Sinningia allagophylla(Gesneriaceae): in vitro cultivation of a native plant of the Brazilian cerrado

We used axillary buds as initial explants for hormone interaction studies required for in vitro cultivation of S. allagophylla. Callus production was achieved on gelled Murashige & Skoog medium (MS) supplemented with indole-3-acetic acid (IAA= 0.1 and 0.5 mg.l­1 alone or combined with 6 benzylaminopurine) (BA= 0.01 and 0.1 mg.l-1). A hormone balance between IAA and BA that would encourage shoot bud development was not found. Nodal segments from axenic cultures grown in the presence of cytokinin (0.1 mg.1­1 of BA) without any auxin on MS medium with half-strength macronutrients were used as a standard explant source for subsequent experiments on optimum mineral culture media composition for S. allagophylla in vitro cultivation. We found that explants kept in vitro on gelled Gamborg et al. (B5) mineral composition culture medium showed better shoot and specially root growth than on MS medium. Comparisons of the ammonium and nitrate ratios of MS and B5 media indicate that B5 medium has a substantial reduced ammonium ion when compared to MS medium, as well as a lower total nitrogen level. The growth response pattern obtained in vitro may be evidence of the adaptation of this species to soils of poor mineral composition as found in the Brazilian cerrado, as well as an indication that nitrogen levels play a key role for S. allagophylla growth.

Late-acting self-incompatibility in Capparis retusa (Capparaceae), a species of Chaco woodland in NE Argentina

The reproductive biology of Capparis retusa was studied by means of controlled pollination experiments and fluorescence microscopy observations of post-pollination events. Self-pollinated flowers mostly failed to form fruits despite the fact that self-pollen tubes grew to the ovary and penetrated ovules within 24 h. Since embryo development to globular stage was observed in some self-fertilized ovules it was concluded that control of self-fertilization in this species occurs by some kind of post-zygotic mechanism.

High irradiance and temperature inhibit the germination of spores of the fern Rumohra adiantiformis (Forst.) Ching (Dryopteridaceae)

Rumohra adiantiformis (Forst.) Ching is a fern (Dryopteridaceae) which is used to compose floral arrangements. Fertile fronds were harvested in the "Permanently Protected Area" of Ilha Comprida, São Paulo, Brazil. Sterilized spores were germinated in Mohr liquid medium modified by Dyer. The effect of 72%, 54%, 17% and 9% of total irradiance on germination under field conditions, was analyzed. Experiments were carried out in March (I), April (II) and August of 2000 (III). Under 54% and 72% of total irradiance in Experiment I (March) the germination was completely inhibited and partially inhibited under 72% of total irradiance in Experiment II (April). The lowest mean germination time () was observed under 9% of total irradiance in Experiments II (11.62 days) and III (8.80 days) respectively, followed by 17% in Experiment III (10.12 days) and 9% of total irradiance in the Experiment I (11.62 days ). The effect of temperatures of 15 ± 1, 20 ± 1, 25 ± 1 and 30 ± 1 ºC on germination was also analyzed. The lowest mean germination time (7.93 days) was observed at 25 ± 1 °C followed by 20 ± 1 °C. The highest mean germination time was observed at 15 ± 1 °C (12.10 days) followed by 30 ± 1 °C (10.63 days), which inhibited germination. The germination of R. adiantiformis was photoinhibited by high irradiance and partially inhibited by the highest temperature tested.

Effect of nitrate and ammonium on the growth and protein concentration of Microcystis viridis Lemmermann (Cyanobacteria)

Cyanobacteria are a very important group in aquatic systems, particularly in eutrophic waters. Therefore studies about their success in the environment are essential. Many hypotheses have tried to explain the dominance of Cyanobacteria, and several emphasized the importance of various nitrogen sources for the success of the group. In this study, we measured the effect of ammonium and nitrate on the growth and protein concentration of Microcystis viridis (Cyanobacteria). This species is well-known because bloom formation in eutrophic waters. The study was carried out, in experimental batch cultures, using the WC medium with different nitrogen sources: ammonium, nitrate, ammonium + nitrate (50% ammonium + 50% nitrate) and ammonium at different concentrations (to test for possible NH4+ toxicity). Protein, ammonium and nitrate concentrations were measured at end of each experiment, whereas samples for cell counts were taken daily. Results showed that Microcystis viridis grew faster with ammonium (µ = 0.393 day-1) than with nitrate (µ = 0.263 day-1) and ammonium + nitrate (µ = 0.325 day-1). This pattern is explained by the metabolism of ammonium that presents higher uptake and assimilation rates than nitrate. Maximum cell concentration, however, was higher in the ammonium + nitrate treatment, followed by nitrate treatment. Higher protein concentration were observed in the treatment with nitrate. In the ammonium toxicity test, no difference between the control and NH4+ at 50% was found. Thus, the ammonium concentrations used in these experiments were not toxic. Our results suggest that Cyanobacteria is able to grow on both nitrogen sources even if ammonium may allow faster growth and bloom development.

Seed predation of Virola bicuhyba (Schott) Warb. (Myristicaceae) in the Atlantic forest of south-eastern Brazil

Seed predation of Virola bicuhyba (Myristicaceae) was studied in an area of Atlantic forest in south-eastern Brazil, with the objective of testing the Janzen-Connell model. The predation of seeds was evaluated at three different distances from the parent tree for two classes of predators: invertebrates and vertebrates. The method of exclosure plots (closed plots) and open control plots was used, distributed at 5, 15 and 25 m from the trunk of each adult fruiting tree of V. bicuhyba. In Experiment 1, 1,200 seeds were used and, in Experiment 2, 1,440 seeds. Both experiments did not agree with Janzen-Connell model, as seed predation by invertebrates and vertebrates was independent of the distance from the parent tree. Seed predation rate was high, however the impact of predation by vertebrates was higher than by invertebrates, indicating that it is the main cause of seed mortality.

Frugivory on Margaritaria nobilis L.f. (Euphorbiaceae): poor investment and mimetism

Dehiscent fruits of Euphorbiaceae usually have two stages of seed dispersal, autochory followed by myrmecochory. Two stages of Margaritaria nobilis seed dispersal were described, the first stage autochoric followed by ornithocoric. Their dehiscent fruits are green and after they detached from the tree crown and fall on the ground, they open and expose blue metallic cocas. We studied the seed dispersal system of Margaritaria nobilis in a semi-deciduous forest in Brazil. In 80 h of focal observations, we recorded only 12 visits of frugivores, however the thrush Turdus leucomelas was the only frugivore that swallowed the fruits on the tree crown. Pitylus fuliginosus (Fringilidae) and Pionus maximiliani (Psittacidae) were mainly pulp eaters, dropping the seeds below the tree. On the forest floor, after fruits dehiscence, jays (Cyanocorax chrysops), guans (Penelope superciliaris), doves (Geotrygon montana) and collared-peccaries (Pecari tajacu) were observed eating the blue diaspores of M. nobilis. Experiments in captivity showed that scaly-headed parrots (Pionus maximiliani), toco toucans (Ramphastos toco), jays (Cyanochorax chrysops), and guans (Penelope superciliaris) consumed the fruits and did not prey on the seeds before consumption. The seeds collected from the feces did not germinate in spite of the high viability. The two stages of seed dispersal in M. nobilis resembles the dispersal strategies of some mimetic species. However M. nobilis seeds are associated with an endocarp, it showed low investment in nutrients, and consistent with this hypothesis, M. nobilis shared important characteristics with mimetic fruits, such as bright color display, long seed dormancy and protection by secondary compounds.

Multiple dormancy and maternal effect on Miconia ferruginata (Melastomataceae) seed germination, Serra de Caldas Novas, Goiás, Brazil

Species with mostly asexual reproduction are interesting subjects for germination studies since variation would be more easily linked to environmental factors. Miconia ferruginata DC. is an apomictic treelet in Brazilian cerrado areas on rocky outcrops. Germination of seeds collected from individuals occurring in the Serra de Caldas Novas State Park, Goiás, was studied in three experiments under controlled conditions. Germination characteristics differed among individuals and were correlated with altitude and soil Al content. Seeds from plants growing at lower altitudes, with lower soil aluminium content, presented malformed seeds with absence of embryo which rendered lower, but better synchronized germination. The nested analysis showed that from the total variance, 78.14% for germinability, 54.56% for uncertainty of the germination process, and 68.30% for the quantity of seeds without embryo was attributed to the altitudinal effect. Individuals nested within altitude contributed up to 16.93% for the total variance. It means that there is low variability among individuals of the same altitude and high variability among individuals from different points of the slope, making clear that for the studied population the environmental effect is stronger than the genetic component to determine the seed quality. The testa of the seeds provides a mechanical dormancy which seems to be associated also with phenolic compounds, which help to disperse germination through time. Photoblastism was also registered for seeds of this species.

Pollination and fruit set in pumpkin (Cucurbita pepo) by honey bees

Species of Cucurbitaceae are cultivated worldwide and are depend on bee pollination for fruit set. Field and lab experiments were conducted at Cornell University, Ithaca, NY, during 1996 and 1997 to determine "Howden" pumpkin (Cucurbita pepo L.) pollen removal and deposition by honeybees and factors relating to male flower attractiveness. Several parameters were evaluated in flowers at anthesis: (1) removal of pollen from anthers by honey bees, (2) pollen deposition on the stigma by honey bees, (3) amount of pollen on the body of honey bees, (4) fruit set after bee pollination, and (5) male flower nectary's pores and flower attractiveness. Honey bees carried between 1,050 to 3,990 pollen grains and 13,765 were removed from an anther after one visit. The amount of pollen deposited on the stigma by the honey bees varied according to the number of visits, from 53 grains with one visit, to 1,253 grains with 12 visits, and the mean number of grains in each visit varied from 53 to 230 grains. The percentage of established fruits was higher (100%) when the flowers received 12 visits of Apis mellifera, corresponding to a load 1,253 pollen grains. The attractiveness of the male flower for pollen and nectar collection was increased by the degree of opening of the access pore to the nectary in the flower.

Surface studies of limestones and dolostones : characterisation using various techniques and batch dissolution experiments with hydrochloric acid solutions

In this thesis, stepwise titration with hydrochloric acid was used to obtain chemical reactivities and dissolution rates of ground limestones and dolostones of varying geological backgrounds (sedimentary, metamorphic or magmatic). Two different ways of conducting the calculations were used: 1) a first order mathematical model was used to calculate extrapolated initial reactivities (and dissolution rates) at pH 4, and 2) a second order mathematical model was used to acquire integrated mean specific chemical reaction constants (and dissolution rates) at pH 5. The calculations of the reactivities and dissolution rates were based on rate of change of pH and particle size distributions of the sample powders obtained by laser diffraction. The initial dissolution rates at pH 4 were repeatedly higher than previously reported literature values, whereas the dissolution rates at pH 5 were consistent with former observations. Reactivities and dissolution rates varied substantially for dolostones, whereas for limestones and calcareous rocks, the variation can be primarily explained by relatively large sample standard deviations. A list of the dolostone samples in a decreasing order of initial reactivity at pH 4 is: 1) metamorphic dolostones with calcite/dolomite ratio higher than about 6% 2) sedimentary dolostones without calcite 3) metamorphic dolostones with calcite/dolomite ratio lower than about 6% The reactivities and dissolution rates were accompanied by a wide range of experimental techniques to characterise the samples, to reveal how different rocks changed during the dissolution process, and to find out which factors had an influence on their chemical reactivities. An emphasis was put on chemical and morphological changes taking place at the surfaces of the particles via X-ray Photoelectron Spectroscopy (XPS) and Scanning Electron Microscopy (SEM). Supporting chemical information was obtained with X-Ray Fluorescence (XRF) measurements of the samples, and Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) and Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) measurements of the solutions used in the reactivity experiments. Information on mineral (modal) compositions and their occurrence was provided by X-Ray Diffraction (XRD), Energy Dispersive X-ray analysis (EDX) and studying thin sections with a petrographic microscope. BET (Brunauer, Emmet, Teller) surface areas were determined from nitrogen physisorption data. Factors increasing chemical reactivity of dolostones and calcareous rocks were found to be sedimentary origin, higher calcite concentration and smaller quartz concentration. Also, it is assumed that finer grain size and larger BET surface areas increase the reactivity although no certain correlation was found in this thesis. Atomic concentrations did not correlate with the reactivities. Sedimentary dolostones, unlike metamorphic ones, were found to have porous surface structures after dissolution. In addition, conventional (XPS) and synchrotron based (HRXPS) X-ray Photoelectron Spectroscopy were used to study bonding environments on calcite and dolomite surfaces. Both samples are insulators, which is why neutralisation measures such as electron flood gun and a conductive mask were used. Surface core level shifts of 0.7 ± 0.1 eV for Ca 2p spectrum of calcite and 0.75 ± 0.05 eV for Mg 2p and Ca 3s spectra of dolomite were obtained. Some satellite features of Ca 2p, C 1s and O 1s spectra have been suggested to be bulk plasmons. The origin of carbide bonds was suggested to be beam assisted interaction with hydrocarbons found on the surface. The results presented in this thesis are of particular importance for choosing raw materials for wet Flue Gas Desulphurisation (FGD) and construction industry. Wet FGD benefits from high reactivity, whereas construction industry can take advantage of slow reactivity of carbonate rocks often used in the facades of fine buildings. Information on chemical bonding environments may help to create more accurate models for water-rock interactions of carbonates.

Looking for the GAP effect in manual responses and the role of contextual influences in reaction time experiments

When the offset of a visual stimulus (GAP condition) precedes the onset of a target, saccadic reaction times are reduced in relation to the condition with no offset (overlap condition) - the GAP effect. However, the existence of the GAP effect for manual responses is still controversial. In two experiments using both simple (Experiment 1, N = 18) and choice key-press procedures (Experiment 2, N = 12), we looked for the GAP effect in manual responses and investigated possible contextual influences on it. Participants were asked to respond to the imperative stimulus that would occur under different experimental contexts, created by varying the array of warning-stimulus intervals (0, 300 and 1000 ms) and conditions (GAP and overlap): i) intervals and conditions were randomized throughout the experiment; ii) conditions were run in different blocks and intervals were randomized; iii) intervals were run in different blocks and conditions were randomized. Our data showed that no GAP effect was obtained for any manipulation. The predictability of stimulus occurrence produced the strongest influence on response latencies. In Experiment 1, simple manual responses were shorter when the intervals were blocked (247 ms, P < 0.001) in relation to the other two contexts (274 and 279 ms). Despite the use of choice key-press procedures, Experiment 2 produced a similar pattern of results. A discussion addressing the critical conditions to obtain the GAP effect for distinct motor responses is presented. In short, our data stress the relevance of the temporal allocation of attention for behavioral performance.

The terrestrial Gastropoda Megalobulimus abbreviatus as a useful model for nociceptive experiments: effects of morphine and naloxone on thermal avoidance behavior

We describe the behavior of the snail Megalobulimus abbreviatus upon receiving thermal stimuli and the effects of pretreatment with morphine and naloxone on behavior after a thermal stimulus, in order to establish a useful model for nociceptive experiments. Snails submitted to non-functional (22ºC) and non-thermal hot-plate stress (30ºC) only displayed exploratory behavior. However, the animals submitted to a thermal stimulus (50ºC) displayed biphasic avoidance behavior. Latency was measured from the time the animal was placed on the hot plate to the time when the animal lifted the head-foot complex 1 cm from the substrate, indicating aversive thermal behavior. Other animals were pretreated with morphine (5, 10, 20 mg/kg) or naloxone (2.5, 5.0, 7.5 mg/kg) 15 min prior to receiving a thermal stimulus (50ºC; N = 9 in each group). The results (means ± SD) showed an extremely significant difference in response latency between the group treated with 20 mg/kg morphine (63.18 ± 14.47 s) and the other experimental groups (P < 0.001). With 2.5 mg/kg (16.26 ± 3.19 s), 5.0 mg/kg (11.53 ± 1.64 s) and 7.5 mg/kg naloxone (7.38 ± 1.6 s), there was a significant, not dose-dependent decrease in latency compared to the control (33.44 ± 8.53 s) and saline groups (29.1 ± 9.91 s). No statistically significant difference was found between the naloxone-treated groups. With naloxone plus morphine, there was a significant decrease in latency when compared to all other groups (minimum 64% in the saline group and maximum 83.2% decrease in the morphine group). These results provide evidence of the involvement of endogenous opioid peptides in the control of thermal withdrawal behavior in this snail, and reveal a stereotyped and reproducible avoidance behavior for this snail species, which could be studied in other pharmacological and neurophysiological studies.

Scaling analysis of small break experiments in PACTEL

The purpose of this thesis is to study the scalability of small break LOCA experiments. The study is performed on the experimental data, as well as on the results of thermal hydraulic computation performed on TRACE code. The SBLOCA experiments were performed on PACTEL facility situated at LUT. The temporal scaling of the results was done by relating the total coolant mass in the system with the initial break mass flow and using the quotient to scale the experiment time. The results showed many similarities in the behaviour of pressure and break mass flow between the experiments.

Optimal and Robust Designs of Step-stress Accelerated Life Testing Experiments for Proportional Hazards Models

Accelerated life testing (ALT) is widely used to obtain reliability information about a product within a limited time frame. The Cox s proportional hazards (PH) model is often utilized for reliability prediction. My master thesis research focuses on designing accelerated life testing experiments for reliability estimation. We consider multiple step-stress ALT plans with censoring. The optimal stress levels and times of changing the stress levels are investigated. We discuss the optimal designs under three optimality criteria. They are D-, A- and Q-optimal designs. We note that the classical designs are optimal only if the model assumed is correct. Due to the nature of prediction made from ALT experimental data, attained under the stress levels higher than the normal condition, extrapolation is encountered. In such case, the assumed model cannot be tested. Therefore, for possible imprecision in the assumed PH model, the method of construction for robust designs is also explored.

Manual of the botany of the Northern United States : including the distriet east of the Mississipi and North of North Carolina and Tennessee

UANL