle-cell trait and carry parasites that have pfdhfr resistant alleles lose their protection against severe malaria as compared to children who have normal haemoglobin and are infected with parasites with these resistant alleles.

Understanding the factors and processes that influence intraspecific genetic variation are essential to better understand evolutionary processes. In this research, I examined patterns of gene flow and their effects on the distribution of genetic variation and spatial genetic structuring at different spatial scales. I used a combination of population genetics, spatial analysis, morphometrics and phylogeography in order to understand the patterns of genetic variation and their resulting phenotypic variations in a semi-aquatic species, the muskrat (Ondatra zibethicus).

The genetic structure of oyster drill Thais clavigera along the west coast of Taiwan were assayed by starch gel electrophoresis. Factors of locality¡]i.e. Shainsan, Taisi, Budai, Chiku¡^,maturity¡]i.e. mature, immature¡^ and sampling time¡]i.e.1999.7~2000.3, 2000.11¡^ were analyzed to evaluate their effect on drill¡¥s genetic structure . Four of the eleven investigated enzyme loci were polymorphic , i.e. Ark, Lap-1, Lap-2, and Pgm-1. Among the four populations , the mean heterozygosity¡]H¡^and genetic distances¡]D¡^ ranged from 0.100 to 0.129 and from 0.0005 to 0.0029, respectively. Therefore, T. clavigera along the west coast of Taiwan belongs to the same population. However, differentiation among populations still existed which was contributed by Ark locus. According to the degree of differentiation, the four populations were ranked as Shainsan¡BBudai¡BTaisi and Chiku . A similar trend was also observed in the four populations, as the drill size increased, the mean heterozygosity and the heterozygosity of Ark locus increased. Sampling time had no significant effect on genetic structure of populations. It is suggested that environmental factors¡]e.g. environmental quality and topography¡^and drill¡¦s reproductive ecology may be important in determining the genetic structure of populations.

Thesis (MSc)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Deciphering patterns of intraspecies population genetic structuring in commercially important shark species is essential for an integrated fisheries management approach to conservation of regional biodiversity. The common smoothhound shark, Mustelus mustelus, is an overexploited, commercially and recreationally important shark species in South Africa. Considering the vulnerable status of the common smoothhound shark and due to very limited available genetic information, this study aimed to develop molecular markers, assess patterns of genetic diversity and population connectivity along the South African coast using multilocus data generated from 12 microsatellite markers and the mitochondrial gene, NADH dehydrogenase subunit 4 (ND4). The cross-species amplification of microsatellites proved useful for genetic diversity and population genetic analysis of the common smoothhound shark. These microsatellites could aid in the molecular characterisation of other endemic and cosmopolitan species and provide valuable tools for the conservation of potentially threatened or exploited shark species. For the microsatellite data, moderate levels of genetic diversity based on the heterozygosity, allelic richness and haplotype diversity were found in a total of 144 individuals sampled across eight study populations. Estimates for pairwise population differentiation, F-statistics, AMOVA and factorial correspondence analysis (FCA) indicated significant genetic structure within and between west- and east coast populations. Additionally, Bayesian clustering analyses detected two putative ancestral gene pools, supporting the presence of a biogeographic barrier at the Cape Agulhas region and therefore genetic discontinuity between the Indian and Atlantic Ocean samples. On the contrary, mitochondrial data indicated that common smoothhound shark is genetically homogenous with substantial interoceanic gene flow. Such conflicting signals found between nuclear and mitochondrial DNA (mitonuclear discordance) can be attributed to a number of factors and could simply be due to the inherent differences in marker properties or an indication of sex biased dispersal. Despite an indication of an expanding common smoothhound shark population based on both marker types, a contemporary genetic bottleneck may have gone undetected as genetic divergence was very low in some of the study populations. Nonetheless, contemporary restriction to gene flow and historical demographics such as range expansion are proposed as the most likely forces explaining genetic structure in present-day common smoothhound sharks in South Africa. For future sustainable exploitation of common smoothhound shark, the possible existence of two genetically differentiated populations and observed asymmetric gene flow along the South African coast should be taken into consideration. It is also recommended that in the future further evaluations of finescale genetic structure and seasonal migration patterns in this commercially important species are conducted in order to allow integration of this knowledge into existing fisheries management practices. / AFRIKAANSE OPSOMMING: Die ontsyfering van patrone van intraspesie populasie genetiese struktuur in kommersieel belangrike haai spesies is noodsaaklik vir 'n geïntegreerde bestuursbenadering tot visserue en bewaring van plaaslike biodiversiteit. Die hondhaai, Mustelus mustelus, is 'n oorbenutte, kommersiële en sporthengelary belangrike haai spesie in Suid-Afrika. Met inagneming van die kwesbare status van die hondhaai en as gevolg van baie beperkte beskikbare genetiese inligting, het hierdie studie gepoog om molekulêre merkers te ontwikkel, asook om die patrone van genetiese diversiteit en populasie struktuur te ondersoek langs die Suid- Afrikaanse kus deur middel van multilokus data gegenereer uit 12 mikrosatelliet merkers en die mitokondriale geen, NADH dehidrogenase subeenheid 4 (ND4). Die kruis-spesie amplifisering van mikrosatelliete was nuttig vir genetiese diversiteit en populasie genetiese analise van die hondhaai. Hierdie mikrosatelliete kan moontlik help met die molekulêre karakterisering in ander inheemse en kosmopolitaanse spesies en kan as waardevolle hulpmiddels dien in die bewaring van potensieel bedreigde en oorbenutte haai spesies. Vir die mikrosatelliet data is matige vlakke van genetiese diversiteit gevind gebaseer op heterosigositeit, alleliese rykheid en haplotipe diversiteit gevind in 'n totaal van 144 individue getoets oor agt studie populasies. Skattings vir paarsgewyse populasie differensiasie, Fstatistieke, AMOVA en faktoriale ooreenstemming analise het betekenisvolle genetiese struktuur aangedui binne en tussen wes- en ooskus populasies. Daarbenewens, het Bayesian groepering analise twee potensiele voorvaderlike geenpoele waargeneem, ter ondersteuning van die teenwoordigheid van 'n biogeografiese versperring by die Cape Agulhas gebied en dus genetiese diskontinuïteit tussen die Indiese en Atlantiese Oseaan monsters. In teenstelling het die mitokondriale data aangedui dat hierdie haai spesie geneties homogeen is met aansienlike interoseaniese geenvloei. Sulke teenstrydige tekens tussen kern en mitokondriale DNS (mitokern onenigheid) kan toegeskryf word aan 'n aantal faktore en kan eenvoudig wees as gevolg van die inherente verskille in merker eienskappe of 'n aanduiding van geslags sydigeverspreiding. Ten spyte van 'n aanduiding van 'n groeiende hondhaai populasie gebaseer op beide merker tipes, kon 'n hedendaagse genetiese bottelnek onopgemerk gegaan het aangesien genetiese divergensie baie laag was in sommige van die studie populasies. Nietemin, hedendaagse restriksie van geenvloei en historiese demografie soos verbreding van reeks voorkoming word voorgestel as die mees waarskynlike dryfkragte wat genetiese struktuur in die hedendaagse hondhaaie in Suid-Afrika verduidelik. Vir toekomstige volhoubare benutting van die spesie, moet die moontlike bestaan van twee geneties verskillende populasies en waargenome asimmetriese geenvloei langs die Suid-Afrikaanse kus in ag geneem word. Vir die toekoms word dit ook aanbeveel dat verdere evaluerings van fyn-skaal genetiese struktuur en seisoenale migrasie patrone in hierdie kommersiël belangrike spesie uitgevoer word om die integrasie van hierdie kennis in die bestaande bestuur van visserye praktyke toe te laat. / National Research Foundation (NRF)

Dispersal is a fundamental process that shapes many aspects of population ecology and evolution. Dramatic local population declines brought about by anthropogenic-driven changes to ecological processes are unfortunately becoming ubiquitous and increasing the urgency to understand dispersal behaviour in many species. For species where direct methods of tracking movement and dispersal are unsuitable, other indirect methods need to be employed to understand and characterise these behaviours. The steep population decline, due to predation, at the largest EU population of Leach’s storm-petrels Oceanodroma leucorhoa (St Kilda, Scotland), and the potential part immigrant birds have in buffering this threat, highlighted the need to understand dispersal and connectivity of widely spaced colonies in this species. The implications for the persistence of local colonies and the wider population of Leach’s storm-petrels could then be assessed. A population genetic analysis of 9 colonies across the North Pacific and Atlantic distribution, using two DNA markers (mtDNA control region and microsatellites), revealed ocean basin differentiation (Global ST = 0.32 P <0.0001, Global FST = 0.04, P <0.0001) but also identified a migrant individual in the Pacific population (STRUCTURE migrant assignment). The Atlantic population was found to be genetically homogenous, with patterns of historical and contemporary gene flow, indicating that long-distance effective dispersal is prevalent in Leach’s storm-petrels within the ocean basin. Bayesian stable isotope analysis of carbon and nitrogen (13C & 15N) provides evidence for natal dispersal as an important dispersal mechanism, and reveals movement of immature birds between colonies during the breeding season as a likely mechanism to promote inter-colony exchange and gene flow. Stable isotope comparison also identified mixing on wintering grounds as another possible influence on dispersal. The potential for immigrant birds to offset the loss caused by predation at the St Kilda colony is supported by these studies, and will likely help the persistence of the colony in the short-term. However, future viability is debateable considering the evidence for both avian and mammalian predation. This research provides a better understanding of the extent and mechanism of dispersal in the Leach’s storm-petrel, which is important to predict the potential impact of environmental change and, where possible, implement effective population management for this species.

En Méditerranée, les zones rocheuses photophiles sont principalement peuplées par des espèces du genre Cystoseira C. Agardh. Ces espèces, ingénieures d’écosystèmes, forment des forêts qui structurent l’habitat benthique des étages infralittoral et circalittoral. Ces espèces (sauf C. compressa) sont protégées par les conventions de Berne depuis 1979 et de Barcelone (1992). Pourtant aucun plan de conservation n’a encore été mis en place. L’objectif de cette thèse était de i) connaître la répartition de ces espèces le long des littoraux PACA et Corse, ii) comprendre les capacités de résilience de C. amentacea var. stricta, iii) comprendre la diversité et la structure génétique des populations de C. amentacea. L’étude de la répartition des espèces a mis en évidence l’abondance de C. amentacea et la forte régression des espèces de petits fonds i.e. C. barbata, C. crinita et C. foeniculacea f. tenuiramosa. L’étude de résilience a montré que C. amentacea est capable de recoloniser son milieu après une perturbation aiguë. Cette recolonisation suit le modèle de facilitation Enfin l’étude de génétique des populations a mis en évidence une forte structuration inter et intra-populations ainsi qu’une forte consanguinité au sein de ces populations. L’absence d’IBD sur l’ensemble de l’échantillonnage révèle la possibilité de dispersion sur de longues distances. Ces résultats n’ont pas permis de développer un plan de gestion pour ces espèces. Cependant, ils ont apporté un certain nombre de connaissances supplémentaires sur l’écologie des espèces, ils ont amorcé le travail de génétique des populations pour le genre Cystoseira et surtout ils ont permis d’identifier les axes de recherches à développer en priorité dans les futures études. Cette thèse confirme la nécessité d’une approche pluridisciplinaire en biologie de la conservation. / In the Mediterranean Sea, rocky shores are mainly inhabited by Cystoseira species. These canopy forming algae are engineers of the ecosystem. All species except for C. compressa, are protected by two international conventions, Berne (1979) and Barcelona (1992). Despite their protected status, no management plans have been developed yet. This work focused on 4 species: C. amentacea var. stricta, C. barbata, C. crinita and C. foeniculacea f. tenuiramosa. The aim of the thesis was to i) gather information about the distribution of these species and highlight the main threats and source of their disappearance along PACA and Corsican coasts, ii) Understand the resilience capacities of C. amentacea after a partial destruction of its habitat and iii) understand the diversity and the genetic structure of populations of C. amentacea along PACA and Corsican shores. The distribution of C. amentacea appears to be still important. On the contrary, the situation is more critical for species of shallow waters like C. barbata, C. crinita and C. foeniculacea f. tenuiramosa. C. amentacea showed good capacity of resilience after the perturbation. The recolonization follows the facilitation model. Finally the genetic study highlights a strong genetic structure within and among populations of C. amentacea. The within populations structure might be due to high inbreeding. No IBD was revealed among the all dataset which means that long distance dispersal might be possible for C. amentacea. The results obtained here were not sufficient to elaborate a management plan for Cystoseira species. But they completed the ecological knowledge we have on them, they brought the first large scale genetic results and they identified on what future studies must focused on.This thesis confirms the necessity to adopt a multidisciplinary approach in conservation biology.

O Brasil é o país mais rico do mundo em espécies de psitacídeos (cerca de 74), sendo 17 delas ameaçadas de extinção. Entre elas está a arara-azul (Anodorhynchus hyacinthinus) que é considerada vulnerável e pode se tornar ameaçada num futuro próximo, em conseqüência do intenso tráfico ilegal e perda do seu habitat. No presente estudo estimamos os níveis de variabilidade e caracterizamos a estrutura genética de populações naturais de A. hyacinthinus. Analisamos 10 locos de microssatélites de 98 indivíduos e seqüências concatenadas de genes mitocondriais (ND5, citocromo-b e ND2; 2123 pb total) de 80 indivíduos. O índice de diversidade genética foi considerado baixo em relação a outras espécies de psitacídeos. Além disso, os índices RST e a análise bayesiana dos dados de microssatélites indicaram moderada estruturação genética entre indivíduos de quatro regiões geográficas (Pantanal norte, Pantanal sul, norte e nordeste), mas os índices de FST indicaram diferenciação somente entre três regiões (norte e nordeste sem diferenciação). A estruturação entre essas três regiões foi congruente com a forte estruturação genética apontada pelos índices de FST e pela rede de haplótipos das seqüências mitocondriais. Baseado nos dados mitocondriais o tempo de divergência entre os grupos genéticos de A. hyacinthinus foi estimado em 16 a 42 mil anos atrás, o que corresponde ao final do Pleistoceno. Ainda, os resultados apontaram para uma população demograficamente estável ao longo do tempo, o que pode indicar que a baixa variabilidade pode ser uma característica da espécie. Entretanto, a rede de haplótipos apresenta forma em estrela com alguns haplótipos de baixa freqüência, o que pode indicar expansão recente, principalmente para região nordeste. Baseado nos dados de estruturação genética populacional, foi possivel indicar a possível origem de indivíduos apreendidos e sem procedência conhecida, o que é importante para realizar ações preventivas de repressão e fiscalização. Adicionalmente, foram analisados sete locos de microssatélites de filhotes amostrados no mesmo ninho (mesma estação reprodutiva, estações reprodutivas consecutivas e estações alternadas) em duas regiões do Pantanal. Os resultados sugerem que a espécie é predominantemente monogâmica estrita, mas há pelo menos 12,5% de paternidade extra-par e 6,5% de parasitismo de ninho. Além disso, foram confirmados dados obtidos em campo de que muitos casais utilizam o mesmo ninho em anos consecutivos e alternados. Finalmente, padronizamos a sexagem molecular de amostras de penas de muda. Concluindo, os resultados genéticos obtidos nesse trabalho trazem informações sobre os processos envolvidos na história evolutiva dessa espécie, além de contribuir com informações sobre o comportamento reprodutivo das araras-azuis proporcionando mais subsídios para elaboração de programas de conservação. / Brazil has the highest number of parrot species in the world (about 74), 17 of them endangered. Among them is the hyacinth macaw (Anodorhynchus hyacinthinus), which is considered vulnerable and could become endangered in the near future, due to the intense illegal traffic and loss of habitat. In this study we estimated levels of variability and characterized the genetic structure of natural populations of hyacinth macaws. We analyzed 10 microsatellite loci from 98 individuals and concatenated sequences of mitochondrial genes (ND5, cytochrome b and ND2, 2,123 bp total) from 80 individuals. The genetic diversity index was low compared to those from other species of parrots. In addition, RST indeces and Bayesian analysis of microsatellite data showed moderate genetic structure among individuals of four regions in Brazil (north Pantanal, south Pantanal, north and northeast), but FST indeces indicate differentiation only between three regions (north and northeast without differentiation). This is in accordance with the strong genetic structure indicated by FST indeces and haplotype network based on mitochondrial sequences. Based on the mitochondrial data, the time of divergence of the genetic groups of hyacinth macaws was estimated to have occurred 16 to 42 thousand years ago, which corresponds to the late Pleistocene. Still, the results suggest that the population has been demographically stable over time, which may indicate that the low variability levels may be a characteristic of the species. However, the haplotype network presents a star shape, which indicate recent expansion, specially in the northeast. Additionally, given the population genetic structure data, it was possible to identify the most probable region of origin of apprehended individuals, this information is important to plan preventive and repressive control. Additionally, we analyzed seven microsatellite loci of chicks sampled in the same nest (same breeding season, alternate breeding seasons and consecutive seasons) in two regions of the Pantanal. The results suggest that the species is predominantly monogamous, but there is at least 12.5% of extra-pair paternity and 6.5% of brood parasitism. Furthermore, the genetic data is congruent with field observations that suggest that many couples return to the same nest in consecutive and alternative breeding seasons. Finally, we standardized for a molecular sexing protocol for molten feathers. In conclusion, the genetic results obtained in this study provide information about the processes involved in the evolutionary history and the reproductive behavior of hyacinth macaws that may help plan conservation actions.

La compréhension de la structure génétique et de la dynamique des populations, ainsi que les mécanismes moléculaires régissant les interactions entre l’hôte et le pathogène sont des éléments clefs pour la gestion des maladies. Au cours de cette étude nous avons cherché à développer dans un premier temps des marqueurs microsatellites à partir de données issues du séquençage d’un isolat de Ganoderma boninense. Ces marqueurs microsatellites nous ont permis d’étudier la structuration génétique et l’histoire démographique de G. boninense. Le génotypage d’un sous-échantillonnage issus de missions de récolte en Malaisie et à Sumatra nous a permis de mettre en évidence deux groupes principaux au sein de l’Asie du Sud-Est. Nous avons ensuite concentré notre attention sur la région regroupant la Malaisie péninsulaire et l’île de Sumatra, zone historique de développement de la culture industrielle du palmier à huile et d’apparition de la pourriture basale du stipe due à G. boninense, qui semble former une seule population. Nous avons examiné à une échelle géographique plus restreinte cette zone géographique afin de mettre en évidence une potentielle sous structure au sein de cette population. En testant l’effet du fond génétique du palmier d’origine de chaque individu, du nombre de génération de palmier précédent le moment de la récolte de l’individu, ou de la distance géographique sur une possible sous-structure des individus au sein de la zone historique, nous avons mis en évidence qu’aucune sous-structure génétique n’émergeait. En revanche, La comparaison de plusieurs scénarios d’évolution démographique a permis de mettre en évidence un phénomène d’expansion très ancien bien antérieur au début du développement de la culture industrielle du palmier à huile. Pour finir, la comparaison des données transcriptomiques entre des isolats agressifs et non agressifs a permis de souligner la présence d’une centaine de gènes différentiellement exprimés possédant une annotation fonctionnelle. Les résultats de ces deux approches pourront permettre une meilleure gestion de la maladie ainsi que l’amélioration des programmes développement et de gestion des résistances. / The understanding of genetic structuration and population dynamic, as well as the molecular mechanisms ruling host/pathogen interaction, are key elements for disease management. During this study, as first step, we were looking to develop microsatellites markers from genomic data obtained from sequencing of a Ganoderma boninense pure strain. Those markers allowed studying genetic structuration and demographic history of G. boninense. Genotyping of a subset of samples from sampling mission in Malaysia and Sumatra have highlighted two main groups in South-East Asia. Then, we focused on a region gathering peninsular Malaysia and Sumatra together, as it is both historical region of industrial oil palm culture development and the first region of basal stem rot observation caused by G. boninense, and to appear as a single population. We examine this region at a lower scale in order to highlight a potential genetic substructure in this population. We tested for effect of genetic tree background of each sample, number of planting generation before sampling and geographical distance between sample in order to observe a potential correlation between genetic substructure and one of those factors. As no correlation appeared, we concluded that this population does not have a genetic substructure. On the other hand, the comparison between several demographic evolution scenarios have shown a strong support for a past expansion event further back in time from the beginning of industrial oil palm culture development. To conclude, the exploration of transcriptomic data between strains owning aggressive or non-aggressive profile showed the differential expression of a hundred genes owning a functional annotation. Results from both approaches will allow the development of better disease management and a better resistance selection and management program.