Orientador: Carlos Ducatti / Banca: José Roberto Sartori / Banca: Alfredo Sampaio Carrijo / Resumo: Este trabalho objetivou determinar o turnover do carbono-13 no Pectoralis major, quilha, tíbia e fígado de frangos de corte em diferentes períodos de crescimento, utilizando a técnica de isótopos estáveis. Foram utilizados 184 pintos de corte, machos, Cobb, distribuídos ao acaso em sete tratamentos com dias de substituição das dietas distintos: 0; 7; 14; 21; 28; 35; 42 dias de idade, denominados: T0-14, T7-21, T14-28, T21-35, T28-42, T35-49 e T42-56 respectivamente. Para mensurar o turnover do carbono-13 (diluição isotópica) nos tecidos em determinado intervalo de tempo, foram utilizados o modelo sigmoidal de regressão de Boltzmann ou função exponencial do tempo. Os valores de meias-vidas foram: 2,5; 2,8; 2,9; 4,9; 5,1; 5,5 e 6,0 dias para músculo peitoral; 2,7; 2,9; 3,1; 4,6; 5,3; 4,1 e 8,0 dias para quilha; 4,4; 3,6; 5,4; 5,8; 4,3; 5,2 e 6,3 dias para tíbia e 1,3; 1,9; 1,4; 1,5; 1,9; 1,7 e 2,1 dias para fígado, nos tratamentos T0-14, T7-21, T14-28, T21-35, T28-42, T35-49 e T42-56 respectivamente. Dessa forma, a meia-vida dos tecidos estudados, em geral, aumentou de acordo com o avanço da idade de avaliação. Dentre os tecidos estudados, a tíbia apresentou maiores valores de meia-vida, indicando lenta incorporação do carbono-13 e o fígado, os menores, sugerindo rápida incorporação. De acordo com os resultados deste estudo, pode-se concluir que a tíbia apresenta turnover e metabolismo lento indicando o sinal isotópico das dietas iniciais da criação dos frangos de corte; e o fígado, turnover e metabolismo rápido, indicado para refletir a dieta próxima da fase de avaliação. / Abstract: This study aimed to evaluate carbon-13 turnover on Pectoralis major, keel, tibia and liver of poultry at different life stages using the carbon stable isotopes technique. In this trial, 216 male chick lings (Cobb) were allocated in 36 galvanized iron cages with gutter-like feeders and nipple-like water dispensers. The trial was conduced in a totally random distribution consisting on seven treatments with distinct days for diet substitution (0, 7, 14, 21, 28, 35, 42 days old). Either Boltzmann sigmoid regression or the exponential time function was used in order to measure the carbon-13 turnover (isotopic dilution) in a specific period of time. The values of half-life was: 2.5, 2.8, 2.9, 4.9, 5.1, 5.5 and 6.0 days for muscle tissue, 2.7, 2.9, 3.1, 4.6, 5.3, 4.1 and 8.0 days for keel, 4.4, 3.6, 5.4, 5.8, 4.3, 5.2 and 6.3 days for tibia; and 1.3, 1.9, 1.4, 1.5, 1.9, 1.7 and 2.1 days for liver, on T1, T2, T3, T4, T5, T6 and T7 treatments, respectively. Generally, the half-life of the studied tissues increased along with the increase on the evaluated ages. The half-life of tibia was higher than the other tissues, indicating the slow carbon- 13 incorporation. The half-life of liver presented the smallest value indicating fast incorporation of carbon-13. According to the results of this study, it is possible to conclude that tibia has slow metabolism and it may indicate the isotopic signature of initial broiler diets, and the liver, fast metabolism tissue, is appropriated to reflect the diets fed at a closer evaluation period. / Mestre

New pedologic, carbon isotope and phytolith analyses along with stratigraphic correlations to nearby archaeological sites record the spatial and temporal distribution of past vegetation patterns in the Sac Valley Archaeological District of southwest Missouri. Radiocarbon ages obtained from a soil core along Bear Creek, CB5, are related to those from Hajic et al. (1998, 2000) indicating that the CB5 location contains correlative middle Rodgers Shelter submember deposits. This relationship also indicates that sedimentation was approximately two times greater at the CB5 locality than at the Big Eddy (23CE426) archaeological site providing much higher temporal resolution for the alluvial history as well as the vegetation proxies during the early to middle Holocene. Most midcontinent climate proxy records include indications of an early Holocene warm period when prairie replaced forests, then a cooler period in which trees dominated the landscape, followed by a warmer middle Holocene period when prairie vegetation was dominant. However, the CB5 δ13C profile of mixed C3/C4 vegetation indicates either that the vegetation at this location was not as sensitive to climate change or that this location was buffered from other influences, e.g. fires, which were critical to the expansion of prairie vegetation. On the other hand, the phytolith assemblages at CB5 indicate that there were periods with abundant C4 grasses even though the δ13C values indicate a dominance of C3 vegetation. This indicates that in the mixed forest/prairie ecotone interpretations of past vegetation from either carbon isotopes or phytolith assemblages alone may not accurately reflect patterns of vegetation. A new core, DDY-KR2, was obtained from the Big Eddy (23CE426) archaeological site and a finer resolution of δ13C values at Big Eddy increased the detail about alluvial activity and revealed subtle changes in the vegetation. The vegetation types suggested by the δ13C values for DDY-KR2 are reflected in the phytolith assemblages validating their usefulness in reconstructing local vegetation history.

The stable isotopic composition of groundwater within a watershed in eastern Iowa was studied in order to understand how water moves through the system. Samples were gathered using multiple observation wells and pore water samplers and then analyzed to determine the δ18O and δ2H of each sample. Shallow pore water is much more variable in its isotopic composition than deeper water and seems to be more greatly affected by evapotranspiration, whereas groundwater below the water table appears to show a stable isotopic signature suggesting the integration of multiple rain events. Other samples of similar depths across the slope of a hill were also used to observe differences across the area. By observing changes over time in the signatures of these samples, it can be seen that the crest of the hill is most greatly influenced by infiltration from precipitation while the side of the hill is influenced more by throughflow. By combining stable isotope analyses, knowledge of the medium through which the water is moving and the general mechanics of a watershed, a more advanced understanding of how water interacts with and moves through the ground can be gained.

Psychotrophic bacteria produce heat-stable lipase, protease, and phospholipase. Pervious studies indicate the production of multiple enzymes in several strains of Pseudomonas fluorescens, but conclusive evidence is lacking. The influence of culture conditions on the production and thermostability of phospholipase, protease, and lipase was investigated in 17 raw milk and environmental isolates. Production and thermostability of the enzymes were influenced by strain, stage of growth, and the culture medium. Cross-reactivity of antibodies raised to a purified protease and a commercial lipase indicated the immunological diversity of the enzymes. Protease purification was undertaken to investigate the production of multiple proteases within a single strain. A single monomeric protease with a molecular weight of 52 kDa was purified from P. fluorescens RO98. Biochemical characterization of the enzyme revealed that it was a zinc-metallo acidprotease with pH and temperature optima of 5.0 35°C, respectively. The enzyme was thermostable with a D55 of 41 min and a D62.5 of 18 h. Although sensitive assays exist for proteases, they are not suitable for detection of protease activity in milk in the presence of milk proteins. Existing immunoassays approach the required sensitivity but take about 6 h and cannot distinguish between active and inactive enzyme. An immunoassay that can be completed within 2 h and that can detect and distinguish both total and active enzyme was explored. The ratio of these two forms gives insight into the history of the milk. The ability of the purified protease to hydrolyze hydrophobic peptides associated with bitterness in Cheddar cheese was also investigated. Results demonstrated that the protease had the potential to debitter Cheddar cheese because it was able to hydrolyze the bitter peptides commonly found during aging. Two lipolytic enzymes with molecular weights of 50 (Pf-lip1) and 12 kDa (Pf-lip2) were purified from P. fluorescens RO98. Differences were observed in their biochemical properties. D62.5-values of 12.7 and 29.9 h were determined for Pf-lip1 and Pf-lip2, respectively. Pf-lip1 preferred longer chain length fatty acids, and Pf-lip2 preferred shorter chain length substrates. Pf-lip1 hydrolyzed milk fat and emulsified triolein, but Pf-lip2 did not, indicating that the latter was an esterase. This information is of significance to the dairy industry because activity tests that assay both the lipolytic enzymes need to be used in order to direct raw milk to short shelf-life products during processing and ensure quality of long shelf-life products

The reliability of foraminifera as stratigraphic index fossils, and as isotopic proxies of marine environments, is based on the assumption that the fossil concepts represent uniform species, responding consistently to their ambient environments. Understanding sources of uncertainty is, therefore, critical. In this dissertation, I explore a potential bias in the application of planktonic foraminifera utilized extensively for Cenozoic paleo-reconstruction and, to a lesser extent, biostratigraphy: the Globigerinoides ruber-elongatus plexus (‘plexus’ meaning a complex network of interconnected members). Taxonomic revisions since 1826 have resulted in the merging of multiple Globigerinoides species names under one general designation (“Globigerinoides ruber”), the implications of which are now under scrutiny. These “morphotypes” of G. ruber have been shown to incorporate stable isotopes and trace elements in seawater dissimilarly, and correspond to multiple genetic species, some of which occupy different environments. Various criteria exist to sub-divide, group, or distinguish members of the Globigerinoides plexus, most notably the recurring use of Globigerinoides elongatus as a less spherical, less symmetrical counterpart to G. ruber. But the efficacy of these various taxonomic criteria has not been tested quantitatively. Most rely on the traits of visually distinctive “end-members,” while specimens in the morphological “transitional zone” are left to an observer’s subjective interpretation. This prevents quantification in census counts, and may lead to erroneous geochemical analyses. Furthermore, molecular clock estimates suggest that the G. elongatus species evolved significantly later than G. ruber, affecting its potential as a biomarker. In this dissertation, I examine the potential of a minimal-criteria system for classifying Globigerinoides-type morphologies using only three conditions: final chamber compression, final chamber asymmetry, and aperture compression. Morphometric analyses on specimens grouped according to this new system allow us to assess to what degree visual classification reflects morphospace discontinuity. Armed with this information, I then explore potential isotopic offsets between members of the Globigerinoides plexus, and its use in reconstructing regional differences in climate or habitat influences in the Gulf of Mexico and Caribbean basins. Finally, having shown that G. ruber and G. elongatus can be reliably visually distinguished, I tracked the species’ fossil presence individually in a deep core from the South China Sea, and confirmed the presence of G. ruber in the South China Sea through the late Miocene, and G. elongatus through the Pleistocene. While it is believed that neither species was traced to its true first occurrence (FO), the relative FO of G. ruber was shown to be 4–5 Ma before G. elongatus

Cova des Pas de Vallgornera (CPV) is morphologically and scientifically, the most prominent karst feature of Mallorca Island (Spain). It consists of over 74 km of passages developed within two carbonate lithologies (reef front and back reef facies) of Upper Miocene age. Two distinct cave patterns are recognized, both tightly controlled by the type of facies; spongework mazes and collapse chambers are characteristic for the reef front unit, whereas linear, fracture-guided galleries develop in the back reef carbonates. CPV is abundantly decorated with a variety of speleothems. The overarching goal of this study is to provide evidence towards the cave's major speleogenetic pathways using diagnostic mineral assemblages and a set of micro- and macro-scale morphological features. In particular, this research investigates whether hypogene processes were/are the main driving force in CPV's speleogenesis and how mineral assemblages and cave morphologies help untangle the complex evolution of the cave. The mineralogy of speleothems (crusts, nodules, crystals, earthy masses) deposited in the vicinity of hypogene features in the reef front is characterized by the presence of aragonite, ankerite, huntite, clay minerals, and quartz. In the back reef limestone, however, the dominant mineral is dolomite, along with aragonite, celestine, huntite, clay minerals, and quartz. Calcite is by far the most ubiquitous mineral throughout the cave. A total of twenty-six minerals were identified throughout the cave; the highest diversity (20 species) is around or inside typical hypogene features occurring within the back reef limestone, whereas in the seaward part only 13 minerals occur. Detailed macroscopic and scanning electron micro-chemical analysis and imaging have permitted the investigation of textural relationships between the minerals associated with vents, rims, and corrosion cupolas above them. These studies along with morphological and stable isotope analyses (δ13C, δ18O, δ34S), confirm that not all minerals are connected with a hypogene stage in the cave evolution, and furthermore, none of them appears to be sulfuric acid by-products. Instead, the mineral assemblages documented in speleothems from CPV clearly support at least three speleogenetic pathways, namely seacoast mixing, ascending of warm groundwaters (basal recharge), and vadose (meteoric recharge). In summary, we have shown that cave minerals (in association with particular cave morphologies) in CPV hold the keys to understanding mineral-forming processes, conditions, and events, allowing establishing their relationship with various speleogenetic pathways.

The aim of this thesis is to develop faster method for stable model computation of non-stratified logic programs and study its efficiency. It focuses mainly on the stable model and weak well founded semantics of logic programs. We propose an approach to compute stable models by where we first transform the logic program using paraconsistent relational model, then we compute the weak-well founded model which is used to generate a set of models consisting of the true and unknown values, which are tested for stability. We perform some experiments to test the efficiency of our approach which incurs overhead to eliminate negative values against a Naïve method of stable model computation.

An improved ability to assess whether individuals have been added through immigration or natality and lost through emigration or mortality could alleviate several problems in population ecology. Fortunately, advances in stable isotope techniques now allow the movements of individuals to be retraced from tissue values and provide an opportunity to link information about the origins of individuals with demographic rates so that questions about the significance of dispersal can be assessed. I used such an approach by combining feather isotope information with demographic rates derived from capture-mark-recapture of individual mallards (Anas platyrhynchos) breeding in the Canadian aspen parklands, at multiple spatiotemporal scales, to answer questions about population persistence, settling patterns by dispersers, and the fitness of immigrant birds relative to residents.<p> Feather isotope (ä34S, äD, ä15N, and ä13C) values from an independent sample of flightless mallard ducklings sampled from across the mid-continent breeding range was used to validate an existing model used for origin assignments. Spatial resolution analysis within the mid-continent mallard breeding range generally showed a loss in prediction when attempting to assign individuals to more narrowly separated geographic origins among boreal, aspen parkland and prairie regions. For feather äD, spatial resolution may be limited by temporal patterns of local climatic events that produce variability in consumer tissue values. Thus, the use of multiple feather isotope signals would provide more reliable information about the origin of individuals for addressing questions about long-distance dispersal in yearling mallards.<p> Demographic rescue in an apparent population sink near Minnedosa, Manitoba, Canada, was due to elevated survival rates from a highly productive group of nesting female mallards using nest tunnels (i.e., an artificial nesting structure) and recruitment of yearling females having natal origins within the aspen parklands. There was little evidence that immigration by yearling females dispersing long-distances was important to annual population growth rates. Consistently high annual survival rates of adult females using nest tunnels lowered the recruitment rates needed for population stability. While tunnel-origin and within-region recruitment of yearling females were nearly equally important to local population growth rate, fine-scale limitations of isotopic origin assignments prevented further assessment of where recruits originated from within the aspen parkland region.<p> Factors related to breeding area settling patterns of yearling females are not well understood despite implications to local population dynamics. The likelihood that immigrant yearling females would settle in a parkland breeding area was positively correlated with local breeding-pair density and the amount of perennial nest cover, but was negatively correlated with the amount of wetlands. Although these relationships were not well estimated, they are most consistent a hypothesis that females were attracted to breeding sites by conspecific cues rather than avoidance. Immigrants comprised an average of 9% (range: 0 39% over 22 sites) of yearling recruits; most had natal origins in the U.S. prairie pothole region but a non-trivial number originated from the boreal forest, indicating a high degree of connectedness among breeding regions resulting from long-distance natal dispersal.<p> One of the most frequent explanations for strong site fidelity in breeding female ducks is that females benefit from site familiarity. However, evidence for differential reproductive success between immigrant and resident yearling females was weak, On sites with favourable wetland conditions and low breeding-pair densities immigrant females were more likely to breed and nest successfully than were residents whereas under opposite wetland and pair conditions, resident females were favoured. Thus, the costs and benefits of a natal dispersal decision seemed to vary with social context and environmental conditions, and further work is needed to clarify these processes.

Surface mining of oil sands in northern Alberta requires stripping of saline-sodic shale overburden, which is typically placed in large upland overburden dumps. Due to the chemical nature of this shale, engineered soil covers must be constructed over the shale to support the growth of forest vegetation. A research site on South Bison Hill (SBH), a shale overburden dump at the Syncrude Canada Ltd. Mildred Lake Mine, has been used by researchers over the past decade to study the performance of a reclamation cover. This study was undertaken to improve the understanding of salt and moisture dynamics in the cover-shale system. In particular, the objective of this study was to develop an estimate of the net percolation rate through the cover soil and into the shale overburden. Stable isotope (ä2H and ä18O) measurements obtained from the pore water of soil samples were used to develop stable isotope profiles at various sampling locations along the slope and plateau of the SBH. Simulated profiles were then generated using 2D, finite element numerical modelling software and compared to the measured profiles. Model parameters were obtained from testing and the work of previous researchers. The model results revealed that the net percolation is greatest (32-50 mm/yr) for the plateau and mid-slope bench sample locations. Net percolation rates for sample locations on the slope were lower at 0-12 mm/yr. The results from the stable isotope modelling were utilized in a SO42- transport model to ascertain if calculated net percolation rates could explain measured salinity profiles. This modelling exercise revealed that calculated SO42- profiles are highly dependent on the assumed SO42- production rates in the shale, which is primarily attributed to pyrite oxidation. The model results showed the isotope-based net percolation rates could explain the measured SO42-profiles for a reasonable range SO42- production rates. The SO42- production rates calculated in the model were greatest for the plateau and mid-slope bench locations and lesser for the sloped locations. The model also showed that the mass of SO42- removed by interflow was minimal compared to the mass generated by pyrite oxidation and that net percolation is the dominant flushing mechanism at net percolation rates of 8 mm/yr or more.

Surface mining of oil sands in northern Alberta requires stripping of saline-sodic shale overburden, which is typically placed in large upland overburden dumps. Due to the chemical nature of this shale, engineered soil covers must be constructed over the shale to support the growth of forest vegetation. A research site on South Bison Hill (SBH), a shale overburden dump at the Syncrude Canada Ltd. Mildred Lake Mine, has been used by researchers over the past decade to study the performance of a reclamation cover. This study was undertaken to improve the understanding of salt and moisture dynamics in the cover-shale system. In particular, the objective of this study was to develop an estimate of the net percolation rate through the cover soil and into the shale overburden. Stable isotope (ä2H and ä18O) measurements obtained from the pore water of soil samples were used to develop stable isotope profiles at various sampling locations along the slope and plateau of the SBH. Simulated profiles were then generated using 2D, finite element numerical modelling software and compared to the measured profiles. Model parameters were obtained from testing and the work of previous researchers. The model results revealed that the net percolation is greatest (32-50 mm/yr) for the plateau and mid-slope bench sample locations. Net percolation rates for sample locations on the slope were lower at 0-12 mm/yr. The results from the stable isotope modelling were utilized in a SO42- transport model to ascertain if calculated net percolation rates could explain measured salinity profiles. This modelling exercise revealed that calculated SO42- profiles are highly dependent on the assumed SO42- production rates in the shale, which is primarily attributed to pyrite oxidation. The model results showed the isotope-based net percolation rates could explain the measured SO42-profiles for a reasonable range SO42- production rates. The SO42- production rates calculated in the model were greatest for the plateau and mid-slope bench locations and lesser for the sloped locations. The model also showed that the mass of SO42- removed by interflow was minimal compared to the mass generated by pyrite oxidation and that net percolation is the dominant flushing mechanism at net percolation rates of 8 mm/yr or more.