The carnivores of Madagascar are likely the least studied of the world's carnivores, thus little is known about threats to their persistence. I provide the first long-term assessment of Madagascar's rainforest carnivore community, including: 1) how multiple forms of habitat degradation (i.e., fragmentation, exotic carnivores, human encroachment, and hunting) affect native and exotic carnivore occupancy; 2) how native and exotic carnivore temporal activity overlap and how body size and niche explain these patterns; 3) how native and exotic carnivores spatially co-occur across the landscape and which variables explain these relationships; and 4) how native and exotic carnivores and humans co-occur with lemurs across Madagascar's largest protected landscape: the Masoala-Makira landscape. From 2008 to 2013 I photographically sampled carnivores and conducted line-transect surveys of lemurs at seven study sites with varying degrees of degradation and human encroachment, including repeat surveys of two sites. As degradation increased, exotic carnivores showed increases in activity and occupancy while endemic carnivore, small mammal, and lemur occupancy and/or activity decreased. Wild/feral cats (Felis sp.) and dogs (Canis familiaris) had higher occupancy (0.37 ± SE 0.08 and 0.61 ± SE 0.07, respectively) than half of the endemic carnivore species across the landscape. Additionally, exotic carnivores had both direct and indirect negative effects on native carnivore occupancy. For example, spotted fanaloka (Fossa fossana) occupancy (0.70 ± SE 0.07) was negatively impacted by both wild/feral cat (beta = -2.65) and Indian civets (beta = -1.20). My results revealed intense pressure from hunting (ex. n = 31 fosa Cryptoprocta ferox consumed per year from 2005-2011 across four villages), including evidence that hunters target intact forest where native carnivore and lemur occupancy and/or activity are highest. I found evidence of high temporal overlap between native and exotic carnivores (ex. temporal overlap between brown-tail vontsira Salanoia concolor and dogs is 0.88), including fosa (Cryptoprocta ferox) avoiding dogs and humans across all seasons. However, I found no evidence of body size or correlates of ecological niche explaining temporal overlap among carnivores. Estimates of spatial co-occurrence among native and exotic carnivores in rainforest habitat revealed strong evidence that native and exotic carnivores occur together less often than expected and that exotic carnivores may be replacing native carnivores in forests close to human settlements. For example, falanouc show a strong increase in occupancy when dogs are absent (0.69 ± SE 0.11) compared to when they are present (0.23 ± SE 0.05). Finally, the two-species interaction occupancy models for carnivores and lemurs, revealed a higher number of interactions among species across contiguous forest where carnivore and lemur occupancy were highest. These various anthropogenic pressures and their effects on carnivore and lemur populations, particularly increases in exotic carnivores and hunting, have wide-ranging, global implications and demand effective management plans to target the influx of exotic carnivores and unsustainable hunting affecting carnivore and primate populations across Madagascar and worldwide. / Ph. D.

Madagascar is one of the world's top conservation priorities due to the intense anthropogenic pressures on its diverse and endemic wildlife. There have been very few studies conducted in the largest protected area complex in Madagascar, the Masoala-Makira landscape (northeastern Madagascar). My goal was to examine the response of ground-dwelling forest birds, tenrecs (Lipotyphla: Tenrecidae) and lemurs to habitat degradation and the presence of exotic predators, and monitor population trends at resurveyed sites from 2008 to 2013. Using camera trap surveys and distance sampling, we observed 26 bird species (n = 4,083 observations), three spiny tenrec species (n = 244 observations) and 12 lemur species (n = 1,172 observations). Out of 13 focal species (seven bird, three tenrec and three lemur species), seven had higher point estimates of occupancy or density at intact forests when compared to intermediately degraded or degraded forest sites. Common tenrecs (Tenrec ecaudatus) and cathemeral lemurs changed their activity patterns, becoming more nocturnal in degraded forests. Feral cat (Felis sp.) trap success was negatively related to the detection of three bird species (red-breasted coua, Coua serriana; scaly ground-roller, Geobiastes squamiger; and Madagascar crested ibis, Lophotibis cristata). At two resurveyed sites (S02 and S05), out of 19 and 17 species, only four and eight species did not show consistent declines in occupancy or encounter rates, respectively, over a six-year period. This research highlights the urgent need for immediate conservation action in the Masoala-Makira protected area complex in order to protect one of the world's biodiversity hotspots. / Master of Science

Harlequin bugs are orange and black aggregation pheromone emitting stink bug pests, specifically of cole crops such as kale, broccoli and collards. This nearly loyal crop preference makes an interesting challenge for trapping them and helping farmers predict pest severity. Harlequin bugs can be found in much of North America, and are a serious problem in the southeastern United States. Presumably their persistence into northern regions is limited by extreme winters. In 2014 and 2015 the arctic polar vortex extended into mid-latitudes bringing a blanket of sustained sub-freezing temperatures to much of the United States. We used these events to determine effects of extreme winter weather on harlequin bug survival. In both years we observed nearly identical low temperatures of -15oC and this linked to high (80-96%) harlequin bug mortality. In the lab we measured exact lethal freezing temperatures in harlequin bugs (i.e. supercooling points) to see if a physiological metric could be used to predict overwinter survival. Harlequin bug adults froze and died at -10.4oC, and similarly, their larger juvenile stages freeze at -11.0oC. Freshly hatched harlequin bugs and unhatched eggs froze at considerably lower temperatures with eggs forming ice crystals at -23.2oC and recent hatches at -21.6oC. Now with an understanding of how harlequin bugs likely survive winter extreme, we can then work on developing a trap to tally their populations in the spring and predict summer and fall pest severity. In the lab and field, harlequin bug adults and large nymphs were more likely found on green and black colors, and statistically less frequently on yellow, white, purple or red colors with the exception of adult females, which were most attracted to red and green in the lab, but green and black in the field. To increase harlequin bug attraction to and termination at traps square corrugated plastic panels were wrapped with an insecticide netting and baited with harlequin bug aggregation pheromone, murgantiol. Bugs were effectively drawn to the panels, with green panels having significantly more dead harlequin bugs and fewer dead beneficial lady beetles (Coleoptera: Coccinellidae) at their base than yellow panels. Thus, green was chosen as the ideal trap color to use for another field experiment that evaluated three trap types -- a corrugated plastic square panel, pyramidal trap, and ramp trap -- each with three lure treatments, murgantiol alone or murgantiol plus a low or high rate of mustard oil. More bugs were killed with the pyramidal trap than with the panel trap or the ramp trap, and more bugs were killed at traps containing murgantiol combined with benzyl isothiocyanate than at those with murgantiol alone. This research demonstrated that with the proper visual elements and odors, harlequin bugs can be drawn to traps and effectively killed after contact with insecticide-incorporated netting. / MSLFS / Harlequin bugs are orange and black aggregation pheromone emitting stink bug pests, specifically of cole crops such as kale, broccoli and collards. This nearly loyal crop preference makes an interesting challenge for trapping them and helping farmers predict pest severity. Harlequin bugs can be found in much of North America, and are a serious problem in the southeastern United States. Presumably their persistence into northern regions is limited by extreme winters. In 2014 and 2015 the arctic polar vortex extended into mid-latitudes bringing a blanket of sustained sub-freezing temperatures to much of the United States. We used these events to determine effects of extreme winter weather on harlequin bug survival. In both years we observed nearly identical low temperatures of -15℃ and this linked to high (80-96%) harlequin bug mortality. In the lab we measured exact lethal freezing temperatures in harlequin bugs (i.e. supercooling points) to see if a physiological metric could be used to predict overwinter survival. Harlequin bug adults froze and died at -10.4℃, and similarly, their larger juvenile stages freeze at -11.0℃. Freshly hatched harlequin bugs and unhatched eggs froze at considerably lower temperatures with eggs forming ice crystals at -23.2℃ and recent hatches at -21.6℃. Now with an understanding of how harlequin bugs likely survive winter extreme, we can then work on developing a trap to tally their populations in the spring and predict summer and fall pest severity. In the lab and field, harlequin bug adults and large nymphs were more likely found on green and black colors, and statistically less frequently on yellow, white, purple or red colors with the exception of adult females, which were most attracted to red and green in the lab, but green and black in the field. To increase harlequin bug attraction to and termination at traps square corrugated plastic panels were wrapped with an insecticide netting and baited with harlequin bug aggregation pheromone, murgantiol. Bugs were effectively drawn to the panels, with green panels having significantly more dead harlequin bugs and fewer dead beneficial lady beetles (Coleoptera: Coccinellidae) at their base than yellow panels. Thus, green was chosen as the ideal trap color to use for another field experiment that evaluated three trap types – a corrugated plastic square panel, pyramidal trap, and ramp trap – each with three lure treatments, murgantiol alone or murgantiol plus a low or high rate of mustard oil. More bugs were killed with the pyramidal trap than with the panel trap or the ramp trap, and more bugs were killed at traps containing murgantiol combined with benzyl isothiocyanate than at those with murgantiol alone. This research demonstrated that with the proper visual elements and odors, harlequin bugs can be drawn to traps and effectively killed after contact with insecticide-incorporated netting.

The potential control of cucumber beetles, Acalymma vittatum (Fab.) and Diabrotica undecimpunctata howardi (Barber)(both Coleoptera: Chrysomelidae) two major pests of cucurbits, was assessed in a cucurbit agroecosystem by using buckwheat (Fagopyrum esculentum Moench) border habitat to attract the natural enemies, Celatoria diabroticae (Shimer) and Celatoria setosa (Coquillett) (both Diptera: Tachinidae) and Chauliognathus pennsylvanicus (Deg.) (Coleoptera: Cantharidae). Five different plants were seeded in the border and buckwheat dominated. Four natural enemy groups were also included as indicators of the attractiveness of the floral border: The Order Diptera; the Families Tachinidae and Syrphidae of Diptera; and the Order Hymenoptera. In 1995-1996, rows of squash, Cucurbita pepo (L.) var. melopepo (Alef.) ‘Seneca Prolific’, and cucumbers, Cucumis sativa (L.) ‘Arkansas Littleleaf’, were planted perpendicular to floral border habitats. Sticky traps and modified Malaise traps on transects at intervals from the border were used to monitor insect numbers. Insect counts and yields of cucurbits were analyzed using analysis of variance with contrasts for linear and quadratic effects and regression model fitting. Borders were strongly attractive to Diptera and moderately attractive to C. pennsylvanicus, Syrphidae, Tachinidae, and Hymenoptera. C. setosa and C. diabroticae counts were too low to analyze and borders habitats did not have any meaningful effect on yields. The border conserved populations of Diptera, leatherwings, Hymenoptera, and tachinids on some dates and may be useful with economic thresholds for pest management because of an observed gradient of insect movement. / Master of Science

Within the "forbidden" range of electron energies between the valence and conduction bands of titanium dioxide, crystal lattice irregularities lead to the formation of electron trapping sites. These sites are known as shallow trap states, where "shallow" refers to the close energy proximity of those features to the bottom of the semiconductor conduction band. For wide bandgap semiconductors like titanium dioxide, shallow electron traps are the principle route for thermal excitation of electrons into the conduction band. The studies described here employ a novel infrared spectroscopic approach to determine the energy of shallow electron traps in titanium dioxide nanoparticles. Mobile electrons within the conduction band of semiconductors are known to absorb infrared radiation. As those electrons absorb the infrared photons, transitions within the continuum of the conduction band produce a broad spectral signal across the entire mid-infrared range. A Mathematical expression based upon Fermi–Dirac statistics was derived to correlate the temperature of the particles to the population of charge carriers, as measured through the infrared absorbance. The primary variable of interest in the Fermi – Dirac expression is the energy difference between the shallow trap states and the conduction band. Fitting data sets consisting of titanium dioxide nanoparticle temperatures and their associated infrared spectra, over a defined frequency range, to the Fermi–Dirac expression is used to determine the shallow electron trap state energy. / Master of Science

Gravimetric analysis was used to determine the release rates and longevities of several designs of pheromone dispensers for mating disruption of leafrollers and codling moth, Cydia pomonella (Linneaus). Release rates were described by linear equations for at least four months, but by the end of the season release rates tended to become erratic. Biocontrol’s red-brown codling moth dispenser lasted for up to four months, and one application of the dispenser in early May should control codling moth for the entire season in Virginia apple orchards. Ecogen and Hercon leafroller dispensers lasted for a shorter time than the codling moth dispensers and would require two applications per season to provide the best control of their target pests. In a commercial northern Virginia apple orchard, the effects of pheromone trap height and pheromone dispenser height on captures of the tufted apple bud moth, Platynota idaeusalis (Walker), were studied. Low traps (2.0 m) were more sensitive than high traps (4.5 m) for monitoring tufted apple bud moth. The number of moths caught in 1994 and 1995 in a two-hectare mating disruption plot with pheromone dispensers placed in the upper third of the tree was not significantly different from the number caught in a plot with pheromone dispensers placed at head height. Fruit damage was very high in both pheromone plots in 1994, but by 1995 it appeared that mating disruption was able to reduce fruit damage due to leafroller larvae. / Master of Science

No / N-Linked glycans were ionized from several matrices with a Shimadzu-Biotech AXIMA-QIT matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometer. [M+Na]+ ions were produced from all matrices and were accompanied by varying amounts of in-source fragmentation products. The least fragmentation was produced by 2,5-dihydroxybenzoic acid and the most by -cyano-4-hydroxycinnamic acid and 6-aza-2-thiothymine. Sialic acid loss was extensive but could be prevented by formation of methyl esters. Fragmentation produced typical low-energy-type spectra dominated by ions formed by glycosidic cleavages. MSn spectra (n = 3 and 4) were used to probe the pathways leading to the major diagnostic ions. Thus, for example, an ion that was formed by loss of the core GlcNAc residues and the 3-antenna was confirmed as being formed by a B/Y rather than a C/Z mechanism. The proposed structures of several cross-ring cleavage ions were confirmed and it was shown that MS3 spectra could be obtained from as little as 10 fmol of glycan

No description available.

O objetivo deste trabalho foi caracterizar a formação e progressão de lesões periapicais induzidas experimentalmente em dentes de camundongos knockout para receptores toll-like 2 (TLR2 KO) comparados a animais wild-type (WT). As lesões periapicais foram induzidas nos primeiros molares inferiores de 28 camundongos WT e de 27 camundongos TLR2 KO. Decorridos 7, 21 e 42 dias da indução da lesão periapical, os animais foram submetidos à eutanásia em câmara de CO2, as mandíbulas foram removidas e submetidas ao processamento histotécnico. A seguir, cortes representativos foram corados com hematoxilina e eosina (HE), para descrição do tecido pulpar e das regiões apical e periapical, em microscopia óptica convencional, e mensuração da área das lesões periapicais, em microscopia de fluorescência. Espécimes sequenciais foram avaliados por meio de: histoenzimologia para a atividade da TRAP, para identificação de osteoclastos; coloração de Brown & Brenn, para localização de bactérias; e imunoistoquímica, para identificação de marcadores da osteoclastogênese (RANK, RANKL, OPG). Os resultados numéricos obtidos da análise morfométrica da extensão da área das lesões periapicais e do número de osteoclastos foram submetidos à análise estatística por meio dos testes não-paramétricos de Mann-Whitney e Kruskal-Wallis, utilizando o software SAS (Statistical Analysis System) for Windows versão 9.1.3. O nível de significância adotado foi de 5%. Os resultados da coloração de Brown & Brenn e Imunoistoquímica foram expressos de maneira qualitativa. O grupo de animais WT apresentou diferença significante na extensão da área das lesões periapicais e no número de osteoclastos entre os períodos experimentais de 7 e 42 dias (p<0,05) e entre 21 e 42 dias (p<0.05). Por outro lado, no grupo de animais TLR2 KO, as diferenças para a extensão da área das lesões periapicais e número de osteoclastos foram encontradas entre os períodos experimentais de 7 e 21 dias (p<0,05) e entre 7 e 42 dias (p<0,05). Quando os períodos dos grupos foram comparados entre si, foram encontradas diferenças estatísticas entre todos os períodos experimentais, tanto para a análise morfométrica da extensão da área das lesões periapicais, quanto para o número de ostoclastos (p<0,05). A análise descritiva do tecido pulpar e das regiões apical e periapical, por meio da coloração de HE, bem como da localização das bactérias, por meio da coloração de Brown & Brenn, não mostrou diferenças entre os dois grupos de animais. Com relação à Imunoistoquímica, as marcações foram semelhantes entre os dois grupos de animais, exceto para as marcações de RANK, as quais não foram encontradas nas lesões periapicais do grupo de animais TLR2 KO. A partir das metodologias empregadas e dos resultados obtidos pode-se concluir que na ausência do TLR2, os animais desenvolveram lesões periapicais significantemente maiores (com maior presença de osteoclastos) quando comparados aos animais WT, sugerindo o importante papel desse receptor na resposta imune e inflamatória do organismo no sentido de combater a infecção do sistema de canais radiculares e dos tecidos perirradiculares. / The aim of the present study was to characterize the formation and progression of periapical lesions experimentally induced in the teeth of toll-like receptors 2 knockout (TLR2 KO) mice compared to wild-type (WT) mice. Periapical lesions were induced in the lower first molars of 28 WT and 27 TLR2 KO mice. After 7, 21 and 42 of periapical lesion induction, the animals were euthanized in a CO2 chamber, and the mandibles were removed and subjected to histotechnical processing. Representative histological sections were stained with hematoxylin and eosin (HE) for description of the features of the pulp tissue and the apical and periapical regions under conventional optical microscopy, and for determination of the size of the periapical lesions under fluorescence microscopy. Sequential specimens were evaluated by: TRAP histo-enzymology for identification de osteoclasts; Brown & Brenn staining for localization of bacteria; and immunohistochemistry for identification of osteoclastogenesis markers (RANK, RANKL, OPG). Data from the morphometric evaluation of the size of periapical lesions and the number of osteoclasts were subjected to statistical analysis by the nonparametric Mann-Whitney and Kruskal-Wallis tests, using the SAS (Statistical Analysis System) software for Windows version 9.1.3. A significance level of 5% was set for all analyses. Data from the Brown & Brenn staining and immunohistochemical analysis were displayed qualitatively. The group of WT mice presented statistically significant difference in the periapical lesion size and number of osteoclasts between the 7- and 42-day experimental periods (p<0.05) as well as between 21 and 42 days (p<0.05). On the other hand, in the group of TLR2 KO mice, significant differences in the periapical lesion size and number of osteoclasts were found between the 7- and 21-day experimental periods (p<0.05) as well as between 7 and 42 days (p<0.05). Comparison of the periods within each group revealed statistically significant differences among all experimental periods for the morphometric evaluation of the size of the periapical lesions and number of osteoclasts (p<0.05). Descriptive analysis of pulp tissue and apical and periapical regions by HE staining and localization of bacteria by Brown & Brenn staining did not show significant differences between the two groups of animals. The immunohistochemical results showed similar immunostaining in both groups of animals, except for RANK expression, which was not observed in the periapical lesions of the TLR2 KO mice. Based on the employed methodology and the obtained results it may be concluded that in the silence of TLR2, the animals developed superior size of periapical lesions (with higher presence of osteoclasts) compared to WT animals, suggesting the important role of this receptor during the immune and inflammatory response against the infection of root canal system and periapical tissues.

Die Telomere sind spezialisierte DNA-Protein-Komplexe, die sich an den Enden der Chromosomen der eukaryotischen Zellen befinden. Die Telomerase ist ein Ribonukleoprotein, welches für die vollständige Replikation der Telomere bei den meisten Eukaryoten verantwortlich ist. Die katalytische Untereinheit des Enzyms (hTERT beim Menschen) besitzt Reverse-Transkriptase-Aktivität, und nutzt eine integrierte RNA (hTR beim Menschen) als Template, um Telomer-Wiederholungssequenzen an den Enden der Chromosomen zu synthetisieren. Die Telomerase ist in den meisten normalen humanen somatischen Zellen unterdrückt. In den meisten Krebszellen jedoch, stellt die Reaktivierung der Telomerase zur Beibehaltung der Telomerlänge eine Voraussetzung für deren unbegrenztes Wachstumspotential dar. Im Rahmen dieser Arbeit sollte ein neuer, einfacher und selektiver Assay für den Nachweis der humanen Telomerase entwickelt werden. In dem neuen Assay sollten die beiden Kernkomponenten des Enzyms, die Protein-Untereinheit und die RNA, die Targets sein. Der Test ist in seiner Grundstruktur wie folgt aufgebaut : 1. Immobilisierung der Telomerase über die hTERT an eine Festphase, beschichtet mit Phosphorothioat-modifizierten (PS) Oligonukleotiden oder Heparin. Zusammen mit der Telomerase werden bei diesem Schritt die Heparin-bindenden Proteine, die in der Probe enthalten sind, an die Festphase gebunden. 2. Spezifischer Nachweis der hTR. Zur Detektion der hTR wird ein Oligonukleotid-Ligations-Assay (OLA) oder eine Reverse-Transkriptase-PCR (RT-PCR) eingesetzt. In der optimierten Endversion wurde zur Immobilisierung des Enzyms eine Festphase, beschichtet mit PS-Oligonukleotiden, verwendet. Die hTR wurde mittels RT-PCR nachgewiesen. Mit dem neuen Assay wurden erfolgreich 75 Tumorzellen detektiert. / Telomeres are specialized DNA-Protein structures located at the ends of linear eukaryotic chromosomes. Telomerase is a ribonucleoprotein, which is responsible for the complete replication of the telomeres in most eukaryotes. The catalytic reverse transcriptase protein subunit (hTERT in humans) of the nucleoprotein uses an integral RNA (hTR in humans) as a template for the addition of telomeric repeat sequences to the ends of chromosomes. Telomerase is repressed in most normal human somatic cells, while the reactivation of telomerase to maintain telomere length is necessary for the unlimited growth potential of most human cancer cells. The aim of this work was the development of a new, simple and selective assay for the detection of human telomerase. The targets of the new assay were the two core subunits of the enzyme : hTERT and hTR. The test comprises two principal steps : 1. Immobilization of the telomerase via the hTERT subunit on a solid phase, coated with heparin or phosphorothioate-modified (PS) oligonucleotides. In this step telomerase is bound together with the heparin-binding proteins of the analysed sample to the surface. 2. Specific detection of the hTR. For the detection of the hTR an oligonucleotide ligation assay (OLA) or a reverse transcriptase PCR (RT-PCR) was used. In the optimized final version of the assay a PS-coated solid phase was used for the immobilization of the enzyme. Reverse transcriptase PCR was applied for detection of the hTR. 75 tumor cells were successfully detected with the new assay.