7 resultados para 300701 Physiology and Genetics
em AMS Tesi di Dottorato - Alm@DL - Università di Bologna
Resumo:
The hydrogen production in the green microalga Chlamydomonas reinhardtii was evaluated by means of a detailed physiological and biotechnological study. First, a wide screening of the hydrogen productivity was done on 22 strains of C. reinhardtii, most of which mutated at the level of the D1 protein. The screening revealed for the first time that mutations upon the D1 protein may result on an increased hydrogen production. Indeed, productions ranged between 0 and more than 500 mL hydrogen per liter of culture (Torzillo, Scoma et al., 2007a), the highest producer (L159I-N230Y) being up to 5 times more performant than the strain cc124 widely adopted in literature (Torzillo, Scoma, et al., 2007b). Improved productivities by D1 protein mutants were generally a result of high photosynthetic capabilities counteracted by high respiration rates. Optimization of culture conditions were addressed according to the results of the physiological study of selected strains. In a first step, the photobioreactor (PBR) was provided with a multiple-impeller stirring system designed, developed and tested by us, using the strain cc124. It was found that the impeller system was effectively able to induce regular and turbulent mixing, which led to improved photosynthetic yields by means of light/dark cycles. Moreover, improved mixing regime sustained higher respiration rates, compared to what obtained with the commonly used stir bar mixing system. As far as the results of the initial screening phase are considered, both these factors are relevant to the hydrogen production. Indeed, very high energy conversion efficiencies (light to hydrogen) were obtained with the impeller device, prooving that our PBR was a good tool to both improve and study photosynthetic processes (Giannelli, Scoma et al., 2009). In the second part of the optimization, an accurate analysis of all the positive features of the high performance strain L159I-N230Y pointed out, respect to the WT, it has: (1) a larger chlorophyll optical cross-section; (2) a higher electron transfer rate by PSII; (3) a higher respiration rate; (4) a higher efficiency of utilization of the hydrogenase; (5) a higher starch synthesis capability; (6) a higher per cell D1 protein amount; (7) a higher zeaxanthin synthesis capability (Torzillo, Scoma et al., 2009). These information were gathered with those obtained with the impeller mixing device to find out the best culture conditions to optimize productivity with strain L159I-N230Y. The main aim was to sustain as long as possible the direct PSII contribution, which leads to hydrogen production without net CO2 release. Finally, an outstanding maximum rate of 11.1 ± 1.0 mL/L/h was reached and maintained for 21.8 ± 7.7 hours, when the effective photochemical efficiency of PSII (ΔF/F'm) underwent a last drop to zero. If expressed in terms of chl (24.0 ± 2.2 µmoles/mg chl/h), these rates of production are 4 times higher than what reported in literature to date (Scoma et al., 2010a submitted). DCMU addition experiments confirmed the key role played by PSII in sustaining such rates. On the other hand, experiments carried out in similar conditions with the control strain cc124 showed an improved final productivity, but no constant PSII direct contribution. These results showed that, aside from fermentation processes, if proper conditions are supplied to selected strains, hydrogen production can be substantially enhanced by means of biophotolysis. A last study on the physiology of the process was carried out with the mutant IL. Although able to express and very efficiently utilize the hydrogenase enzyme, this strain was unable to produce hydrogen when sulfur deprived. However, in a specific set of experiments this goal was finally reached, pointing out that other than (1) a state 1-2 transition of the photosynthetic apparatus, (2) starch storage and (3) anaerobiosis establishment, a timely transition to the hydrogen production is also needed in sulfur deprivation to induce the process before energy reserves are driven towards other processes necessary for the survival of the cell. This information turned out to be crucial when moving outdoor for the hydrogen production in a tubular horizontal 50-liter PBR under sunlight radiation. First attempts with laboratory grown cultures showed that no hydrogen production under sulfur starvation can be induced if a previous adaptation of the culture is not pursued outdoor. Indeed, in these conditions the hydrogen production under direct sunlight radiation with C. reinhardtii was finally achieved for the first time in literature (Scoma et al., 2010b submitted). Experiments were also made to optimize productivity in outdoor conditions, with respect to the light dilution within the culture layers. Finally, a brief study of the anaerobic metabolism of C. reinhardtii during hydrogen oxidation has been carried out. This study represents a good integration to the understanding of the complex interplay of pathways that operate concomitantly in this microalga.
Resumo:
Weaning is a crucial period in the management of piglets. In modern piggeries economic interest make weaning age decrease more and more and the detrimental consequences of weaning have as much importance as earlier the weaning occurs. The risk of development of post-weaning diarrhea (PWD) in piglets is high and PWD is the cause of serious economic losses in pig herds. In the past the supplementation of the feed given after weaning with growth promoters antibiotics, in order to keep PWD under control, used to be a common practice, but their usage has been banned in EU since 2006. This measure led to the investigation of alternative suitable feed supplements that would be reasonably efficient in protecting and sustaining animal health and performance. Aim of this thesis was to evaluate the effect of some different alternatives to growth-promoters antibiotics on weaning piglets and to assess if some of them could be considered as valuables options to replace auxinic in animal feeding. The study is composed by four experimental trials. The first one aims to identify mechanisms involved in the auxinic effects of antibiotics in the diets; the following three evaluate the addition butyric acid, tryptophan, and nitrate as alternative to in-feed antimicrobials. Although some results are controversial, it appears from the data presented that the alternatives to in-feed antibiotics considered may exert positive effects on some zootechnical and health parameters on piglet in the post-weaning period. Anyway, the mechanism of action and the interaction with microbiota of such additives should be investigated inside out because many effects remains poorly understood.
Resumo:
This PhD Thesis includes five main parts on diverse topics. The first two parts deal with the trophic ecology of wolves in Italy consequently to a recent increase of wild ungulates abundance. Data on wolf diet across time highlighted how wild ungulates are important food resource for wolves in Italy. Increasing wolf population, increasing numbers of wild ungulates and decreasing livestock consume are mitigating wolf-man conflicts in Italy in the near future. In the third part, non-invasive genetic sampling techniques were used to obtain genotypes and genders of about 400 wolves. Thus, wolf packs were genetically reconstructed using diverse population genetic and parentage software. Combining the results on pack structure and genetic relatedness with sampling locations, home ranges of wolf packs and dispersal patterns were identified. These results, particularly important for the conservation management of wolves in Italy, illustrated detailed information that can be retrieved from genetic identification of individuals. In the fourth part, wolf locations were combined with environmental information obtained as GIS-layers. Modern species distribution models (niche models) were applied to infer potential wolf distribution and predation risk. From the resulting distribution maps, information pastures with the highest risk of depredation were derived. This is particularly relevant as it allows identifying those areas under danger of carnivore attack on livestock. Finally, in the fifth part, habitat suitability models were combined with landscape genetic analysis. On one side landscape genetic analyses on the Italian wolves provided new information on the dynamics and connectivity of the population and, on the other side, a profound analysis of the effects that habitat suitability methods had on the parameterization of landscape genetic analyses was carried out to contributed significantly to landscape genetic theory.
Resumo:
In organic and biodynamic vineyards, canopy management practices should be carefully and timely modulated, particularly in a context of climate change, for successfully achieving balanced plants, ventilated and exposed berries, elevated grape and wine quality. In 2013 and 2014, characterized by contrasting climatic conditions, the implications of post-veraison (late) or pea-size trimming, post-veraison or pre-harvest late defoliations and shoot-positioning (post-veraison) were assessed against long-shoots non treated controls, under field conditions on organically-cultivated cv. Sangiovese. The key agronomic and enological relevance of late trimming and defoliations clearly emerged in both seasons. Berry skin phenolics (e.g. anthocyanins, flavonols) increased markedly, without changes in technological parameters. In case of early trimming, such positive effects were observed only in 2013. Maintaining long shoots for shading decreased anthocyanins, flavonols and total phenolics concentrations and promoted the production of compact bunches. Experimental data strongly designated late trimming, a practice proved to contain yield and bunch compactness, as a valuable alternative to cluster thinning. Late trimming, defoliations and shoot positioning reduced the severity of Botrytis cluster rot. The highest levels of berry skins phenolic compounds in late trimmed and defoliated plants could have contributed control the severity of this pathogen. The enological benefits induced by late trimming and defoliations and shoot positioning emerged in both young and aged wines. For the first time, cell cultures from cv. Sangiovese berry tissues were obtained and enabled to investigate, in controlled conditions, the relations between mechanisms regulating secondary metabolism in grapevine cells and changes induced by environmental and agronomic factors. The Doctoral Dissertation strongly highlights the need to consider, for a proper interpretation of the multiple modifications induced by canopy management strategies, physiological mechanisms other than the canonic source-sink relationships, in particular their impact on the vine hormonal status.
Resumo:
The objective of this thesis was to study the response mechanisms of grapevine to Fe-deficiency and to potential Fe chlorosis prevention strategies. The results show that the presence of bicarbonate in the nutrient solution shifted the activity of PEPC and TCA cycle enzymes and the accumulation/translocation of organic acids in roots of Fe-deprived plants. The rootstock 140 Ruggeri displayed a typical behavior of calcicole plants under bicarbonate stress. The Fe chlorosis susceptible rootstock 101-14 reacted to a prolonged Fe-deficiency reducing the root activity of PEPC and MDH. Noteworthy, it accumulates high levels of citric acid in roots, indicating a low capacity to utilizing, transporting and/or exudating organic acids into the rhizosfere. In contrast, 110 Richter rootstock is capable to maintain an active metabolism of organic acids in roots, accumulating them to a lesser extent than 101-14. Similarly to 101-14, SO4 genotype displays a strong decrease of mechanisms associated to Fe chlorosis tolerance (PEPC and MDH enzymes). Nevertheless it is able to avoid excessive accumulation of citric acid in roots, similar as 110 Richter rootstock. Intercropping with Festuca rubra increased leaf chlorophyll content and net photosynthesis. In addition, intercropping reduces the activity of PEPC in roots, similary to Fe-chelate supply. Applications of NH4+ with nitrification inhibitor prevents efficiently Fe-deficiency, increases chlorophyll content, and induces similar root biochemical responses as Fe-EDDHA. Without the addition of nitrification inhibitors, the effectiveness of NH4+ supply on Fe chlorosis prevention resulted significantly lower. The aspects intertwined in this investigation highlight the complexity of Fe physiology and the fine metabolic tuning of grapevine genotypes to Fe availability and soil-related environmental factors. The experimental evidences reveal the need to carry out future researches on Fe nutrition maintaining a continous flow of knowledge between theoretical and agronomical perspectives for fully supporting the efforts devoted to convert science into practice.
Resumo:
During the last few years, several methods have been proposed in order to study and to evaluate characteristic properties of the human skin by using non-invasive approaches. Mostly, these methods cover aspects related to either dermatology, to analyze skin physiology and to evaluate the effectiveness of medical treatments in skin diseases, or dermocosmetics and cosmetic science to evaluate, for example, the effectiveness of anti-aging treatments. To these purposes a routine approach must be followed. Although very accurate and high resolution measurements can be achieved by using conventional methods, such as optical or mechanical profilometry for example, their use is quite limited primarily to the high cost of the instrumentation required, which in turn is usually cumbersome, highlighting some of the limitations for a routine based analysis. This thesis aims to investigate the feasibility of a noninvasive skin characterization system based on the analysis of capacitive images of the skin surface. The system relies on a CMOS portable capacitive device which gives 50 micron/pixel resolution capacitance map of the skin micro-relief. In order to extract characteristic features of the skin topography, image analysis techniques, such as watershed segmentation and wavelet analysis, have been used to detect the main structures of interest: wrinkles and plateau of the typical micro-relief pattern. In order to validate the method, the features extracted from a dataset of skin capacitive images acquired during dermatological examinations of a healthy group of volunteers have been compared with the age of the subjects involved, showing good correlation with the skin ageing effect. Detailed analysis of the output of the capacitive sensor compared with optical profilometry of silicone replica of the same skin area has revealed potentiality and some limitations of this technology. Also, applications to follow-up studies, as needed to objectively evaluate the effectiveness of treatments in a routine manner, are discussed.