Wolbachia é uma bactéria intracelular encontrada tanto nos tecidos somáticos quanto nos reprodutivos de diversas espécies de artrópodes e nematódeos. Estudos filogenéticos baseados nos genes 16S e ftsZ indicaram que o gênero Wolbachia congrega seis supergrupos taxonômicos (\"A\" a \"F\"). Infecções por Wolbachia têm sido associadas a diversas alterações na reprodução de seus hospedeiros, p. exemplo, a incompatibilidade citoplasmática (IC), partenogênese, feminização de machos genéticos e morte dos machos. A identificação das diferentes cepas da bactéria é mais precisa quando a análise por locos múltiplos (MLST) é aplicada. Infecção por Wolbachia foi descrita em diversas espécies de moscas-das-frutas da familia Tephritidae, Bactrocera ascita, Rhagoletis cerasi, Ceratitis capitata, nas quais a bactéria induz a incompatibilidade citoplasmática. No gênero Anastrepha, endêmico do Continente Americano, infecção por Wolbachia foi descrita em várias espécies pela análise do gene wsp, existindo também a indicação de que IC mediada por Wolbachia ocorra em duas espécies do grupo fraterculus. A ocorrência de IC aliada à sugestão do emprego da Wolbachia em programas de controle populacional das moscas-das-frutas, impõem a necessidade de uma caracterização mais precisa das diferentes cepas da Wolbachia. No presente trabalho foram amplificados e sequenciados fragmentos dos genes gatB, coxA, hcpA, ftsZ e fbpA, que integram a metodologia de MLST (\"Multiloci Sequence Typing\") e do gene wsp da Wolbachia. Foram analisadas amostras populacionais do complexo de espécies crípticas de Anastrepha fraterculus do Brasil e da Argentina, Peru, Equador, Colômbia, Guatemala e México, além de amostras de Anastrepha obliqua do Brasil. Todas as amostras estavam infectadas com Wolbachia do supergrupo \"A\". Para os cinco genes, foram encontrados haplótipos únicos e outros já descritos anteriormente, determinando, assim, os alelos de cada um presentes nas amostras. O conjunto de cinco alelos de cada amostra determinou a linhagem da bactéria que estava presente. Comparação entre as análises filogenéticas das sequências de cada um dos genes isoladamente, mostrou discordância nas relações entre os alelos e amostras populacionais. As sequências dos cinco genes concatenadas, com 2079 pb, foram analisadas tendo sido encontrados 20 linhagens, com distâncias variando de 0,001 a 0,058. A análise filogenética isolou as linhagens de Wolbachia obtidas das amostras de Anastrepha em clados distintos, demonstrando que diferentes linhagens estão presentes nesses hospedeiros e regiões geográficas. Mostrou, também, que pode ocorrer mais que uma cepa de Wolbachia em uma mesma amostra populacional. Uma das linhagens foi detectada em duas espécies do complexo fraterculus e é, também, a mais comumente encontrada (ST1) em diferentes organismos. As sequências do wsp tinham cerca de 500 pb, tendo sido encontradas 22 sequências distintas. O nível de variabilidade de nucleotídeos não é uniforme ao longo do gene, formando um padrão com quatro regiões hipervariáveis, \"HVRs\". As distâncias genéticas entre os haplótipos de wsp mostrou uma variação de 0,001 a 0,235. Foram observadas evidências de recombinação intragência entre os haplótipos do gene wsp. A análise filogenética também isolou os haplótipos de Wolbachia em clados distintos, porém, em contraste com o MLST, a árvore do gene wsp, não suporta os grupos monofiléticos gerados pelo MLST. Os resultados mostram que linhagens similares de Wolbachia estão disseminadas por vasta extensão do Continente Americano, além da presença de linhagens específicas em determinadas áreas geográficas. Análises de ovários e testículos de indivíduos infectados e não infectados (curados por tratamento térmico) de A. sp. 1 e de A. obliqua foram feitas para avaliar possíveis efeitos da Wolbachia nesses hospedeiros. A análise das preparações dos ovários, coradas pelo DAPI, não mostrou diferenças perceptíveis nesta análise morfológica entre fêmeas infectadas e não infectadas, de ambas as espécies. A produção de espermatozoides aumenta progressivamente durante alguns dias, após a emergência das imagos, e cai nos dias seguintes. A análise da produção de espermatozoides pelos machos infectados e pelos curados mostrou que as diferenças entre eles não foram significativas, em ambas as espécies de hospedeiros. Foram feitas estimativas da fecundidade de fêmeas infectadas e não infectadas, de ambas as espécies. Mostrou-se que fêmeas infectadas são mais fecundas que as não infectadas em A. sp.1, mas mostram fecundidade similar em A. obliqua. As taxas de eclosão de larvas foram também estimadas em cruzamentos intraespecíficos compatíveis (fêmeas infectadas ou não cruzadas com machos não infectados) e cruzamentos incompatíveis (fêmeas não infectadas cruzadas com machos infectados) de ambas as espécies. A fertilidade foi significativamente mais elevada entre os ovos produzidos pelas fêmeas infectadas, de ambas as espécies. Foi observado que machos infectados, em ambas as espécies, estão relacionados com os cruzamentos onde ocorreram as taxas mais altas de eclosão. Analisando os cruzamentos incompatíveis, foi demonstrada a presença de incompatibilidade citoplasmática (IC), como seria esperado pela atuação da Wolbachia. Foi mostrado um alto valor para os índices de IC em A. sp,1 (IC= 54,01%) e em A. obliqua (IC = 66,2%). Os resultados sugerem que podem existir relações mutualísticas insipientes da Wolbachia com suas espécies de Anastrepha hospedeiras / Wolbachia is an intracellular bacteria found in somatic and in the reproductive tissues of various arthropods and nematodes. Phylogenetic studies based on 16S and ftsZ genes indicated the existence of six Wolbachia taxonomic supergroups (\"A\" through \"F\"). Infection of Wolbachia have been linked to several changes in the reproduction of their hosts, like cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males and male killing. T Wolbachia infection has been described in several species of fruit flies of the family Tephritidae, like Bactrocera ascita, Rhagoletis cerasi, Ceratitis capitata, in which the bacteria induces cytoplasmic incompatibility. In Anastrepha, endemic to the American Continent, Wolbachia infection has been described in several species by analysis of the wsp gene, and there is also indications that Wolbachia-mediated CI occurs in two species of the fraterculus group. The occurrence of CI coupled with the suggestion of the use of Wolbachia in population supression programs, impose the need for a more precise characterization of the different strains of Wolbachia. The identification of the different strains of the bacteria is most accurate when the methodology of multiple loci (MLST) is applied. In this study fragments of genes gatB, coxA, hcpA, ftsZ and fbpA, integrating the methodology MLST, and of wsp gene were amplified and sequenced. Population samples of the Anastrepha fraterculus.complex of cryptic species from Brasil, Argentina, Peru, Ecuador, Colombia, Guatemala and Mexico, and samples of A. oblique from Basil were analysed. All samples were infected with supergroup \"A\" Wolbachia. For each of the five MLST genes, unique as well already known haplotypes were found. Phylogenetic analyses of each gene isolated showed incongruences in the relationships among haplotypes and population samples. The concatenated sequences of the five genes, with 2079 bp, were analyzed and 20 haplotypes were found, with distances ranging from 0.001 to 0.058. Phylogenetic analysis of Wolbachia isolated haplotypes into distinct clades, demonstrating that different strains of Wolbachia were present in these hosts, and in distinct geographic areas. Hosts specific haplotypes were found as well as more than one strain of Wolbachia was found in given population samples. A haplotypes (ST1) was detected in two species of the complex and is also the most commonly found in different organisms. Twenty two different sequences of about 500 bp were found for the wsp gene. The level of nucleotide variability is not uniform along the gene, forming a pattern with four hypervariable regions, HVRs. Genetic distances between haplotypes showed a variation from 0.001 to 0.235. Phylogenetic analysis of the haplotypes also isolated Wolbachia into distinct clades, but in contrast to the MLST, the tree formed by wsp gene does not support the monophily of some groups. The data show that strains of Wolbachia are disseminated along the American Continent, and also that there are specific strains in determined geographic areas. Analyses of ovaries and testes from infected and non infected (cured by heat treatment) individuals of A. sp. 1 and A. obliqua were made in search of possible effects of Wolbachia on its hosts. Ovaries from infected and cured females of both species, stained by DAPI, showed no visible differences in this morphological analysis. The production of sperms increases during few days after ermergence and drops out later one. Analysis of infected and cured males showed that the production of sperms were not significant between them, for both the host species. Fecundity of infected females of A, sp.1 was significantly higher than that of cured females, but was similar in A. obliqua, Egg hatching was scored in compatible intraspecies crosses and also in incompatibles crosses, of both species. Fertility was significntly higher for infected females of both species. Infected males of both species were found associated to crosses in which the higher egg hatching was observed. Analyses of incompatible crosses showed that CI occurred at high rates in A. sp.1 (CI = 54.01%) and in A. obliqua (CI = 66.2%). The data suggest that an incipient mutualism may be present in the relationships of Wolbachia and its Anastrepha hosts

Wolbachia é uma bactéria intracelular encontrada tanto nos tecidos somáticos quanto nos reprodutivos de diversas espécies de artrópodes e nematódeos. Estudos filogenéticos baseados nos genes 16S e ftsZ indicaram que o gênero Wolbachia congrega seis supergrupos taxonômicos (\"A\" a \"F\"). Infecções por Wolbachia têm sido associadas a diversas alterações na reprodução de seus hospedeiros, p. exemplo, a incompatibilidade citoplasmática (IC), partenogênese, feminização de machos genéticos e morte dos machos. A identificação das diferentes cepas da bactéria é mais precisa quando a análise por locos múltiplos (MLST) é aplicada. Infecção por Wolbachia foi descrita em diversas espécies de moscas-das-frutas da familia Tephritidae, Bactrocera ascita, Rhagoletis cerasi, Ceratitis capitata, nas quais a bactéria induz a incompatibilidade citoplasmática. No gênero Anastrepha, endêmico do Continente Americano, infecção por Wolbachia foi descrita em várias espécies pela análise do gene wsp, existindo também a indicação de que IC mediada por Wolbachia ocorra em duas espécies do grupo fraterculus. A ocorrência de IC aliada à sugestão do emprego da Wolbachia em programas de controle populacional das moscas-das-frutas, impõem a necessidade de uma caracterização mais precisa das diferentes cepas da Wolbachia. No presente trabalho foram amplificados e sequenciados fragmentos dos genes gatB, coxA, hcpA, ftsZ e fbpA, que integram a metodologia de MLST (\"Multiloci Sequence Typing\") e do gene wsp da Wolbachia. Foram analisadas amostras populacionais do complexo de espécies crípticas de Anastrepha fraterculus do Brasil e da Argentina, Peru, Equador, Colômbia, Guatemala e México, além de amostras de Anastrepha obliqua do Brasil. Todas as amostras estavam infectadas com Wolbachia do supergrupo \"A\". Para os cinco genes, foram encontrados haplótipos únicos e outros já descritos anteriormente, determinando, assim, os alelos de cada um presentes nas amostras. O conjunto de cinco alelos de cada amostra determinou a linhagem da bactéria que estava presente. Comparação entre as análises filogenéticas das sequências de cada um dos genes isoladamente, mostrou discordância nas relações entre os alelos e amostras populacionais. As sequências dos cinco genes concatenadas, com 2079 pb, foram analisadas tendo sido encontrados 20 linhagens, com distâncias variando de 0,001 a 0,058. A análise filogenética isolou as linhagens de Wolbachia obtidas das amostras de Anastrepha em clados distintos, demonstrando que diferentes linhagens estão presentes nesses hospedeiros e regiões geográficas. Mostrou, também, que pode ocorrer mais que uma cepa de Wolbachia em uma mesma amostra populacional. Uma das linhagens foi detectada em duas espécies do complexo fraterculus e é, também, a mais comumente encontrada (ST1) em diferentes organismos. As sequências do wsp tinham cerca de 500 pb, tendo sido encontradas 22 sequências distintas. O nível de variabilidade de nucleotídeos não é uniforme ao longo do gene, formando um padrão com quatro regiões hipervariáveis, \"HVRs\". As distâncias genéticas entre os haplótipos de wsp mostrou uma variação de 0,001 a 0,235. Foram observadas evidências de recombinação intragência entre os haplótipos do gene wsp. A análise filogenética também isolou os haplótipos de Wolbachia em clados distintos, porém, em contraste com o MLST, a árvore do gene wsp, não suporta os grupos monofiléticos gerados pelo MLST. Os resultados mostram que linhagens similares de Wolbachia estão disseminadas por vasta extensão do Continente Americano, além da presença de linhagens específicas em determinadas áreas geográficas. Análises de ovários e testículos de indivíduos infectados e não infectados (curados por tratamento térmico) de A. sp. 1 e de A. obliqua foram feitas para avaliar possíveis efeitos da Wolbachia nesses hospedeiros. A análise das preparações dos ovários, coradas pelo DAPI, não mostrou diferenças perceptíveis nesta análise morfológica entre fêmeas infectadas e não infectadas, de ambas as espécies. A produção de espermatozoides aumenta progressivamente durante alguns dias, após a emergência das imagos, e cai nos dias seguintes. A análise da produção de espermatozoides pelos machos infectados e pelos curados mostrou que as diferenças entre eles não foram significativas, em ambas as espécies de hospedeiros. Foram feitas estimativas da fecundidade de fêmeas infectadas e não infectadas, de ambas as espécies. Mostrou-se que fêmeas infectadas são mais fecundas que as não infectadas em A. sp.1, mas mostram fecundidade similar em A. obliqua. As taxas de eclosão de larvas foram também estimadas em cruzamentos intraespecíficos compatíveis (fêmeas infectadas ou não cruzadas com machos não infectados) e cruzamentos incompatíveis (fêmeas não infectadas cruzadas com machos infectados) de ambas as espécies. A fertilidade foi significativamente mais elevada entre os ovos produzidos pelas fêmeas infectadas, de ambas as espécies. Foi observado que machos infectados, em ambas as espécies, estão relacionados com os cruzamentos onde ocorreram as taxas mais altas de eclosão. Analisando os cruzamentos incompatíveis, foi demonstrada a presença de incompatibilidade citoplasmática (IC), como seria esperado pela atuação da Wolbachia. Foi mostrado um alto valor para os índices de IC em A. sp,1 (IC= 54,01%) e em A. obliqua (IC = 66,2%). Os resultados sugerem que podem existir relações mutualísticas insipientes da Wolbachia com suas espécies de Anastrepha hospedeiras / Wolbachia is an intracellular bacteria found in somatic and in the reproductive tissues of various arthropods and nematodes. Phylogenetic studies based on 16S and ftsZ genes indicated the existence of six Wolbachia taxonomic supergroups (\"A\" through \"F\"). Infection of Wolbachia have been linked to several changes in the reproduction of their hosts, like cytoplasmic incompatibility (CI), parthenogenesis, feminization of genetic males and male killing. T Wolbachia infection has been described in several species of fruit flies of the family Tephritidae, like Bactrocera ascita, Rhagoletis cerasi, Ceratitis capitata, in which the bacteria induces cytoplasmic incompatibility. In Anastrepha, endemic to the American Continent, Wolbachia infection has been described in several species by analysis of the wsp gene, and there is also indications that Wolbachia-mediated CI occurs in two species of the fraterculus group. The occurrence of CI coupled with the suggestion of the use of Wolbachia in population supression programs, impose the need for a more precise characterization of the different strains of Wolbachia. The identification of the different strains of the bacteria is most accurate when the methodology of multiple loci (MLST) is applied. In this study fragments of genes gatB, coxA, hcpA, ftsZ and fbpA, integrating the methodology MLST, and of wsp gene were amplified and sequenced. Population samples of the Anastrepha fraterculus.complex of cryptic species from Brasil, Argentina, Peru, Ecuador, Colombia, Guatemala and Mexico, and samples of A. oblique from Basil were analysed. All samples were infected with supergroup \"A\" Wolbachia. For each of the five MLST genes, unique as well already known haplotypes were found. Phylogenetic analyses of each gene isolated showed incongruences in the relationships among haplotypes and population samples. The concatenated sequences of the five genes, with 2079 bp, were analyzed and 20 haplotypes were found, with distances ranging from 0.001 to 0.058. Phylogenetic analysis of Wolbachia isolated haplotypes into distinct clades, demonstrating that different strains of Wolbachia were present in these hosts, and in distinct geographic areas. Hosts specific haplotypes were found as well as more than one strain of Wolbachia was found in given population samples. A haplotypes (ST1) was detected in two species of the complex and is also the most commonly found in different organisms. Twenty two different sequences of about 500 bp were found for the wsp gene. The level of nucleotide variability is not uniform along the gene, forming a pattern with four hypervariable regions, HVRs. Genetic distances between haplotypes showed a variation from 0.001 to 0.235. Phylogenetic analysis of the haplotypes also isolated Wolbachia into distinct clades, but in contrast to the MLST, the tree formed by wsp gene does not support the monophily of some groups. The data show that strains of Wolbachia are disseminated along the American Continent, and also that there are specific strains in determined geographic areas. Analyses of ovaries and testes from infected and non infected (cured by heat treatment) individuals of A. sp. 1 and A. obliqua were made in search of possible effects of Wolbachia on its hosts. Ovaries from infected and cured females of both species, stained by DAPI, showed no visible differences in this morphological analysis. The production of sperms increases during few days after ermergence and drops out later one. Analysis of infected and cured males showed that the production of sperms were not significant between them, for both the host species. Fecundity of infected females of A, sp.1 was significantly higher than that of cured females, but was similar in A. obliqua, Egg hatching was scored in compatible intraspecies crosses and also in incompatibles crosses, of both species. Fertility was significntly higher for infected females of both species. Infected males of both species were found associated to crosses in which the higher egg hatching was observed. Analyses of incompatible crosses showed that CI occurred at high rates in A. sp.1 (CI = 54.01%) and in A. obliqua (CI = 66.2%). The data suggest that an incipient mutualism may be present in the relationships of Wolbachia and its Anastrepha hosts

Despite high volumes of manure production in the Shenandoah Valley, there are still areas of forage production in the Shenandoah Valley that are nutrient deficient, because manures have traditionally been applied to higher value row crops. Furthermore, anecdotal reports have suggested that application of poultry litter to pastureland may increase the abundance of weeds. This study was conducted to compare the effects of poultry litter and inorganic fertilizers on soil fertility, aboveground botanical composition and soil seed bank composition of established mixed naturalized pastures. Two sites each had the following treatments applied for two consecutive years: 1) split application of litter: 3.36 Mg ha-1 in spring and 3.36 Mg ha-1 in summer; 2) inorganic fertilizer at same N, P and K rates as Trt. No. 1; 3) single application of litter: 6.72 Mg ha-1 in spring; 4) inorganic fertilizer at same N, P and K rates as Trt. No. 3; 5) unfertilized control. Forage yield, soil pH, Mehlich 1 P (M1P), P balance, water soluble phosphorus (WSP), shifts in aboveground botanical composition, seed bank composition, and species richness (SPR) were monitored during this experiment. In addition, we examined whether poultry litter carries germinable weed seeds that can be transferred to forage systems after application. On average, soil pH was 0.2 higher for litter than chemical fertilizer treatments, but this difference was not significant. Mehlich 1 P and WSP both increased throughout the two years, but no significant differences were found amongst split and single application or litter and inorganic fertilizers. Phosphorus balance analysis indicated that forage yield did not remove adequate P to prevent environmental concerns, when using N based applications. Fertilization increased yield 3.5 Mg ha-1 on average and yield responses to application timing and fertilizer types were similar. Desirable species abundance increased significantly (linear regression, P< 0.05) over time in both fertilizer and poultry litter application treatments. Furthermore at one site, Shenandoah County, poultry litter application actually reduced the number of weedy species in plots (P=0.04). Weedy plant abundance did not differ at either site, and no significant change in species richness (number of species/area) was observed at Rockbridge County for any treatment. No treatment effects were found for seed bank species richness and number of germinated seeds (m-2). No seedlings germinated directly from poultry litter collected from several sites in Virginia. Poultry litter was comparable to inorganic fertilizer in terms of increasing soil WSP, M1P, forage yield, and above ground composition of desirable forage species. Timing of application made no difference. Additionally, our results indicate that poultry litter has no viable seed and does not increase weed abundance in mixed naturalized pastures. / Master of Science

Detta examensarbete handlar om förbättringsmöjligheterna av kommunikationen mellan entreprenören och projektören. Dagens byggbransch är under en stor förändring till BIM och det är då extra viktigt att kommunikationen fungerar mellan parterna så samarbetet går så smidigt och problemfritt som möjligt för bästa resultat. Ett förbättrat samarbete skapar en bättre relation mellan företagen samt kan hålla ner kostnader och tid för projektet. För att komma fram till det bästa resultatet på arbetet har intervjuer gjorts med både entreprenören samt projektören i projektet Stenkumla - Dunsjö. Entreprenören i projektet är Skanska och projektören är WSP. De flesta intervjuer som har gjorts har varit med entreprenörerna för att ta reda på vad det är de saknar. Resultatet från intervjuerna har sedan diskuterats med projektörerna för att se om de visste om de synpunkter som entreprenörerna hade och om de hade en lösning. Efter intervjuerna visade det sig att de flesta entreprenörerna ansåg att en pusselbit som saknades var utbildning, de ansåg att de inte hade fått tillräckligt med utbildning för att till exempel använda modellen på bästa sätt. Eftersom BIM är ett nytt arbetssätt där modellen är en central del ansågs denna punkt viktig. Det kom upp fler synpunkter från entreprenören där vissa synpunkter endast handlade om brist på kommunikation och där många handlade om bristen på utbildning. Resultatet kan leda till att företagen öppnar fler dialoger och sätter mer tid till lyssna vad som kan förbättras. / This degree project is about improving the possibilities of communication between the entrepreneur and the projector. Today's construction industry is in a major change to BIM, and it is therefore particularly important that the communication works between the parties as cooperation goes as smoothly and seamlessly as possible for best results. An enhanced cooperation to create a better relationship between the companies and can keep down costs and time to the project. To arrive at the best result of the study interviews with both the contractor and the designer of the project Stenkumla – Dunsjö were made. The major entrepreneur of the project is Skanska and the projector isWSP. Most of the interviews that have been done have been with entrepreneur to find out what they are missing. The result of the interviews have then been discussed with the projector to see if they knew about the comments that the entrepreneurs had and if they had a solution for it already. After the interviews it turned out that most of the entrepreneurs felt that a piece of the puzzle that was missing were education, they felt that they had not received sufficient education on how to use the model in the best way. Since BIM is a new approach in which the model is a central part it was considered important. It came up more comments from the entrepreneur where the comments only were about lack of communication where many were about the lack of education. The result can lead companies to open more dialogues and puts more time to listen to what can be improved.

This research focused on the reproductive incompatibility and genetic differences between the two strains of plum curculio, Conotrachelus nenuphar (Herbst). Two molecular markers served as the basis for the strain distribution analysis of plum curculio and Wolbachia symbiont. One marker is the partial mitochondrial cytochrome oxidase gene subunit I (mtCOI) of plum curculio. Another marker is the Wolbachia Surface Protein (wsp) gene of Wolbachia associated with plum curculio. First, the reproductive compatibility of cross-populations mating in plum curculio was studied during the summers of 2004 and 2006. The results confirmed the reproductive incompatibility among plum curculio geographic populations. A unidirectional incompatibility was revealed in an approximate north and south transect of the range of plum curculio (4 x 4 two factorial design: NY, VA, FL, and WV): there was a significant low fertility in WV males mated with NY (40%) and VA (29%) females. The Florida population showed a different pattern: FL males have a significantly lower fertility with VA (46%) and WV (37%) females while FL females were compatible with all males from the four populations. The results of experiment 2 indicated that within the northern geographic area populations (3 x 3 two factorial design: NY, MA, and NJ) were compatible with each other. An opposite unidirectional reproductive incompatibility was revealed in the combination of NJ males with FL females, which showed a significant low fertility (47%). A bi-directional incompatibility occurred between FL and WV reciprocal cross mating. FL males mated with WV females (26%) and WV males mated with FL females (21%) both have the significant low fertility compared to fertility of within their population matings. The genetic diversity among plum curculio populations from different geographic locations was investigated using the partial mtCOI gene. A total of 50 samples from 10 populations were sequenced. PCR products were 863 bp in length. A total of 23 unique sequence haplotypes were found in the 50 samples tested. Haplotype G (n = 5), L (n = 12) and T (n = 13) comprised 60% of 50 samples. The nucleotide distances between those haplotypes ranged from 0.12% to 4.87%. Genetic distances between northern and southern group plum curculios range from 4.17% to 4.87%. Two distinct major clades were found, using three different phylogenetic analyses: 1) neighbor joining (NJ), 2) maximum-parsimony (MP), and 3) maximum-likelihood (ML). 100% bootstraps support the northern clade and the southern clade was strongly supported (100/100/86, NJ/MP/ML) as well. The mid-southern subclade within the southern clade was also strongly supported (70/82/71, NJ/MP/ML) and the far-southern subclade was supported in NJ tree (81%) but was not resovled in MP and ML trees. The mid-southern subclade included haplotypes from two NJ, Washington, VA (Ra), Blacksburg, VA (BL) and 50% of WV populations and the far-southern subclade included haplotypes from FL, GA, Whitethorne, VA (Ke), Troutville, VA (Bo) and another 50% of WV populations. The results suggested that the northern and the southern clade could correspond with the northern and southern strains, respectively, of plum curculio. In this study, the mtCOI sequence was highly informative as a molecular marker in that it was useful to distinguish C. nenuphar from northern and from southern geographic locations in the eastern United States. However, the number of generations per year of several geographic populations within the southern clade still needs to be determined. The distribution of Wolbachia infection associated with plum curculio strains was investigated. 91 of 93 samples were infected by Wolbachia. Three unique Wolbachia strains were identified. The strains wCne1 and wCne2 (593 bp) were 97% identical, and their sequences were both 84% identical with wCne3 (590 bp). The wsp sequence of wCne1 was 99% identical to Wolbachia sequenced from the neotropical beetle, Chelymorpha alternans Boheman (Keller et al. 2004). The wCne2 sequence was 98.5% identical to the flower bug, Orius nagaii Yasunaga (Miura and Tagami, unpublished). The wCne3 sequence was 100% identical to Wolbachia sequenced from the tephritid fruit fly, Dacus destillatoria (Jamnongluk et al. 2000) and the ant, Formica exsecta (Reuter and Keller 2003). PCR - Restriction Fragment Length Polymorphism (RFLP) was used for superinfection detection. Of 93 samples, 15 (16.1%), 21 (22.6%), 19 (20.4%), 36 (38.7%) samples were infected by wCne1, wCne2, wCne1 plus wCne2, and wCne3, respectively. Only two (2.2%) samples had no infection. The wCne3 strain was always present as a single infection. Therefore, current results suggest that Wolbachia strains approximate the distribution of plum curculio strains: the northern strain is infected with wCne1 and wCne2 strains in supergroup B, the southern strain is infected with wCne3 strain in supergroup A and the mid-Atlantic region is the convergence area. Compared with the haplotype distribution of plum curculio mtCOI gene, there was a closer relation of the mid-southern PC clade to the far-southern clade than to the northern clade. However, Wolbachia symbionts in mid-southern PC are more closely related to those in northern PC than to those in far-southern PC. The relationship of Wolabchia infection with reproductive incompatibility between plum curculio populations is also discussed. / Ph. D.

L'évolution des technologies actuelles permet de traiter un grand nombre d'expériences (ou de simulations) et d'envisager un nombre important de paramètres. Cette situation conduit à des matrices de grande, voire très grande, dimension et nécessite le développement de nouveaux outils pour évaluer et visualiser ces données et, le cas échéant, en réduire la dimension. L'évaluation de la qualité de l'information apportée par l'ensemble de points constituant une base de données ou un plan d'expériences peut se faire au travers de critères basés sur des calculs de distance, qui renseigneront sur l'uniformité de la répartition dans l'espace multidimensionnel. Parmi les méthodes de visualisation, l'Analyse en Composantes Curvilignes a l'avantage de projeter des données en grande dimension dans un espace bidimensionnel en préservant la topologie locale, ce qui peut aider à détecter des amas de points ou des zones lacunaires. La réduction de dimension s'appuie sur une sélection judicieuse de sous-ensembles de points ou de variables, via des algorithmes. Les performances de ces méthodes ont été évaluées sur des cas d'étude issus des études QSAR, de la spectroscopie et de la simulation numérique. / Technological progress has now made many experiments (or simulations) possible, along with taking into account a large number of parameters, which result in (very) high-dimensional matrix requiring the development of new tools to assess and visualize the data and, if necessary, to reduce the dimension. The quality of the information provided by all points of a database or an experimental design can be assessed using criteria based on distances that will inform about the uniformity of repartition in a multidimensional space. Among the visualization methods, Curvilinear Component Analysis has the advantage of projecting high-dimensional data in a two-dimensional space with respect to the local topology. This also enables the detection of clusters of points or gaps. The dimensional reduction is based on a judicious selection of subsets of points or variables, via accurate algorithms. The performance of these methods was assessed on case studies of QSAR, spectroscopy and numeric simulation.

This thesis aims to enable improved urban sustainability in India and has therefore been conducted at WSP’s office in Delhi. The objective has been to chart the Indian characteristics as well as to identify difficulties regarding urban sustainability. The purpose and objective have been accomplished through the implementation of three research questions. The questions have been answered by a literary review of existing theories and a complementary document analysis. Furthermore, a case study of a new development in India with long-term sustainability in focus of the design has been conducted. For an Indian city to achieve a sustainable urban development, five pillars of sustainability have been identified; political, physical, ecological, social and economic. The ecological, social and economic are pillars from the common definition of sustainability, however they have different meaning and focus in the Indian context. The physical and political pillars are therefore characteristic for the Indian urban development. The physical pillar is added in the Indian context since short term planning and focus on profit is dominating the building industry. The demand for maintenance is because of that larger than the supply which results in a need for more emphasis on the physical built environment. The political pillar is applied because of the concerns for the value and quality of governance actions. It affects the four other pillars since the government should provide guidance, both with instructions and by executions, which is not always apparent. The Indian government has introduced several strategies in order to achieve sustainability in Indian cities. Rating tools for sustainability, such as Leed and Griha, together with the planning of spatial city forms, such as compact city form and mixed land use, which are the main procedures. Benefits in terms of lower interest rates on loans and a quicker clearance are given to developers who intend to build sustainably. Though the many theories and strategies seem ambitious, they are not always as effective when translated into practice. This is partly because of the lack of follow-up and partly the clients focus on quick profit. Thus, India is facing many challenges in order to reach a sustainable urban development. Together with urbanisation and growth in population, corruption is the main challenge since many other follows. These are lack of awareness in the field of sustainability, short term planning and the focus on profit as well as public safety and poverty. If India is serious about developing its cities sustainably, it is significant for the Indian government to take an inspiring role in using and promoting sustainability.

Hydroxyapatite coatings have been used as a barrier between a metal core of replacement and living tissue for long time in medicine. Plasma spray is a typical technology for preparation these coatings. This thesis concerns a new plasma spray technology with hybrid system of arc stabilization (WSP-H). Simultaneously it uses an innovative method of feeding the initial material in a form of water suspension (SPPS). The aim of this thesis was to produce water solutions of hydroxyapatite and make coatings with WSP-H. These coatings were characterized in terms of their morfology, chemical and phase composition. Their biocompatibility was tested by a cultivation and an analysis of RAW 264.7 and MG-63 cells. The coatings made by SPPS have a very rough surface which is suitable for biological application. The biocompatibility testing discovered that the coatings will by tolerated in a human body very well. This method of production of hydroxyapatite coatings could be a good replacement of older methods after a better optimalisation of the processing parameters and after reaching purity of coating.

Syfte: Undersöka vilka möjligheter som finns, för projekteringsledaren, att optimera tidplaneringen i projekteringsskedet och på så sätt dra större fördel av att arbeta med BIM.   Frågeställningar: Hur ser det planerade och faktiska aktivitetsflödet, eventuellt även informationsflödet, ut i några av WSP Managements redan utförda BIM-projekt? Vilka åsikter har olika roller i projekten om detta? Vilka problemområden upplever projekteringsledare att det finns i projekteringsskedet i ett BIM-projekt? Finns tendenser till flaskhalsar i vissa projekteringsfaser? Vilka tankar finns i branschen när det gäller möjligheten att tidplanera bättre i projekteringsskedet för att kunna dra större fördel av BIM? Går det att förena tidplanering av projekteringsskedet i BIM-projekt med andra redan inarbetade metoder?   Metod: Insamlandet av fakta till detta examensarbete har skett genom  en induktiv litteraturstudie samt intervjuer av kvalitativt slag.   Slutsatser: De resultat som framkommit under examensarbetets gång tyder på att optimerad tidplanering av projekteringsskedet i ett BIM-projekt gör att det går att dra större fördel av att arbeta med BIM.   Flera av de problem och flaskhalsar som nämns i intervjuerna visar att dessa kan förhindras med bättre tidplanering. Framförallt är det förarbetet, där nytta, mål och BIM-användning identifieras som framhävs som viktigt. Andra förutsättningar som nämns är högre engagemang från projekteringsledaren, att denne bör komma in i ett tidigt skede i projektet och bör samarbeta med någon som har stor kunskap och erfarenhet av BIM.   Tidplanen för projekteringsskedet i ett BIM-projekt skiljer sig från dagens tidplan på så sätt att det är informationsleveranserna som kommer styra hur de olika aktiviteterna planeras istället för de handlingar som skapas idag. Samma metoder och program som används idag kan användas för tidplanering av ett BIM-projekt. Behovet av att bryta ner projektet i mindre delar för att studera relationen mellan de olika aktiviteterna och på så sätt skapa en fungerade tidplan kvarstår. / Purpose: Explore the possibilities to optimize the scheduling in the design phase to obtain greater benefits from working with BIM.   Problems: What is the planned and actual work flow, possibly including the information flow, in some of WSP Management’s conducted BIM-projects? What are the opinions about this, from the different roles in the projects? Which problem areas are the managers of the design phase experiencing in the design phase of a BIM-project? Are there signs of bottlenecks in some of the stages? What are the thoughts in the industry regarding the possibility to improve the time scheduling of the design phase in order obtain greater benefits from working with BIM. Is it possible to use already established methods for the time scheduling of BIM-projects?   Methods: The collection of data for this thesis has been done through an inductive literature review and qualitative interviews.   Conclusions: The results from this thesis indicate that optimizing of scheduling leads to greater benefits from working with BIM.   Several of the problems and bottle necks that are mentioned in the interviews shows that they could be prevented with better time scheduling. Above all it's the initial work, where the BIM goals and Uses are identified, that are mentioned as important. Other conditions mentioned are higher commitment from the design manager, that this person should come in at an early stage in the project and collaborate with someone who has extensive knowledge and experience of BIM.   The schedule for the design phase in a BIM-project will be distinguished from today’s schedule in the way that the information delivery will control how the various activities are planned, instead of documents. The same  methods and programs that are used today can be used for the scheduling of a BIM-project. The need for a break-down structure in order to study the relationship between the various activities still exists in order to make a functioning time schedule.

Cette thèse propose une méthodologie pour des études en simulation numérique en grande dimension. Elle se décline en différentes étapes : construction de plan d'expériences approprié, analyse de sensibilité et modélisation par surface de réponse. Les plans d'expériences adaptés à la simulation numérique sont les "Space Filling Designs", qui visent à répartir uniformément les points dans l'espace des variables d'entrée. Nous proposons l'algorithme WSP pour construire ces plans, rapidement, avec de bons critères d'uniformité, même en grande dimension. Ces travaux proposent la construction d'un plan polyvalent, qui sera utilisé pour les différentes étapes de l'étude : de l'analyse de sensibilité aux surfaces de réponse. L'analyse de sensibilité sera réalisée avec une approche innovante sur les points de ce plan, pour détecter le sous-ensemble de variables d'entrée réellement influentes. Basée sur le principe de la méthode de Morris, cette approche permet de hiérarchiser les variables d'entrée selon leurs effets. Le plan initial est ensuite "replié" dans le sous-espace des variables d'entrée les plus influentes, ce qui nécessite au préalable une étude pour vérifier l'uniformité de la répartition des points dans l'espace réduit et ainsi détecter d'éventuels amas et/ou lacunes. Ainsi, après réparation, ce plan est utilisé pour l'étape ultime : étude de surfaces de réponse. Nous avons alors choisi d'utiliser l'approche des Support Vector Regression, indépendante de la dimension et rapide dans sa mise en place. Obtenant des résultats comparables à l'approche classique (Krigeage), cette technique semble prometteuse pour étudier des phénomènes complexes en grande dimension. / This thesis proposes a methodology of study in numeric simulation for high dimensions. There are several steps in this methodology : setting up an experimental design, performing sensitivity analysis, then using response surface for modelling. In numeric simulation, we use a Space Filling Design that scatters the points in the entire domain. The construction of an experimental design in high dimensions must be efficient, with good uniformity properties. Moreover, this construction must be fast. We propose using the WSP algorithm to construct such an experimental design. This design is then used in all steps of the methodology, making it a versatile design, from sensitivity analysis to modelling. A sensitivity analysis allows identifying the influent factors. Adapting the Morris method principle, this approach classifies the inputs into three groups according to their effects. Then, the experimental design is folded over in the subspace of the influent inputs. This action can modify the uniformity properties of the experimental design by creating possible gaps and clusters. So, it is necessary to repair it by removing clusters and filling gaps. We propose a step-by-step approach to offer suitable repairing for each experimental design. Then, the repaired design is used for the final step: modelling from the response surface. We consider a Support Vector Machines method because dimension does not affect the construction. Easy to construct and with good results, similar to the results obtained by Kriging, the Support Vector Regression method is an alternative method for the study of complex phenomena in high dimensions.